WO2007082471A1 - Anti-infective compound, preparation method thereof and use thereof - Google Patents

Anti-infective compound, preparation method thereof and use thereof Download PDF

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WO2007082471A1
WO2007082471A1 PCT/CN2007/000179 CN2007000179W WO2007082471A1 WO 2007082471 A1 WO2007082471 A1 WO 2007082471A1 CN 2007000179 W CN2007000179 W CN 2007000179W WO 2007082471 A1 WO2007082471 A1 WO 2007082471A1
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compound
infective
water
added
piperazinyl
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PCT/CN2007/000179
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French (fr)
Chinese (zh)
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Mao Chen
Shaoxuan Zhu
Xuebin Liu
Lizhen Zheng
Shuwen Xu
Danqing Liu
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Guangzhou Baiyunshan Pharmaceutical Co., Ltd. Guangzhou Baiyunshan Pharmaceutical Factory
Guangzhou Pharmaceutical Industrial Research Institute
Guangzhou Baiyunshan Pharmaceutical Co., Ltd. Guangzhou Baiyunshan Chemical Pharmaceutical Factory
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Priority to CNA2006100330280A priority patent/CN101003540A/en
Application filed by Guangzhou Baiyunshan Pharmaceutical Co., Ltd. Guangzhou Baiyunshan Pharmaceutical Factory, Guangzhou Pharmaceutical Industrial Research Institute, Guangzhou Baiyunshan Pharmaceutical Co., Ltd. Guangzhou Baiyunshan Chemical Pharmaceutical Factory filed Critical Guangzhou Baiyunshan Pharmaceutical Co., Ltd. Guangzhou Baiyunshan Pharmaceutical Factory
Publication of WO2007082471A1 publication Critical patent/WO2007082471A1/en

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    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07DHETEROCYCLIC COMPOUNDS
    • C07D513/00Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00
    • C07D513/02Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00 in which the condensed system contains two hetero rings
    • C07D513/04Ortho-condensed systems
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics

Abstract

Anti-infective compound, preparation method thereof and use thereof. The present invention disclosed a crystal of 6-fluoro-1-methyl-4-oxo-7-(1-piperazinyl)-4H-[1,3]thiazeto[3,2-a]quinoline-3-carboxylic acid (II) methanesulfonate, which is separated out by solvent crystallization following the reaction of compound (II) with pharmaceutical methanesulfonic acid using water or water containing organic solvent as solvents. The obtained compound has a definitive structure. It is stable and has confirmed anti-bacterial effect. The water solubility of compound (II) has been improved by this way, and it is convenient to make appropriate dosage forms for clinic use to expand the extension of clinic use. By preparing the methanesulfonate of compound (II), the toxicity of the active compound (II) has been reduced and the safety of its clinic use has been improved.

Description

一种抗感染化合物及其制备方法和用途 技术领域 本发明涉及一种喹诺酮类抗感染的药物, 具体涉及 6-氟- 1-甲基- 4 - 氧代 -7- (1-哌嗪基 ) - 4H- [1, 3] 硫氮杂环丁垸并 [3, 2- a]喹啉- 3-羧酸甲 磺酸盐和制备方法及其用途。 背景技术 喹诺酮类药物是近年来迅速发展起来的抗菌药物, 具有抗菌谱广、 抗 菌力强、 结构简单、 给药方便, 与其它常用抗菌药物无交叉耐药性, 疗效 价格比高等优势, 因而愈来愈受到各国的重视, 成为竞相生产和应用的热 点药品。 已上市的喹诺酮产品有几十个品种, 是抗感染药物中幵发最活跃 的领域之一。  FIELD OF THE INVENTION The present invention relates to a quinolone anti-infective drug, in particular to 6-fluoro-1-methyl-4-oxo-7-(1-piperazinyl) - 4H- [1, 3] thiazepine-[3,2-a]quinoline-3-carboxylic acid mesylate, preparation method and use thereof. BACKGROUND OF THE INVENTION Quinolones are antibacterial drugs that have been rapidly developed in recent years. They have broad antibacterial spectrum, strong antibacterial activity, simple structure, convenient administration, no cross-resistance with other commonly used antibacterial drugs, and high cost-effectiveness. It has become the hot spot drug for competing production and application. There are dozens of quinolone products on the market, and it is one of the most active areas of anti-infective drugs.
6-氟- 1-甲基 -4-氧代 -7- (1_哌嗪基 ) - 4H- [1, 3]硫氮杂环丁烷并 [3, 2 a] 喹啉 -3-羧酸 (UFX, NM394)是一种抗感染效果非常显著的喹诺酮类药物, 美 国化学文摘 CA中的 108 :p94537d; 111 : pl94791n; 113 : p231357q; 116 :p 41483s ; 124 : pr8660q; 128 : 123459a等文献报道了其对革兰氏阳性菌和 革兰氏阴性菌具有广谱抗菌作用,特别是对金黄色葡萄球菌、肺炎链球菌、 粪肠球菌、 粘液沙雷菌、 绿脓杆菌等细菌显示出强大的抗菌效果。 日本新 药公司与明治制果公司共同开发研制了以 6-氟 -1-甲基- 4-氧代 -7- (1-哌 嗪基) - 4H- [1, 3] 硫氮杂环丁烷并 [3, 2-a]喹啉- 3-羧酸 II为活性体的前 药的口服制剂, 药名为 Prulifloxacin。 由于化合物 II (NM394)不溶于水, 所以影响其药物的使用。 6-fluoro- 1-methyl-4-oxo-7-(1-piperazinyl)-4H-[1,3]thiazetino[3,2 a]quinoline-3-carboxylate Acid (UFX, NM394) is a quinolone with a very significant anti-infective effect, 108 in the Chemical Abstracts CA, p94537d , 111: pl94791n , 113: p231357q , 116: p 41483s, 124: pr8660q , 128: 123459a, etc. It has been reported in the literature that it has a broad-spectrum antibacterial effect against Gram-positive bacteria and Gram-negative bacteria, especially for bacteria such as Staphylococcus aureus, Streptococcus pneumoniae, Enterococcus faecalis, Serratia marcescens, and Pseudomonas aeruginosa. Strong antibacterial effect. Japan New Drug Company and Meiji Fruit Co., Ltd. jointly developed 6-fluoro-1-methyl-4-oxo-7-(1-piperazinyl)-4H-[1,3]thiazetidine And [3, 2- a ] quinoline-3-carboxylic acid II is an oral preparation of a prodrug of the active substance, and the drug name is Prulifloxacin. Since Compound II (NM394) is insoluble in water, it affects the use of its drug.
