TW422694B - Compositions for treatment of diabetic complications - Google Patents

Abstract

Use of 6-desaturated n-6 fatty acids, especially gammalinolenic acid (GLA), dihomogammalinolenic acid (DGLA) or arachidonic acid (AA), together with a pharmaceutically acceptable material reducing intracellular levels of sorbitol in the body, particularly an aldose reductase inhibitor, in the treatment of (including prophylactic treatment), and in the preparation of medicaments for the treatment of (including prophylactic treatment), the long-term complications of diabetes mellitus. Pharmaceutical compositions of said materials. The ascorbate esters of 6-desaturated n-6 fatty acids (other than GLA or DGLA) per se.

Description

u , A7 一__________. .。B7_______ 五、發明説明（】） 發明背景 糖尿病是常見的疾病，通常分爲胰島素依賴型及非胰島 素依賴型二類。這二型糖尿病都可以控制飲食處理，第— 型還需使用胰島素，第二型還需使用各種藥物。但是，雖 則醫生及病人本人可相當滿意地控制血中葡萄糖的變化， 卻不能預防由此病所引起的對組織的長期的損害，這類損 害可有多種形式，但主要的是對眼睛的損害（視網膜病變） ，對神經的損害（神經病變），對腎臟的損害（腎臟病變）， 及對心血管系統的損害。有些病人的這類損害只限於一個 系統或另一系統，但多數病人有二種或多種損害同時發生 ’這顯ΤΠ"其基本機轉是相你的。 現有多種減少或預防此類損害的方法，此類損害總稱之 爲糖尿病長期併發症。本發明有關於其中二種方法。 一種方法係基於必需脂肪酸（E F Α)的代謝，即阻斷主要 食物EFA ’亞油酸，轉化成其第—代謝物，广-亞麻酸 (GLA)，即下表1中所示順序的第一，6·去飽和的，心 6EFA(於此文中’ ’6 -去飽和EFA'—詞意爲6 -去飽和步驟 之外的EFAs，並不必然表示ef A的6-位上有雙鍵）：- ----^---^---政------ΪΤ------.^ (請先閱讀背面之注意事項再填窝本1) 經濟部中央標準局貝工消費合作社印製 -4 - 本紙張·尺度逋用中國國家標準（CNS ) A4規格（210Χ297公釐） 422694 A7 B7 五、發明説明（2 1 8 :2 Δ -9，12 (亞油酸） Δ - 6去飽和酶 1 8 :3 Λ -6, 9, 12 (r -亞桊酸） 20 :3 Δ - 8 , 11，14 (二高 r -亞爲酸） 加長 1 △ - 5去飽和酶 20:4 Λ -5, 8, 11，14 (花生四烯酸） k 22 :4 Δ -7, 10, 13，16 (腎上腺酸）u, A7 a __________... B7_______ 5. Description of the invention () Background of the Invention Diabetes is a common disease and is generally divided into insulin-dependent and non-insulin-dependent types. Both type 2 diabetes can be controlled by diet. Type 1 also requires insulin, and type 2 requires various drugs. However, although doctors and patients can control the changes in blood glucose quite satisfactorily, they cannot prevent the long-term damage to the tissue caused by this disease. Such damage can take many forms, but the main one is eye damage. (Retinopathy), damage to the nerves (neuropathy), damage to the kidneys (nephropathy), and damage to the cardiovascular system. In some patients, this type of damage is limited to one system or another, but most patients have two or more types of damage at the same time. This shows that the basic mechanism is the same. There are many ways to reduce or prevent such damage, which are collectively referred to as long-term complications of diabetes. The present invention relates to two of these methods. One method is based on the metabolism of essential fatty acids (EF A), which blocks the main food EFA 'linoleic acid and converts it into its first metabolite, broad-linolenic acid (GLA), which is the first in the order shown in Table 1 below. One, 6 · desaturated, 6EFA (in this article '' 6-desaturated EFA '-the word means EFAs outside the 6-desaturation step, does not necessarily mean that there is a double bond at the 6-position of ef A ):----- ^ --- ^ --- political ------ ΪΤ ------. ^ (Please read the notes on the back before filling in the book 1) Central Bureau of Standards, Ministry of Economic Affairs Printed by Bei Gong Consumer Cooperative-4-This paper · size is in accordance with Chinese National Standard (CNS) A4 (210 × 297 mm) 422694 A7 B7 V. Description of the invention (2 1 8: 2 Δ -9, 12 (linoleic acid ) Δ-6 desaturase 1 8: 3 Λ -6, 9, 12 (r -arsonic acid) 20: 3 Δ-8, 11,14 (two-high r-sulfinic acid) Lengthen 1 △-5 to Saturase 20: 4 Λ -5, 8, 11, 14 (arachidonic acid) k 22: 4 Δ -7, 10, 13, 16 (adrenal acid)

Δ - 4去飽和酶 I 經濟部中央標準局員工消費合作杜印製 22:5 Δ -4，7，10，13，16 結果是，GLA的代謝物，包括二高•亞麻酸（DGLA) 輿花生四烯酸（AA)，的含量減少。由於此等代謝物是神 經膜及所有組織細胞膜構造的重要成分，也由於此等代謝 物是細胞信號系統成分，如前列線素及二醯基甘油的先質 ，此等化合物的缺乏是會構成損害的。特別是會發生由 DGL A衍生的前列線素e及由花生四烯酸衍生的前列環素 的缺乏，而此二化合物卻是血管舒張劑，爲多種組織維梏 血液流入微循環所必需。 -5* 本紙張尺度適用中國國家標準（CNS > A4規格（2】〇X297公釐） -------_---d------，玎------d. (請先閱讀背面之注意事項再填寫本頁) 經濟部中央標準局員工消費合作社印裂 422β94 B7 五、發明説明（3 ) 於糖尿病合併症中，亞麻酸之轉化成G L A受損害，以 GLA及/或其代謝物DGLA及AA作治療是有益的。糖尿病 物的神經的前列環素含量減少，可用G L A治療得到恢復。 另一方法是以體内細胞中的葡萄糖代謝物，山梨糖醇， 的生成過多引起損害爲基礎。葡萄糖可由醛糖還原酶轉化 成山梨糖醇，但在葡萄糖正常濃度下，此一轉化步驟很慢 ’所以山梨糖醇的含量通常也很低。相反，在葡萄糖濃度 高時，轉化速度轉快，山梨糖醇的含量也增高s此不同含 量對细胞有不同的影響，引起克分子滲透壓的紊亂，並缓 亂細胞备種代謝’包括肌踔循環。此等影響有人認爲是糖 尿病合併症的原因。如果情形確是如此，使用抑制劑阻斷 酶，或者更廣泛地給予減少山梨糖醇含量的物質，在治療 /預防糖麻病合併症上應會是有價値的。此類抑制劑，越 糖還原酶抑制劑，已由許多醫藥公司發展成功。 而且’最近的報告（ehnningham et al.，J. Am. Coil. Nutrition 13,（No. 4) 344-350 (1994)顯示，抗壞血酸在抑 制酶方面與任何醛糖還原酶抑制劑藥物有同等效力。 在研究治療時，發明人等給鼠注射鏈脲菌素 (streptozotocin)，使胰臟分泌胰島素的細胞受損害，產生 神經傳導變慢的效果’發展成糖尿病合併症的模型。此種 慢神經傳導可用胰島素治療恢復正常。