201002345 六、發明說明： 【發明所屬之技術領域】 本發明係關於與人類蛋白酶活化的受體2 (麗)相關 炎症性病症的治療方法’包括炎症性疼痛、手術後刀口疼 痛、神經性疼痛、骨折疼痛、骨質疏鬆骨折疼痛以及痛風性 關節疼痛。 【先前技術】 蛋白酶活化的受體2(PAR2,亦稱為C140受體)是GPCR 1〇 家族的―個以，絲族被胰蛋Μ和肥大細胞蛋白酶類胰 蛋白酶透過胺基酸末端的蛋白裂解，使得暴露的係鎖配體结 合到受體上而導致活化。GPCR家埃有4個成員，其中三個， PARI、PAR3和PAR4為凝血酶受體。pAR4也可為胰蛋白 酶活化。類似於係鎖配體序列的短合成肽已被成功地用來啟 15 動除了 PAR3以外的所有PAR配體。 編碼此四種GPCR家族成員的基因保留於所有物種 中，並具有類似的帶有打斷編碼受體胺基酸末端的單—内含 子結構。小鼠和人類PAR2 83%是同源的，但是在胰蛋白^ 裂解點内為高度保留的。PAR表現廣泛，且已涉及到諸如體 2〇 内穩態、組織修復、血管生成和發炎等重要過程。 人類？八112(8£(3 1〇]^0:1-2)的核酸和胺基酸序列在美 國第5,629，174、5,716,789、6,297,026號專利以及美國第 2003/0018184號專利公開案中均有描述。PAR2抗體在例如 美國第5,874,400號專利和歐洲第〇 8〇4 452 B1號專利中均 201002345 有描述。使用PAR2抗體的方法在美國第2007/0237759號專 利公開書中有描述。 【發明内容】 在第一方面’本發明特徵在於利用一種能夠抑制PAR2 蛋白裂解並阻斷PAR2活化的PAR2拮抗劑來抑制PAR2活 性的方法。在一個具體實施例中，該方法包括對患有與pAR2 相關的病症或能夠藉由PAR2的抑制而改善或減輕的病症之 患者投與一療效量的PAR2拮抗劑。此類病症或障礙包括如 炎症和/或疼痛。炎症性病症包括，例如胃腸系統或氣道的 發炎，風濕性疾病、骨關節炎、與炎症相關的疼痛、手術後 刀口疼痛、神經痛、骨折疼痛、骨質疏鬆性骨折疼痛，以及 痛風性關節疼痛。 可用於本發明的方法中的PAR2拮抗劑是一種能夠阻斷 人類PAR2活性的分子，包括藉由阻斷受體末端蛋白裂解 而阻斷PAR2啟動的分子、阻斷裂解後受體活化的分子，或 抑制PAR2表現的分子。PAR2拮抗劑可為一種抗體、抗體 可結合抗原的片段、肽體、可溶的「誘餌」受體，或反義或 siRNA分子。在一個較佳的具體實施例中，pAR2拮抗劑為 種抗體或抗體可結合抗原的片段。PAR2抗體可能是完整 的人類抗體或部分為人類抗體或抗體可結合抗原的片段。在 個更佳的具體貫施例中，PAR2拮抗劑為一個完整人類抗 體或抗體可結合抗原的片段。 本發明的治療方法包括將PAR2作為單一藥劑投與，或 201002345 與弟一種治療劑一起投與。第二種治療劑可為例如一種或多 種白細胞介素-1 (IL-1)抑制劑，抗癲癇藥物，如卡巴番定 (gabapentain ) ° 在一個較佳的具體實施例中，本發明之特徵在於一種利 用抗-PAR2抗體或一個抗體可結合抗原的片段治療人類受 試者神經性疼痛的方法。如下面實驗一節中所介紹的，在一 個動物模型中，利用抗-PAR2抗體拮抗劑顯著地減輕了長期 熱敏疼痛和機械性觸誘發疼痛。 在第二方面，本發明之特徵在於蛋白酶活化的受體2 (PAR2)拮抗劑用來製造用於治療、抑制或改善炎症症狀或疾 病的藥物之用途，其中炎症症狀或疾病為類風濕性關節炎或 骨關節炎。在一個較佳的具體實施例中，PAR2拮抗劑是一 個抗人類PAR2抗體或一抗體可結合抗原的片段；較佳的是 抗-PAR2抗體或完整人類抗體的片段。 在第三方面，本發明之特徵在於一種蛋白酶活化的受體 2 (PAR2)拮抗劑用來製造治療、抑制或改善疼痛的藥物，其 中疼痛為炎症性疼痛、手術後刀口疼痛、神經性疼痛、骨折 疼痛、骨質疏鬆性骨折疼痛、骨癌疼痛和痛風性關節疼痛。 在一個較佳的具體實施例中，治療的炎症性疼痛為與腸易激 症候群相關的疼痛。在一個較佳的具體實施例中’ PAR2拮 抗劑是一個抗人類PAR2抗體或一個抗體可結合抗原的片 段；首選的是抗_ PAR2抗體或完整人類抗體的片段。 在PAR2拮抗劑用途的特定具體實施例中，PAR2被用 作第一治療劑，與一或多種另外治療劑，例如另一個PAR2 5 201002345 拮抗劑、一個細胞因子抑制劑，如一個白細胞介素（I 抑制劑（例如列洛西普或阿那白滞素）；一個IL_ i 8抑制劍1 ·) 一個IL-6抑制劑，如一個抗或抗_iL6受體抗體；—们 抗癲癇藥物’例如卡巴番定；一個NGF抑制劑，如—個打 -NGF抗體；低劑量秋水仙鹼；阿司匹林；或其他Nsaid 類固醇，例如醋酸潑尼松、曱氨喋呤；低劑量環孢黴素A . 一種TNF抑制劑；和/或協同治療劑，例如尿酸合成抑制巧 (別嘌呤醇）；尿酸分泌啟動劑（羧苯磺胺、苯磺唑_和/或 苯溴馬隆）和/或皮質類固醇。 詳細說明 在說明本發明的方法之前’應當理解，本發明不限於所 描述的特定方法和實驗條件，因為這些方法和條件可能變 化。還應當理解，此處所用的術語僅用於說明特定實施例的 目的’並非意在加以限制，因為本發明的範圍僅為所附的申 請專利範圍所限制。 除非上下文内容另有明確規定，否則’本說明以及所附 的專利申請範圍中所用的單數形式「a」、「an」和「the」 包括複數形式。因此，「一個方法」包括此處所述類型的一 個或更多的方法和/或步驟，和/或對熟悉本領域的技術人員 閱讀了本披露書後將變得很明顯的一個或更多的方法和/或 步驟。 除非他處另有定義，此處所用的所有技術和科學術語與 一般熟悉本發明所屬領域的技術人員的通常理解具有同樣 201002345 的含義。儘管任何與此處所述的方法和材料類 法和材料在本發明的實施和試驗中均可採用，下 首選的方法和材料。 面也明的疋 定義 人類蛋白酶-啟動的受體2(「PAR2j) 列SEQ Π3 N0:1編碼的蛋白質和具有胺基酸為 NO:2的蛋白質。PAR2也稱為「c14〇受體」。 山 此處所用的術語「PAR2拮抗劑」係指— 10 活性或表現的分子。 個抑制PAR2 此處所用的術語「抗體」係指由四個多肽鏈和由二护 物鍵互聯的兩個重鏈⑻和兩個輕鏈（L)組成的[疫球 蛋白分子。每個重鏈由一個重鏈可變區域（縮寫為 或VH)和一個重鏈不變區域組成。重鏈不變區由三個域 cm、CH2和CH3組成。每個輕鏈由一個輕鏈可變區域（^ 寫為LCVR或Vl)和一個輕鏈不變區域組成。輕鏈不變區 =一個域CL組成。VH和VL區可進一步分成稱為互補決 疋區（CDR)的高變區，其中散佈有保存性更好的稱為框架201002345 VI. INSTRUCTIONS OF THE INVENTION: TECHNICAL FIELD OF THE INVENTION The present invention relates to a method for treating an inflammatory disorder associated with human protease-activated receptor 2 (Li), including inflammatory pain, postoperative scalp pain, neuropathic pain, Fracture pain, osteoporotic fracture pain, and gouty joint pain. [Prior Art] Protease-activated receptor 2 (PAR2, also known as C140 receptor) is a protein of the GPCR 1〇 family, which is blocked by trypsin and mast cell protease trypsin. Cleavage causes the exposed tethered ligand to bind to the receptor resulting in activation. GPCR has four members, three of which, PARI, PAR3 and PAR4 are thrombin receptors. pAR4 can also be trypsin activated. Short synthetic peptides similar to the tethered ligand sequences have been successfully used to initiate all PAR ligands except PAR3. The genes encoding these four GPCR family members are retained in all species and have a single-intron structure with a disrupted receptor amino acid terminus. Mouse and human PAR2 are 83% homologous but highly retained within the trypsin cleavage site. PAR is widely performed and has involved important processes such as homeostasis, tissue repair, angiogenesis, and inflammation. Humanity? The nucleic acid and amino acid sequences of the eight 112 (8 £ (3 1 〇) ^ 0: 1-2) are described in U.S. Patent