NM394除了不溶于水, 影响其药物的使用外, 其静脉注射毒性大也是影 响其药用的原因, 据文献报道 UFX (匪 394)静脉给药毒性很大。 工 shida S 于 1996 年报导大鼠静注 匪 394 四周的毒性研究结果, 雄性和雌性 Sprague- Drwlog大鼠, 分别静注剂量为 3、 10和 30mg/kg的匪 394, 为期 四周, 10、 30mg/kg剂量的大鼠的水消耗量和尿排量明显增加, 尿沉淀物 中发现结晶性物质和小上皮细胞。 30mg/kg剂量的大鼠出现尿浊现象, 10 In addition to being insoluble in water, NM394 affects the use of its drugs. The high toxicity of intravenous injection is also the cause of its medicinal use. According to the literature, UFX (匪394) is highly toxic. In 2009, Shida S reported the results of a toxicity study of 394 weeks of intravenous infusion in rats. Male and female Sprague-Drwlog rats were given intravenously at doses of 3, 10, and 30 mg/kg, for a period of four weeks, 10, 30 mg. The water consumption and urinary excretion of the rats in the /kg dose were significantly increased, and crystalline substances and small epithelial cells were found in the urine precipitate. Urine turbidity occurred in rats at 30 m g / kg dose, 10
1 1
确 认 本 和 30mg/kg剂量组血 Y -球蛋白减少, 10和 30mg/kg组小鼠血尿素氮和肌 酐增加, 这表明肾功能已受到损害, 此外, 10 和 30mg/kg剂量组大鼠发 生病理学的改变, 如管状肾病变, 同时发现结晶状物质; 30mg/kg剂量组 大鼠肾和盲肠的重量增加; 上述的变化除血 Y -球蛋白外, 都是可逆的, 3mg/kg剂量组没有发现明显的问题, 所以應 394对大鼠的 N0AEL应该是 3mg/kg。 Confirmation In the 30 mg/kg dose group, blood Y-globulin decreased, and in the 10 and 30 mg/kg groups, blood urea nitrogen and creatinine increased, indicating that renal function has been impaired. In addition, pathology occurred in rats in the 10 and 30 mg/kg dose groups. Changes, such as tubular nephropathy, and the discovery of crystalline substances; the weight of the kidney and cecum of the 30 mg/kg dose group increased; the above changes were reversible except for blood Y-globulin, and the 3 mg/kg dose group did not. An obvious problem was found, so the N0AEL of 394 pairs of rats should be 3 mg/kg.
上述研究清楚说明, 没有不良反应的剂量是 3mg/kg, 按剂量转换规律 相当于约 35 mg/70kg人, 这是一个很低的剂量, 我们知普利沙星的正常 剂量为 200-60 mg /日 (以丽 394计) , 这一结果说明 NM394静脉给药毒 性太大, 没有临床应用的前景。 The above studies clearly indicated that the dose without adverse reactions was 3 mg/kg, and the dose conversion rule was equivalent to about 35 m g / 70 kg, which is a very low dose. We know that the normal dose of prisafloxacin is 200-60 mg. / Day (in 394 liters), this result indicates that NM394 intravenous toxicity is too much, there is no prospect of clinical application.
尽管 EP0315828和 US4, 843, 070中公开了 6-氟- 1-甲基- 4-氧代- 7- (1- 哌嗪基 ) 4H- [1, 3]硫氮杂环丁烷并 [3, 2-a]喹啉- 3-羧酸及其制备方法, 也 指出了其某些可药用的盐类, 但没有披露如何得到水溶性良好的 6-氟- 1- 甲基- 4-氧代 -7- (1-哌嗪基 ) - 4H- [1, 3]硫氮杂环丁烷并 [3, 2-a]喹啉 _3 -羧 酸衍生物, 因而其应用范围大受限制。 发明内容  6-Fluoro-1-methyl-4-oxo-7-(1-piperazinyl) 4H-[1,3]thiazetetane[3] is disclosed in EP 0 315 828 and US Pat. No. 4,843,070. , 2-a] quinoline-3-carboxylic acid and its preparation method, also pointed out some pharmaceutically acceptable salts thereof, but did not disclose how to obtain 6-fluoro-1-methyl- 4- Oxo-7-(1-piperazinyl)-4H-[1,3]thiazetino[3,2-a]quinoline-3-carboxylic acid derivative, thus its application range is greatly affected limit. Summary of the invention
本发明的目的是提供一种易溶于水的、 能注射给药且毒性低、 安全性 好的抗感染喹诺酮类化合物及该化合物的制备方法和用途。  SUMMARY OF THE INVENTION An object of the present invention is to provide an anti-infective quinolone compound which is easily soluble in water, can be administered by injection, has low toxicity and is safe, and a preparation method and use thereof.
