200407153 玖、發明說明： 技術領域 本發明係有關使用a)環鳥嘌呤核甞單磷酸（cGMP)之特 有的磷酸二酯酶第5型（PDE5)抑制劑及b)血管收縮素轉化 酶（ACE)抑制劑之組合，以治療心血管及代謝疾病，特別 是高血壓。 先前技術 血壓（B P)係由一些單獨或合併的血液動力參數所定義。 心臟收縮血壓（systolic blood pressure, SBP)為心臟收縮時 所得到的尖峰動脈血壓。心臟舒張血壓（diastolic Mood pressure，DBP)為心臟放鬆時所得到的最小動脈血壓。SBP 與DBP之間的差係定義為脈搏壓（pulse pressure，PP)。 高血壓或升高的BP已定義為至少為140 mmHg的SBP及/ 或至少為90 mmHg的DBP。根據此定義，已開發國家中高 血壓的盛行率為成年人口的大約20%，攀升至60歲或更高 齡者的大約60至70%，然而這些高血壓患者大部分在非臨 床環境測量時具有正常的B P。6 0 %較年長的高血壓人口部 分具有獨立性收縮性高血壓（ISH)，換言之，他們具有升高 的SBP和正常的DBP。而高血壓則與中風、心肌梗塞、心 房微顫、心臟衰竭、末梢血管疾病和腎功能障礙的風險增 加有關（Fagard，RH; Am. J. Geriatric Cardiology 11(1)， 23-28，2002; Brown, MJ and Haycock，S; Drugs 59(Suppl 2)， 1-12, 2000)。 高血壓之病態生理學為持續討論的主題。雖然一般均贊 85169.doc 200407153 同高血壓為心臟輸出量和末梢血管阻力之 果，但是大部分的高血壓患者具有異常的心：的結 加的末梢阻力，因此並不確定要先改變：及增 (B⑽⑽，G等人；BMJ 322,9l2 9i6，扇… 固參數 儘管在各種藥理分類上可取得大量的藥物 劑、α·腎上腺素性拮㈣m腺素性指 尿 道阻斷劑、血管收縮辛鍊仆陆^以、 ^、句通 ，.猫素轉化酶抑制劑和血管收縮 抗劑，但仍無法滿足對於有效治療高血壓之要求。t 可阻斷腎素-血管收縮素-酸固嗣系統之血管 ACE抑制劑係推萬作A古 、力作的 效"視為= 一線治療。該抑制劑有 性。最常見的副作用，據報導有 括二 發生咳嗽。其他較不常報導的副作用包 括珍、血管性水腫、高血_症和功能性腎衰竭。 二::酸二酿酶第5型為環^票呤核誓單餐之特有的磷酸 二二ΓΓ抑制劑會降低eGMP的水解速率，因此能使 : 用。已發現此等抑制劑可用於治療男性*** 功把不良0 發明内容 一 Λ樣，本發明係提供使用包括a)pDE5抑制劑及 卩制心組合，製造用以治療疾病（特収心血管及 代谢疾病，更詳言之為高血壓）之藥劑。 實施方式 :本文中所使用之術語，“治療，，包括姑息性治療、根治 ’《療和預防性治療。“高血壓”包括特徵為超常態血壓的 85169.doc 200407153 所有疾病，例如原發性高血壓、肺動脈高血壓、繼發性高 血壓、收縮性高血壓、與糖尿病有關之高血壓、與 動脈粥:大硬化有關之高血壓和腎血管性高血壓以及進二 步擴展至已知升高血壓為風險因素的情況。因此，“治療高 血壓”包括治療或預防由高血壓引起的併發症和其他有： 的共病現象，包括鬱血性心臟衰竭、绞魅、中風、青光 艮及^力此不良（包括腎衰竭）。代謝疾病特別包括代謝症 候群（亦已知為X综合症）、糖尿病及葡萄糖耐受不良，包括 其併發症，例如糖尿病性視網膜病變和糖尿病性神經病 變。 乂下PDE5抑制劑及ACE抑制劑之組合，包含特有的 PDE5抑制劑及特有的織抑制劑之組合，稱之為°本發明之 組合。 j發明之組合比單獨使用pDE5抑制劑或ace抑制劑户 療高血壓時，具有更有效、更強勁、較低毒性或具有其: 更多期望性質的優點。 以下PDE5抑制劑”表示用於本發明之pDE5抑制劑，包 括PDE5抑制劑的所有醫藥可容許之鹽、溶劑合物和二素= 形體。同樣地’ “ACE抑制劑”表示用於本發明之ace抑= 劑，包括ACE抑制劑的所有醫藥可容許之鹽、溶劑合物和 同素異形體。 PDE5抑制劑及ACE抑制劑之合適度可利用文獻方去， 估其藥效和選擇性，接著根據標準藥劑實務評估其毒性砰 樂物動力學（吸收、代謝、分布和***），等輕易地加以測 85169.doc 200407153 定。適合的化合物為具有藥效和選擇性，在治療劑量下沒 有顯著毒性作用，且較佳為口服以後具有生物有效性者。 可將藥效定義為ICm值，其為抑制5〇%之酵素活性所需 要的化合物濃度。可使用後文所述之PDE5測定法，測$ PDE5抑制劑的！(:⑼值。該PDE5抑制劑對於？£^5酶較佳具 有低於1 00 nM，更佳為低於50 nM的IC5〇值。200407153 发明 Description of the invention: TECHNICAL FIELD The present invention relates to the use of a) a phosphoguanylate nuclear phosphonium monophosphate (cGMP) -specific phosphodiesterase type 5 (PDE5) inhibitor and b) angiotensin converting enzyme (ACE). A combination of inhibitors to treat cardiovascular and metabolic diseases, especially hypertension. Prior art Blood pressure (BP) is defined by a number of individual or combined hemodynamic parameters. The systolic blood pressure (SBP) is the peak arterial blood pressure obtained when the heart contracts. Diastolic Mood pressure (DBP) is the minimum arterial blood pressure obtained when the heart relaxes. The difference between SBP and DBP is defined as pulse pressure (PP). Hypertension or elevated BP has been defined as SBP of at least 140 mmHg and / or DBP of at least 90 mmHg. According to this definition, the prevalence of hypertension in developed countries is about 20% of the adult population and climbs to about 60 to 70% of those aged 60 or older. However, most of these patients with hypertension have normal conditions when measured in nonclinical settings. BP. 60% of the older hypertensive population have independent systolic hypertension (ISH), in other words, they have elevated SBP and normal DBP. Hypertension is associated with an increased risk of stroke, myocardial infarction, atrial microfibrillation, heart failure, peripheral vascular disease, and renal dysfunction (Fagard, RH; Am. J. Geriatric Cardiology 11 (1), 23-28, 2002; Brown, MJ and Haycock, S; Drugs 59 (Suppl 2), 1-12, 2000). The pathophysiology of hypertension is the subject of ongoing discussion. Although 85169.doc 200407153 and hypertension are generally the result of cardiac output and peripheral vascular resistance, most patients with hypertension have abnormal heart: combined with peripheral resistance, so they are not sure to change first: and Zeng (B⑽⑽, G et al .; BMJ 322, 9l2 9i6, fan ... Solid parameters Although a large number of medicinal agents can be obtained in various pharmacological classifications, α · adrenergic antagonist m adrenergic refers to urethral blocker, vasoconstriction chain chain 陆 ^ It can be used to treat feline converting enzyme inhibitors and vasoconstriction agents, but it still cannot meet the requirements for effective treatment of hypertension. T Can block the vascular ACE of the renin-angiotensin-acid fixation system Inhibitors promote the effectiveness of Wanzuo Agu, masterpieces " deemed = first-line treatment. The inhibitor is sexual. The most common side effects, including cough, are reported. Other less frequently reported side effects include rare, vascular Edema, hypertensive disorder, and functional renal failure. 2 :: Acid Digerase Type 5 is a unique dibiphosphate phosphate ΓΓ inhibitor that reduces the hydrolysis rate of eGMP, which can reduce the hydrolysis rate of eGMP. Make use of These inhibitors have been found to be useful in the treatment of male erectile dysfunction. SUMMARY OF THE INVENTION The present invention provides a combination of a) pDE5 inhibitor and control heart, for the manufacture of diseases (specific cardiovascular and metabolic) Disease, and more specifically, hypertension). Embodiments: As used herein, the term "treatment, including palliative care, radical cure" and "prophylactic and preventive treatment." "Hypertension" includes 85169.doc 200407153 all diseases characterized by hypernormal blood pressure, such as primary Hypertension, pulmonary hypertension, secondary hypertension, systolic hypertension, hypertension associated with diabetes, atherosclerosis: hypertension associated with sclerosis and renal vascular hypertension, and further expansion to known rises Hypertension is a risk factor. Therefore, "treatment of hypertension" includes the treatment or prevention of complications caused by hypertension and other comorbidities that include: congestive heart failure, colic, stroke, blue light, and ^ Poor health (including renal failure). Metabolic diseases include, in particular, metabolic syndrome (also known as X syndrome), diabetes, and glucose intolerance, including complications such as diabetic retinopathy and diabetic neuropathy. 