也可用各種形式的 -亞油酸治療使恢復正常，或用不同的醛糖還原酶抑制 劑治療使恢復正常。 發明人等已建成劑量曲線，顯示以G L A及酶抑制劑恢復 本紙張尺度適用中國國家標準（CNS ) Λ4規格（210X297公產） ^ - -裝 訂^ (請先聞讀背面之注意事項再填寫本頁) 第84109228號專利申請案 中文説明書修正頁(貼年3月）五、發明説明（4) 4226 B4 A7 B7 經濟部中央標孪局負工消費合作社印製Δ-4 Desaturase I Central Government Bureau of Standards, Ministry of Economic Affairs, Consumer Consumption Cooperation, Du printed 22: 5 Δ -4, 7, 10, 13, 16 The result is that GLA metabolites, including digoline • linolenic acid (DGLA) Decreased arachidonic acid (AA). Since these metabolites are important components of the structure of nerve membranes and cell membranes of all tissues, and because these metabolites are components of the cell signaling system, such as precursors of prostaglandin and diglycerin, the lack of these compounds may constitute Damage. In particular, deficiency of prostaglandin e derived from DGL A and prostacyclin derived from arachidonic acid will occur, but these two compounds are vasodilators and are necessary for various tissues to maintain blood flow into the microcirculation. -5 * This paper size applies to Chinese national standards (CNS > A4 size (2) × 297mm) -------_--- d ------, 玎 ------ d. (Please read the notes on the back before filling this page) Printed by the Consumers Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 422β94 B7 V. Description of the invention (3) In the case of diabetic comorbidities, the conversion of linolenic acid into GLA is damaged. GLA and / or its metabolites DGLA and AA are beneficial. Diabetic neuroprostaglandin content is reduced and can be restored with GLA treatment. Another method is the glucose metabolite in the cells in the body, sorbitol Based on the damage caused by excessive production of glucose. Glucose can be converted to sorbitol by aldose reductase, but at normal glucose concentrations, this conversion step is slow ', so the content of sorbitol is usually very low. In contrast, in glucose When the concentration is high, the transformation speed becomes faster, and the content of sorbitol also increases. This different content has different effects on the cells, causing disturbances in osmolarity, and slowing the cell preparation metabolism, including the muscle loop. Affects what some people think is diabetes Causes of comorbidities. If this is the case, using inhibitors to block enzymes, or more broadly administering substances that reduce sorbitol, should be of value in the treatment / prevention of comorbidities. Inhibitors, vibrose reductase inhibitors, have been successfully developed by many pharmaceutical companies. And 'recent reports (ehnningham et al., J. Am. Coil. Nutrition 13, (No. 4) 344-350 (1994) show Ascorbic acid has the same efficacy in inhibiting enzymes as any aldose reductase inhibitor drug. In the study of treatment, the inventors injected rats with streptozotocin, causing the pancreatic insulin-secreting cells to be damaged and nerves produced. The effect of slow conduction has developed into a model of diabetic comorbidities. This slow nerve conduction can be restored to normal by insulin treatment. It can also be restored to normal by various forms of linoleic acid treatment, or treated with different aldose reductase inhibitors The inventors have established a dose curve, which shows that the recovery of this paper with GLA and enzyme inhibitors applies the Chinese National Standard (CNS) Λ4 specification (210X297 (Production) ^--Binding ^ (Please read the precautions on the back before filling this page) No. 84109228 Patent Application Chinese Specification Revision Page (posted in March) V. Invention Description (4) 4226 B4 A7 B7 Economy Printed by the Ministry of Standards and Technology Bureau's Consumer Cooperatives

正常的神經傳導，此實驗詳情已於各文獻敘述過，如N ENormal nerve conduction, details of this experiment have been described in various literatures, such as N E

Cameron，M.A. Corter and S. Robertson 於’'Diabetes" , Vol 40，頁532-39 (1991)所述。主要是，用鏈脲菌素使鼠生 糖尿病，六個星期内不以GLA或其他脂肪酸治療，然後用 脂肪·酸治療2星期，有的使用其他試驗物質，有的不使用其 他試驗物質。與不治療的糖尿病组比較，藥物的效果以對 恢復傳導速度的程度顯示。 本發明工作 發明人等已研究了 GLA及醛糖還原酶抑制劑ZD5522效果 問的相互作用。開始時，每一有效成分都以閥量給予，期使 糖尿病模型的傳導速度增進約5 %。可期望同時給予二種治 療時’最佳能有相加效果，導致增進1 〇 %。然有以幾個實驗 ，視所選確切劑量而定，實際的增進達4 0-80%，較期望的 値大近1.5 - 6倍》這是完全未曾想到的，也是有很大治療意 義的，這可使治療效果大增而同時又使副作用減少，因爲用 較現在所使用的較少的藥物劑量即可達治療效果。使用第二 醛糖還原酶抑制劑WAY 1 2 1 5 09也獲得類似結果。 此外，在使用技壞血酸g旨代替趁糖還原酶抑制劑（A RI s) 之類藥物時，還有只使用天然物質的主要優點，即使大劑 量也具时受性，而且還可以技壞血酸基- GLA等單一化合物 形式使用。 圖式簡單説明 圖1爲各種形式的GLA，包括待宵草油（EP0)、二亞油醯 基-單-r-亞麻醯基-甘油（DLMG)及三-r-亞麻醯基-甘油 本紙疾尺度適用中国S家標準（CNS ) A4規格（2丨0X297公釐） n. m - In - - I I - I—I. ^^1 T* (請先閱讀背面之注意事項再填寫本頁) W至1 補充 第84109228號專利申請案4226 9 中文説明書修正頁(86年3月） 。 