本发明的发明者发现, 将 6-氟- 1-甲基- 4-氧代 -7- (1-哌嗪基) - 4H [1, 3] 硫氮杂环丁烷并 [3, 2- a]喹啉- 3-羧酸与甲磺酸盐反应, 控制反应 条件, 得到如下式 I所示的喹诺酮类化合物结晶盐, 其化学名是 6-氟- 1- 甲基 -4-氧代 -7- (1-哌嗪基 ) - 4H- [1, 3] 硫氮杂环丁烷并 [3, 2- a]喹啉 -3 - 羧酸甲磺酸盐:  The inventors of the present invention found that 6-fluoro-1-methyl-4-oxo-7-(1-piperazinyl)-4H[1,3]thiazetetane[3, 2- a] quinoline-3-carboxylic acid is reacted with methanesulfonate to control the reaction conditions, and a quinolone compound crystal salt represented by the following formula I is obtained, and its chemical name is 6-fluoro-1-methyl-4-oxo -7-(1-Piperazinyl)-4H-[1,3]thiazetino[3,2-a]quinoline-3-carboxylic acid methanesulfonate:
该结晶盐具有意想不到的高水溶性, 从而使得抗感染喹诺酮类化合物的使 用或应用范围大大拓宽。 例如, 由这种结晶盐制备的抗感染药物组合物, 既可以是一种胃肠道给药的片剂、 胶廳剂或颗粒剂, 也可以是非胃肠道给 药的注射剂、 眼用制剂、 耳用制剂、 妇科用制剂或皮肤外用制剂。。 The crystalline salt has an unexpectedly high water solubility, so that the use or application range of the anti-infective quinolone compound is greatly broadened. For example, an anti-infective pharmaceutical composition prepared from such a crystalline salt may be a tablet for gastrointestinal administration, a capsule or granule, or an injection for parenteral administration or an ophthalmic preparation. , otic preparations, gynecological preparations or external preparations for skin. .
化合物 I的结晶粉末用 Cu-Ko 射线测量得到的 X-射线粉末衍射图中 的 2Θ、 d-面间距和相对强度 (1/1。) 具有如下数值 (图谱见附图 1 ):  The 2Θ, d-plane spacing and relative intensity (1/1.) in the X-ray powder diffraction pattern of the crystalline powder of Compound I measured by Cu-Ko ray have the following values (see Figure 1 for the spectrum):
2 Θ d 1/1。  2 Θ d 1/1.
5. 520 15. 99± 0. 2 0. 84-1. 00  5. 520 15. 99± 0. 2 0. 84-1. 00
10. 320 8. 56 + 0. 2 0. 65-0. 78  10. 320 8. 56 + 0. 2 0. 65-0. 78
11. 100 7. 96± 0. 2 0. 83-0. 96  11. 100 7. 96± 0. 2 0. 83-0. 96
14. 040 6. 30 ± 0. 2 0. 42-0. 64  14. 040 6. 30 ± 0. 2 0. 42-0. 64
14, 420 6. 13土 0. 2 0. 24-0. 45  14, 420 6. 13 soil 0. 2 0. 24-0. 45
16. 340 5. 42±0. 2 0. 25-0. 46  16. 340 5. 42±0. 2 0. 25-0. 46
16. 880 5. 24± 0. 2 0. 27-0. 51  16. 880 5. 24± 0. 2 0. 27-0. 51
17. 780 4. 98 + 0. 2 0. 29-0. 49  17. 780 4. 98 + 0. 2 0. 29-0. 49
18. 740 4. 73 ± 0. 1 0. 46-0. 72  18. 740 4. 73 ± 0. 1 0. 46-0. 72
19. 820 4. 47 + 0. 1 0. 39-0. 58  19. 820 4. 47 + 0. 1 0. 39-0. 58
22. 720 3. 91 ±0. 1 0. 35-0. 63  22. 720 3. 91 ±0. 1 0. 35-0. 63
26. 140 3. 40 + 0. 1 0. 52-1. 00  26. 140 3. 40 + 0. 1 0. 52-1. 00
26. 800 3. 32± 0. 1 0. 45-1. 00  26. 800 3. 32± 0. 1 0. 45-1. 00
核磁 NMR (DMS0-d6, 500Hz) 确证了化合物 I结构如上式 I所示。红外 IR (KBr, cm—1)进一步确证了其结构如上式 I所示。 NMR NMR (DMS0-d 6, 500Hz ) confirmed the structure of compound I of formula I as described above. Infrared IR (KBr, cm- 1 ) further confirmed its structure as shown in Formula I above.
用本方法制得的化合物 I的元素分析数据如下:  The elemental analysis data of Compound I prepared by this method are as follows:
本品在水中易溶, 不溶于甲醇、 无水乙醇、 丙酮及乙醚。  This product is soluble in water, insoluble in methanol, absolute ethanol, acetone and ether.
本发明提供的化合物 I的制备方法是通过以下途径实现的: 用水或含 有机溶剂体积百分比为 5〜50%的水作溶液, 于 0〜60°C加入 6-氟 -1-甲基 -4-氧代 -7- (1-哌嗪基 ) - 4H- [1, 3] 硫氮杂环丁烷并 [3, 2- a]喹啉- 3-羧 酸 (化合物 II ) 和甲磺酸, 所用溶液的量为化合物 II重量的 5〜50倍, 搅 拌溶解后, 加入化合物 II重量的 1〜: L00倍的有机溶剂, 即析出化合物 I, 化合物 II和甲磺酸的投料摩尔比为 1 : 0. 8〜1. 5, 其中所说的有机溶剂是 甲醇、 乙醇、 异丙醇、 丙酮、 四氢呋喃中的任意一种。 The preparation method of the compound I provided by the invention is achieved by the following methods: The organic solvent has a volume percentage of 5 to 50% of water as a solution, and 6-fluoro-1-methyl-4-oxo-7-(1-piperazinyl)-4H-[1, is added at 0 to 60 °C. 3] thiazetazo[3,2-a]quinoline-3-carboxylic acid (compound II) and methanesulfonic acid, the amount of the solution used is 5 to 50 times the weight of the compound II, after stirring and dissolving, The organic solvent is methanol and ethanol. The organic solvent is methanol or ethanol. The organic solvent is methanol and ethanol. The organic solvent is methanol and ethanol. Any one of isopropyl alcohol, acetone, and tetrahydrofuran.
本发明还提供了一种抗感染药物组合物, 其以喹诺酮类化合物 I作为 有效成分, 并含有常规药用载体。具体可以是胃肠道给药的片剂、胶囊剂、 颗粒剂, 或是非胃肠道给药的注射剂、 眼用制剂、耳用制剂、妇科用制剂、 皮肤外用制剂。  The present invention also provides an anti-infective pharmaceutical composition comprising a quinolone compound I as an active ingredient and a conventional pharmaceutical carrier. Specifically, it may be a tablet, a capsule, a granule for gastrointestinal administration, or an injection for parenteral administration, an ophthalmic preparation, an otic preparation, a gynecological preparation, or an external preparation for skin.