Nos. 5,629,174, 5, 716, 789, 6, 297, 026, and U.S. Patent No. 2003/0018184. The PAR2 antibody is described in, for example, U.S. Patent No. 5,874,400 and U.S. Patent No. 4,452,452, the entire disclosure of which is incorporated herein by reference. In a first aspect, the invention features a method of inhibiting PAR2 activity using a PAR2 antagonist capable of inhibiting PAR2 protein cleavage and blocking PAR2 activation. In a specific embodiment, the method comprises treating a condition associated with pAR2 A patient having a condition that is ameliorated or ameliorated by inhibition of PAR2 is administered a therapeutic amount of a PAR2 antagonist. Such conditions or disorders include, for example, inflammation and/or pain. Inflammatory disorders include, for example, inflammation of the gastrointestinal system or airways. , rheumatic diseases, osteoarthritis, inflammation-related pain, postoperative knee pain, neuralgia, fracture pain, osteoporotic fracture pain, and gouty joint pain The PAR2 antagonist useful in the method of the present invention is a molecule capable of blocking human PAR2 activity, including a molecule that blocks PAR2 initiation by blocking cleavage of the receptor terminal protein, and a molecule that blocks activation of the receptor after cleavage. Or a molecule that inhibits the expression of PAR2. The PAR2 antagonist can be an antibody, a fragment of an antibody that binds to an antigen, a peptibody, a soluble "bait" receptor, or an antisense or siRNA molecule. In a preferred embodiment Wherein, the pAR2 antagonist is a fragment of an antibody or antibody that binds to an antigen. The PAR2 antibody may be an intact human antibody or a fragment of a human antibody or antibody that binds to an antigen. In a better specific embodiment, PAR2 antagonism The agent is a fragment of an intact human antibody or antibody that binds to the antigen. The method of treatment of the invention comprises administering PAR2 as a single agent, or 201002345 with a therapeutic agent. The second therapeutic agent can be, for example, one or more Interleukin-1 (IL-1) inhibitor, anti-epileptic drug, such as gabapentain ° In a preferred embodiment, the invention A method for treating neuropathic pain in a human subject using an anti-PAR2 antibody or an antibody-binding fragment, as described in the experimental section below, using an anti-PAR2 antibody antagonist in an animal model. Long-term heat-sensitive pain and mechanical-induced pain are alleviated. In a second aspect, the invention features a protease-activated receptor 2 (PAR2) antagonist for use in the manufacture, treatment, inhibition or amelioration of inflammatory symptoms or diseases. The use of a drug, wherein the symptom or disease of inflammation is rheumatoid arthritis or osteoarthritis. In a preferred embodiment, the PAR2 antagonist is an anti-human PAR2 antibody or a fragment to which an antibody binds to an antigen; preferably an anti-PAR2 antibody or a fragment of an intact human antibody. In a third aspect, the invention features a protease activated receptor 2 (PAR2) antagonist for use in the manufacture of a medicament for treating, inhibiting or ameliorating pain, wherein the pain is inflammatory pain, postoperative scalp pain, neuropathic pain, Fracture pain, osteoporotic fracture pain, bone cancer pain, and gouty joint pain. In a preferred embodiment, the treated inflammatory pain is pain associated with irritable bowel syndrome. In a preferred embodiment the 'PAR2 antagonist is an anti-human PAR2 antibody or a fragment to which an antibody binds to an antigen; preferred is a fragment of an anti-PAR2 antibody or an intact human antibody. In a specific embodiment of the use of a PAR2 antagonist, PAR2 is used as a first therapeutic agent with one or more additional therapeutic agents, such as another PAR2 5 201002345 antagonist, a cytokine inhibitor, such as an interleukin ( I inhibitor (such as raloxicept or anakinra); an IL_i 8 inhibits sword 1 ·) an IL-6 inhibitor, such as an anti- or anti-iL6 receptor antibody; - anti-epileptic drugs' For example, carbacyclodine; an NGF inhibitor, such as a dozen-NGF