B7 五、發明説明（5) (tri-GLA)，的劑量反應曲線》此曲線顯示鏈脲菌素-糖尿 病鼠的神經傳導速度向正常恢復的百分比。此曲線顯示以 與G L A相同的毫克/公斤/天的劑量給予時，各種不同來源 的GLA的效果》 圖2顯示以8毫克/公斤/天的GLA，0.25毫克/公斤/天的 2D5522及0.2毫克/公斤/天的WAY 121509作單獨給予或同 時給予時，對正常及糖尿病動物神經傳導的影響。 ZD5522是3，，5’ -二曱基-4'-硝基甲基·項醯基- 2- (2 -甲苯 基）-N-乙醯苯胺。WAY 1 2 1 509是螺-(異喹啉-4( 1H)， 3、吡咯烷）-1，2\3、5'-(2Η) -四酮。此三種化合物的劑量 在單獨給予時，對神經傳導只有小的改進，但以Z D 5 5 2 2 或WAY 121509與GLA共同给予時，對神經傳導所產生的 效果卻比二者相加的效果大得多。 圖3顯示，單獨给予GLA及抗壞血酸及二者同時給予以及 給予單一抗壞血酸基-GLA對糖尿病鼠坐骨神經血流及傳導 速度的影響。在此情形下，二種化合物同時给予所產生的 效果比相加略大，但本身也有價値。但抗壞血酸基-G L A作 爲單一化合物给予時，其作用明顯地較單獨給予GLA或抗 壞血酸二者相加的效果爲佳，也比同時给予GLA及抗壞血 酸爲佳3给予單一化合物抗壞血酸基-G L A之所以明顯的較 佳，可能是因爲同時遞送分子的二種成分至有此需要的位 置的關係。 本發明 這樣，本發明一方面提供醫藥上可接受的组合物，内含 8- 本紙乐尺度逋用中國S家橾準（CNS ) A4規格U丨0X291公t ) I---------钟衣------ΪΤ------.^ (请先閱讀背面之注意事項再填寫本頁) 經濟部中央標準局貝工消資合作社印製 經濟部中央標準局負工消費合作杜印製 4 226 A7 __ B7 _ 五'發明説明（6) 6-去飽和的η·6-脂肪酸，特別是GLA，DGLA或AA，以 及減少體内山梨糖醇在細胞内的含量的物質，特別是醛糖 還原酶抑制劑。較佳的抑制劑是抗壞血酸酯或其複合物形 式’如鹽、酯或其他衍生物，尤其是與脂肪酸所形成的酯 ，但也可用A RI s，，藥物，，。 本發明另一方面係關於使用6 -去飽和的η - 6 -脂昉酸，特 別是GLA’ DGLA或ΑΑ，以及減少體内山梨糖醇在細胞 内的含量的醫藥上可接受的物質，特別是該物質是醛糖還 原酶抑制劑時，以治療（包括預防性治療）長期糖尿病合併 症。如上述的抗壞血酸酯爲佳。一般而言，此等活性物質 可一起給予，但本發明也包括將脂肪酸及降低山梨糖醇含 量的物質分別給予，然在分別給予時要使其有效量同時達 於體内。 在另一方面’本發明係關於治療（包括預防性治療）長期 糖尿病合併症的藥物的製備，製備時可用時使用各活性物 質’也可使用其一種’如係後者，在治療時另—活性成分 也須分別同時給予。 本發明另一方面在提供6 -去飽和的η·6 -脂肪酸（除(^[八 或D G L Α外）的抗壞业酸酿新穎的化合物，特別是抗壞血 酸基-A A ’及使用於其他治療的相同化合物。 是以，本發明特別關於糖尿病合併症的治療，尤其是， 但不限於，神經病變，視網膜病變，腎病變，及心血管病 變；治療係合併給予選自GLA，DGLA或AA的EFA(必需 脂肪酸）及抗壞血酸酯或其他能抑制搭糖還原酶或能降低 本紙張尺度適用中國國家標準（CNS ) A4規格（2〖0.X297公釐） I i I | II ! II^ (請先閲讀背面之注意事項再填寫本頁) 經濟部中央標準局負工消費合作社印製 422694 at __— _B7 五、發明説明（了、 ~. 一 身體山梨糖醇含量的物質。此等活性物質可以分離的調配 物提供，但分離的調配物係同時給予（如果分別給予，至 少須於同時在體内有有效量），也可製成複合劑形；不論 是分離的調配物或複合劑形，都可製成各種合適的劑形。 特別是可使用抗壞血酸基-GLA，抗壞血酸基-Dgla，或 抗壞血酸基- AA。劑形可以是膠囊，鼓，散，微膠囊，乳 液’微脂粒’微膠粒，或任何適於經口給予、經腸給予、 非經腸給予、或局部給予的劑形，這都是精於此技藝者所 熟知的β EF Α的劑量可在，例如，丨〇毫克至克/天的範圍内， 較佳是100毫克至5克/天，更佳是300毫克至2克/天，劑 量單位劑形可含此等劑量或其低倍的量。醛糖還原酶抑制 劑的劑量須適於所選用的成分，此處所示爲包括所有己知 此類藥物的廣範圍内的每天有效量，並非表示每一藥物的 劑量都在這一極端範圍内，即0.01至100毫克/公斤/天， 並請注意，任何能減低葡萄糖轉化成山梨糖醇的物質都可 使用。其例有 ponalrestat，tolrestat，及epalrestat,廣範圍説 ，其劑量都是0·1至20毫克/公斤/天，此爲以70公斤成年 人計算出的對應的量。如係使用抗壞血酸酯，劑量是，例 如，1 〇毫克至5 0克/天，較佳是5 0毫克至2克，此爲在使 用抗壞血酸基-G L Α或其他複合形式時以抗壞血酸計算。 EFA可以是任何醫藥上可接受的形式，可在體内導致可 利用的£ F A。特別合適的形式，除各種抗壞血酸基-E F A 外，包括單-，二-及三酸甘油酯，自由態脂肪酸，膽固醇 -10 - 本紙張尺度適用中國國家榇準（CNS ) A4規格（210X297公釐） ---^-------¾衣------il------t (請先閲讀背面之注意事項再填寫本頁) 經濟部中央標準局員工消費合作杜印裝 422694 A7 !_________B7 五、發明説明（s ) 酷’鹽，醯胺及各種形式的嶙脂質，但不包括醫藥上不能 接受的形式。天然待宵草油或加多DLMG(二亞油醯基-單 -广-亞麻醯基-甘油）含量的待宵草油爲特別方便，或用其 他來源的DLMG °也可使用衍生自EFA的脂肪醇。 以下的實例説明本發明，並以E F A及已知的醛糖還原酶 抑制藥物的组合物説明，再以EFA與抗壞血酯的組合物説 明’最後以抗壞血酸的EFA酯製備説明。ARIs”藥物"在 有合適的化學構造時，可視爲是EFA化合物，此等單一化 合物較佳是使用於分別给予的二種製劑。 -igi二-醛糖還原酶抑制劑藥物組合物 經糖還原酶抑制劑（ARIs)有很多不同的種類，也可能 將來會發現更多種。同樣，也可能發現有些藥物以不同的 機轉抑制山梨糖醇的產生。下面爲使用已知的特定的 ARIs的調配物的實例： 1· 400毫克硬或軟膠囊，含有三酸甘油酯' 自由態脂肪酸 、鹽或其他形式的GLA，DGLA或AA，與含300毫克 ARI的鍵劑ponalrestat共同給予，2粒E F A膠囊及1粒 ponalrestat共同給予，早晚各一次。 2 ·同實例1，然ARI爲tolrestat，每天二次，每次2 0 0毫克 的鍵劑。 3 -同實例1，然ARI爲epalrestat，每天三次’每次150毫 克的錠劑。 4.同實例 1，然用 ADN-138(Ono)，SNK-860(Sanwa)， M16209(Fuji)，ZD5522(Zeneca)或 WAY 12 1 5 09(Wyeth) -a - 本纸張尺度適用中國國家標準（CNS ) A4ft格（210 X 297公釐） ；------I------1T------t {請先閱讀背面之注意事項再填寫本頁) 經濟部中央標隼局貝工消f合作杜印製 422694^7 1>7五、發明説明（9 ) 代替ARI s。 5 _同實例1，然E F A與A RI係製成一劑形，此劑形可以是 膠囊，錠，計量容器或任何精於此技藝者所知的其他合適 的劑形’如微膝囊’散’微脂粒，反向微脂粒（reverse liposomes)或供經腸、經口、非經腸或其他給予途徑給予 的形式。 6. 同實例1，然EFA係調配成200毫克、300毫克、500 毫克、600毫克、800毫克或1克的膠囊，每天給予總劑量 爲200毫克至3克。 實例--抗壞血酸酯組合物 7. 實例1或6中之GLA，DGLA或AA與50毫克至2克抗壞 血酸同時給予，二者製成錠或其他合適的調配物。 8. 使用抗壞血酸基-G L A，如以下法合成者，或抗壞血酸 基- DGLA或抗壞血酸基-AA，使抗壞血酸的給予量如前 述’製成如下的任何劑形：-- (a) 錠劑，含50，100，25 0，500或75 0毫克抗壞血酸基 -GLA’杌壞血酸基- DGLA或抗壞血酸基-AA，或只含此 成分或又含合通的賦形劑。 (b) 軟明膠膠囊或硬膠膠囊，含5〇，1〇〇，250，或5 00毫 克抗壞血酸基-GLA，抗壞血酸基-DGLA，或抗壞如酸基 •AA，溶於加了 GLA或DGLA的自由態脂肪酸内，或溶於 三酸甘油酯内，此三酸甘油酯之一或個基團選自GLA或 DGLA。 (c_)供經口、經腸或非經腸給予的抗壞血酸基-GLA或抗壞 • 12- 本紙^Ϊ適用中國國家標準rCNS)A4規格（210X297公釐) ： — -------1----¾衣------1T------^ {請先閲讀背面之注意事項再填寫本頁) 經濟部中央標準局*:工消費合作杜印製 422b j i a? 42Pe.QA_ B7____ 五、發明説明（10 ) 血酸基-D G L A的乳劍、散劑、液劑、糊劑或溶峰。例如 ，乳劑可用適當的乳化劑，如卵磷脂或燕麥半乳糖脂製備 ，製成最終乳劑含達2 %- 3 0 %重量比的抗壞血酸基_GLa ，抗壞血酸基-DGLA或抗壞血酸基-AA。 (d)供局部使用抗壞血酸基-GLA，抗壞血酸基_DgLAs 抗壞血酸基-AA的膏、霜、洗液、洗髮液或其他合政的調 配物。