TW202233221A - Pde5 inhibitor - Google Patents

Abstract

An object of the present invention is to provide a substance having a novel PDE5 inhibitory effect.

A PDE5 inhibitor according to the present invention contains powder or extract of isaria japonica from silkworm.

Description

PDE5抑制劑 PDE5 inhibitors

本發明係揭示一種關於磷酸二酯酶(PDE：Phosphodiesterase)5抑制劑的技術。 The present invention discloses a technology related to phosphodiesterase (PDE: Phosphodiesterase) 5 inhibitor.

已知具有PDE5抑制作用之物質可有效地預防或改善尿路障礙及肺動脈性肺高血壓症(非專利文獻1)。亦已知具有PDE5抑制作用之源自天然的物質(專利文獻1及2)。 Substances having a PDE5 inhibitory effect are known to be effective in preventing or improving urinary tract disorders and pulmonary arterial pulmonary hypertension (Non-Patent Document 1). Naturally derived substances having a PDE5 inhibitory effect are also known (Patent Documents 1 and 2).

[先前技術文獻] [Prior Art Literature]

[專利文獻] [Patent Literature]

[專利文獻1]日本特開2019-34920 [Patent Document 1] Japanese Patent Laid-Open No. 2019-34920

[專利文獻2]日本特開2018-154610 [Patent Document 2] Japanese Patent Laid-Open No. 2018-154610

[非專利文獻] [Non-patent literature]

[非專利文獻1]日藥理誌，154，265-269，2019 [Non-Patent Document 1] Japanese Pharmacological Journal, 154, 265-269, 2019

本發明之一課題為提供一種新穎的具有PDE5抑制作用之物質。 An object of the present invention is to provide a novel substance with PDE5 inhibitory effect.

為了解決上述課題經過重複精心研究之結果，發現蠶蟲草具有PDE5抑制作用。根據所述之發現進一步重複地研討，進而提供以下述為代表之發明。 In order to solve the above problem, after repeated and careful research, it was found that Cordyceps silkworm has a PDE5 inhibitory effect. Based on the above-mentioned findings, we have further repeated studies and provided inventions represented by the following.

項1 item 1

一種PDE5抑制劑，係含有蠶蟲草的粉末或萃取物。 A PDE5 inhibitor containing powder or extract of Cordyceps sinensis.

項2 item 2

如項1所記載之PDE5抑制劑，係飲食品組成物之形態。 The PDE5 inhibitor as described in Item 1 is in the form of a food and drink composition.

項3 item 3

如項1或2所記載之PDE5抑制劑，係排尿障礙及/或肺動脈性肺高血壓症之預防或改善用者。 The PDE5 inhibitor as described in item 1 or 2 is used for the prevention or improvement of urination disorders and/or pulmonary arterial pulmonary hypertension.

項4 item 4

一種蠶蟲草的粉末或萃取物之用途，係用以製造PDE5抑制劑。 The use of a powder or extract of Cordyceps sinensis is used to manufacture a PDE5 inhibitor.

項5 item 5

一種蠶蟲草的粉末或萃取物之用途，係用以製造排尿障礙及/或肺動脈性肺高血壓症之預防或改善用組成物。 A powder or extract of Cordyceps sinensis is used to manufacture a composition for preventing or improving urination disorders and/or pulmonary arterial pulmonary hypertension.

項6 item 6

一種方法，係對必須抑制PDE5的對象投予蠶蟲草的粉末或萃取物。 In one method, powder or extract of Cordyceps sinensis is administered to a subject who must inhibit PDE5.

項7 item 7

如項6所記載之方法，其中，對象係必須預防或改善排尿障礙及/或肺動脈性肺高血壓症。 The method according to item 6, wherein the subject needs to prevent or improve urination disorders and/or pulmonary arterial pulmonary hypertension.

項8 item 8

一種蠶蟲草的粉末或萃取物，係使用於PDE5之抑制。 A powder or extract of Cordyceps sinensis is used for PDE5 inhibition.

項9 item 9

一種蠶蟲草的粉末或萃取物，係使用於排尿障礙及/或肺動脈性肺高血壓症之預防或改善。 A powder or extract of Cordyceps sinensis is used for the prevention or improvement of urination disorder and/or pulmonary arterial pulmonary hypertension.