antibody; low-dose colchicine; aspirin; or other Nsaid steroids, such as prednisone acetate, amidoxime; low-dose cyclosporine A. A TNF inhibitor; and/or a synergistic therapeutic agent, such as uric acid synthesis inhibition (allopurinol); a uric acid secretion promoter (carboxybenzenesulfonamide, benzoxazole- and/or benzbromarone) and/or a corticosteroid. DETAILED DESCRIPTION OF THE INVENTION Before the method of the present invention is described, it should be understood that the present invention is not limited to the specific methods and experimental conditions described, as these methods and conditions may vary. It is also understood that the terminology used herein is for the purpose of the description of the invention, and is not intended to The singular forms "a", "an", and "the" are used in the <RTI ID=0.0> </ RTI> </ RTI> <RTIgt; Thus, "a method" includes one or more of the methods and/or steps of the type described herein, and/or one or more that will become apparent to those skilled in the art after reading this disclosure. Method and / or steps. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as the meaning of 201002345, as commonly understood by one of ordinary skill in the art to which the invention pertains. Although any of the methods and materials methods and materials described herein can be employed in the practice and testing of the present invention, the preferred methods and materials are preferred.也 疋 疋 definition Human protease-activated receptor 2 ("PAR2j" column SEQ Π 3 N0:1 protein and protein with amino acid NO: 2. PAR2 is also known as "c14 〇 receptor". Mountain The term "PAR2 antagonist" as used herein refers to a molecule that is -10 active or expressed. Inhibition of PAR2 The term "antibody" as used herein refers to a [globulin] molecule consisting of four polypeptide chains and two heavy chains (8) and two light chains (L) interconnected by a two-protective bond. Each heavy chain consists of a heavy chain variable region (abbreviated as or VH) and a heavy chain invariant region. The heavy chain constant region consists of three domains cm, CH2 and CH3. Each light chain consists of a light chain variable region (written as LCVR or Vl) and a light chain invariant region. Light chain constant area = a domain CL composition. The VH and VL regions can be further divided into hypervariable regions called complementary ruling regions (CDRs), where the scatter is better preserved.
區（FR)的區域。每個VH和VL由三個CDR和四個FR 組成，按下面的從胺基酸末端到羧基末端的次序排列： FR1、CDR1、FR2、CDR2、FR3、CDR3、FR4。 中和」或「阻斷」抗體係指與PAR2結合造成街PAR2 生物活性抑制的抗體。這一對PAR2的生物活性抑制可透過 測篁一個或多個PAR2生物活性指標評估。這些pAR2生物 201002345 活性指標可藉由此技術中已知的一個或多個標準的體外或 體内分析來評估（見下面的實例）。 此處所用的術語一個抗體的「抗原結合部分」（或簡稱 「抗體部分」或「抗體片段」）係指一個或多個保留特異結 5 合到一個抗原（如PAR2)上的能力的抗體片段。業已證明， 一個抗體的抗原結合功能可由全長抗體的片段來完成。「抗 原結合部分」這一術語所包含的結合片段的實例包括⑴一 個Fab片段，即一個由VL、VH、CL和CH1域組成的單價 片段；（ii) 一個F(ab')2片段，即一個由鉸鏈區的一個二硫化 ίο 物鍵連接的兩個Fab片段組成的二價片段；（iii) 一個由VH 和CH1域組成的Fd片段；（iv)—個由一個抗體的單臂VL 和VH域組成的Fv片段；（v) —個由VH域組成的dAb片 段（Ward et al. (1989) Nature 241:544-546);以及一個孤立的 CDR。此外，儘管Fv片段的兩個域VL和VH是由不同的 15 基因編碼的，它們可以透過重組方法，由一個合成的連接劑 連接在一起成為一個單獨的蛋白質鏈，其中VL和VH區配 對形成單價分子（稱為單鏈Fv (scFv);參見例如Bird et al. (1988) Science 242:423-426; *Hustonetal.(1988)Proc. Natl. Acad. Sci. USA 85:5879-5883 )。這樣的單鏈抗體也被包 20 含在一個抗體的「抗原結合部分」的範圍以内。其他形式的 單鏈抗體’如雙特異抗體也包括在内。雙特異抗體是二價雙 特異抗體，其中VH和VL域表現在一個肽單鏈上，但使用 一個太短以致不允許同一鏈上的兩個域配對的連接劑，從而 迫使這兩個域與另一個鏈上的互補域配對並形成兩個抗原 8 201002345 結合點（參見例如 Holliger et al. (1993) Proc. Natl. Acad Sci. USA 90:6444-6448; Poljak et al. (1994) Structure 2:1121-1123)。 此處所用的術語「人類抗體」意在包括得自人類種系免 5 疫球蛋白序列可變和恆定區域的抗體。本發明的人類抗體可 能包括未被人類種系免疫球蛋白序列編碼的胺基酸殘基（例 如由體外隨機或點特異變異或體内體細胞突變引入的變 異），如CDR，尤其是CDR3。但是，此處所用的術語「人 類抗體」無意包括CDR序列得自另外的哺乳動物物種，如 10 小鼠’然後植入到人類框架序列的抗體。 術口吾療效1」係指能夠達到陳述的特定目的之PAR2 拮抗劑的濃度或量。例如，一個抗_伙尺2抗體或其抗體的抗 原結合片段的「療效量」可由經驗確定。此外，「有效治療 里」係指能夠達到陳述的治療效果的pAR2拮抗劑，如一個 15 抗-PAR2抗體或抗原結合片段的濃度或量。這個量也可由經 驗確定。 醫藥組成物和治療用途 蘿與提供了—種包含如上所述的人類PAR2拮抗劑禾 =可接受的载體之醫藥組錢。由本發明的腦拮拍 :盥分組成的醫藥組成物可用作預防和/或治療-'、 有關的疾病或病症，如炎症性疼痛的藥劑。 形劑= 將與結ί到調物中的適當載體， '、 起#又與，以提供改善的轉移、遞送和而: 20 201002345 受度等。多種適當調配物可在所有藥劑化學家所熟知的雷氏 藥學大全（Remington's Pharmaceutical Sciences (15th ed. Mack Publishing Company, Easton，PA)中找到。