乂The combination of the PDE5 inhibitor and the ACE inhibitor includes a combination of a unique PDE5 inhibitor and a unique weaving inhibitor, which is referred to as the present invention. The combination of the invention is more effective, more powerful, less toxic, or has the advantages of: more desirable properties than when pDE5 inhibitor or ace inhibitor is used alone to treat hypertension. The following PDE5 inhibitor "means The pDE5 inhibitors used in the present invention include all pharmaceutically acceptable salts, solvates, and dioxins of the PDE5 inhibitor. Similarly, "ACE inhibitor" means an ace inhibitor used in the present invention, including all pharmaceutically acceptable salts, solvates and allotropes of ACE inhibitors. The appropriateness of PDE5 inhibitors and ACE inhibitors can be evaluated in the literature to evaluate their efficacy and selectivity, and then to evaluate their toxic and bolus kinetics (absorption, metabolism, distribution and excretion) according to standard pharmaceutical practices, etc. Tested at 85169.doc 200407153. Suitable compounds are those that are pharmacologically effective and selective, have no significant toxic effects at therapeutic doses, and are preferably those that are biologically effective after oral administration. The efficacy can be defined as the ICm value, which is the concentration of the compound required to inhibit 50% of the enzyme activity. You can use the PDE5 assay described later to test for PDE5 inhibitors! (: ⑼ value. The PDE5 inhibitor preferably has an IC50 value of less than 100 nM, and more preferably less than 50 nM for a? 5 enzyme.
選擇性比可由熟習此項技術者藉由所關注的特定酶之 對應iCw值的比而輕易地加以測定。pDE3酶和pDE4酶的 ICM值可使用已建立的文獻方法加以測定，參見BaiiardsA 等人；Journal of Urology 159, 2164-2171，1998。 PDE5抑制劑較佳具有對於PDE5酶的選擇性。該抑制劑 較佳具有對於PDE5超過PDE3大於100 ,更佳為大於3〇〇的 選擇性。該PDE5抑制劑更佳具有超過PDE3和pDE4兩者大 於100，更佳為大於3〇〇的選擇性。 該PDE5抑制劑對於pDE5較佳具有低於丨〇〇 nM的lC5〇值 及超過PDE3大於1〇〇倍的選擇性。 口服生物有效性係指能到達全身循環的口服藥比例。決 足藥物之口服生物有效性的因素為溶解作用、膜滲透性及 肝臟清除作用。代表性為首先使用活體外技術，然後使用 活體内技術的篩選串級來測定口服生物有效性。 溶解作用’係藉由胃腸道（GIT)的水分使藥物溶解，可在 適當的pH值下進行模擬GIT的活體外溶解度實驗而加以預 測。PDE5抑制劑較佳具有5〇//§/1111的最小溶解度。溶解度 可藉由此項技術中已知的標準步驟，例如Upinski CA等 85169.doc 200407153 人；Adv· Rug Deliv· Rev· 23屮3)，弘25, 199?所述者來加 以測定。 膜滲透性係指化合物通過GIT的細胞。親脂性為預測該 膜滲透性之關鍵特性，其係利用有機溶劑和緩衝劑藉由活 體外的Log D7.4測量來加以測定。該pDE5抑制劑較佳具有· -2至+4，更佳為]至+3的L〇g D? 4。L〇g 〇可藉由此項技術 中已知的標準步驟，例如Stopher，D*McClean，S;J.Pharm. Pharmacol· 42(2)，144, 1990所述者來加以測定。 貝貝上會加上細胞單層測定法（例如Cac〇2)，以預測在排 出轉運體（例如P-醣蛋白，所謂的Cac〇2通量）存在時的適合 膜滲透性。該PDE5抑制劑較佳具有大於，更 佳為大於5χ 10 6cm.s-1的Caco2通量。Caco2通量值可藉由此 項技術中已知的標準步驟，例如Artursson，P和]viagnusson， C; J.Pharm.Sci，79(7)，595-600, 1990 所述者來加以測定。 代謝穩定性係針對在吸收過程中代謝化合物或是肝臟 在吸收後直接代謝化合物所提供GIT的能力：首度效應。 例如微粒體、肝細胞等測定系統可預測代謝不安定性。該 PDE5抑制劑較佳於測定系統中顯示代謝穩定性，該測定系 統相當於低於〇 · 5的肝抽取物。測定系統與資料操作的實例 見述於Obach，RS ; Curr. Opin. Drug Disc. Devel. 4⑴， 36-44，2001 及 Shibata，Y等人；Dmg Met. Disp. 28(12)， 1518-1523, 2000 。 由於上述步驟的相互作用，因此可藉由在動物中的活體 内實驗，得到藥物具有在人體内之口服生物有效性的進一 85169.doc -10- 200407153 步支持。絕對生物有效性係藉由個別口服該化合物或口服 該化合物之混合物來加以測定。對於絕對測定（％ 口服生物 有效性）而言，也可採用靜脈路徑。動物體内之口服生物有 效性的評估實例可見於Ward，KW等人；Drug Met. Disp. 29(1)，82-87，2001 ; Berman，J等人；J. Med· Chem. 40(6)， 827_829, 1997及 Han KS和 Lee，MG ; Drug Met. Disp. 27(2)， 221-226, 1999 ° 本發明使用的PDE5抑制劑實例為： 歐洲專利申請案第0463756號、第0526004號及已公開之 國際專利申請案第93/06 104號、第98/49166號、第99/543 33 號、第00/24745號、第01/27112號和第01/27 11 3號所揭示的 吡唑并[4,3-d]嘧啶-7-酮類；歐洲專利申請案第0995750 號、第0995 75 1號及已公開之國際專利申請案第9：3/〇7149 號所揭示的吡唑并[3,4-d]嘧啶-4-酮類；已公開之國際專利 申請案第01/1 8004號、第02/00660號及第02/59126號所揭示 的p比唆并[4,3-d]喊違類；已公開之國際專利申請案第 9 3 /12 0 9 5號所揭示的。奎哇淋-4-酮類；已公開之國際專利申 請案第94/05661號所揭示的p比淀并[3，2_d]11密淀_心酮類；歐 洲專利中請案第109271 8號及已公開之國際專利申請案第 94/00453號所揭示的嗓呤嗣類；已公開之國際專利申請 案第95/19978號所揭示的成呼并[2’，1’：6，1]?比淀并[3,4-13] 吲哚二酮類；歐洲專利申請案第1〇92719號及已公開 之國際專利申請案第99/24433號所揭示的味σ坐并 [5 l-f][l 2 4]***-酮類；已公開之國際專利申請案第 85169.doc -11 - 200407153 93/07 124號所揭示的二環化合物& R〇teUa Dp等人；1他(1· Chem· 43(7)，125 7-1263，2000所揭示的咪峻并峻峻4酮 類。 該等已公開之國際專利申請案及期刊論文之内容，特別 是其中申請專利範圍之治療活性化合物的通式及例示化 合物’其全部以引用的方式併入本文中。 本發明使用之PDE5抑制劑的更多實例包括·· 4-溴-5-(吡 啶基甲基胺基）-6-[3-(4-氯苯基）-丙氧基]-3(2H)嗒畊嗣； 1-[4-[(1，3 -冬并一 1:7惡戊-5-基甲基）胺基]-6 -氯-2 -峻峻淋基 (quinozolinyl)]-4-六氫说啶-羧酸，單鈉鹽；（ + )-順 -5,63,7,9,9,9&-六氫-2-[4-(三氟甲基）-苯基甲基-5-甲基-環戊 -4,5]味吨并[2,1-b]嗓吟-4(3H)酮；咬洛西林（furazlocillin); 順-2-己基-5-曱基-3,4,5,6&，7,8,9,9&-八氫環戊[4,5]-味17坐并 [2，1-b]嘌呤-4-酮；3-乙醯基-1-(2·氯苯甲基）-2-丙基峭哚-6-羧酸鹽；3-乙醯基-1-(2-氯苯甲基）-2-丙基⑼哚-6-羧酸鹽； 4-溴-5-(3-吡啶基甲基胺基）-6-(3-(4-氣苯基）-丙氧 基）-3-(2H)嗒畊酮；1-甲基-5-(5-嗎啉基乙醯基-2-正丙氧基 苯基）-3-正丙基-1，6 -二氯-7Η-ρ比吐并（4,3-d)喃淀-7 -嗣； 1-[4-[(1，3-苯并二p惡茂-5-基甲基）胺基]-6-氯-2-ρ奎峻琳 基]-4-六氫峨淀-複酸，單鋼鹽；Pharmaprojects(世界藥物 研究和開發狀況訊息資料庫）第45 1 6號（Glaxo Wellcome); Pharmaprojects 第 5 05 1 號（Bayer) ; Pharmaprojects 第 5 064 號 (Kyowa Hakko ;參見世界專利第96/26940號）； Pharmaprojects 第 5069 號（Schering Plough) ； GF-196960 85169.doc 200407153 (Glaxo Wellcome) ; E-8010和 E-4010 (Eisai) ; Bay-38-3045 和 38-9456 (Bayer)及 Sch-5 1866。 本發明使用之較佳的PDE5抑制劑包括： 5-[2-乙氧基-5-(4-甲基-1-哌畊基磺醯基）苯基]-1-甲基 -3-正丙基-1，6-二氫-7H-吡唑并[4,3-d]嘧啶-7-酮（西地那 非），亦已知為1-[[3-(6,7-二氫-1-甲基-7-氧代-3-丙基-1H-吡唑并[4,3-d]嘧啶-5-基）-4-乙氧基苯基]磺醯基]-4-甲基哌 畊（參見歐洲專利申請案第0463756號）； 5-(2-乙氧基-5-嗎琳基乙酿基苯基）-1-甲基-3-正丙基 -I,6-二氫-7H-吡唑并[4,3_d]嘧啶_7_酮（參見歐洲專利第 0526004號）； 3 -乙基-5-[5-(4-乙基哌畊-1-基磺醯基）-2-正丙氧基苯 基]-2七比啶-2-基）甲基-2,6-二氫-7H-吡唑并[4,3-d]嘧啶-7-酮（參見世界專利第98/49 166號）； 3 -乙基-5-[5-(4-乙基喊畊-1-基續醯基）-2-(2-甲氧基乙氧 基）吡啶-3-基]-2-(吡啶-2-基）甲基-2,6-二氫-7H-吡唑并 [4,34]嘧啶-7-酮（參見世界專利第99/54333號）； ( + )-3-乙基-5-[5_(4-乙基哌畊_1_基磺醯基）-2-(2-甲氧基 -l(R)-甲基乙氧基）吡啶-3-基]-2-甲基-2,6-二氫-7H-吡唑并 [4,3-d]嘧啶-7-酮，亦已知為3_乙基_5-{5-[4_乙基哌啩_丨_基 績驢基]_2_([(lR)-2_甲氧基“-甲基乙基]氧基）吡咬_3· 基卜2-甲基-2,6-二氫- 7H-吡唑并[4,3-d]嘧啶-7-酮（參見世 界專利第99/M333號）； 5-[2-乙氧基-5-(4-乙基哌畊4 —基磺醯基）吡啶基] 85169.doc -13 - 200407153 乙基-2-[2-甲氧基乙基]_2,6-二氫_7H-吡唑并[4,3_d]嘧啶一 酉同，亦已知4 H6-乙氧基_5-[3-乙基-6,7-二氫-2-(2-甲氧基 乙基）-7_氧代-2H-吡唑并[4,3_d]嘧啶-5-基卜弘吡啶基磺醯 基卜4-乙基哌畊（參見世界專利申請案第〇ι/2?ιΐ3號，實施 例8); 5 [2-兴丁氧基_5-(4-乙基哌畊-卜基磺醯基）吡啶 基>3-乙基-2-(1-甲基六氫吡啶基）_2，卜二氫_7H-吡唑并 [4,3-d]嘧啶酮（參見世界專利第〇1/27113號，實施例15); 5_〇乙氧基-5-(4-乙基哌畊基磺醯基）吡啶-3_基 乙基-2-苯基-2,6-二氫-7H-吡唑并[4,3-d]嘧啶_7_酮（參見世 界專利第01/27113號，實施例66); 5·(5·乙醯基-2-丙氧基—3-吡啶基）-3-乙基-2-(1-異丙基-3-吖丁哫基）-2,6-二氫-7H-吡唑并[4,3-d]嘧啶-7-酮（參見世界 專利第01/27 112號，實施例124); 5-(5-乙醯基-2-丁氧基-3-吡啶基）-3-乙基-2-(1-乙基-3-吖 丁啶基）_2,6-二氫-7Ή-吡唑并[4,3-d]嘧啶_7_酮（參見世界專 利第01/27112號，實施例132); (611，12&11)-2,3,6,7，12，123-六氫-2-甲基-6-(3,4-亞甲基二 氧苯基）-ΐ7比畊并[2’，1’：6, l]p比啶并[3,4-b] 4丨嗓-1,4-二酮（他 達那非’ IC-351)’即已公開之國際專利申請案第95/19978 號實施例78和95的化合物，以及實施例1、3、7和8的化合 物； 2-[2 -乙氧基- 5- (4 -乙基-喊p井-1_基-1-績g盛基）-苯基]-5 -甲 基-7-丙基-311-咪吐并[5，1-：^][1，2,4]三峻-4-酮（瓦地那非）， 85169.doc -14- 200407153 5F已知為1-[[3-(3,4_二氫I甲基I氧代_7_丙基咪唑并 [5，l-f]-二唑_2_基乙氧基苯基]磺醯基]·‘乙基哌畊，即 已公開之國際專利申請案第99/2^33號實施例20、19、337 和3 3 6的化合物； [7-(3_氯_4_甲氧基苯甲基胺基）-1-甲基-3-丙基-1H-吡唑 并[4,3-d]嘧哫_5_基甲氧基]乙酸（參見世界專利第〇2/59126 號，實施例1); 4- （4-氯苯甲基）胺基-6,7,8-三甲氧基喹唑啉（已公開之國 際專利申請案第93/〇7124號（EISAI)之實施例11);及 7,8-一氣-8_氧代-6-[2-丙氧基苯基]-1仏咪峻并[4,5*^]0套 口坐琳和1-[3-[1-[(4-氟苯基）甲基]-7,8-二氫-8-氧代-11"1-咪唾 并[4,5-g]喹唑啉-6-基]-4-丙氧基苯基]碳醯胺（R0teiia dp等 人；J· Med· Chem· 43(7)，125 7-1263, 2000的化合物 3和 14)。 本發明使用之更佳的PDE5抑制劑係選自該組群及其醫 藥可容許之鹽： 5_[2-乙氧基-5-(4-甲基-1-成p井基績醯基）苯基]甲基 -3-正丙基-1，6-二氫- 比吐并[4,3-d]喃淀-7-酮（西地那 非）； (611，12&11)-2,3,6,7，12，12^六氫-2-甲基-6-(3,4-亞甲基二 氧苯基）-吡啡并[2’，1 ’：6,1]吡啶并[3,4-b]叫丨哚-1，4-二酮㈠也 達那非，IC-351); 2-[2-乙氧基-5-(4-乙基-哌啡-1-基-1-磺醯基）-苯基；μ5_甲 基-7-丙基-3士咪唑并[5，1-][1[1，2,4]***-4-酮（瓦地那非）； 5- [2 -乙氧基-5-(4-乙基口瓜_-1-基績酿基）ρ比淀-3-基]_3一 85169.doc -15 - 200407153 乙基-2-[2-甲氧基乙基]、2，心二 酮；及 氫-7H-吡唑并[4,3-d]嘧啶_7 5-(5-乙醯基-2-丁氧基卜其 、 比哫基）_3·乙基-2-(1-乙基吖 丁呢基）-2,6_二氫-7Η-Μ 并[4,3_d],n酉同。 