本發明提供具有PDE5抑制作用之物質。一實施形態中，係提供一種利用PDE5抑制作用而預防或改善排尿障礙及/或肺動脈性肺高血壓症之手段。 The present invention provides substances with PDE5 inhibitory effect. In one embodiment, a means for preventing or ameliorating dysuria and/or pulmonary arterial pulmonary hypertension using PDE5 inhibition is provided.

圖1係表示針對蠶蟲草測定PDE5抑制活性之結果。「試樣(Sample)」係表示蠶蟲草之熱水萃取物。「受質(Substrate)」係表示cGMP。 Fig. 1 shows the results of measuring the PDE5 inhibitory activity against Cordyceps sinensis. "Sample" means the hot water extract of Cordyceps sinensis. "Substrate" means cGMP.

PDE5抑制劑較佳含有蠶蟲草的粉末或其萃取物。蠶蟲草係寄生於蠶並生育之細腳擬青黴(P.tenuipes，亦稱為「冬蟲夏草」)。蠶蟲草亦可使用市售者及栽培者之任一者。蠶蟲草之栽培方法係任意，例如可舉出：將蠶的幼蟲或蠶蛹加熱滅菌處理後，將冬蟲夏草之菌植菌進行育成的方法。再者，可舉出日本特開2005-237201所揭示之方法。具體而言，可將形成繭之前的蠶的幼蟲或蠶蛹煮沸並使其乾燥，將此蠶的乾燥粉末50至90重量%，及剩餘部分為由豆類、穀類、 海藻類或菇類之乾燥粉末的1種或2種以上構成的食物乾燥粉末混合，於此中添加培養液並混練，將此鋪在育成箱的底部做成培養基，將此培養基封入植菌袋並進行加熱滅菌處理後，對培養基接種冬蟲夏草的菌並進行育成。 The PDE5 inhibitor preferably contains a powder of Cordyceps sinensis or an extract thereof. Cordyceps silkworm is a parasitic species of Paecilomyces tenuipes (P. tenuipes, also known as "Cordyceps sinensis") which is parasitic on silkworms. Cordyceps silkworm can also use any of a commercial one and a cultivator. The cultivation method of silkworm Cordyceps is arbitrary, for example, the method of growing the fungus of Cordyceps sinensis after heat sterilization of silkworm larvae or silkworm chrysalis is mentioned. Furthermore, the method disclosed in Japanese Patent Application Laid-Open No. 2005-237201 can be mentioned. Specifically, the larvae or pupa of silkworms before cocoon formation can be boiled and dried, and 50 to 90% by weight of the dried powder of the silkworms and the remainder are made of beans, cereals, One or two or more kinds of dry powder of seaweed or mushrooms are mixed with dry food powder, and the culture medium is added and kneaded. The medium is spread on the bottom of the growing box to form a medium, and the medium is sealed in a bacterial planting bag. And after heat sterilization treatment, the culture medium is inoculated with the fungus of Cordyceps sinensis and grown.

在一實施形態中，蠶蟲草較佳為粉末的形態。蠶蟲草的粉末係可使蠶蟲草凍結乾燥，並進行粉碎而獲得。蠶蟲草之粉末係可為僅將蠶蟲草之子實體粉末化成者，較佳為將子實體及宿主(例如，蠶)粉末化而成者。 In one embodiment, Cordyceps silkworm is preferably in the form of powder. The powder of Cordyceps sinensis can be obtained by freeze-drying and pulverizing Cordyceps sinensis. The powder of Cordyceps sinensis may be obtained by powdering only the fruiting body of Cordyceps sinensis, preferably by powdering the fruiting body and the host (eg, silkworm).