這些調配物 包括，如粉劑、糊劑、油膏、凝膠、蠟劑、油劑、脂類、含 5 有泡囊的脂（陽離子或陰離子）、DNA偶聯物、無水吸收性 糊劑、水包油或油包水乳劑、乳膠狀碳蠟（各種分子量的聚 乙二醇）、半固體狀凝膠以及含有聚乙二醇的半固體狀混合 物。任何上述混合物都可適用於本發明的治療或療法，前提 是調配物中的活性成分不得為調配物所滅活，且調配物在生 10 理上與施用路徑相容並具有耐受性。有關藥物化學家熟悉的 賦形劑和載體的其他資訊，請參見p〇well et al.Area of the area (FR). Each VH and VL consists of three CDRs and four FRs, arranged in the order from amino acid end to carboxy terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. A neutralizing or "blocking" anti-system refers to an antibody that binds to PAR2 and causes inhibition of the biological activity of PAR2. This inhibition of biological activity of PAR2 can be assessed by measuring one or more PAR2 biological activity indicators. These pAR2 organism 201002345 activity indicators can be assessed by one or more standard in vitro or in vivo assays known in the art (see examples below). The term "antigen-binding portion" (or simply "antibody portion" or "antibody fragment") of an antibody as used herein refers to one or more antibody fragments that retain the ability to bind to an antigen (eg, PAR2). . It has been demonstrated that the antigen binding function of an antibody can be accomplished by fragments of full length antibodies. Examples of the binding fragment encompassed by the term "antigen-binding portion" include (1) a Fab fragment, a monovalent fragment consisting of VL, VH, CL and CH1 domains; (ii) an F(ab')2 fragment, ie a bivalent fragment consisting of two Fab fragments joined by a disulfide linkage of the hinge region; (iii) an Fd fragment consisting of a VH and CH1 domain; (iv) a one-arm VL of an antibody An Fv fragment consisting of a VH domain; (v) a dAb fragment consisting of a VH domain (Ward et al. (1989) Nature 241:544-546); and an isolated CDR. Furthermore, although the two domains VL and VH of the Fv fragment are encoded by different 15 genes, they can be joined together by a synthetic linker to form a single protein chain, in which the VL and VH regions are paired. Monovalent molecules (referred to as single-chain Fv (scFv); see, eg, Bird et al. (1988) Science 242: 423-426; * Hustonetal. (1988) Proc. Natl. Acad. Sci. USA 85: 5879-5883). Such a single-chain antibody is also contained within the range of the "antigen-binding portion" of one antibody. Other forms of single chain antibodies such as bispecific antibodies are also included. A bispecific antibody is a bivalent, bispecific antibody in which the VH and VL domains are expressed on a single peptide chain, but using a linker that is too short to allow pairing of the two domains on the same chain, thereby forcing the two domains to The complementary domains on the other strand pair and form two antigens 8 201002345 binding sites (see, eg, Holliger et al. (1993) Proc. Natl. Acad Sci. USA 90:6444-6448; Poljak et al. (1994) Structure 2 :1121-1123). The term "human antibody" as used herein is intended to include antibodies derived from variable and constant regions of the human germline. Human antibodies of the invention may include amino acid residues that are not encoded by human germline immunoglobulin sequences (e.g., variants introduced by random or point-specific mutations in vitro or in vivo somatic mutations), such as CDRs, particularly CDR3. However, the term "human antibody" as used herein is not intended to include antibodies whose CDR sequences are obtained from another mammalian species, such as 10 mice' and then implanted into human framework sequences. Oral efficacy 1" refers to the concentration or amount of a PAR2 antagonist that is capable of achieving the stated purpose. For example, the "effect amount" of an anti-binding antibody of an anti-stick 2 antibody or an antibody thereof can be determined empirically. Further, "effective treatment" refers to a concentration or amount of a pAR2 antagonist, such as a 15 anti-PAR2 antibody or antigen-binding fragment, capable of achieving the stated therapeutic effect. This amount can also be determined empirically. Pharmaceutical Compositions and Therapeutic Uses Lori provides a pharmaceutical group containing a human PAR2 antagonist as described above = an acceptable carrier. The brain composition consisting of the brain of the present invention can be