该组合物, 可用于胃肠道给药,如制成胶囊, 片剂或颗粒剂, 是以喹 诺酮类化合物 I与常规的药理上可相容的赋形剂, 如淀粉, 麦芽糖, 蔗糖, 碳酸钙或磷酸钙; 粘合剂如淀粉, 阿拉伯胶, 羧甲基纤维素, 羟丙基纤维 素, 结晶纤维素; 润滑剂如硬脂酸镁或滑石粉; 分解剂如羧甲基钙或滑石 粉相混合, 用常规的制剂方法制备。 该组合物通常是这样制取的, 将化合 物 I与至少一种上述的载体或赋形剂相混合, 化合物 I占整个组合物的比 率一般为 0. 1〜100% ( W/W ) 。  The composition can be used for gastrointestinal administration, such as capsules, tablets or granules, and is a quinolone compound I and a conventional pharmacologically compatible excipient such as starch, maltose, sucrose, carbonic acid. Calcium or calcium phosphate; binders such as starch, gum arabic, carboxymethyl cellulose, hydroxypropyl cellulose, crystalline cellulose; lubricants such as magnesium stearate or talc; decomposers such as carboxymethyl calcium or talc The powder phase is mixed and prepared by a conventional formulation method. 1〜100% ( W / W ) The ratio of the compound I to the total composition is generally 0. 1~100% (W / W).
本发明提供的化合物 I的抗感染药物组合物, 也可用于非胃肠道给 药, 如制成注射剂、'眼用制剂、 耳用制剂、 妇科用制剂、 皮肤外用制剂等。 化合物 I占整个组合物的比率一般为 0. 1〜100% ( W/W ) 。  The anti-infective pharmaceutical composition of the compound I provided by the present invention can also be used for parenteral administration such as injections, ophthalmic preparations, otic preparations, gynecological preparations, external preparations for skin and the like. The ratio of the compound I to the entire composition is generally from 0.1 to 100% (W/W).
本发明具有以下优点:  The invention has the following advantages:
1、 提供一个有优良抗感染作用的喹诺酮类化合物, 其结构确定, 性 质稳定, 在水中易溶, 可以方便地制备成各种适宜的剂型用于临床。  1. Provide a quinolone compound with excellent anti-infective effect. Its structure is determined, its properties are stable, and it is easily soluble in water. It can be conveniently prepared into various suitable dosage forms for clinical use.
2、 本发明的制备生产工艺简便、 合理, 生产成本低。  2. The preparation and production process of the invention is simple and reasonable, and the production cost is low.
3、 增加了抗感染药物制剂新品种, 改善了活性药物化合物 ( Π ) 即 NM394 的水溶性, 血管刺激, 肌肉剌激试验表明本发明化合物适用于肌肉注射和 静脉注射, 扩大临床应用范围。  3. New varieties of anti-infective drug preparations have been added, and the water-soluble, vascular-stimulated, muscle-stimulating test of the active drug compound (Π), NM394, has been improved, indicating that the compound of the present invention is suitable for intramuscular injection and intravenous injection, and expands the clinical application range.
4、 通过制备化合物 ( II ) 即 NM394的甲磺酸盐, 降低了活性物质 NM394 的毒性, 增加了临床用药安全性。 下面通过试验例说明本发明的有益效果。 4. By preparing compound (II), that is, the mesylate salt of NM394, the toxicity of the active substance NM394 is lowered, and the safety of clinical use is increased. The beneficial effects of the present invention will be described below by way of test examples.
试验例 1 : 本发明所提供的化合物 I的稳定性影响因素实验  Test Example 1 : Experiment on the influence factors of the stability of the compound I provided by the present invention
试验样品: 见下面实施例 2所提供的样品 01, 为本发明所提供的化合 试验方法:  Test sample: See sample 01 provided in Example 2 below, which is the compound test method provided by the present invention:
1、 质量测定方法  1, quality measurement method
1 )、 含量测定方法: 照高效液相色谱法 (中国药典 2005年二部附录 V D) 测定。  1), content determination method: According to high performance liquid chromatography (Chinese Pharmacopoeia 2005 two appendix V D) determination.
色谱条件与系统适用性试验 : 用十八烷基硅烷键合硅胶为填充剂; 乙 腈- 0. 005mol/l磷酸二氢钠溶液 (取 0. 005mol/l磷酸二氢钠溶液 100ml , 加三乙胺 0. 2 ml , 用磷酸调节 pH值至 6. 0) (40: 60 ) 为流动相; 流速为 1. 0ml/分钟; 检测波长为 278nm。 理论板数按 6_氟 -1-甲基- 4-氧代 -7- (1- 哌嗪基 ) - 4H- [1, 3] 硫氮杂环丁烷并 [3, 2- a]喹啉- 3-羧酸计应不低于 2000; 6-氟 -1-甲基- 4-氧代- 7- (1-哌嗪基) - 4H- [1, 3] 硫氮杂环丁烷并 [3, 2-a]喹啉- 3-羧酸峰与相邻杂质峰的分离度应符合要求。  Chromatographic conditions and system suitability test: using octadecylsilane bonded silica as a filler; acetonitrile - 0. 005mol / l sodium dihydrogen phosphate solution (take 0. 005mol / l sodium dihydrogen phosphate solution 100ml, plus three The amine is 0. 2 ml, and the pH is adjusted to 6. 0) (40: 60) with a mobile phase; the flow rate is 1.0 ml/min; and the detection wavelength is 278 nm. The number of theoretical plates is 6_fluoro-1-methyl-4-oxo-7-(1-piperazinyl)-4H-[1,3]thiazetino[3,2- a]quina The carboxylic acid-3-carboxylic acid meter should not be less than 2000; 6-fluoro-1-methyl-4-oxo-7-(1-piperazinyl)-4H-[1,3]thiazetidine The resolution of the [3, 2-a] quinoline-3-carboxylic acid peak and the adjacent impurity peak should meet the requirements.