在一實施形態中，蠶蟲草較佳為萃取物的型態。為了獲得萃取物所使用之方法及條件為任意。例如，可直接將蠶蟲草進行榨取或使用適當的溶劑進行萃取，或在將蠶蟲草進一步切斷、粉碎及/或乾燥後，進行榨取或使用適當的溶劑進行萃取。溶劑之種類沒有特別的限制，可列舉例如：水；甲醇、乙醇、異丙醇等低級醇；丙二醇、1,3-丁二醇等多元醇；丙酮等酮類；二***、二噁烷、乙腈；乙酸乙酯等酯類；二甲基亞碸、二甲苯、苯、氯仿等。在一實施形態中較佳的溶劑係水，較佳為熱水(60℃以上、70℃以上、80℃以上、或90℃以上)。 In one embodiment, the Cordyceps silkworm is preferably in the form of an extract. The method and conditions used to obtain the extract are arbitrary. For example, Cordyceps sinensis can be directly extracted or extracted with an appropriate solvent, or after further cutting, pulverization and/or drying of Cordyceps sinensis, it can be extracted or extracted with an appropriate solvent. The type of solvent is not particularly limited, for example, water; lower alcohols such as methanol, ethanol, and isopropanol; polyhydric alcohols such as propylene glycol and 1,3-butanediol; ketones such as acetone; diethyl ether, dioxane, Acetonitrile; Esters such as ethyl acetate; Dimethyl sulfite, xylene, benzene, chloroform, etc. The preferred solvent-based water in one embodiment is hot water (60°C or higher, 70°C or higher, 80°C or higher, or 90°C or higher).

在一實施形態中，蠶蟲草之萃取物較佳是藉由下述方法獲得者。亦即，較佳係可於蠶蟲草的乾燥粉末添加熱水，在60℃以上80℃以下的溫度下攪拌預定的時間(例如，1小時以上5小時以下)，進行過濾並將所得之濾液乾燥而獲得萃取物。乾燥粉末與熱水的混合比率為任意，例如，相對於乾燥粉末1重量份，熱水可設為5至20重量份、或7至15重量份。 In one embodiment, the extract of Cordyceps sinensis is preferably obtained by the following method. That is, it is preferable to add hot water to the dry powder of Cordyceps sinensis, stir at a temperature of 60°C or more and 80°C or less for a predetermined time (for example, 1 hour or more and 5 hours or less), filter and dry the obtained filtrate. to obtain the extract. The mixing ratio of the dry powder and the hot water is arbitrary. For example, the hot water can be set to 5 to 20 parts by weight, or 7 to 15 parts by weight with respect to 1 part by weight of the dry powder.

蠶蟲草的粉末或萃取物的水分含量係沒有特別的限制，例如，可設為20重量%以下、10重量%以下、8重量%以下、或5重量%以下。 The moisture content of the powder or extract of Cordyceps sinensis is not particularly limited, but can be, for example, 20 wt % or less, 10 wt % or less, 8 wt % or less, or 5 wt % or less.

PDE5抑制劑係可為僅由蠶蟲草的粉末或其萃取物所構成者，亦可為進一步含有任意的其他成分者。其他成分可列舉例如：賦形劑、結合劑、崩解 劑、潤滑劑、濕潤劑、流動化劑、保存劑、界面活性劑、穩定劑、稀釋劑、溶解劑、等張化劑、殺菌劑、防腐劑、調味劑、矯臭劑、著色劑、香料等。其他成分亦可選擇其他具有PDE5抑制活性之成分或其他生理活性成分。PDE5抑制劑之形態係任意，例如可作成為：粉末狀、顆粒狀、錠劑狀、膠囊狀、或液體狀。 The PDE5 inhibitor may be composed of only the powder of Cordyceps sinensis or its extract, or may further contain any other components. Other ingredients include, for example: excipients, binding agents, disintegration Agents, lubricants, wetting agents, fluidizing agents, preservatives, surfactants, stabilizers, diluents, dissolving agents, isotonic agents, bactericides, preservatives, flavoring agents, odorants, colorants, fragrances, etc. . Other ingredients can also be selected from other ingredients with PDE5 inhibitory activity or other physiologically active ingredients. The form of the PDE5 inhibitor is arbitrary, and for example, it can be in the form of powder, granules, lozenges, capsules, or liquids.