Cameron, M.A. Corter and S. Robertson, as described in '' Diabetes ", Vol 40, pp. 532-39 (1991). Mainly, rats were diabetic with streptozotocin, and were not treated with GLA or other fatty acids for six weeks, and then treated with fat and acid for two weeks. Some used other test substances and some did not use other test substances. Compared to the untreated diabetic group, the effect of the drug is shown to the extent that it restores conduction velocity. The work of the present invention The inventors have studied the interaction between GLA and the effect of aldose reductase inhibitor ZD5522. At the beginning, each active ingredient is given in a valence to increase the conduction speed of the diabetes model by about 5%. It can be expected that when two treatments are given at the same time, the best effect will be additive, leading to an increase of 10%. However, there are several experiments, depending on the exact dose chosen. The actual increase is 40-80%, which is nearly 1.5-6 times larger than the expected》. This is completely unexpected and has great therapeutic significance. This can greatly increase the therapeutic effect while reducing the side effects, because the therapeutic effect can be achieved with less drug dose than currently used. Similar results were obtained with the second aldose reductase inhibitor WAY 1 2 1 5 09. In addition, when using ascorbic acid g instead of drugs such as sugar reductase inhibitors (A RI s), there is also the main advantage of using only natural substances. Even large doses are time-sensitive and can also be used. Used as a single compound such as ascorbyl-GLA. Brief description of the drawing Figure 1 shows various forms of GLA, including sage oil (EP0), linoleyl-mono-r-linoleyl-glycerol (DLMG), and tri-r-linoleyl-glycerin paper Disease standards are applicable to China Standards (CNS) A4 specifications (2 丨 0X297 mm) n. M-In--II-I—I. ^^ 1 T * (Please read the precautions on the back before filling this page) W to 1 Supplement No. 84109228 Patent Application 4226 9 Revised Chinese Manual (March 86). B7 V. Description of the invention (5) (tri-GLA), dose-response curve This curve shows the percentage of neurotransmission rate of normal streptozotocin-glycosuric rats. This curve shows the effects of GLA from various sources when given at the same mg / kg / day dose as GLA. Figure 2 shows GLA at 8 mg / kg / day, 2D5522 at 0.25 mg / kg / day, and 0.2 mg. The effect of WAY 121509 / kg / day on nerve conduction in normal and diabetic animals when given alone or simultaneously. ZD5522 is 3,5'-difluorenyl-4'-nitromethyl · n-fluorenyl-2- (2-tolyl) -N-ethylanilide. WAY 1 2 1 509 is spiro- (isoquinoline-4 (1H), 3, pyrrolidine) -1, 2 \ 3, 5 '-(2Η) -tetraone. The doses of these three compounds have only a small improvement in nerve conduction when given alone, but when given in combination with ZD 5 5 2 2 or WAY 121509 and GLA, the effect on nerve conduction is greater than the combined effect Much more. Figure 3 shows the effects of GLA and ascorbic acid alone and simultaneous administration of both and single ascorbyl-GLA on blood flow and conduction velocity of sciatic nerve in diabetic rats. In this case, the simultaneous administration of the two compounds has a slightly greater effect than the addition, but it is also valuable in itself. However, the effect of ascorbyl-GLA as a single compound is significantly better than that of GLA alone or the combination of ascorbic acid, and it is better than the simultaneous administration of GLA and ascorbic acid. It may be better because the two components of the molecule are simultaneously delivered to the position where it is needed. The present invention is so. On the one hand, the present invention provides a pharmaceutically acceptable composition, which contains 8-Paper scale, using China S Standard (CNS) A4 specification U 丨 0X291g t) I ------- --Zhongyi ------ ΪΤ ------. ^ (Please read the notes on the back before filling out this page) Printed by the Central Standards Bureau of the Ministry of Economic Affairs Industrial and consumer cooperation Du Yin 4 226 A7 __ B7 _ Five 'invention description (6) 6-desaturated η · 6-fatty acids, especially GLA, DGLA or AA, and reduce the content of sorbitol in the body in the cell Substances, especially aldose reductase inhibitors. Preferred inhibitors are in the form of ascorbate or a complex thereof such as a salt, an ester or other derivative, especially an ester formed with a fatty acid, but ARIs, drugs, and the like can also be used. Another aspect of the present invention relates to the use of 6-desaturated η-6-lipidic acid, especially GLA 'DGLA or AA, and a pharmaceutically acceptable substance that reduces the intracellular content of sorbitol in the body, in particular When the substance is an aldose reductase inhibitor, it is used to treat (including preventive treatment) long-term diabetes complications. Ascorbic acid esters as described above are preferred. In general, these active substances can be administered together, but the present invention also includes the administration of fatty acids and sorbitol-reducing