特佳的PDE5抑制劑為％「2 氫 勹〕乙虱基_5·(4_甲基_丨_哌 磺醯基）苯基]-1_甲基正 — 土 止丙基_1，6-_氫_7H_吡唑并[4 3 喊淀_7_酉同（西地那非）（亦已知為1-[[3-(6,7_二氫小甲基7 似-3-丙基-m心并[4,3♦密淀_5_基）-4_乙氧基^基] 石黃驗基]-4-甲基哌畊）及其醫疏 /、酋木可谷卉又鹽。西地那非檸 酸鹽為較佳之鹽。 =發明使用之織抑制劑包括直接作用之ACE抑制劑及 其前藥兩者，包括阿拉普利、阿林普利'阿提普利、苯那 普利、苯那普利拉、西隆普利、卡托普利、西拉普利、西 拉普利拉、得拉普利、依那普利、依那普利拉、福辛普利' 咪達普利”?丨哚普利、賴苯普利、賴諾普利、莫西普利、 莫維晋利、潘他普利、培哚普利、喹那普利、喳那普利拉、 雷米普利、阮提普利'螺普利、替莫普利、壬肽抗壓素、 群；!? 9利及佐务普利。此外，該A c E抑制劑可為“雙重 ACE/NEP抑制劑”，亦即能抑制ACE和中性内肽酶（^加^^ endopeptidase，NEP)兩者的化合物，例如奥馬曲拉、法西 口木曲、米沙晋利、桑帕曲拉、BMS-189921、MDL-100240 及Ζ13752Α。 用以治療高血壓之PDE5抑制劑及ACE抑制劑的較佳組 合為： 85l69.doc -16- 200407153 西地那非和喹那普利鹽酸鹽； 西地那非和苯那普利鹽酸鹽； 西地那非和卡托普利； 西地那非和依那普利順丁烯二酸鹽； 西地那非和福辛普利； 西地那非和賴諾普利； 西地那非和莫西普利； 西地那非和雷米普利； 西地那非和群多普利； 他達那非和喳那普利鹽酸鹽； 他達那非和苯那普利鹽酸鹽； 他達那非和卡托普利； 他達那非和依那普利順丁烯二酸鹽； 他達那非和福辛普利； 他達那非和賴諾普利； 他達那非和莫西普利； 他達那非和雷米普利； 他達那非和群多普利； 瓦地那非和喹那普利鹽酸鹽； 瓦地那非和苯那普利鹽酸鹽； 瓦地那非和卡托普利； 瓦地那非和依那普利順丁烯二酸鹽； 瓦地那非和福辛普利； 瓦地那非和賴諾普利； 85169.doc 200407153 瓦地那非和莫西普利； 瓦地那非和雷米普利；及 瓦地那非和群多普利。 本發明之醫藥組合在治療包括心血管及代謝疾病的疾 病方面有用，且此等組合亦可用於治療其他疾病（例如血栓 症），及治療進行過經皮冠狀動脈腔内成形術的患者（PTC A 後的患者）。 所治療之心血管異常較佳為高血壓、鬱血性心臟衰竭、 絞痛症、中風或腎衰竭。此心血管異常更佳為原發性鬲血 壓、肺動脈高血壓、繼發性高血壓、獨立性收縮性高血壓、 與糖尿病有關之高血壓、與動脈粥狀硬化有關之高血壓、 腎血管性高血壓、鬱血性心臟衰竭、絞痛症、中風或腎衰 竭。於特佳之具體實施例中，所治療之異常為原發性高血 壓。於另一特佳之具體實施例中，所治療之異常為肺動脈 高血壓。於另一特佳之具體實施例中，所治療之異常為繼 發性高血壓。於另一特佳之具體實施例中，所治療之異常 為獨立性收縮性高血壓。於另一特佳之具體實施例中，所 治療之異常為與糖尿病有關之高血壓。於另一特佳之具體 實施例中，所治療之異常為與動脈粥狀硬化有關之高血 壓。於另一特佳之具體實施例中，所治療之異常為腎血管 性高血壓。 所治療之代謝疾病較佳為葡萄糖耐受不良或糖尿病，包 括其併發症，例如糖尿病性視網膜病變和糖尿病性神經病 變。此代謝疾病更佳為葡萄糖耐受不良、第1型糖尿病、 85169.doc -18 - 200407153 非騰島素依賴之第2型糖尿病或胰島素依賴之第2 病。 本發明之组合可單獨投藥，但—般係選擇與所欲投藥路 徑和標準藥劑實務有關的適當醫藥賦形劑、稀釋劑或載 體，以混合物形式進行投藥。 例如本發明之組合可用錠劑、膠囊、多重微粒、凝膠、 薄膜、栓劑、酏劑、溶液或懸浮液的形式，經口、經頰或 經舌下進行投藥’其可含有調味劑或著色劑，及用於立即 釋放、延遲釋放、緩慢釋放、持續釋放、脈衝釋放或控制 釋放的應用上。本發明之組合也可以快速分散或快速溶解 的劑型或以高能分散形式或以包覆粒子進行投藥。適合的 配方可視需要呈包覆或未包覆形式。 此類固體醫藥組合物，例如錠劑，可含有賦形劑（如微晶 纖維素、乳糖、擰檬酸鈉、碳酸鈣、磷酸氫二躬、甘胺酸 和殿粉（較佳為玉米澱粉、馬鈴薯殿粉或樹薯殿粉）），崩解 劑（如殿粉乙醇酸鈉、交聯叛甲基纖維素鈉和特有的複合石夕 酸鹽）及造粒黏結劑（如聚乙締P比洛垸酮、經丙基甲基纖維 素（HPMC)、羥丙基纖維素（HPC)、蔗糖、明膠和***膠）。 此外，可包括潤滑劑，例如硬脂酸鎂、硬脂酸、二十二酸 甘油i旨和滑石。 下列配方實例僅作為說明之用，而非意圖限制本發明之 範疇。活性成分表示本發明之組合。 配方1 : 使用下列成分製備錠劑： 85169.doc -19- 200407153 將活性成分（50 mg)與纖維素（微晶）、二氧切、硬脂酸 (發煙）進行摻合並且將該混合物壓縮形成鍵劑。 配方2 : 可將活性成分（100叫)與等滲透壓生理食鹽水（1000 ml) 組合製備靜脈注射配方。 、該錠劑係藉由標準方法(例如直接壓縮法或者濕式或乾 式造粒法）加以製備。錠芯可包覆適當的包衣。 相似種類的固體組合物也可用來作為明膠或HPMC膠囊 中的填无物。就這-點而言，較佳的賦形劑包括乳糖、殿 粉、纖維素、乳糖或高分子量聚乙二醇。對於水性懸浮液 及/或驰劑而言，可將觸5抑制劑及仰抑制劑與各種甜 味劑或調味劑、有色物質或染料’與乳化劑及/或懸浮劑以 及與例如水、乙醇、丙-醢、 内一知甘油及其組合之稀釋劑加以 組合。 、，緩慢釋放及脈衝釋放的劑型可含有立即釋放之劑型所 評述的職形劑和作為釋放速率調節劑之額外的賦形劑，咳 等賦形劑可包覆在裝置的主體上及/或包含在裝置的主體 4中。釋放速率調節劑包括（但不只限於）幾丙基甲基纖維 素、甲基纖維素、幾甲基纖維素鋼、乙基纖維素、乙酸纖 維素、聚氧化乙缔、三仙膠、卡波姆、具銨基之甲基丙缔 酸酯共聚物、氫化萬麻油、巴西棕櫚蠟、石蠟、乙酸酞酸 纖:素、酉太酸幾丙基甲基纖維素、甲基丙缔酸酿共聚物及 其此合物。緩慢釋放及脈衝釋放的劑型可含有釋放速率調 即賦形劑 <-種或其組合。釋放速率調節賦形劑可同時存 85169.doc -20- 200407153 劑型上（即表面或 在於劑型内部（即基質内部），及/或存在於 包衣上）。 決速刀政劑或快速溶解劑配方（fddFS)可含有下列成 分：：斯巴甜、醋磺内酯鉀、檸檬酸、交聯羧甲基纖維素 納:交聯聚維酮、二抗壞血酸、丙埽酸乙醋、乙基纖維素、 明膠、歿丙基甲基纖維素、硬脂酸鎂、甘露糖醇、甲基丙 缔酸甲酯、薄荷調味劑、聚乙二醇、高溫燃燒二氧化矽、 二氧化矽、澱粉乙醇酸鈉、硬脂醯反丁烯二酸鈉、山梨醇、 木糖醇。本文中使用的分散或溶解係描述？〇〇1^取決於所 使用之原料藥的溶解度，亦即，在原料藥不溶處可製備快 速分散劑型，而在原料藥可溶處可製備快速溶解劑型。 本發明之組合也可以腸外的方式投藥，例如海綿體内、 靜脈内、動脈内、腹膜内、脊椎内、（腦或心）室内、尿道 内、胸骨内、顱内、肌内或皮下的方式，或者可藉由輸液 或無針注射技術予以投藥。對於此種腸外投藥而言，本發 明之組合最好以無菌水溶液形式加以使用，該無菌水溶液 可含有其他物質，例如使溶液與血液為等滲透壓的足夠職 類或葡萄糖。必要時，該無菌水溶液應適當進行緩衝（較佳 為緩衝至3至9的pH值）。藉由熟習此項技術者所熟知的標 準製藥技術，可在無菌條件下輕易完成適當之腸外配方的 製劑。 下列劑量和本文中的其他劑量係針對具有大約65至70 kg體重範圍的平均人類患者而言。熟習此項技術者能輕易 地決定體重落於此範圍之外的患者（例如兒童或年長者）所 85169.doc 200407153 需要的劑量。 在此種配方中，本發明之組合的劑量取決於該組合之藥 效’但是對於一日投藥三次而言，可預計該劑量在1至5〇〇 mg之PDE5抑制劑及丨至1〇〇呵之ACE抑制劑的範圍内。較 佳的劑量在10至1〇〇 mg(例如1〇、25、50和100 mg)之PDE5 抑制劑及5至50 mg(例如5、10、25和50 mg)之ACE抑制劑 的範圍内，該劑量可一日投藥一次、兩次或三次（較佳為一 次）。然而精確劑量係由開藥醫師所決定，並取決於患者的 年齡和體重及症狀的嚴重程度。 對於人類患者之經口和腸外投藥而言，本發明之組合的 母日劑虽通常為5至500 mg/kg(單次或分次劑量）。 因此錠劑或膠囊可含有5 mgS25〇 mg(例如1〇至丨⑼mg) 的本發明之組合，以適當地一次以一或二或更多個進行投 藥。無論如何，醫師會決定最適合個別患者的實際劑量， 孩劑量會隨著年齡、體重和反應而改變。上述劑量為例示 的平均情形。而在本發明之範疇内，當然可以存在應為較 高或較低劑量的個別案例。熟習此項技術者應了解如有必 要或需要時（亦即prn)可以單次劑量服用本發明之組合。應 了解本又中關於治療的全部參考資料係包括急性治療（需 要時採取）及慢性治療（長期持續治療）。 本發明之組合也可以鼻内或藉由吸入的方式進行投 藥，而以乾粉吸入器的形式或是由加壓容器、泵、噴霧機、 霧化器或噴霧器，在使用或沒有使用適當推進劑（例如一氣 一氟甲k、二氯氟甲烷、二氯四氟乙烷、氟烷（如I，〗1 85169.doc -22- 200407153 四氣乙淀（HFA 134A[商標])或1，1，1，2,3,3,3-七氟丙烷（111^ 227EA[商標]))、二氧化碳或其他適合氣體）的情況下，產 生氣落膠喷霧，以便於輸送。於加壓氣溶膠的情形中，可 藉由提供輸送經過計量之量的閥來決定劑量單位。該加壓 容器、系、噴霧機、霧化器或噴霧器可容納活性化合物溶 液或懸浮液，例如使用乙醇與推進劑之混合物作為溶劑， 居/合劑可額外含有潤滑劑，如去水山梨醇三油酸酯。用於 吸入為或吹入器的膠囊或（例如由明膠所製成者）可配製成 含有本發明之組合和適當乾粉基質（如乳糖或澱粉）的乾粉 混合物。 將氣溶膠或乾粉配方以較佳方式配置，使輸送給患者之 每個經過計量的劑量或“puff，，含有i炖至5〇 mg的本發明之 組合。氣落膠的總每日劑量在i处至50 mg的範圍内，該氣 /合膠可一整天投藥一次或者更頻繁以分次劑量予以投藥。 或者，本發明之組合可以栓劑或***栓的形式進行投 藥，或是可以凝膠、水凝膠、洗劑、溶液、乳霜、軟膏或 撒布h彳的开> 式加以局部塗敷。本發明之組合也可以在皮膚 上或穿透皮膚的方式投藥，例如使用皮膚貼布、沉積或皮 下〉王射。本發明之組合也可經由肺臟或直腸路徑進行投 藥。 對於局部塗敷皮膚而言，可將本發明之組合配製成含有 活性化合物的適當軟膏，該活性化合物係懸浮或溶解在例 如具有下列一或多種的混合物中：礦物油、液態石蠟脂、 白石蠟脂、丙二醇、聚氧乙烯聚氧丙晞化合物、乳化蠟和 85169.doc -23 - 200407153 ::。’可將本發明之組合配製成懸浮或溶解在例如下 列-或夕種的混合物中的適當洗劑或乳霜：礦物 ；：梨醇單硬脂酸酿、丙二醇、液態謂、聚山梨酸酷60 病㈣、棕櫚醇、2_辛基十H苯甲醇和水。 2可使用本發明之組合與環糊精結合。已知環糊精可和 樂物分子形成包含複合物或非包含複合物。形成藥物-環糊 精複合物可變更藥物分子的溶解度、溶解速率、生物有效 性及/或安定性。藥物-環糊精複合物-般可用於大部分的 劑型及投藥路徑。使藥物與環糊精複合的另一種方式可使 用㈣助添加劑，例如載體、稀釋劑或助溶劑。最常使用 的疋α -、召-和r _環糊精，其適當實例見述於已公開之國 際專利中請案帛91/llm號1 9碰5職^ 98/55148 號0 口服本發明之組合為較佳路徑，而且最方便。在受藥者 有呑嗓異常或是口服後藥物吸收不良的情況中，此藥物可 經腸外、經舌下或經頰進行投藥。 在三重療法中的第三劑可為第二PDE54Ace抑制劑，或 者可選 >自第三種醫藥組群。例如，可為中性内肽酶抑制 劑、血官收縮素11受體拮抗劑、鈣通道阻斷劑（如氨氯地 平）、降血脂藥物（如阿托發司他汀）、万·阻斷劑（亦即石-腎上腺素受體拮抗劑）或利尿劑。 應了解本發明係涵蓋下列進一步的態樣，且第一態樣之 上述特有的具體實施例係擴展至此等態樣·· i)用以治療高血壓之本發明醫藥組合（用於同時、分別或 85169.doc -24 - 200407153 循序投藥）； ii) 用以治療高血壓之套組’該套組包括：a)含有 抑制劑之第一醫藥組合物；b)含有ACE抑制劑之第二醫藥 組合物；及c )此爭組合物之各咨， i i i) 治#患者高血壓之方法’包括用有效量的本發明之 組合治療該患者。 測定法 適用於本發明之較佳的化合物為具有藥效和選擇性的 PDE5抑制劑。在活體外對於環鳥嘌呤核苷3’，5、單蹲酸 (cGMP)及環腺嘌呤核苷3’，5’-單磷酸（cAMP)之磷酸二騎轉 的PDE抑制活性可藉由測量其IC5〇值（酵素活性達50%抑制 所需要的化合物濃度）加以測定。 所需要的PDE酶實質上藉由Thompson, WJ等人； Biochemistry 18(23)，5228-5237，1979，如同 Ballard SA等人；j Urology 159(6)，2164-2171，1998所述方法的改良，可分離自各 種來源，包括人類海綿體、人類和兔子的血小板、人類心 室、人類骨骼肌、牛視網膜。尤其是cGMP特有的PDE5及 cGMP抑制的cAMP PDE3可得自人類海绵體組織、人類血 小板或免子的血小板；cGMP刺激的PDE2係得自人類海綿 體；鈣/攜鈣素（Ca/CAM)依賴的PDE1係得自人類心室； c AMP特有的PDE4係得自人類骨骼肌；以及受光體PDE6係 得自牛視網膜。磷酸二酯酶7至11可產自轉染至SF9細胞之 全長人類重組克隆。 測定法可利用 Thompson WJ 及 Appleman MM ; Biochemistry 85169.doc -25- 200407153 10(2)，311-316,1971，實質上如同 Ballard SA等人；J· Urology 159(6)，2164-2171，1998所述“批次”方法的改良，或是使用 Amersham pic在商品碼TRKQ7090/7100之下所述之協定的 改良，利用直接偵測[3H]所標記之AMP/GMP的閃燦標記測 定法來進行。總之，對於閃爍標記測定法而言，係藉由測 定在改變抑制劑濃度及低受質，（在〜1/3 Km或更低的濃度 下，未標記比[3H]所標記之cGMP或cAMP的比例為3: 1)存 在時的定量酵素來研究PDE抑制劑的效果，以至於 1C5〇 s Ki。利用測定緩衝液使最終測定體積達到1〇〇 w [20mM Tris-HCl pH 7.4，5 mM MgCl2，1 mg/ml牛血清白 蛋白]。利用酵素起始反應，於30°C下培養30至60 min，以 提供< 3 0%的受質週轉率，然後以5 〇 w的石夕酸妃SPA珠粒 (分別含有3 mM未標記的PDE 9和11環狀核苷酸）終止反 應。重新密封培養亚並搖動20 min，在黑暗中使該珠粒沉 降30 min之後，接著在TopCount讀盤儀（packard，Meriden， CT)上计數，將放射性單位轉換成未受到抑制之對照（丨〇〇%) 的 ％活性，利用 Microsoft Excel extensi〇n的‘Fit Curve(擬合 曲線）’對抑制劑的濃度作圖，得到抑制劑的IC^值。 動物實驗 在人類高血壓的動物模式中，使用依那普利作為代表性 的ACE抑制劑及3_乙基_5_[5_(4·乙基哌呼小基磺醯 基）-2-(2-甲氧基乙氧基）_口比淀_3_基]_2七比咬·2-基）甲基 2’6一氫7^1叶匕4并[4,3-〇1]<1密呢_7-嗣（已公開之國際專利 第99/54333號實施例4的化人物、你、Α化士 ]化口物）作為代表性的PDE5抑制 85169.doc -26- 200407153 劑以顯示本發明之組合的功效。 