used as a medicament for preventing and/or treating -', related diseases or conditions such as inflammatory pain. The agent = will be combined with the appropriate carrier, ', and #, to provide improved transfer, delivery and while: 20 201002345 degree and so on. A variety of suitable formulations can be found in Remington's Pharmaceutical Sciences (15th ed. Mack Publishing Company, Easton, PA), which is well known to all pharmaceutical chemists. These formulations include, for example, powders, pastes, ointments, Gels, waxes, oils, lipids, lipids containing 5 vesicles (cationic or anionic), DNA conjugates, anhydrous absorbent pastes, oil-in-water or water-in-oil emulsions, latex-like carbon waxes ( Various molecular weight polyethylene glycols), semi-solid gels, and semi-solid mixtures containing polyethylene glycol. Any of the above mixtures may be suitable for use in the treatment or therapy of the present invention, provided that the active ingredient in the formulation is not The formulation is inactivated and the formulation is compatible and tolerant in the route of administration. For additional information on excipients and carriers familiar to pharmaceutical chemists, see p〇well et al.
Compendium of excipients for parenteral formulations”（非腸 道調配物賦形劑便覽）PDA，1998,】pharm Sci Techn〇i 52.238-311以及其中引用的文獻。 15 劑量隨被投與的受試者年齡和體格大小、目標疾病、病 症、使用路徑等不同可有變化。當]?八112拮抗劑用 來預防和 /或^台療諸如成人患者的炎症性疼痛時，通常用單劑量靜脈 給藥較有利，劑量為每公斤體重大約〇 〇1到大約2〇毫克， 較佳的是每公斤體重大約〇丨到大约1〇毫克，更佳的是每 2。 公,體重大約(U到大約5毫克，每天大約i到5次，較佳 的疋大約1到3次。在其他非腸道施用和口服施用中，該梓 抗劑可按照上所給的劑量的相應射投與。#病症特別嚴^ 時’可根據病情增加劑量。 已知有各種藥物遞送系統可用來施用本發明的醫藥紐 10 201002345 成物，例如包裹在能夠表現變異病毒、受體介導的胞吞作用 的脂質體、微顆粒、微膠囊、重組細胞中（參見如WuandWu, 1987, J. Biol. Chem. 262:4429 4432)。藥物引入的方法包括 但不限於皮内、肌肉内、腹膜内、靜脈、皮下、鼻内、硬腦 5 膜，和口腔專途徑。該醫藥組成物可採用任何方便的路徑施 用，如灌注或靜脈團注，透過上皮和黏膜襯層（如口腔粘膜、 直腸和小腸粘膜）並可與其他生物活性藥劑一起施用。施用 可為全身性或局部施用。 在另一個具體實施例中，該醫藥組成物可用小囊，尤其 ίο 疋月曰質體遞送（參見如Langer, 1990，Science 249:1527-1533 )。在另一個具體實施例中，該醫藥組成物可 以一個緩釋系統遞送。在一個具體實施例中，可使用一個泵 (參見 Langer, supra; Sefton (1987) CRC Crit. Ref. Biomed.Compendium of excipients for parenteral formulations PDA, 1998, pharm Sci Techn〇i 52.238-311 and references cited therein. 15 Dosage with subject age and physique of the subject being administered The size, target disease, condition, route of use, etc. may vary. When the 8112 antagonist is used to prevent and/or treat inflammatory pain such as an adult patient, it is usually advantageous to administer a single dose intravenously. The dosage is from about 1 to about 2 mg per kg of body weight, preferably about 1 mg per kg of body weight, more preferably about 2. Male, body weight (U to about 5 mg per day) About i to 5 times, preferably about 1 to 3 times. In other parenteral and oral administrations, the antagonist can be administered in accordance with the dose given above. 'The dose can be increased according to the condition. Various drug delivery systems are known for administering the drug of the invention, such as liposomes, microparticles capable of exhibiting variant viruses, receptor-mediated endocytosis. , microcapsules, recombinant cells (see, eg, WuandWu, 1987, J. Biol. Chem. 262: 4429 4432). Methods of drug introduction include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, The hard brain 5 membrane, and the oral route. The pharmaceutical composition can be administered by any convenient route, such as perfusion or intravenous bolus, through the epithelium and mucosal lining (such as the oral mucosa, rectum and small intestine mucosa) and with other organisms. The active agent is administered together. Administration can be systemic or topical. In another embodiment, the pharmaceutical composition can be delivered by a small sac, particularly 疋 疋 曰 ( (see, eg, Langer, 1990, Science 249: 1527- 1533). In another embodiment, the pharmaceutical composition can be delivered in a sustained release system. In one embodiment, a pump can be used (see Langer, supra; Sefton (1987) CRC Crit. Ref. Biomed.