substances separately, but the effective amounts are simultaneously achieved in the body when administered separately. In another aspect, the invention relates to the preparation of a medicament for the treatment (including prophylactic treatment) of long-term diabetic comorbidities, and each active substance can be used when the preparation is available, or one of the active substances can be used. The ingredients must also be given separately. Another aspect of the present invention is to provide novel compounds of ascorbic acid that provide 6-desaturated η · 6-fatty acids (except (^ [eight or DGL Α), especially ascorbyl-AA ', and use in other treatments. Therefore, the present invention is particularly related to the treatment of diabetic comorbidities, especially, but not limited to, neuropathy, retinopathy, nephropathy, and cardiovascular disease; the treatment system is combined with administration of a compound selected from GLA, DGLA or AA EFA (essential fatty acids) and ascorbate or other can inhibit sugar reductase or reduce the size of this paper Applicable Chinese National Standard (CNS) A4 specification (2 〖0.X297mm） I i I | II! II ^ (Please Read the notes on the back before filling out this page) Printed by the Central Standards Bureau of the Ministry of Economic Affairs and Consumer Cooperatives 422694 at __— _B7 V. Description of the invention (~, ~. A body of sorbitol content. These active substances can Separate formulations are provided, but the separate formulations are given at the same time (if administered separately, at least at the same time there is an effective amount in the body), they can also be made into a compound dosage form; whether it is a separate formulation Compound dosage forms can be made into various suitable dosage forms. In particular, ascorbic acid-GLA, ascorbic acid-Dgla, or ascorbic acid-AA can be used. The dosage form can be capsule, drum, powder, microcapsule, emulsion Lipids' micelles, or any dosage form suitable for oral, enteral, parenteral, or topical administration are all dosages of β EF Α that are well known to those skilled in the art, For example, in the range of 0 mg to g / day, preferably 100 mg to 5 g / day, and more preferably 300 mg to 2 g / day, the dosage unit dosage form may contain such doses or a lower multiple thereof. The dosage of the aldose reductase inhibitor must be suitable for the selected ingredients. The wide range of daily effective amounts including all known drugs are shown here. It does not mean that the dosage of each drug is at this extreme. Range, 0.01 to 100 mg / kg / day, and please note that any substance that can reduce the conversion of glucose to sorbitol can be used. Examples are ponalrestat, tolrestat, and epalrestat. 0 · 1 to 20 mg / kg / day, this is to 7 The corresponding amount calculated by an adult of 0 kg. If ascorbate is used, the dosage is, for example, 10 mg to 50 g / day, preferably 50 mg to 2 g. This is the ascorbyl-GL A or other complex forms are calculated as ascorbic acid. EFA can be any pharmaceutically acceptable form that can result in available FA in the body. Particularly suitable forms include mono-, di-, and di-ascorbyl-EFA -And triglycerides, free-state fatty acids, cholesterol-10-This paper size is applicable to China National Standard (CNS) A4 (210X297 mm) --- ^ ------- ¾ Clothing ---- --il ------ t (Please read the notes on the back before filling out this page) Consumer Co-operation of the Central Bureau of Standards of the Ministry of Economic Affairs Du printed 422694 A7! _________ B7 V. Description of the invention (s) Cool 'salt, 醯Amines and various forms of phospholipids, but excluding medically unacceptable forms. Natural nightgrass oil or more DLMG (dilinoleyl-mono-canton-linolepine-glycerol) content is particularly convenient, or DLMG from other sources can also be used. Derived from EFA Fatty alcohol. The following examples illustrate the present invention, and are illustrated by the composition of EFA and a known aldose reductase inhibitory drug, followed by the composition of EFA and ascorbate, and finally the preparation by EFA ester of ascorbic acid. "ARIs" drugs "can be regarded as EFA compounds when there is a suitable chemical structure, these single compounds are preferably used in two preparations administered separately. -Igi di-aldose reductase inhibitor pharmaceutical composition via sugar There are many different types of reductase inhibitors (ARIs), and more may be discovered in the future. Similarly, some drugs may be found to inhibit sorbitol production with different mechanisms. The following is the use of known specific ARIs Examples of formulations: 1. 