動物 自發性高血壓大鼠（SHR)為人類高血壓之廣泛使用的動 物模式。根據已公開的方法（Gardiner, SM等人；Br· J· Pharmacol. 13 2(8)，1625-1629，2001)，利用都卜勒流動探 頭作為工具測量雄性SHRs(20至22週大）之腸繫膜、後軀和 腎臟的血流、主動脈血壓及心跳速率。 藥物 整個實驗期間以0.4 mL/h的速率將依那普利（7.5//g/mL) 、PDE5抑制劑（200 //g/mL)及依那普利與PDE5抑制劑（7.5 /zg/mL + 200 vg/mL)之組合的溶液進行輸液。對照動物接受 化合物載體；利用鹽酸調整至pH 4的等滲透壓生理食鹽水。 協定 記錄基線的血液動力參數。使動物（n = 7或8/組）任意排 列，然後利用藥物溶液連續輸液歷時4天進行治療。在實 驗期間，每天監測血液動力參數之變化達7小時。下表中 係提出以和載體反應的差異所表示的概括結果。 治療 伊那普利 PDE5 抑制劑 組合 平均BP之整體變化(mmHg) -2.4 -12.1 -17.8 腸繫膜傳導率之變化(％) +22.4 +22.1 +48.1 腎臟傳導率之變化(％) +14.2 -0.8 +34.2 主動脈傳導率之變化(％) +3.7 +19.8 +30.1 該結果，特別是對於腎臟傳導率而言，係顯示組合此兩 種藥劑可產生大於其個別效果之總和的效果。 85169.doc -27 -The selectivity ratio can be easily determined by those skilled in the art by the ratio of the corresponding iCw values of the particular enzyme of interest. The ICM values of pDE3 and pDE4 enzymes can be determined using established literature methods, see BaiiardsA et al .; Journal of Urology 159, 2164-2171, 1998. The PDE5 inhibitor is preferably selective for the PDE5 enzyme. The inhibitor preferably has a selectivity for PDE5 over PDE3 over 100, more preferably over 300. The PDE5 inhibitor more preferably has a selectivity greater than 100, more preferably greater than 300, over both PDE3 and pDE4. The PDE5 inhibitor preferably has a C50 value of less than 1000 nM and a selectivity greater than 100-fold over PDE3. Oral bioavailability refers to the proportion of oral medicines that reach systemic circulation. Factors that determine the oral bioavailability of a drug are lysis, membrane permeability, and liver clearance. Representative is the use of in vitro techniques first and then the in vitro technique of screening cascades to determine oral bioavailability. Solubility 'is to dissolve the drug by the water in the gastrointestinal tract (GIT), and it can be predicted by performing an in vitro solubility test simulating GIT at an appropriate pH value. The PDE5 inhibitor preferably has a minimum solubility of 50 // § / 1111. Solubility can be determined by standard procedures known in the art, such as those described by Upinski CA et al. 85169.doc 200407153; Adv. Rug Deliv. Rev. 23 屮 3), Hong 25, 199 ?. Membrane permeability refers to cells through which the compound passes through the GIT. Lipophilicity is a key property to predict the permeability of the membrane, and it is determined by in vitro Log D7.4 measurement using organic solvents and buffers. The pDE5 inhibitor preferably has a Lo g D? 4 of -2 to +4, more preferably] to +3. L0g0 can be determined by standard procedures known in the art, such as those described by Stopher, D * McClean, S; J. Pharm. Pharmacol. 42 (2), 144, 1990. Cellular monolayer assays (such as CacO2) are added to Babe to predict appropriate membrane permeability in the presence of excreted transporters (such as P-glycoprotein, the so-called CacO2 flux). The PDE5 inhibitor preferably has a Caco2 flux greater than, more preferably greater than 5x10 6 cm.s-1. Caco2 flux values can be determined by standard procedures known in the art, such as those described in Artursson, P. and Viagnusson, C; J. Pharm. Sci. 79 (7), 595-600, 1990. Metabolic stability refers to the ability of the compound to metabolize GIT provided by the compound during absorption or the liver directly metabolizes the compound after absorption: the first effect. Assay systems such as microsomes and hepatocytes can predict metabolic instability. The PDE5 inhibitor preferably exhibits metabolic stability in an assay system which is equivalent to a liver extract of less than 0.5. Examples of measurement systems and data operations are described in Obach, RS; Curr. Opin. Drug Disc. Devel. 4A, 36-44, 2001 and Shibata, Y et al .; Dmg Met. Disp. 28 (12), 1518-1523 , 2000. Due to the interaction of the above steps, in vivo experiments in animals can be used to obtain further support for the drug's oral bioavailability in humans. 85169.doc -10- 200407153 Absolute bioavailability is determined by taking the compound orally or a mixture of the compounds orally. For absolute determinations (% oral bioavailability), the intravenous route can also be used. Examples of evaluation of oral bioavailability in animals can be found in Ward, KW et al .; Drug Met. Disp. 29 (1), 82-87, 2001; Berman, J et al .; J. Med. Chem. 40 (6 ), 827_829, 1997 and Han KS and Lee, MG; Drug Met. Disp. 27 (2), 221-226, 1999 ° Examples of PDE5 inhibitors used in the present invention are: European Patent Applications No. 0463756, No. 0526004 And published international patent applications Nos. 93/06 104, 98/49166, 99/543 33, 00/24745, 01/27112 and 01/27 11 3 Pyrazolo [4,3-d] pyrimidin-7-ones; pyrazoles disclosed in European Patent Application Nos. 0995750, 0995 75 1 and Published International Patent Application No. 9: 3 / 〇7149 Zolo [3,4-d] pyrimidin-4-ones; p-pyridine disclosed in published international patent applications Nos. 01/1 8004, 02/00660, and 02/59126 , 3-d] shout violation; disclosed in the published International Patent Application No. 9 3/12 0 95. Quinalin-4-ones; p-pyrido [3,2_d] 11 dense lakes_cardiones disclosed in International Patent Application No. 94/05661; European Patent Application No. 109271 8 And the larynx class disclosed in Published International Patent Application No. 94/00453; the confluence disclosed in Published International Patent Application No. 95/19978 [2 ', 1': 6,1] • Biyodo [3,4-13] indole diones; the flavor σ disclosed in European Patent Application No. 1092719 and Published International Patent Application No. 99/24433 [5 lf] [l 2 4] Triazole-ones; Bicyclic compounds disclosed in published international patent application Nos. 85169.doc -11-200407153 93/07 124 & RoteUa Dp et al; 1 he (1 · Chem · 43 (7), 125 7-1263, 2000 Mijun and Junjun 4 ketones disclosed. The contents of these published international patent applications and journal articles, especially the therapeutic activity in the scope of patent applications The general formula of the compounds and the exemplified compounds are incorporated herein by reference in their entirety. More examples of PDE5 inhibitors used in the present invention include 4-bromo-5- (pyridylmethylamino) ) -6- [3- (4-Chlorophenyl) -propoxy] -3 (2H) Dageng; 1- [4-[(1,3 -Dongfangyi 1: 7oxapent-5- Methyl) amino] -6-chloro-2 -quinozolinyl] -4-hexahydropyridine-carboxylic acid, monosodium salt; (+)-cis-5,63,7,9 , 9,9 & -hexahydro-2- [4- (trifluoromethyl) -phenylmethyl-5-methyl-cyclopentane-4,5] -4 (3H) ketone; furazlocillin; cis-2-hexyl-5-fluorenyl-3,4,5,6 &, 7,8,9,9 & -octahydrocyclopenta [4, 5]-味 17 sitting and [2,1-b] purin-4-one; 3-ethylamidino-1- (2.chlorobenzyl) -2-propylandol-6-carboxylate; 3-ethylfluorenyl-1- (2-chlorobenzyl) -2-propyloxindole-6-carboxylate; 4-bromo-5- (3-pyridylmethylamino) -6- ( 3- (4-Gaphenyl) -propoxy) -3- (2H) dacrotonone; 1-methyl-5- (5-morpholinylacetamido-2-n-propoxyphenyl) -3-n-propyl-1,6-dichloro-7Η-ρ than tetra (4,3-d) ranodo-7- 嗣; 1- [4-[(1,3-benzodi-p-oxine [Mocen-5-ylmethyl) amino] -6-chloro-2-ρ-quinolynyl] -4-hexahydroeodo-polyacid, single steel salt; Pharmaprojects (World Information Database on Drug Research and Development Status ) No. 45 1 6 (Glaxo Wellcome); Pharmaprojects No. 5 05 1 (Bayer); Pharmaprojects No. 5 064 (Kyowa Hakko; see World Patent No. 96/26940); Pharmaprojects No. 5069 (Schering Plough); GF-196960 85169.doc 200407153 (Glaxo Wellcome); E-8010 and E-4010 (Eisai); Bay-38-3045 and 38-9456 (Bayer) and Sch-5 1866. The preferred PDE5 inhibitors used in the present invention include: 5- [2-ethoxy-5- (4-methyl-1-piperidylsulfonyl) phenyl] -1-methyl-3-n Propyl-1,6-dihydro-7H-pyrazolo [4,3-d] pyrimidin-7-one (sildenafil), also known as 1-[[3- (6,7-di Hydrogen-1-methyl-7-oxo-3-propyl-1H-pyrazolo [4,3-d] pyrimidin-5-yl) -4-ethoxyphenyl] sulfonyl] -4 -Methylpiperin (see European Patent Application No. 0463756); 5- (2-ethoxy-5-morpholinylethenylphenyl) -1-methyl-3-n-propyl-I, 6-dihydro-7H-pyrazolo [4,3_d] pyrimidin-7_one (see European Patent No. 0526004); 3-ethyl-5- [5- (4-ethylpiperin-1-yl Sulfonyl) -2-n-propoxyphenyl] -2heptapyridin-2-yl) methyl-2,6-dihydro-7H-pyrazolo [4,3-d] pyrimidin-7- Ketones (see World Patent No. 98/49 166); 3-ethyl-5- [5- (4-ethylsulfonyl-1-ylcontinyl) -2- (2-methoxyethoxy ) Pyridin-3-yl] -2- (pyridin-2-yl) methyl-2,6-dihydro-7H-pyrazolo [4,34] pyrimidin-7-one (see World Patent No. 99/54333 No.); (+)-3-ethyl-5- [5_ (4-ethylpiperin_1_ylsulfonyl) -2- (2-methoxy-l (R) -methylethyl ) Pyridin-3-yl] -2-methyl-2,6-dihydro-7H-pyrazolo [4,3-d] pyrimidin-7-one, also known as 3-ethyl-5 {5- [4_ethylpiperazine_ 丨 _jiji donkeyyl] _2 _ ([(lR) -2_methoxy “-methylethyl] oxy) pyridine_3 · jib 2-methyl -2,6-dihydro-7H-pyrazolo [4,3-d] pyrimidin-7-one (see World Patent No. 99 / M333); 5- [2-ethoxy-5- (4 -Ethylpiperazine 4-ylsulfonyl) pyridyl] 85169.doc -13-200407153 ethyl-2- [2-methoxyethyl] _2,6-dihydro_7H-pyrazolo [4 , 3_d] pyrimidine is the same, 4 H6-ethoxy_5- [3-ethyl-6,7-dihydro-2- (2-methoxyethyl) -7_oxo- 2H-pyrazolo [4,3_d] pyrimidin-5-ylpyridinylsulfonylsulfonyl 4-ethylpigen (see World Patent Application No. 0/2/2, Example 8); 5 [2-Xingbutoxy_5- (4-ethylpiperazine-bulfylsulfonyl) pyridyl> 3-ethyl-2- (1-methylhexahydropyridyl) _2, dihydro 7H-pyrazolo [4,3-d] pyrimidone (see World Patent No. 01/27113, Example 15); 5_ethoxy-5- (4-ethylpiperidinylsulfonium Yl) pyridine-3_ylethyl-2-phenyl-2,6-dihydro-7H-pyrazolo [4,3-d] pyrimidine_7 Ketone (see World Patent No. 01/27113, Example 66); 5 · (5 · Ethyl-2-propoxy-3-pyridyl) -3-ethyl-2- (1-isopropyl Group-3-azetidinyl) -2,6-dihydro-7H-pyrazolo [4,3-d] pyrimidin-7-one (see World Patent No. 01/27 112, Example 124); 5- (5-ethylfluorenyl-2-butoxy-3-pyridyl) -3-ethyl-2- (1-ethyl-3-azetidinyl) _2,6-dihydro-7fluorene-pyrazolo [ 4,3-d] pyrimidin-7_one (see World Patent No. 01/27112, Example 132); (611,12 & 11) -2,3,6,7,12,123-hexahydro-2 -Methyl-6- (3,4-methylenedioxyphenyl) -pyrene 7 bipyrido [2 ', 1': 6, l] p than pyrido [3,4-b] 4 丨1,4-diketone (tadanafil 'IC-351)' is the compound of Examples 78 and 95, and the compounds of Examples 1, 3, 7 and 8 in published International Patent Application No. 95/19978 ; 2- [2-Ethoxy- 5- (4-ethyl-amino-p-well-l-yl-l-g-g-Singyl) -phenyl] -5 -methyl-7-propyl-311- Mido [5,1-: ^] [1,2,4] Trijun-4-one (Vadinafil), 85169.doc -14- 200407153 5F is known as 1-[[3- (3 , 4_dihydro I methyl I oxo_7_propylimidazo [5, lf] -diazole_2_ylethyl Oxyphenyl] sulfonyl] · 'ethylpiperin, the compounds of Examples 20, 19, 337, and 3 3 6 of Published International Patent Application No. 99/2 ^ 33; [7- (3 _Chloro_4_methoxybenzylamino) -1-methyl-3-propyl-1H-pyrazolo [4,3-d] pyrimidin_5_ylmethoxy] acetic acid (see World Patent No. 02/59126, Example 1); 4- (4-chlorobenzyl) amino-6,7,8-trimethoxyquinazoline (published International Patent Application No. 93 / 〇7124 (EISAI) Example 11); and 7,8-monogas-8_oxo-6- [2-propoxyphenyl] -1 amidol and [4,5 * ^] 0 sets Mizuprin and 1- [3- [1-[(4-fluorophenyl) methyl] -7,8-dihydro-8-oxo-11 " 1-imidosulfa [4,5-g] Quinazolin-6-yl] -4-propoxyphenyl] carbamidine (Roteiia dp et al .; J. Med. Chem. 43 (7), 125 7-1263, compounds 3 and 14 of 2000). A more preferred PDE5 inhibitor used in the present invention is selected from the group and its pharmaceutically acceptable salts: 5_ [2-ethoxy-5- (4-methyl-1- to p-stilbenzyl) Phenyl] methyl-3-n-propyl-1,6-dihydro-bitalo [4,3-d] nanodo-7-one (sildenafil); (611,12 & 11)- 2,3,6,7,12,12 ^ hexahydro-2-methyl-6- (3,4-methylenedioxyphenyl) -pyrino [2 ', 1': 6,1] Pyrido [3,4-b] is called indole-1,4-diketone and also danafil, IC-351); 2- [2-ethoxy-5- (4-ethyl-piperidine- 1-yl-1-sulfonyl) -phenyl; μ5-methyl-7-propyl-3 simidazo [5,1-] [1 [1,2,4] triazol-4-one ( (Vardenafil); 5- [2-ethoxy-5- (4-ethylmouth guar-1-anyl group) ρ Bidian-3-yl] _3-85169.doc -15-200407153 Ethyl-2- [2-methoxyethyl], 2, cardiodione; and hydrogen-7H-pyrazolo [4,3-d] pyrimidine_7 5- (5-ethylfluorenyl-2- Butoxy group, fluorenyl) -3.ethyl-2- (1-ethylazetnyl) -2,6_dihydro-7Η-M and [4,3_d], n are the same. A particularly good PDE5 inhibitor is% "2-Hydroxypyrene] ethylenyl_5 · (4_methyl_ 丨 _piperazinyl) phenyl] -1_methyl-n-ethoxypropyl-1,6 -_Hydrogen_7H_Pyrazolo [4 3 chanting lake_7_Identical (sildenafil) (also known as 1-[[3- (6,7_dihydro small methyl 7 like -3 -Propyl-moxo [4,3 ♦ dense lake_5_yl) -4_ethoxy ^ yl] chrysanthemum] -4-methylpigenol) and its medicine Gu Hui salt. Sildenafil citrate is the preferred salt. = The weaving inhibitors used in the invention include both direct-acting ACE inhibitors and their prodrugs, including Alappril and Alimpril 'Ati Puli, Benazepril, Benazepril, Silompril, Captopril, Silapri, Silapura, Delapril, Enalapril, Enalapril , Fosinopril, 'Midapril "? 