Eng. 14:201)。在另一個實施例中，可使用聚合物（參見如 15 Medical Applications of Controlled Release (受控釋放的醫 學應用）（1974) Langer and Wise (eds.), CRC Pres” Boca Raton, Florida; Controlled Drug Bioavailability (受控的藥物 生物可用度），Drug Product Design and Performance, (1984) Smolen and Ball (eds.)，Wiley，New York)。在還有一個實施 20 例中，可在該醫藥組成物的目標處放置一個受控釋放系統， 因此僅需要全身劑量的一部分（參見如Goodson, in Medical Applications of Controlled Release (1984) supra, vol. 2, pp. 115-138)。其他受控釋放系統的討論見Langer (1990, Science 249:1527-1533)。 11 201002345 口腔施用該組成物之實例包括固體或液體劑量形式，具 體來說有片劑（包括糖衣丸和薄膜包衣片劑）、丸劑、顆粒、 粉劑、膠囊（包括軟膠囊）、糖漿、乳劑、懸浮劑等。這樣 的組合物是以公開的方法製造的並包含製藥領域常用的一 5 種載體、一種稀釋劑或一種賦形劑。對片劑來說，該載體或 賦形劑的實例有乳糖、澱粉、硬脂酸鎂等。 注射劑可包括靜脈、皮下、皮内和肌肉注射劑、滴注等。 這些注射製劑可用公開的方法製備。注射製劑例如可用常規 製備注射劑的方法將拮抗劑或其鹽溶於、懸浮於或乳化在無 10 菌水介質或油介質中來製備。注射劑的水介質有，例如生理 鹽水、一個含有葡萄糖和其他輔助成分的等滲溶液，可和一 個適當的增溶劑如醇（如乙醇）、多元醇（如丙二醇、聚乙 二醇）、非離子表面活性劑[如聚山梨醇酯80、氫化蓖麻油的 HCO-50 (聚氧乙烯（50莫耳）加成物）]等結合使用。所用 15 的油介質有，如芝麻油、大豆油，可和一個增溶劑如苯曱酸 苄酯、苯曱醇結合使用。這樣製備的注射劑最好封裝在一個 適當的安瓿瓶中。用於直腸施用的栓劑可將上述拮抗劑分子 或其鹽與常規栓劑基質混合而製備。 有利的是，上述用於口服或經腸施用的該醫藥組成物可 20 製備成一個能容納一劑活性成份的單位劑量形式。這樣的單 位劑量形式包括，如片劑、丸劑、膠囊、注射劑（安瓿）、 栓劑等。例如，每個單位劑量形式，尤其是注射劑，一般包 含5到500毫克抗-PAR2抗體或抗體片段，其他劑量形式首 選的是包含大約5到100毫克和大約10到250毫克的上述 12 201002345 抗體。 如Λ所-述，本發明進一步提供了 -種預防或治療與 相關或涉及PAR2的疾錢病症㈣法，該方法包括 ^而要的Μ者投與-枝量的種域PAR2括抗 劑。 治療適應难_ t明提供其巾PAR2為對需衫㈣钱者很有用的 Γ「II。例如，該方法能夠治療、預防、治癒、緩解、改 10 15 種疾病、障礙、病症或病（「病症」）。根據 3明二台，症的特徵是隱2不適當的表現和活性的 3二火^2拮抗劑治療的具體的醫療狀況或疾病包括 ，和包括胃潰瘍和十二指腸潰瘍；氣道的炎症性病 慢性阻塞性肺病等等；風濕病，包括諸如成人 =二：祕關節炎、硬皮病;系統性紅斑狼瘡;痛風; 月關即火，^濕性多肌痛；血清陰性脊柱關節病，包括強直 !·生脊柱k和萊特爾氏病、銀屑病關節炎和慢性萊姆關節炎； 斯提爾病和與類風濕性關節炎有關的眼色素層炎；因隨意肌 =其他肌肉的發炎而造成的疾病，包括皮肌炎、包涵體肌 炎、多肌炎和淋巴血管肌脂瘤。其他適應症包括諸如乳癌 (Adriaenssens et al. (2008) Cancer Res 68:346-51 )。 在一個更特定的治療用途中，抗-PAR2抗體或抗體片段 對治療與各種病症相關的疼痛很有用，包括癌症疼痛、關節 13 20 201002345 炎疼痛、糖尿病神經病變、帶狀皰疹神經痛、偏頭痛。 組合治療 在本發明治療方法的特定具體實施例中’患有與痛風有 關的關節痛的受試者係以拮抗劑如抗-PAR2抗體或抗體片 段以及第二治療藥劑的組合治療。該第二治療藥劑較佳的是 白細胞介素-1 (IL-1)拮抗劑，如 rilonacept (“IL-lTrap”; Regeneron Pharmaceuticals, Inc.)。適當的第二治療藥劑可為 選自由下列所組成之群組的一個或多個藥劑：ril〇nacept、阿 那白滯素、一個從人類正-丨受體拮抗劑（IL1Ra)非糖基化 10 的重組子、一個抗-IL-1抗體或一個抗 IL_18藥物，如 IL-18BP或衍生物、一個IL_18酸性磷酸酶、抗_凡_18、抗_ IL-18R1或抗-IL-18Racp。可與NGF抗體或抗體片段及另外 的適當治療劑組合使用的其他組合治療包括，例如低劑量秋 水仙鹼、阿司匹林、類固醇，如醋酸潑尼松、氨甲喋呤，低 劑量環孢黴素A、TNF抑制劑、其他炎症性抑制劑^:半胱 氨酸蛋白酶-1、P38、IKK1/2、CTLA-4Ig、抗_比_6 或抗_IL6Ra 等’和/或共治療藥物’如尿酸合成抑制劑來抑制尿酸在體 内的積聚’如射呤醇、尿酸分泌啟動子來加速體内積聚的 尿酸的分泌，例如，羧苯雜、笨0叫/或苯漠馬 尿酸分泌啟動子的實例；皮質類固醇，以 、 炎藥物（NSMD)。 Λ及其他非類固醇抗 在各種具體貫施例中，組合治療包括# 所述的兩種、三種、四種、五種、六種：試者投與此處 更多種PA T? 9 :略 動劑或拮抗劑。當使用很多分子或治療昧β 1 π M t ’卓個分子或治療 14 20 201002345 可以任何次序、Eng. 14:201). In another embodiment, a polymer can be used (see, eg, 15 Medical Applications of Controlled Release (1974) Langer and Wise (eds.), CRC Pres" Boca Raton, Florida; Controlled Drug Bioavailability (controlled drug bioavailability), Drug Product Design and Performance, (1984) Smolen and Ball (eds.), Wiley, New York). In another implementation of 20 cases, the target of the pharmaceutical composition is available. A controlled release system is placed, so only a portion of the systemic dose is required (see, eg, Goodson, in Medical Applications of Controlled Release (1984) supra, vol. 2, pp. 115-138). For a discussion of other controlled release systems, see Langer (1990, Science 249: 1527-1533). 11 201002345 Examples of oral administration of the composition include solid or liquid dosage forms, in particular tablets (including dragees and film-coated tablets), pills, granules, powders , capsules (including soft capsules), syrups, emulsions, suspensions, etc. Such compositions are manufactured by published methods and are commonly used in the pharmaceutical field. A carrier, a diluent or an excipient. Examples of the carrier or excipient are lactose, starch, magnesium stearate, etc. The injection may include intravenous, subcutaneous, intradermal and Intramuscular injection, instillation, etc. These injection preparations can be prepared by a publicly available method. The injection preparation can be prepared, for example, by dissolving, suspending or emulsifying the antagonist or its salt in a 10-free aqueous medium or an oil medium by a conventional method for preparing an injection. The aqueous medium for injection is, for example, physiological saline, an isotonic solution containing glucose and other auxiliary ingredients, and a suitable solubilizing agent such as an alcohol (e.g., ethanol), a polyhydric alcohol (e.g., propylene glycol, polyethylene glycol), and a non-aqueous solution. Ionic surfactants [such