400 mg hard or soft capsules containing triglycerides' free-state fatty acids, salts or other forms of GLA, DGLA or AA, co-administered with ponalrestat, a bond containing 300 mg of ARI, 2 capsules EFA capsules and 1 capsulerestat are given together, one morning and one evening. 2 · Same as Example 1, then ARI is tolrestat, twice daily, 200 mg each time. 3-Same as Example 1, but ARI is epalrestat, daily Three '150 mg tablets each time. 4. Same as in Example 1, but using ADN-138 (Ono), SNK-860 (Sanwa), M16209 (Fuji), ZD5522 (Zeneca) or WAY 12 1 5 09 (Wyeth) -a-This paper size applies to Chinese national standards (CNS) A4ft (210 X 297 mm); ------ I ------ 1T ------ t {Please read the notes on the back before filling this page) Central Ministry of Economic Affairs Cooperating with the Standards Bureau, Laboratories, and Printing 422694 ^ 7 1 &7; V. Description of the Invention (9) Replaces ARI s. 5 _ Same as Example 1, but EFA and A RI are made into a dosage form, this dosage form can be Is a capsule, tablet, metering container or any other suitable dosage form known to those skilled in the art, such as micro knee capsules, loose liposomes, reverse liposomes, or for enteral, oral , Parenteral or other forms of administration. 6. Same as Example 1, but EFA is formulated into 200 mg, 300 mg, 500 mg, 600 mg, 800 mg or 1 g capsules, giving a total daily dose of 200 mg To 3 g. Example-Ascorbate Composition 7. The GLA, DGLA or AA in Example 1 or 6 was given simultaneously with 50 mg to 2 g of ascorbic acid, and both were made into tablets or other suitable formulations. 8. Use of ascorbic acid -GLA is synthesized as follows, or ascorbyl-DGLA or ascorbyl-AA, so that the amount of ascorbic acid administered is as described above. What dosage form:-(a) Lozenges, containing 50, 100, 25 0, 500 or 7500 mg ascorbyl-GLA '杌 ascorbyl-DGLA or ascorbyl-AA, or contain only this ingredient or Contains excipients. (b) Soft gelatin capsules or hard gelatin capsules containing 50, 100, 250, or 500 mg ascorbyl-GLA, ascorbyl-DGLA, or ascorbic acid-AA, dissolved in GLA or Within the free fatty acids of DGLA, or dissolved in triglycerides, one or more of the triglycerides is selected from GLA or DGLA. (c_) Ascorbic acid-GLA or ascorbic acid for oral, enteral or parenteral administration • 12- This paper ^ ΪApplies to Chinese National Standard rCNS) A4 (210X297 mm):-------- 1 ---- ¾ 衣 ------ 1T ------ ^ {Please read the notes on the back before filling this page) Central Standards Bureau of the Ministry of Economic Affairs *: Industrial and consumer cooperation Du printed 422b jia? 42Pe.QA_ B7____ V. Description of the invention (10) Blood acid-DGLA infant sword, powder, liquid, paste or peak dissolution. For example, the emulsion can be prepared with an appropriate emulsifier, such as lecithin or oat galactolipid, to make a final emulsion containing ascorbyl group-GLa, ascorbyl-DGLA or ascorbyl-AA in an amount of 2% to 30% by weight. (d) Ascorbic acid-GLA, ascorbic acid-DgLAs ascorbic acid-AA creams, creams, lotions, shampoos or other suitable formulations for topical use.

實例--抗壞血酸基-EFAs的合成 9.抗壞血酸基-GLA 合成時’將氣化氫氣（2.0克）於0°C通入Ν,Ν-二甲基乙 醯胺（26.5毫升）内。於此形成的泥樣物中加抗壞血酸 (9.69克）於二氯甲烷（13.25毫升）内之泥樣物中，此混合 物於0°C攪拌至成溶液。於此溶液中，於〇6C在氮氣下費時 4小時以上加z，z ’ z-十八-6,9,12-三烯醯氣（氣化gl A 酸）（1 4 8克），此混合物於上述溫度靜置1 8小時，再於室 溫靜置1小時。冷至0 °C時，加醋酸乙酯（2 0 0毫升）及水 (1 0 0毫升），將此混合物攪拌1小時D有機層用鹽水 (5x100毫升）洗，乾燥（Nhsc»4)並於5〇°C/1〇毫米汞柱 然後於50°C/0.1毫米汞柱蒸發4小時，製得抗壞血酸_6_ [(z，z ’ z)_十八_6，9 ’ i2_三烯酸酯](製得u.25克， 產出率S8%)，爲灰黃色堠樣。Example-Synthesis of Ascorbyl-EFAs 9. Synthesis of Ascorbyl-GLA ′ Gasified hydrogen (2.0 g) was passed into Ν, Ν-dimethylacetamide (26.5 ml) at 0 ° C. Ascorbic acid (9.69 g) was added to the mud sample formed in the mud sample in methylene chloride (13.25 ml), and the mixture was stirred at 0 ° C until a solution was obtained. In this solution, z, z'z-octadecene-6,9,12-triene tritium (gasified gl A acid) (14.8 g) was added at 0 ° C under nitrogen for more than 4 hours. The mixture was allowed to stand at the above temperature for 18 hours and then at room temperature for 1 hour. When cooled to 0 ° C, ethyl acetate (200 ml) and water (100 ml) were added, and the mixture was stirred for 1 hour. The organic layer was washed with brine (5x100 ml), dried (Nhsc »4) and Ascorbic acid_6 _ [(z, z'z) _octadecane-6,9'i2_triene was prepared by evaporation at 50 ° C / 10mm mercury for 4 hours at 50 ° C / 0.1mm mercury. Ester] (prepared u.25 g, yield S8%), grayish yellow tincture.