丨 Dopril, Rifepril, Rainopril, Mocepril, Movitin, Pantapril, Perindopril, Quinapril , Jainapril, Ramipril, Ruantipril, Lopipril, Temopril, Noni-Peptide Compressor, Qun; 9? Li and Sapulpril. In addition, the AC inhibitor may be a "dual ACE / NEP inhibitor", that is, a compound capable of inhibiting both ACE and neutral endopeptidase (NEP), such as omatrala, Faxikou Muqu, Misha Jinli, Sampatra, BMS-189921, MDL-100240 and Z13752Α. The preferred combination of PDE5 inhibitor and ACE inhibitor for treating hypertension is: 85l69.doc -16- 200407153 sildenafil and quinapril hydrochloride; sildenafil and benazepril hydrochloride Salt; sildenafil and captopril; sildenafil and enalapril maleate; sildenafil and fosinopril; sildenafil and lisinopril; sildenafil Nelfinil and moxipril; sildenafil and ramipril; sildenafil and trondopril; tadalafil and janapril hydrochloride; tadalafil and benazepril Hydrochloride; tadalafil and captopril; tadalafil and enalapril maleate; tadalafil and fosinopril; tadalafil and lisinopril; Tadalafil and Moxipril; Tadalafil and Ramipril; Tadalafil and Trendopril; Vardenafil and Quinapril Hydrochloride; Vardenafil and Benazeri Pride hydrochloride; Vardenafil and captopril; Vardenafil and enalapril maleates; Vardenafil and fosinopril; Vardenafil and lisinopril 85169.doc 200407153 Vardenafil and Moxipril Vardenafil and ramipril; and Vardenafil and trandolapril. The pharmaceutical combination of the present invention is useful in the treatment of diseases including cardiovascular and metabolic diseases, and these combinations can also be used to treat other diseases (such as thrombosis) and to treat patients who have undergone percutaneous coronary angioplasty (PTC Patient after A). The cardiovascular abnormality to be treated is preferably hypertension, congestive heart failure, colic, stroke or renal failure. This cardiovascular abnormality is more preferably primary iliac blood pressure, pulmonary hypertension, secondary hypertension, independent systolic hypertension, diabetes-related hypertension, atherosclerosis-related hypertension, and renal vascularity. High blood pressure, congestive heart failure, colic, stroke or kidney failure. In a particularly preferred embodiment, the abnormality to be treated is primary hypertensive blood pressure. In another particularly preferred embodiment, the abnormality treated is pulmonary hypertension. In another particularly preferred embodiment, the abnormality treated is secondary hypertension. In another particularly preferred embodiment, the abnormality treated is independent systolic hypertension. In another particularly preferred embodiment, the abnormality treated is hypertension associated with diabetes. In another particularly preferred embodiment, the abnormality to be treated is high blood pressure associated with atherosclerosis. In another particularly preferred embodiment, the abnormality treated is renal vascular hypertension. The metabolic disease to be treated is preferably glucose intolerance or diabetes, including complications such as diabetic retinopathy and diabetic neuropathy. This metabolic disease is more preferably glucose intolerance, type 1 diabetes, 85169.doc -18-200407153 non-tonguelin dependent type 2 diabetes or insulin dependent second disease. The combination of the present invention can be administered alone, but generally the appropriate pharmaceutical excipients, diluents or carriers are selected for administration in the form of a mixture, which is related to the desired route of administration and standard pharmaceutical practice. For example, the combination of the present invention can be administered orally, buccally or sublingually in the form of lozenges, capsules, multiparticulates, gels, films, suppositories, elixirs, solutions or suspensions, which may contain flavoring or coloring Agents, and applications for immediate, delayed, slow, sustained, pulsed, or controlled release. The combination of the present invention can also be administered in a fast-dispersing or fast-dissolving dosage form or in a high-energy dispersion form or as coated particles. Suitable formulations are available in coated or uncoated form as required. Such solid pharmaceutical compositions, such as lozenges, may contain excipients such as microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, dihydrogen phosphate, glycine, and corn flour (preferably corn starch). , Potato powder or cassava powder)), disintegrants (such as sodium powder sodium glycolate, cross-linked methylcellulose sodium and unique compound lysates) and granulating adhesives (such as polyethylene glycol) P-biloxone, via propyl methylcellulose (HPMC), hydroxypropyl cellulose (HPC), sucrose, gelatin and gum arabic). In addition, lubricants such as magnesium stearate, stearic acid, glyceryl behenate, and talc may be included. The following formulation examples are for illustrative purposes only and are not intended to limit the scope of the invention. The active ingredient represents a combination according to the invention. Formulation 1: Preparation of lozenges using the following ingredients: 85169.doc -19- 200407153 Blend the active ingredient (50 mg) with cellulose (microcrystalline), dioxin, stearic acid (smoke) and mix the mixture Compression to form a bonding agent. Formula 2: The active ingredient (100 tablets) can be combined with isotonic saline (1000 ml) to prepare an intravenous injection formula. The tablets are prepared by standard methods (such as direct compression or wet or dry granulation). The core can be coated with a suitable coating. Similar kinds of solid compositions can also be used as fillers in gelatin or HPMC capsules. In this regard, preferred excipients include lactose, starch, cellulose, lactose or high molecular weight polyethylene glycols. For aqueous suspensions and / or agents, contact inhibitors and inhibitors can be used with various sweeteners or flavoring agents, colored substances or dyes' with emulsifiers and / or suspending agents and with, for example, water, ethanol , Propane-fluorene, glycerol and its diluent. The slow-release and pulsatile-release dosage forms may contain the excipients described in the immediate release dosage form and additional excipients as release rate modifiers. Excipients such as cough may be coated on the body of the device and / or Contained in the main body 4 of the device. Release rate modifiers include, but are not limited to, isopropyl methylcellulose, methyl cellulose, isomethyl cellulose steel, ethyl cellulose, cellulose acetate, polyoxyethylene, Sanxian gum, and Carbo Methacrylate, methyl acrylate copolymer with ammonium group, hydrogenated sesame oil, carnauba wax, paraffin wax, cellulose acetate phthalate: cellulose, acetic acid polypropyl cellulose, methyl acrylic acid copolymer And this combination. Slow-release and pulsed-release dosage forms may contain release rate-adjusting excipients <-or a combination thereof. Release rate-regulating excipients can be stored simultaneously on the dosage form (i.e., on the surface or inside the dosage form (i.e., inside the matrix), and / or on the coating). The speed-defining agent or fast dissolving agent formula (fddFS) may contain the following ingredients: Spartame, potassium acetolactone, citric acid, croscarmellose sodium: crospovidone, diascorbic acid, Ethyl propionate, ethyl cellulose, gelatin, propyl methyl cellulose, magnesium stearate, mannitol, methyl methyl acrylate, mint flavor, polyethylene glycol, high temperature combustion Silicon oxide, silicon dioxide, sodium starch glycolate, sodium stearyl fumarate, sorbitol, xylitol. What is the dispersion or dissolution system used in this article? 