as polysorbate 80, hydrogenated castor oil HCO-50 (polyoxyethylene (50 mol) adduct)], etc. are used in combination. The oil medium used is 15, such as sesame oil, soybean oil, It can be used in combination with a solubilizing agent such as benzyl benzoate or benzofuran. The injection prepared in this way is preferably packaged in a suitable ampoule. The suppository for rectal administration can be sterilized. Molecules or its salt with conventional suppository bases prepared by mixing. Advantageously, the above-described pharmaceutical composition for oral or parenteral administration may be prepared as a 20 to accommodate an active ingredient in unit dosage form. Such unit dosage forms include, for example, tablets, pills, capsules, injections (ampoules), suppositories, and the like. For example, each unit dosage form, especially an injection, will generally contain from 5 to 500 mg of anti-PAR2 antibody or antibody fragment, and other dosage forms will preferably comprise from about 5 to 100 mg and from about 10 to 250 mg of the above 12 201002345 antibody. As described in the above, the present invention further provides a method for preventing or treating a disease associated with or involving PAR2 (IV), which method comprises the administration of a seed-area PAR2 antagonist. Treatment adaptation is difficult _ t Ming provides its towel PAR2 is useful for those who need shirts (4). For example, this method can treat, prevent, cure, alleviate, change 10 diseases, disorders, illnesses or diseases (" Illness"). According to 3 Ming 2, the symptoms are characterized by inappropriate 2 inappropriate performance and activity of the 3 2 fire ^ 2 antagonist treatment of specific medical conditions or diseases including, and including gastric ulcer and duodenal ulcer; airway inflammatory disease chronic obstructive Pulmonary diseases, etc.; rheumatism, including such as adult = two: secret arthritis, scleroderma; systemic lupus erythematosus; gout; moon off is fire, ^ wet polymyalgia; seronegative spondyloarthropathy, including rigidity! Spine k and Lyttle's disease, psoriatic arthritis and chronic Lyme arthritis; Styrian disease and uveitis associated with rheumatoid arthritis; caused by voluntary muscles = inflammation of other muscles Diseases, including dermatomyositis, inclusion body myositis, polymyositis, and lymphatic angiomyolipoma. Other indications include, for example, breast cancer (Adriaenssens et al. (2008) Cancer Res 68:346-51). In a more specific therapeutic use, anti-PAR2 antibodies or antibody fragments are useful for treating pain associated with a variety of conditions, including cancer pain, joints, 13 20 201002345, inflammatory pain, diabetic neuropathy, herpes zoster neuralgia, partial headache. Combination Therapy In a particular embodiment of the method of treatment of the invention, a subject having joint pain associated with gout is treated with a combination of an antagonist such as an anti-PAR2 antibody or antibody fragment and a second therapeutic agent. The second therapeutic agent is preferably an interleukin-1 (IL-1) antagonist such as rilonacept ("IL-lTrap"; Regeneron Pharmaceuticals, Inc.). A suitable second therapeutic agent can be one or more agents selected from the group consisting of ril〇nacept, anakinra, and a non-glycosylation from a human ortho-receptor antagonist (IL1Ra). A recombinant of 10, an anti-IL-1 antibody or an anti-IL_18 drug, such as IL-18BP or a derivative, an IL-18 acid phosphatase, an anti-IL18, an anti-IL-18R1 or an anti-IL-18 Racp. Other combination therapies that can be used in combination with NGF antibodies or antibody fragments and additional suitable therapeutic agents include, for example, low doses of colchicine, aspirin, steroids such as prednisone acetate, methotrexate, low dose cyclosporine A, TNF inhibition Agents, other inflammatory inhibitors ^: caspase-1, P38, IKK1/2, CTLA-4Ig, anti-specific ratio _6 or anti-IL6Ra, etc. and / or co-therapeutic drugs such as uric acid synthesis inhibitors To inhibit the accumulation of uric acid in the body 'such as sterol, uric acid secretion promoter to accelerate the accumulation of uric acid in the body, for example, examples of carboxybenzene, stupid 0 or benzoic acid secretion promoter; cortex Steroids, inflammatory drugs (NSMD). Λ and other non-steroidal anti-inhibitions In various specific embodiments, the combination therapy includes two, three, four, five, and six as described in #: The tester casts more PA T? 9 : Agent or antagonist. When using many molecules or treating 昧β 1 π M t ‘one molecule or treatment 14 20 201002345 can be in any order,