抗壞血酸基-GLA 將乾氣化氫氣（5.4份，克）於（-2)-2°C在攪拌下通入無 水二甲基6酿胺（7 1.25份，毫升）及乾二氣甲烷（4〇份， (諳先閲讀背面之注意事項再填寫本頁) _裝.Ascorbic acid-GLA Pass dry gasified hydrogen (5.4 parts, g) at (-2) -2 ° C while stirring. Anhydrous dimethyl 6 amine (7 1.25 parts, ml) and dry digas methane (4 〇 copies, (谙 read the notes on the back before filling in this page) _ installed.

rIT 本紙張尺度適用中賴家標準（CNS)八4規格（21()><297公釐） ~ — 經濟部中失標率局員工消费合作社印聚 U 辦 A7 _B7------- 五、發明説明（11) 毫升）之混合物内a於此混合物中加抗壞血酸（2 6 0 5份’ 克），此混合物攪拌至成澄清溶液。於（-”^冗用彳^小時 在氮氣下滴加z，z ’ z -十八-6 ’ 9 ’ 12 -二稀'酿氣亞麻 醯氯）（3 9.6份，克），再攪拌1 - 2小時後’將混合物於（_ 2)-2°C靜置20_24小時。眞空（25-3〇flC/2〇-30毫巴）除去 二氣甲烷，於此形成之泥樣物t在攪拌下加水（250份，毫 升）。加飽和鹽水（2 5 0份，毫升），此混合物於氮氣下強烈 攪拌10-15分鐘。靜置10-15分鐘後，分離出頂層半固體 產物。移除水層，如上述用水及鹽水再處理4次。將產物 溶於醋酸乙酯（250份，毫升）内，用水（2x50份，毫升）萃 取並乾燥（無水硫酸鈉）。眞空（35-40 °C/ 20-30毫巴）除去 醋酸乙酯，將殘餘物溶於乙醇（250份，毫升）内。眞空 (35-40 °C/ 20-30 毫巴，然後70-75 °C/ 0.1-0.5 毫巴）除去 乙醇，製得抗壞血酸- 6(z，z，z -十八-6，9，12 -三烯酸 酯），抗壞血酸基-GLA，爲灰黃色蠟樣。rIT This paper standard is applicable to China Laijia Standard (CNS) 8-4 specification (21 () > < 297 mm) ~ — The Office of Consumers Cooperatives of the Ministry of Economic Affairs of the Bureau of Standards and Loss Cooperatives Printing and Printing Office A7 _B7 ----- -V. Description of the invention (11) ml) A mixture of ascorbic acid (2 605 parts' g) was added to this mixture, and the mixture was stirred until a clear solution was obtained. Add z, z'z -eighteen-6'9'12-two dilute'gas-flaxed linseed chloride (-) (3 9.6 parts, grams) under (-"^ redundantly over ^^ hours under nitrogen, and stir for 1 more -After 2 hours', the mixture is left to stand at (_ 2) -2 ° C for 20-24 hours. The digas methane is removed by emptying (25-3〇flC / 20-30 mbar), and the mud sample formed here is at Water (250 parts, ml) was added with stirring. Saturated brine (250 parts, ml) was added. The mixture was stirred vigorously under nitrogen for 10-15 minutes. After standing for 10-15 minutes, the top semi-solid product was separated. The water layer was removed and treated with water and brine 4 times as described above. The product was dissolved in ethyl acetate (250 parts, ml), extracted with water (2x50 parts, ml) and dried (anhydrous sodium sulfate). Empty (35-40) ° C / 20-30 mbar) remove ethyl acetate and dissolve the residue in ethanol (250 parts, ml). Empty (35-40 ° C / 20-30 mbar, then 70-75 ° C / 0.1 -0.5 mbar) was removed from ethanol to prepare ascorbic acid-6 (z, z, z-octadecyl-6,9,12-trienoate), ascorbic acid-GLA, as a gray-yellow waxy sample.

11.抗壞血酸基-DGLA 以同樣方法，但用等量的z，z，z -二十-8，11，14·三 烯醯氣（二高-r -亞麻醯氣）代替Γ -亞麻醯氣，製得抗壞血 酸- 6- (ζ，ζ，ζ-二十-8 ’ 11，14-三烯酸酯），抗壞血酸 基- DGLA，爲灰黃色蝶樣。 1Ζ抗壞血酸基-ΑΑ 以同樣方法，但用等量的ζ，ζ，ζ_二十-5，8，11，1 4 -四 烯酿氣（花生四烯醯氣）代替亞麻醯氣，製得抗壞血酸_ 6-(z ’ ζ ’ ζ ， ζ_ 二十 _5 ， 8 ， η ’ 14•四晞酸醋 ） ， 抗壞血酸基 _ A A ’爲灰黃色壞樣。 -14 - 本紙張尺度朗巾® g家標準（CNS_) Μ規格（210X297公釐） ----；--^----¾------ίτ------^ (請先閱讀背面之注意事項再填寫本頁)11. Ascorbyl-DGLA In the same way, but the same amount of z, z, z-twenty-8,11,14 · triene radon (two high-r-linseed radon) instead of Γ-linseed radon. Ascorbic acid-6- (ζ, ζ, ζ-twenty-8'11,14-trienoate) and ascorbic acid group-DGLA were obtained, which was a gray-yellow butterfly. 1Z ascorbic acid-AA is prepared in the same way, but using the same amount of ζ, ζ, ζ_20-5,8,11,1 4 -tetrane gas (arachidene tritium) instead of linseed tritium to obtain Ascorbic acid 6- (z'ζ'ζ, ζ_twenty_5, 8, η'14 • tetraacetic acid vinegar), ascorbic acid group_AA 'is grayish yellow. -14-This paper size Long towel® g standard (CNS_) M specification (210X297 mm) ----;-^ ---- ¾ ------ ίτ ------ ^ ( (Please read the notes on the back before filling out this page)

Claims (1)