〇〇 ^^ Depending on the solubility of the drug substance used, that is, fast-dispersing dosage forms can be prepared where the drug substance is insoluble, and fast-dissolving dosage forms can be prepared where the drug substance is soluble. The combination of the present invention can also be administered parenterally, such as intracorporeal, intravenous, intraarterial, intraperitoneal, intraspinal, (brain or heart), intraurethral, intrasternal, intracranial, intramuscular or subcutaneous Method, or can be administered by infusion or needleless injection techniques. For such parenteral administration, the combination of the present invention is preferably used in the form of a sterile aqueous solution, which may contain other substances such as a sufficient grade to make the solution and blood isotonic, or glucose. If necessary, the sterile aqueous solution should be appropriately buffered (preferably to a pH of 3 to 9). Proper parenteral formulations can be easily prepared under sterile conditions by standard pharmaceutical techniques familiar to those skilled in the art. The following doses and others herein are for an average human patient having a weight range of approximately 65 to 70 kg. Those skilled in the art can easily determine the dosage required by patients (such as children or the elderly) whose weight falls outside this range 85169.doc 200407153. In such a formulation, the dosage of the combination of the present invention depends on the efficacy of the combination ', but for three times a day administration, it is expected that the dosage will be from 1 to 500 mg of PDE5 inhibitor and from 1 to 100. Well within the scope of ACE inhibitors. Preferred doses are in the range of 10 to 100 mg (e.g. 10, 25, 50 and 100 mg) of PDE5 inhibitors and 5 to 50 mg (e.g. 5, 10, 25 and 50 mg) of ACE inhibitors This dose can be administered once, twice or three times a day (preferably once). However, the exact dose is determined by the prescribing physician and depends on the age and weight of the patient and the severity of the symptoms. For oral and parenteral administration in human patients, the parent-day dosage of the combination of the present invention is usually 5 to 500 mg / kg (single or divided doses). The lozenge or capsule may therefore contain 5 mg of S25 mg (e.g. 10 to ⑼ mg) of the combination of the invention, suitably administered in one or two or more at a time. Regardless, the physician will determine the actual dose that is best for the individual patient, and the child dose will vary with age, weight and response. The above doses are exemplified on average. In the context of the present invention, there can of course be individual cases which should be higher or lower doses. Those skilled in the art will understand that the combination of the present invention can be taken in a single dose if necessary or needed (i.e. prn). It should be understood that all references to treatment in this booklet include acute treatment (taken when needed) and chronic treatment (long-term continuous treatment). The combination of the present invention can also be administered intranasally or by inhalation, and in the form of a dry powder inhaler or by a pressurized container, pump, sprayer, nebulizer or sprayer, with or without the use of a suitable propellant (Such as monogas, monofluoromethane, dichlorofluoromethane, dichlorotetrafluoroethane, halothane (such as I, 〖1 85169.doc -22- 200407153, Siqi Yidian (HFA 134A [trademark]), or 1,1 1,2,3,3,3-Heptafluoropropane (111 ^ 227EA [Trademark])), carbon dioxide or other suitable gas), a gas-falling gel spray is generated to facilitate transportation. In the case of pressurized aerosols, the dosage unit may be determined by providing a valve that delivers a metered amount. The pressurized container, system, sprayer, nebulizer or atomizer can contain a solution or suspension of the active compound, for example, a mixture of ethanol and a propellant is used as a solvent, and the mixture may additionally contain a lubricant such as sorbitan trihydrate. Oleate. Capsules for inhalers or insufflators (for example, made of gelatin) or a dry powder mixture containing the combination of the present invention and a suitable dry powder base such as lactose or starch can be formulated. The aerosol or dry powder formulation is configured in a preferred manner such that each metered dose or "puff," which is delivered to the patient, contains a combination of the invention stewed to 50 mg. The total daily dose of air-fall gum is between Within the range of i to 50 mg, the gas / gum can be administered once a day or more frequently in divided doses. Alternatively, the combination of the present invention can be administered in the form of suppositories or vaginal suppositories, or can be coagulated. Glue, hydrogel, lotion, solution, cream, ointment, or spread &> application topically. The combination of the present invention can also be administered on the skin or through the skin, such as using a skin patch Cloth, deposition or subcutaneous> Wang She. The combination of the present invention can also be administered via the lung or rectal route. For topical application to the skin, the combination of the present invention can be formulated into a suitable ointment containing the active compound, which The system is suspended or dissolved in, for example, a mixture having one or more of the following: mineral oil, liquid paraffin, white paraffin, propylene glycol, polyoxyethylene polyoxypropylene compound, emulsified wax, and 85169.doc -23-200407153 ::. 'The combination of the present invention can be formulated as a suitable lotion or cream suspended or dissolved in a mixture of, for example, the following: or minerals: sorbitol monostearate, propylene glycol , Liquid syrup, polysorbate 60 sickness, palmityl, 2-octyl decaH benzyl alcohol and water. 