一種分子治療，a molecular treatment,
【實施方式】 、在任何有效時長内施用’例如同時、順次、 ，治療方法為，在第一個療程用 ’或在其他治療完成後，才開始進行第二個療 第二個療程開始之間的時間長 &何時長’例如’數秒、數分鐘、 10 實例L神經性疼痛的治療。 PA R 2被發現能夠增加疼痛傳播並介導腸易激症候群的 内臟疼痛。PAR2在幾個組織中表現，包括疼痛初級傳入神 經元。PAR2在這些感覺神經元中的表現與兩個已知的疼痛 感受調諧子，降鈣素基因相關的肽（CGRP)和p物質（sp) 15 位於相同位置。PAR2激動劑已被證明能夠刺激SP和CGRp 在组織中的釋放’誘發長時間的熱敏疼痛和機械性觸誘發疼 痛，並升高脊柱FOS蛋白質（一個確立的疼痛感受神經元 啟動標記物）。 進行了實驗來測試PAR-2對神經痛的抑制效果。對 20 C57BL/6雄性小鼠進行塞爾托玆（Seltzer)神經痛模型和一個 抗-PAR-2抗體進行實驗。透過用7-0絲缝合線繞每只小鼠— 條腿的髖部神經大約1/3到1/2直徑紮一個很緊的缚線，該 神經痛模型造成了部分神經受傷。手術後’讓小鼠恢復至少 兩天，然後對它們進行至少幾個星期的手術後哈格裏夫斯 15 201002345 (Hargreaves)熱敏疼痛檢驗和von Frey機械性觸誘發疼痛檢 驗研究。在塞爾托玆手術後，在朮後第4天和第7天對小鼠 進行試驗，以便確認已經產生了疼痛。然後，手術後第7天 在進行了行為疼痛測試後，給小鼠皮下注射了抗-PAR2抗 體（50mg/kg; Molino et al. (1998) Arterioscler Thromb Vase Biol 18:825-832或IgG陰性對照抗體g〇〇 mg/kg))。測試結 果列於圖1和圖2中。 【圖式簡單說明】 圖1.哈格裏夫斯檢驗結果。人=1〇〇 mg/kg IgG對照 mAb (n-6);〇=50 mg/kg 抗-PAR2 mAb (n=5); 2(治療）X Π(時間）帶有Bonferroni事後比較檢驗的混合因數 ANOVA;治療效果顯著，p=〇 〇34〇 (* p<〇 〇5; *** p<〇 〇〇1 對 IgG對照）。 圖 2.von Frey 檢驗結果。λ = 1〇〇 mg/kg IgG 對照 mAb (n-6);〇=50 mg/kg 抗-PAR2 mAb (n=6); 2(治療）X ι〇(時間） 帶有Bonferroni事後比較檢驗的混合因數an〇VA ;治療效 果顯著，p=〇.〇〇57。 【主要元件符號說明】 16[Embodiment], in any effective duration, 'for example, at the same time, sequentially, the treatment method is, after the first treatment period' or after other treatments are completed, the second treatment begins. The time between long & when to grow 'for example' seconds, minutes, 10 cases of L neuropathic pain treatment. PA R 2 was found to increase pain transmission and mediate visceral pain in irritable bowel syndrome. PAR2 is expressed in several tissues, including the primary afferent neuron of pain. The expression of PAR2 in these sensory neurons is located at the same position as the two known pain-sensing tuners, the calcitonin gene-related peptide (CGRP) and the p-substance (sp) 15. PAR2 agonists have been shown to stimulate the release of SP and CGRp in tissues' induced long-term heat-sensitive pain and mechanically induced pain, and elevated spinal FOS protein (an established pain-sensitivity neuron initiation marker) . Experiments were conducted to test the inhibitory effect of PAR-2 on neuralgia. Experiments were performed on a 20 C57BL/6 male mouse with a Seltzer neuralgia model and an anti-PAR-2 antibody. The neuralgia model caused partial nerve injury by using a 7-0 silk suture to wrap a very tight line around each mouse-legged hip nerve about 1/3 to 1/2 of the diameter. After surgery, the mice were allowed to recover for at least two days, and then they were subjected to at least several weeks of surgery after Hargreaves 15 201002345 (Hargreaves) thermal pain test and von Frey mechanical tactile pain test. After the Sertoz operation, the mice were tested on the 4th and 7th postoperative days to confirm that pain had developed. Then, after performing a behavioral pain test on the 7th day after surgery, the mice were injected subcutaneously with anti-PAR2 antibody (50 mg/kg; Molino et al. (1998) Arterioscler Thromb Vase Biol 18: 825-832 or IgG negative control Antibody g〇〇mg/kg)). The test results are shown in Figures 1 and 2. [Simple description of the diagram] Figure 1. Hargreaves test results. Human = 1 mg / kg IgG control mAb (n-6); 〇 = 50 mg / kg anti-PAR2 mAb (n = 5); 2 (treatment) X Π (time) with Bonferroni post hoc comparison test mixture Factor ANOVA; therapeutic effect was significant, p = 〇〇 34 〇 (* p < 〇〇 5; *** p < 〇〇〇 1 vs IgG control). Figure 2. von Frey test results. λ = 1〇〇mg/kg IgG control mAb (n-6); 〇=50 mg/kg anti-PAR2 mAb (n=6); 2 (treatment) X ι〇 (time) with Bonferroni post hoc comparison test The mixing factor is 〇VA; the treatment effect is remarkable, p=〇.〇〇57. [Main component symbol description] 16