PDE5抑制劑所含有之蠶蟲草的粉末或其萃取物的量係任意，例如以乾燥重量換算，可設定在0.1%以上100%以下的範圍。含量的下限係例如可設為：1%以上、2%以上、5%以上、10%以上、15%以上、20%以上、30%以上、40%以上、50%以上、60%以上或70%以上，上限可設為99%以下、95%以下、90%以下、80%以下、70%以下、或60%以下。此等下限及上限係可任意地組合。 The amount of the powder of Cordyceps sinensis or its extract contained in the PDE5 inhibitor is arbitrary, but can be set in the range of 0.1% or more and 100% or less in terms of dry weight, for example. The lower limit of the content can be, for example, 1% or more, 2% or more, 5% or more, 10% or more, 15% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, or 70% or more. % or more, and the upper limit can be set to 99% or less, 95% or less, 90% or less, 80% or less, 70% or less, or 60% or less. These lower and upper limits can be arbitrarily combined.

在一實施形態中，PDE5抑制劑較佳為飲食品之形態。飲食品中，不僅包含一般食品，亦包含保健機能食品(具體而言，特定保健用食品、營養機能食品、機能性表示食品)、患者用食品、補給品等。 In one embodiment, the PDE5 inhibitor is preferably in the form of food and drink. Foodstuffs include not only general foods but also health functional foods (specifically, foods for specified health use, nutritional functional foods, and functionally indicated foods), foods for patients, supplements, and the like.

在一實施形態中，PDE5抑制劑之形態係可為：液劑、乳劑、懸浮劑、糖漿劑等液狀形態；散劑、顆粒劑、細粒劑、錠劑、丸劑、膠囊劑(包含硬膠囊、軟膠囊)、***錠(troche)、咀嚼錠、凝膠狀、膏狀、糊狀、慕思狀、片狀等半固形或固形形態。 In one embodiment, the form of the PDE5 inhibitor can be: liquid forms such as liquids, emulsions, suspensions, syrups, etc.; powders, granules, fine granules, lozenges, pills, capsules (including hard capsules); , soft capsule), troche, chewable tablet, gel, paste, paste, mousse, tablet and other semi-solid or solid forms.

一般食品係沒有特別的限制，可列舉例如：飲料類(例如，果汁飲料、蔬菜汁、清涼飲料、酒精飲料、碳酸飲料、運動飲料、滋養強壯飲品、營養飲品等)、菓子類(例如，點心菓子、米菓、巧克力、軟糖、糖果、果凍、布丁、和菓子、西式糕點等)、調味料類(例如，醬油、味噌、味醂風調味料、混合香料、醬、湯、醬料、番茄醬等)、沙拉醬類、麵類(例如，烏龍麵、蕎麥麵、義大利麵 等)、畜肉魚肉加工食品類(例如，香腸、維也納香腸、火腿、漢堡肉等)、乳製品類(優酪乳、牛乳、起司、鮮奶油、奶油等)等。 General food products are not particularly limited, and examples include beverages (for example, fruit juice drinks, vegetable juices, soft drinks, alcoholic drinks, carbonated drinks, sports drinks, nourishing and strong drinks, nutritional drinks, etc.), fruits (for example, desserts) Fruits, rice crackers, chocolate, fudge, candy, jelly, pudding, wagashi, western cakes, etc.), seasonings (for example, soy sauce, miso, mirin seasoning, mixed spice, sauce, soup, sauce, ketchup etc.), salad dressings, noodles (e.g., udon, soba, pasta etc.), processed meat and fish foods (for example, sausage, Vienna sausage, ham, hamburger, etc.), dairy products (yogurt, milk, cheese, fresh cream, cream, etc.).

在一實施形態中，上述PDE5抑制劑係藉由抑制PDE5的活性，而可使用於預防或改善排尿障礙及/或肺動脈性肺高血壓症之用途中。雖不論尿路障礙的原因、類型，惟可舉出伴隨下部尿路症狀之尿路障礙。 In one embodiment, the above-mentioned PDE5 inhibitor can be used for the prevention or improvement of dysuria and/or pulmonary arterial pulmonary hypertension by inhibiting the activity of PDE5. Regardless of the cause or type of urinary tract disorder, urinary tract disorders with lower urinary tract symptoms can be cited.

在一實施形態中，PDE5抑制劑係可製成醫藥之形態。含有PDE5抑制劑之醫藥，係可於蠶蟲草的粉末或萃取物中因應所需而組合藥學上可容許的賦形劑進行調製。 In one embodiment, the PDE5 inhibitor can be formulated into a pharmaceutical form. The medicine containing PDE5 inhibitor can be prepared by combining pharmaceutically acceptable excipients in the powder or extract of Cordyceps sinensis as required.