2 The combination of the present invention can be used in combination with cyclodextrin. It is known that cyclodextrin can form molecular compounds Complexes or non-containing complexes. The formation of drug-cyclodextrin complexes can alter the solubility, rate of dissolution, bioavailability, and / or stability of drug molecules. Drug-cyclodextrin complexes are generally used in most dosage forms And route of administration. Another way to compound a drug with a cyclodextrin can use a co-additive, such as a carrier, diluent, or co-solvent. The most commonly used 疋 α-,-, and r_cyclodextrin, suitable examples thereof See in the published international patent application No. 91 / llm No. 9 9 No. 5 ^ 98/55148 No. 0 Oral administration of the combination of the present invention is the better route, and the most convenient. In patients who have a bad throat or In the case of drug malabsorption after oral administration, this drug can be administered by Parenteral, sublingual, or buccal administration. The third dose in the triple therapy may be a second PDE54Ace inhibitor, or alternatively, from a third pharmaceutical group. For example, it may be a neutral endopeptidase Inhibitors, heparin contractor 11 receptor antagonists, calcium channel blockers (such as amlodipine), lipid-lowering drugs (such as atorvastatin), wan blockers (that is, stone-adrenaline receptors Body antagonists) or diuretics. It should be understood that the present invention covers the following further aspects, and the above-mentioned specific embodiments of the first aspect are extended to these aspects i) the invention for treating hypertension Pharmaceutical combination (for simultaneous, separate or sequential administration of 85169.doc -24-200407153); ii) a kit for treating hypertension 'the kit includes: a) a first pharmaceutical composition containing an inhibitor; b) A second pharmaceutical composition containing an ACE inhibitor; and c) each of the compositions, iii) a method for treating a patient ' s hypertension, comprising treating the patient with an effective amount of a combination of the invention. Assay method A preferred compound suitable for use in the present invention is a PDE5 inhibitor which is pharmacologically effective and selective. The PDE inhibitory activity of cyclic guanosine 3 ', 5, monosquatine (cGMP) and cyclic adenine nucleoside 3', 5'-monophosphate (cAMP) in vitro can be measured in vitro The IC50 value (the concentration of the compound required for 50% inhibition of the enzyme activity) was measured. The required PDE enzyme is substantially improved by the method described by Thompson, WJ, et al .; Biochemistry 18 (23), 5228-5237, 1979, as Ballard SA et al .; j Urology 159 (6), 2164-2171, 1998 Can be isolated from a variety of sources, including human corpora cavernosa, human and rabbit platelets, human ventricles, human skeletal muscle, and bovine retina. In particular, cGMP-specific PDE5 and cGMP-inhibited cAMP PDE3 can be obtained from human corpus cavernosa tissue, human platelets, or immune platelets; cGMP-stimulated PDE2 is derived from human corpus cavernosum; calcium / calcium carrying (Ca / CAM) dependence PDE1 was obtained from human ventricles; cAMP-specific PDE4 was obtained from human skeletal muscle; and photoreceptor PDE6 was obtained from bovine retina. Phosphodiesterase 7 to 11 can be produced from full-length human recombinant clones transfected into SF9 cells. Assays can be made using Thompson WJ and Appleman MM; Biochemistry 85169.doc -25- 200407153 10 (2), 311-316, 1971, essentially as Ballard SA et al .; J. Urology 159 (6), 2164-2171, 1998 An improvement to the "batch" method, or an improvement to the protocol described by Amersham pic under article code TRKQ7090 / 7100, using a flash mark assay that directly detects the AMP / GMP labeled [3H] to get on. In short, for the scintillation labeling assay, the cGMP or cAMP labeled with [3H] is not labeled at a concentration of ~ 1/3 Km or lower by changing the concentration of the inhibitor and the low mass. The ratio is 3: 1) Quantitative enzymes in the presence of PDE inhibitors were studied to the extent of 1C50s Ki. The final measurement volume was 100 w [20 mM Tris-HCl pH 7.4, 5 mM MgCl2, 1 mg / ml bovine serum albumin] using the measurement buffer. The enzyme-initiated reaction was used to incubate at 30 ° C for 30 to 60 min to provide a mass transfer rate of <30%. Then, 50 liters of Aspartate SPA beads (each containing 3 mM unlabeled PDE 9 and 11 cyclic nucleotides) stop the reaction. After resealing the culture subculture and shaking for 20 min, the beads were allowed to settle in the dark for 30 min, and then counted on a TopCount plate reader (packard, Meriden, CT) to convert the radioactive unit into an uninhibited control (丨〇〇%)% activity, using Microsoft Excel extension's "Fit Curve (fitted curve)" plotted against the concentration of the inhibitor to obtain the IC ^ value of the inhibitor. Animal experiments In the animal model of human hypertension, enalapril was used as a representative ACE inhibitor and 3_ethyl_5_ [5_ (4 · ethylpiperidinylsulfonyl) -2- (2 -Methoxyethoxy) _koubito_3_yl] _2 heptabita · 2-yl) methyl 2'6 monohydrogen 7 ^ 1 leaf 4 and [4,3-〇1] < 1 Secret_7- 嗣 (Published character, you, A, and chemist) of Example 4 of Published International Patent No. 99/54333) as a representative PDE5 inhibitor 85169.doc -26- 200407153 Shows the efficacy of the combination of the invention. Animals Spontaneously hypertensive rats (SHR) are widely used animal models of human hypertension. According to published methods (Gardiner, SM et al .; Br · J · Pharmacol. 13 2 (8), 1625-1629, 2001), the Doppler flow probe was used as a tool to measure male SHRs (20 to 22 weeks old). Blood flow to the mesentery, hindquarters, and kidneys, aortic blood pressure, and heart rate. Enalapril (7.5 // g / mL), PDE5 inhibitor (200 // g / mL), Enalapril and PDE5 inhibitor (7.5 / zg / mL + 200 vg / mL). Control animals received the compound carrier; isotonic saline was adjusted to pH 4 with hydrochloric acid. Agreement Record baseline hemodynamic parameters. Animals (n = 7 or 8 / group) were randomly arranged and then treated with continuous infusion of the drug solution for 4 days. During the experiment, changes in hemodynamic parameters were monitored daily for 7 hours. The following table summarizes the results expressed in terms of differences with the support. Overall change in mean BP (mmHg) in combination with treatment of Enalapril PDE5 inhibitor (mmHg) -2.4 -12.1 -17.8 Change in mesenteric conductivity (%) +22.4 +22.1 +48.1 Change in renal conductivity (%) +14.2 -0.8 +34.2 Change in aortic conductivity (%) +3.7 +19.8 +30.1 This result, especially for renal conductivity, shows that combining these two agents can produce an effect greater than the sum of their individual effects. 85169.doc -27-