A8 B8 C8 D8 第84109228號專七 申42案2694 中文申請專利範圍修正本(89年9月） 六、申請專利範圍 1. 一種用於治療糖尿病長期併發症（包括預防性治療）之醫藥 組合物’其係由6-去飽和的n-6-脂肪酸（Εϊγα)，特別是r _ 亞麻酸（GLA)，二高，r -亞麻酸（DGla)或花生四烯酸（AA) ’以及減少體内細胞内山梨糖醇含量的物質，特別是越糖 還原酶抑制劑所組成’其中EFA與減少體内細胞内山梨糖 醇含量的物質之比例範園為1 : !至丨〇 〇 : 1。 2. 根據申請專利範圍第1項用於治療糖尿病長期併發症（包括 預防性治療）之组合物，其中EFA的劑量為1〇毫克至10克〆 天且減少體内細胞内山梨糖醇含量的物質劑量為〇.〇1至 1〇〇毫克/公斤/天。 3‘根據申請專利範園第1或2項之組合物，其中抑制劑是原樣 的抗壞血酸酯或其複合的形式。 4.根據申請專利範圍第3項之组合物，其中抗壞血酸醋係形成 抗壞血酸基-E F A，特別是抗壞血酸基· 〇 L A、抗壞血酸 基-DGLA或抗壞血酸基-AA。 & —種6 -去飽和的n _ 6 -必需脂肪酸（GL A或D G L A除外）的抗 壞血酸酯，特別是抗壞血酸基_AA。 6. —種用於治療糖尿病長期併發症（包括預防性治療）之醫藥 组合物，係6 -去飽和的η - 6 -必需脂肪酸（G L A或D G L A除 外）的抗壞血酸酯’〜特別是抗壞血酸基-A A，其中抗壞血酸 酯劑量為1 0毫克至1 〇克/天。 7- —種用於治療糖尿病長期併發症（包括預防性治療）之醫藥 組合物，其係由6 -去飽和的n _ 6 -必需脂.肪酸，特別是GL A ，DGLA或AA與抗壞血酸酯所組成，該脂肪酸及抗壞血酸 本紙浪凡及通用中國固家標準（CNS ) A4规格（2丨0X297公釐） (請先閲讀背面之注意事項再填寫本頁) 訂. 經濟部中央標率局貝工消費合作社印笨 4226a4 § D8 六、申請專利範圍 酯係以抗壞血酸基_EFA酯型態合併在一起，其中抗壞血酸 基-E F A劑量為1 〇毫克至丨〇克/天。 8. —種用於治療糖尿病長期併發症（包括預防性治療）之醫藥 组合物，其係由6 -去飽和的n _ 6 -脂肪酸，特別是G L A， DGLA或AA，以及抗壞血酸酯所组成，其中EFA與抗壞 血酸酯之比例範圍為1 : 1至1 〇 〇 : i。 9. 根據申请專利範圍第8項用於治療糖尿病長期併發症（包括 預防性治療）之組合物，其中EF A的劑量為1 〇毫克至丨〇克/ 天且抗壞血酸酯的劑量為1 〇毫克至5克/天。 (請先閱讀背面之注意事項再填寫本頁) X 訂- 經濟部中央標準局貝工消費合作社印装 本紙浪尺度適用中國國家梯準（CNS)A4規格（2丨0X297公釐）A8 B8 C8 D8 No. 84109228 No. 42 of Special Application No. 2694 Chinese Patent Application Amendment (September 89) VI. Application for Patent Scope 1. A pharmaceutical composition for treating long-term complications of diabetes (including preventive treatment) 'It consists of 6-desaturated n-6-fatty acids (Eϊγα), especially r_ linolenic acid (GLA), dihomo, r-linolenic acid (DGla) or arachidonic acid (AA)' and reduced body Intracellular sorbitol content substances, in particular composed of glucosyl reductase inhibitors, where the ratio of EFA to substances that reduce intracellular sorbitol content ranges from 1 :! to 丨 〇: 1. 2. The composition for treating long-term complications of diabetes (including preventive treatment) according to the scope of the patent application, wherein the dosage of EFA is 10 mg to 10 g per day and the amount of sorbitol in the body is reduced. The substance dose is from 0.01 to 100 mg / kg / day. 3 ' The composition according to item 1 or 2 of the patent application park, wherein the inhibitor is an ascorbate as it is or a complex form thereof. 4. A composition according to item 3 of the scope of application for a patent, wherein the ascorbic acid vinegar forms ascorbyl-EFA, particularly ascorbyl · OLA, ascorbyl-DGLA or ascorbyl-AA. &-Ascorbyl ester of 6-desaturated n-6-essential fatty acids (except GL A or D G L A), especially ascorbyl group -AA. 6. —A pharmaceutical composition for the treatment of long-term complications of diabetes (including prophylactic treatment), which is 6-desaturated η-6-essential fatty acids (except GLA or DGLA) ascorbate '~ especially ascorbic acid- AA, in which the ascorbate dose is 10 mg to 10 g / day. 7-—A pharmaceutical composition for treating long-term complications of diabetes (including preventive treatment), which is composed of 6-desaturated n_6-essential fats. Fatty acids, especially GL A, DGLA or AA and ascorbic acid Composed of esters, the fatty acid and ascorbic acid, paper Langfan and common Chinese solid standard (CNS) A4 specification (2 丨 0X297 mm) (Please read the notes on the back before filling this page) Order. Central Standards Bureau, Ministry of Economic Affairs Beigong Consumer Cooperatives Co., Ltd. Yinben 4226a4 § D8 6. The scope of patent application The esters are combined together in the form of ascorbyl-EFA ester, where the dosage of ascorbyl-EFA is 10 mg to 10 g / day. 8. A pharmaceutical composition for the treatment of long-term complications of diabetes (including prophylactic treatment), which consists of 6-desaturated n-6 fatty acids, especially GLA, DGLA or AA, and ascorbate, The ratio of EFA to ascorbate ranges from 1: 1 to 100: i. 9. A composition for treating long-term complications of diabetes (including prophylactic treatment) according to item 8 of the scope of the patent application, wherein the dose of EF A is 10 mg to 10 g / day and the dose of ascorbate is 10 mg To 5 g / day. (Please read the precautions on the back before filling this page) Order X-Printed by Shellfish Consumer Cooperatives, Central Bureau of Standards, Ministry of Economic Affairs The paper wave size is applicable to China National Standard (CNS) A4 (2 丨 0X297 mm)