只要可發揮作為目的之效果，則PDE5抑制劑之攝取(投予)量沒有特別的限定，可因應對象者(對象動物)之體格、年齡、症狀、適用形態、使用目的等而適當地設定。例如，每1日的攝取(投予)量，係以體重60kg的成人為基準，以蠶蟲草的粉末或其萃取物的乾燥重量換算，可設定在0.001至8000mg，0.01至5000mg等之範圍。攝取(投予)次數係任意，例如，每1日可單次攝取(投予)亦可複數次攝取(投予)。 The ingestion (administration) amount of the PDE5 inhibitor is not particularly limited as long as the intended effect is exhibited, and can be appropriately set according to the physique, age, symptoms, application form, and purpose of use of the subject (subject animal). For example, the daily intake (administration) amount can be set in the range of 0.001 to 8000 mg, 0.01 to 5000 mg, etc. in terms of the dry weight of the powder of Cordyceps sinensis or its extract based on an adult with a body weight of 60 kg. The frequency of ingestion (administration) is arbitrary, and for example, it may be ingested (administered) once per day or may be ingested (administered) several times.

如上述，蠶蟲草的粉末或其萃取物係由於具有PDE5抑制作用，因此可為了製造PDE5抑制劑而使用蠶蟲草的粉末或萃取物。再者，亦可為了製造排尿障礙及/或肺動脈性肺高血壓症之預防或改善用組成物，而使用蠶蟲草的粉末或萃取物。 As described above, the powder or extract of Cordyceps sinensis has a PDE5 inhibitory effect, so the powder or extract of Cordyceps sinensis can be used for the production of a PDE5 inhibitor. Furthermore, in order to manufacture the composition for prevention or improvement of urination disorder and/or pulmonary arterial pulmonary hypertension, the powder or extract of Cordyceps sinensis can also be used.

如上述，蠶蟲草的粉末或其萃取物係由於具有PDE5抑制作用，因此可對必須抑制PDE5的對象投予蠶蟲草的粉末或萃取物，而提供PDE5抑制作用。再者，藉由抑制PDE5，可預防或改善排尿障礙及/或肺動脈性肺高血壓症， 因此藉由對必須預防或改善排尿障礙及/或肺動脈性肺高血壓症之對象投予蠶蟲草的粉末或萃取物，可進行該預防或改善。 As described above, the powder or extract of Cordyceps sericata has a PDE5 inhibitory effect. Therefore, the powder or extract of Cordyceps can be administered to a subject who needs to inhibit PDE5, thereby providing a PDE5 inhibitory effect. Furthermore, by inhibiting PDE5, urinary disorders and/or pulmonary arterial pulmonary hypertension can be prevented or improved, Therefore, the prevention or improvement can be performed by administering the powder or extract of Cordyceps sinensis to a subject who must prevent or improve urination disorders and/or pulmonary arterial pulmonary hypertension.

[實施例] [Example]

以下，藉由實施例更詳細地說明本發明，但本發明不受到該等所限制。 Hereinafter, the present invention will be described in more detail by way of examples, but the present invention is not limited by these examples.

1.蠶蟲草熱水萃取物 1. Silkworm Cordyceps Hot Water Extract

將以蠶蛹作為宿主而獲得之細腳擬青黴(蠶蟲草)凍結乾燥，藉由混合器粉碎而獲得蠶蟲草粉末。在反應容器中，饋入所得之粉末(42g)及超純水420mL，以70℃攪拌3小時後，藉由進行過濾去除殘渣而獲得溶液。接著，進行濃縮使溶液成為30mL後，藉由進行噴霧乾燥而獲得蠶蟲草熱水萃取物。 Paecilomyces solani (Cordyceps sinensis) obtained by using silkworm chrysalis as a host was freeze-dried, and pulverized by a mixer to obtain Cordyceps sinensis powder. The obtained powder (42 g) and 420 mL of ultrapure water were put into a reaction vessel, and after stirring at 70° C. for 3 hours, the residue was removed by filtration to obtain a solution. Next, after concentrating the solution to 30 mL, spray-drying was performed to obtain a hot water extract of Cordyceps sinensis.