测定方法: 取化合物 I约 10mg, 精密称定, 置 100ml量瓶中, 加流动 相适量, 超声处理使溶解并稀释制成每 lml约含 50μ 的溶液。 精密量取 20μ1 , 注入液相色谱仪, 记录色谱图; 另取 6-氟- 1-甲基 -4-氧代- 7- (1- 哌嗪基) - 4Η- [1, 3] 硫氮杂环丁垸并 [3, 2- a]喹啉- 3-羧酸 (对照品) 约 10mg, 同法测定。按外标法以峰面积计算, 即得。化合物 I中含有 6-氟- 1 - 甲基- 4-氧代 -7- (1-哌嗪基 ) - 4Η- [1, 3] 硫氮杂环丁垸并 [3, 2- a]喹啉- 3- 羧酸为 75〜81%。 Determination method: Take about 10m g of compound I, accurately weigh it, put it into a 100ml volumetric flask, add the appropriate amount of mobile phase, sonicate and dissolve and dilute to make a solution containing about 50μ per lml. Precisely measure 20μ1, inject into the liquid chromatograph, record the chromatogram; take 6-fluoro-1-methyl-4-oxo-7-(1-piperazinyl)-4Η-[1,3] sulfur nitrogen Heterocyclic butyrazino[3,2-a]quinoline-3-carboxylic acid (control) was about 10 mg, determined by the same method. According to the external standard method, the peak area is calculated. Compound I contains 6-fluoro-1- 1 -methyl-4-oxo-7-(1-piperazinyl)-4-yl-[1,3]thiazepine-[3,2- a]quina The porphyrin-3-carboxylic acid is 75 to 81%.
2)样品的 pH值测定方法: 取化合物 I, 加水制成每 10ml水中含有 0. lg 化合物 I的溶液, 依法测定。  2) Determination of pH of the sample: Take Compound I, add water to make a solution containing 0.1 g of Compound I per 10 ml of water, and determine according to law.
3)样品的澄清度及颜色: 取化合物 I, 加水制成每 10ml水中含有 0. lg化 合物 I的溶液, 依法测定。  3) Clarity and color of the sample: Take Compound I, add water to make a solution containing 0.1 g of Compound I per 10 ml of water, and determine according to law.
2、 光照试验  2, light test
将样品 01置于药物光照试验仪下, 光照强度为 4500LX± 500LX, 放置 5, 10天后取样观察色泽、 测定 pH值、 澄清度、 含量。 结果见下表。 样品 时间(天) pH值 色泽 澄清度 含量% Sample 01 was placed under a medical light tester, and the light intensity was 4500 LX ± 500 LX. After 5 and 10 days of storage, the color was measured, and the pH, clarity, and content were measured. See the table below for the results. Sample time (days) pH value color clarity content%
01 0 2. 89 < 1 <1 79. 5  01 0 2. 89 < 1 <1 79. 5
5 2. 90 < 1 < 1 79. 4  5 2. 90 < 1 < 1 79. 4
10 2. 91 < 1 < 1 79. 3  10 2. 91 < 1 < 1 79. 3
试验结果表明, 样品 01在光照条件下基本稳定。  The test results show that sample 01 is substantially stable under illumination conditions.
3、 高温试验 3, high temperature test
将样品 01置于 60Ό的恒温箱中, 放置 5, 10天后取样观察色 泽、 测定 PH值、 澄清度、 含量。 结果见下表。  Sample 01 was placed in a 60-inch incubator, placed for 5, 10 days, and samples were taken to observe the color, pH, clarity, and content. See the table below for the results.
试验结果表明, 样品 01本品在高温放置时含量、 色泽、 澄清度 及 pH值无明显变化, 故本品稳定。  The test results show that the sample 01 has no obvious change in content, color, clarity and pH value when it is placed at high temperature, so the product is stable.
4、 高湿试验  4, high humidity test
将样品 01于 25士 2 °C相对湿度 RH95士 5%的条件下放置, 放 置 5, 10天后取样观察色泽、 测定 pH值、 澄清度、 含量。 结果见下表。  The sample 01 was placed under a condition of 25 ± 2 ° C relative humidity RH 95 ± 5%, and placed for 5, 10 days, and the color was measured, and the pH, clarity, and content were measured. See the table below for the results.
试验结果表明, 本品在高湿放置时含量、 色泽、 澄清度及 pH值 无明显变化, 故本品稳定。  The test results show that the product has no obvious change in content, color, clarity and pH value when placed in high humidity, so the product is stable.
试验 2: 本发明所提供的化合物 I的抑菌试验, 测定 M I C最低浓度。 试验方法: 采取常规平皿二倍稀释法进行体外抑菌试验  Test 2: The bacteriostatic test of the compound I provided by the present invention, the minimum concentration of M I C was determined. Test method: In vitro bacteriostasis test using conventional plate double dilution method
样品 01, 为本发明所提供的化合物 I ; 样品 02, 为化合物 II。 Sample 01, which is a compound I provided by the present invention ; and sample 02, is a compound II.
试验结果: test results:
细菌名称 MIC90值( g/ml)  Bacterial name MIC90 value (g/ml)
样品 02 样品 01 金葡萄球菌 0.5 0.5 Sample 02 sample 01 Staphylococcus aureus 0.5 0.5
肺炎链球菌 2 ^2.2 Streptococcus pneumoniae 2 ^2.2
4 3.9  4 3.9
f少 0.25 0.25  f less 0.25 0.25
绿脓杆菌 2 2.0  Pseudomonas aeruginosa 2 2.0
大肠埃希菌 0.06 0.06  Escherichia coli 0.06 0.06
肺炎克雷伯杆菌 ^0.06 0.07 Klebsiella pneumoniae ^0.06 0.07
0.06 0.06  0.06 0.06
试验结果说明化合物 I对革兰氏阳性菌和革兰氏阴性菌与化合物 II 同样具有广谱抗菌作用, 特别是对绿脓杆菌、 沙雷菌属、 肠杆菌属等革兰 氏阴性菌显示出强大的抗菌效果。  The test results show that Compound I has a broad-spectrum antibacterial effect on Gram-positive bacteria and Gram-negative bacteria and Compound II, especially for Gram-negative bacteria such as Pseudomonas aeruginosa, Serratia, Enterobacter and the like. Strong antibacterial effect.
试验例 3、 连续静脉给药 28天毒性实验: Test Example 3. Continuous intravenous administration 28-day toxicity test:
试验使用 SD大鼠 80只, 按动物体重和性别随机分为 4组, 每组 20 只, 雌雄各半。 设化合物 I ( M394甲磺酸盐) 10、 30、 60mg/kg三个剂 量组和一个空白对照组, 尾静脉注射给药, 每日 1次, 连续给药 4周, 恢 复观察 2周。 每天进行一般状况态观察, 每周统计一次体重、 摄食量, 给 药末期取一半动物采血和收集尿液做血液生化、 电解质、 尿液分析检测, 同时放血处死剖检做大体病理检查, 并取股骨、腕关节软骨、股关节软骨、 脑 (大脑、 小脑、 脑干)、 脊髓 (颈、 胸、 腰段)、 垂体、 胸腺、 甲状腺、 甲状旁腺、 食管、 唾液腺、 胃、 十二指肠、 回肠、 结肠、 肝脏、 胆囊、 肾 脏、 肾上腺、 脾脏、 胰腺、 气管、 肺脏、 主动脉、 心脏、 附睾、 睾丸、 卵 巢、 子宫、 前列腺、 乳腺、 坐骨神经、 膀胱、 眼 (眼科检查发现异常时)、 视神经、 给药局部(尾静脉)、 胸骨(骨和骨髓)、 淋巴结(肠系膜淋巴结) 等用 10%福尔马林固定; 对肝脏、 肾脏、 空肠、 盲肠、 膀胱等脏器先取材, 常规石蜡制片, 切片厚度约 4μιη, Η-Ε染色, OLYMPUS BX40型光学显微镜 观察。 恢复期取余下的动物做上述观测与检测指标。 Eighty SD rats were randomly divided into 4 groups according to animal weight and sex, 20 in each group, half male and half female. Compound I (M394 mesylate) 10, 30, 60 mg / kg three dose groups and a blank control group, tail vein injection, once a day, continuous administration for 4 weeks, resume observation for 2 weeks. The general condition observation was performed every day, and the body weight and food intake were counted once a week. At the end of the administration, half of the animals were collected for blood collection and urine was collected for blood biochemistry, electrolytes, urine analysis, and bloodletting and necropsy were performed for gross pathological examination. Femur, wrist cartilage, femoral articular cartilage, brain (brain, cerebellum, brain stem), spinal cord (neck, chest, lumbar), pituitary, thymus, thyroid, parathyroid, esophagus, salivary gland, stomach, duodenum , ileum, colon, liver, gallbladder, kidney, adrenal gland, spleen, pancreas, trachea, lung, aorta, heart, epididymis, testis, ovary, uterus, prostate, breast, sciatic nerve, bladder, eye (when abnormalities are found in ophthalmology) , optic nerve, local administration (tail vein), sternum (bone and bone marrow), lymph nodes (mesenteric lymph nodes), etc. fixed with 10% formalin; for liver, kidney, jejunum, cecum, bladder and other organs, conventional Paraffin wax, slice thickness about 4μηη, Η-Ε dyeing, OLYMPUS BX40 optical microscopy Observed. During the recovery period, the remaining animals were taken to make the above observation and detection indicators.
试验结果表明:,  The results showed that:,
lOmg/kg'bw剂量组大鼠尿液 pH值略有降低,未见有毒理学意义的变 化。 30 mg/kg*bw剂量组的部分大鼠血清尿素氮有所增加, 雄性大鼠尿液 pH值降低。 60mg/kg*bw剂量组的雄性大鼠血清尿素氮和肌酐明显增加, 尿液 pH值降低, 个别大鼠尿液中可见结晶物质、 股关节和腕关节的轻度 影响; 肾脏脏器系数明显上升, 部分大鼠出现肾小管轻度扩张、 肾小管上 皮轻度变性、 肾脏蛋白管型、 细胞管型及结晶形成。 表明高剂量组的大鼠 肾脏已受到损害, 对肾脏有一定的毒性作用。 在恢复期末, 各给药组动物 血液生化检查及病理组织学检查均未见有毒理学意义的改变,提示 NM394 甲磺酸盐对大鼠引起的毒性反应是可恢复的, 未见延迟性毒性反应。  In the lOmg/kg 'bw dose group, the urine pH was slightly decreased, and no toxicological changes were observed. The serum urea nitrogen of some rats in the 30 mg/kg*bw dose group increased, and the urine pH of male rats decreased. In the 60mg/kg*bw group, the serum urea nitrogen and creatinine were significantly increased, the urine pH was decreased, and the mild effects of crystal material, femoral joint and wrist joint were observed in the urine of individual rats; Ascending, some rats developed mild tubular dilatation, mild degeneration of renal tubular epithelium, renal protein cast, cell tubular type and crystal formation. It indicates that the kidneys of rats in the high-dose group have been damaged and have certain toxic effects on the kidneys. At the end of the recovery period, no toxicological changes were observed in the blood biochemical examination and histopathological examination of the animals in each administration group, suggesting that the toxicity of NM394 mesylate to rats was recoverable, and no delayed toxicity was observed. .