2.PDE5抑制活性之測定 2. Determination of PDE5 inhibitory activity

PDE5抑制活性係使用PDE-Glo(商標)磷酸二酯酶分析(Phosphodiesterase Aassay)(Promega公司製)進行。以使蠶蟲草熱水萃取物的最終濃度成為33μg/ml、11μg/g、3.7μg/ml、1.2μg/ml之方式利用超純水稀釋，將經稀釋者作為試樣使用。酵素係使用人類磷酸二酯酶5A1(Sigma-Aldrich製)。將3100U/μl之PDE 1μl與反應緩衝液205.7μl混合，調製出PDE溶液。將PDE溶液15μl與各試樣10μl混合後使其短暫離心(spin down)2分鐘，在室溫震盪5分鐘。在此混合液25μl中添加並混合20μM之cGMP 25μl，於PCR管各分注8μl。將此載置於振盪機，以室溫、400rpm使其反應。於各管添加檢測緩衝液4μl，進行短暫離心後以室溫振盪20分鐘。將此與Kinase-Glo試藥10μl於384孔盤(Thermo Fisher製)之各孔分注10μl，以室溫進行振盪並進行反應。將使用未添加蠶蟲草熱水萃取物之試樣的試驗例 作為0%對照組，將未添加PDE之試驗例作為100%對照組，測定由蠶蟲草熱水萃取物所致之相對的抑制率。測定係重複4次。將結果示於圖1。 The PDE5 inhibitory activity was carried out using PDE-Glo (trademark) Phosphodiesterase Assay (manufactured by Promega). It was diluted with ultrapure water so that the final concentrations of the hot water extract of Cordyceps sinensis were 33 μg/ml, 11 μg/g, 3.7 μg/ml and 1.2 μg/ml, and the diluted ones were used as samples. As the enzyme, human phosphodiesterase 5A1 (manufactured by Sigma-Aldrich) was used. 1 μl of 3100 U/μl PDE was mixed with 205.7 μl of the reaction buffer to prepare a PDE solution. After mixing 15 μl of the PDE solution with 10 μl of each sample, it was spin down for 2 minutes and shaken at room temperature for 5 minutes. To 25 μl of this mixture, 25 μl of 20 μM cGMP was added and mixed, and 8 μl was dispensed into each PCR tube. This was placed on a shaker and reacted at room temperature and 400 rpm. 4 μl of detection buffer was added to each tube, briefly centrifuged, and then shaken at room temperature for 20 minutes. 10 μl of this and the Kinase-Glo reagent were dispensed into each well of a 384-well plate (manufactured by Thermo Fisher) in 10 μl, and the reaction was performed by shaking at room temperature. A test example using a sample without the addition of a hot water extract of Cordyceps sinensis As a 0% control group, a test example without adding PDE was used as a 100% control group, and the relative inhibition rate by the hot water extract of Cordyceps silkworm was measured. The assay was repeated 4 times. The results are shown in FIG. 1 .

由圖1之結果可確認，在蠶蟲草熱水萃取物中存在PDE5抑制活性，其活性係依存於濃度而上昇。由此結果可認為蠶蟲草係可有效地治療或預防肺動脈性肺高血壓症及排尿障礙等疾病。 From the results in Fig. 1, it was confirmed that PDE5 inhibitory activity exists in the hot water extract of Cordyceps serrata, and the activity increases depending on the concentration. From these results, it is considered that the Cordyceps sinensis system can effectively treat or prevent diseases such as pulmonary arterial pulmonary hypertension and urination disorders.

Claims (3)

一種PDE5抑制劑，係含有蠶蟲草的粉末或萃取物。 A PDE5 inhibitor containing powder or extract of Cordyceps sinensis. 如請求項1所述之PDE5抑制劑，係飲食品組成物之形態。 The PDE5 inhibitor according to claim 1 is in the form of a food and drink composition. 如請求項1或2所述之PDE5抑制劑，係排尿障礙及/或肺動脈性肺高血壓症之預防或改善用者。 The PDE5 inhibitor according to claim 1 or 2 is used for preventing or ameliorating urination disorders and/or pulmonary arterial pulmonary hypertension.

Images