根据以上研究结果, 静脉注射 NM394甲磺酸盐对大鼠的无毒反应剂 量为 30 mg/kg»bW o  Based on the above results, the amount of NM394 mesylate intravenously administered to rats was 30 mg/kg»bW o
本研究结果与 Ishida. S ( 1996年) 报导的大鼠静脉注射 NM394四周 的毒性研究结果不同: NM394 主要表现在肾毒性作用 (10、 30mg/kg-bw 剂量组都出现肾毒性、 血液尿素氮和肌酐明显增加, 但其毒性作用是可逆 的;其无毒作用剂量为 3 mg/kg,bw) ;但 NM394甲磺酸盐 10 mg/kg*bw剂 量组仅出现尿液 pH值降低, 未观察到其他明显的中毒症状, 而同剂量的 NM394 出现肾病变、 尿结晶性物质及血液尿素氮和肌酐明显增加, 这与 NM394甲磺酸盐的 60 mg/kg«bw剂量组的变化基本一致, 而 30mg/kg*bw 的 NM394除上述症状外,还出现尿浊现象和血 γ-球蛋白减少,这与 ΝΜ394 甲磺酸盐的 60 mg/kg«bw剂量组的变化程度接近。  The results of this study are different from those reported by Ishida. S (1996) for intravenous injection of NM394 around rats: NM394 is mainly characterized by nephrotoxicity (10, 30 mg/kg-bw dose group, nephrotoxicity, blood urea nitrogen) And creatinine increased significantly, but its toxic effect is reversible; its non-toxic dose is 3 mg / kg, bw); but NM394 mesylate 10 mg / kg * bw dose group only showed a decrease in urine pH, not Other obvious symptoms of poisoning were observed, and nephropathy, urinary crystalline substances, and blood urea nitrogen and creatinine were significantly increased in the same dose of NM394, which was consistent with the change in the 60 mg/kg «bw dose group of NM394 mesylate. In addition to the above symptoms, NM394 at 30 mg/kg*bw also showed turbidity and decreased blood gamma globulin, which was similar to the change in the 60 mg/kg «bw dose group of ΝΜ394 mesylate.
根据以上研究结果, 静脉注射 NM394甲磺酸盐对大鼠的无毒反应剂 量为 30 mg/kg*bw。 而文献报道的大鼠静脉注射 NM394无毒反应剂量为 3mg/kg-bw, 本发明药物化合物毒性反应大大降低。 试验例 4、 血管剌激、 肌肉刺激实验: According to the above results, intravenous NM394 mesylate salt is a non-toxic reagent for rats. The amount is 30 mg/kg*bw. The non-toxic reaction dose of intravenous NM394 in rats reported in the literature is 3 mg/kg-bw, and the toxicity of the drug compound of the present invention is greatly reduced. Test Example 4, Vaso Stimulation, Muscle Stimulation Experiment:
1、 血管刺激性试验: 被试药物用兔 2 只, 左侧兔耳给予高浓度试液 1. 2mg/mL, 右侧兔耳给予相应低浓度试液 0. 4mg/mL。 另取一只兔作为空 白对照, 左右耳均给予等量氯化钠注射液。 每天注射给药一次, 连续给 3天。末次给药后 48小时剖检。肉眼观察结果表明,匪 394甲磺酸盐高、 低浓度试液则未见剌激性现象, 兔耳血管轮廓较清晰, 兔耳厚薄均匀, 未见明显异常改变。 1. Vascular irritancy test: 2 rabbits were used for the test drug, and the left rabbit ears were given a high concentration test solution 1. 2 mg/mL, and the right rabbit ears were given a corresponding low concentration test solution 0.4 mg/mL. Another rabbit was taken as a blank control, and the right and left ears were given an equal amount of sodium chloride injection. The drug was administered once a day for 3 consecutive days. The necropsy was performed 48 hours after the last administration. The results of macroscopic observation showed that the 高 394 methanesulfonate high and low concentration test solution showed no irritating phenomenon, the rabbit ear blood vessel contour was clear, the rabbit ear thickness was uniform, and no obvious abnormal changes were observed.
2 、 肌肉刺激性试验: 每种被试物用兔 2只, 左侧股肌给予高浓度试 液 1.2mg/mL, 右侧股肌给予相应被试物的低浓度试液 0.4mg/mL。 另取一 只兔作为空白对照, 左右侧股肌均给予等量氯化钠注射液。 每天注射给药 一次, 连续给 3天。末次给药后 48小时剖检。 肉眼观察结果表明, NM394 甲磺酸盐高浓度试液对兔注射部位深层肌肉组织有轻度刺激性, 主要表现 为出现分散性针状出血点, 但试液低浓度未见明显异常改变。 ; 附图说明 2. Muscle irritation test: 2 rabbits were used for each test substance, 1.2 mg/mL of high concentration test solution was administered to the left femoral muscle, and 0.4 mg/mL of low concentration test solution of the corresponding test substance was administered to the right femoral muscle. Another rabbit was used as a blank control, and the right and left lateral femoral muscles were given the same amount of sodium chloride injection. The drug was administered once a day for 3 consecutive days. The necropsy was performed 48 hours after the last administration. Visual observation showed that the NM394 mesylate high concentration test solution was mildly irritating to the deep muscle tissue of the rabbit injection site, mainly showing the occurrence of scattered needle-like bleeding points, but no significant abnormal changes were observed in the low concentration of the test solution. BRIEF DESCRIPTION OF THE DRAWINGS
附图 1是实施例 1提供的化合物 I的 X-射线粉末衍射图谱。  BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is an X-ray powder diffraction pattern of Compound I provided in Example 1.
下面通过实施例进一步阐述本发明的技术方案, 本发明并不仅限于以 下实施例。 具体实施方式  The technical solutions of the present invention are further illustrated by the following examples, and the present invention is not limited to the following embodiments. detailed description
实施例 1  Example 1
100L反应罐中加入 50L水及 5L乙醇, 于 15〜20°C下加入 10Kg化合物 II, 搅拌下滴加甲磺酸 2. 8Kg, 搅拌 30分钟, 加入 0. 5Kg针用活性碳, 搅 拌 15分钟, 过滤至 500L的结晶罐中; 用 10L水洗涤反应罐, 过滤至 500L W 100L 50L reaction tank is added with water and ethanol 5L, 10K g of Compound II was added at 15~20 ° C, added dropwise with stirring methanesulfonic acid 2. 8Kg, stirred for 30 minutes, 0. 5Kg needle with activated carbon, stirred for 15 Minute, filter into 500L crystallization tank; wash the reaction tank with 10L water, filter to 500L W
的结晶罐中。 搅拌下, 结晶罐中滴加约异丙醇 350L, 3小时加完异丙醇, 析出固体。 保温 5〜: UTC搅拌 2小时, 过滤, 用异丙醇 20L洗涤一次, 在 35〜40°C真空干燥。 得 8.7Kg化合物 I。  In the crystallization tank. Under stirring, about 350 L of isopropanol was added dropwise to the crystallization tank, and isopropanol was added over 3 hours to precipitate a solid. Incubation 5~: UTC was stirred for 2 hours, filtered, washed once with 20 L of isopropyl alcohol, and dried under vacuum at 35 to 40 °C. 8.7 Kg of Compound I was obtained.
核磁' HNMR(DMS0-d6, D20 , 500Hz) Nuclear magnetic 'HNMR (DMS0-d 6 , D 2 0 , 500Hz)
红外 IR(KBr,cnT)数据如下: 1698 (m、 vc=Q羧酸)、 1628 (s c=0酮)、 1628-1500 (m、 s、 vc=c (芳环、 芳杂环))、 1457 (m、 Infrared IR (KBr, cnT) data are as follows: 1698 (m, v c = Q carboxylic acid), 1628 (sc = 0 ketone), 1628-1500 (m, s, v c = c (aromatic ring, aromatic heterocyclic) ), 1457 (m,
)、 1397 (m、 SQI1羧酸) ), 1397 (m, S QI1 carboxylic acid)
产品元素分析结果如下:  The product element analysis results are as follows:
分析项目 C H N S 测定值% 52.11 4.86 9.56 7.39  Analytical project C H N S measured value 52.11 4.86 9.56 7.39
52.21 4.79 9.64 7.28 计算值% 52.17 4.81 9.61 7.32 产品用 Cu- Kct 射线测量得到的 X-射线粉末衍射图中的 2 Θ、 d-面间 距和相对强度 1/1。具有如下数值: 52.21 4.79 9.64 7.28 Calculated value 52.17 4.81 9.61 7.32 The product has a 2 Θ, d-plane spacing and a relative intensity of 1/1 in the X-ray powder diffraction pattern measured by Cu-Kct ray. Has the following values:
2 Θ d 1/1。  2 Θ d 1/1.
5.520 15.99 0.84  5.520 15.99 0.84
10.320 8.56 0.74  10.320 8.56 0.74
11.100 • 7.96 0.83  11.100 • 7.96 0.83
14.040 6.30 0.58  14.040 6.30 0.58
14.420 6.13 0.31  14.420 6.13 0.31
16.340 5.42 0.32  16.340 5.42 0.32
16.880 5.24 0.40  16.880 5.24 0.40
17.780 4, 98 0.49  17.780 4, 98 0.49
18.740 4.73 0.46  18.740 4.73 0.46
19.820 4.47 0.46  19.820 4.47 0.46
22.720 3.90 0.38  22.720 3.90 0.38
26.140 3.40 1.00  26.140 3.40 1.00
26.800 3.32 0.74  26.800 3.32 0.74
' 实施例 2  'Example 2
反应罐中加入 50L水及 15L丙酮, 于 10〜15°C下加入 10Kg化合物 II, 搅拌下滴加甲磺酸 2.8Kg, 搅拌 1小时, 加入 0.5Kg针用活性碳, 搅拌 20 分钟, 过滤脱碳; 滤液经 0.22 μ m超滤膜压入无菌室 500L结晶罐。 水 10L 洗涤反应罐及管道, 也压入结晶罐中。 开搅拌, 滴加丙酮 350L, 4小时加 完丙酮, 析出固体。 保温 0〜5°C搅拌 5小时, 过滤, 丙酮 30LX2洗涤两 次, 35〜40°C真空干燥。 得化合物 I的无菌粉末 8, 4Kg。  Add 50L of water and 15L of acetone to the reaction tank, add 10Kg of compound II at 10~15°C, add 2.8Kg of methanesulfonic acid with stirring, stir for 1 hour, add 0.5Kg of needle with activated carbon, stir for 20 minutes, filter off Carbon; The filtrate was pressed into a sterile chamber 500L crystallization tank through a 0.22 μm ultrafiltration membrane. Water 10L Washing the reaction tank and piping, and also pressing into the crystallization tank. After stirring, 350 L of acetone was added dropwise, and acetone was added over 4 hours to precipitate a solid. The mixture was stirred at 0 to 5 ° C for 5 hours, filtered, and washed twice with acetone 30 LX 2 and dried under vacuum at 35 to 40 ° C. A sterile powder of Compound I 8, 8, Kg was obtained.
核磁1 H匪 R(DMS0-ds, D20, 500Hz), 红外 IR(KBr, cm—1), 及 X-射线粉末 衍射数据同例 1。 实施例 3 Nuclear magnetic 1 H匪R (DMS0-d s , D 2 0, 500 Hz), infrared IR (KBr, cm- 1 ), and X-ray powder diffraction data are the same as in Example 1. Example 3
在 100ml三颈瓶中加入 50ml水, 于 0〜5°C下加入 10g化合物 II, 搅拌 下滴加甲磺酸 4. lg, 搅拌 30分钟, 加入 0.5g针用活性碳, 搅拌 15分钟, 过滤脱碳; 10ml水洗涤三颈瓶, 过滤。 合并滤液至 500ml三颈瓶中, 搅拌 下, 滴加乙醇 350ml, 2小时加完混合溶液, 析出固体。 保温 15〜20°C搅 拌 2小时, 过滤, 无水乙醇 20ml洗涤一次, 35〜40°C真空干燥。 得 6. 9g 化合物 I。 Add 50 ml of water to a 100 ml three-necked flask, add 10 g of compound II at 0 to 5 ° C, add 4. l g of methanesulfonic acid with stirring, stir for 30 minutes, add 0.5 g of needle with activated carbon, and stir for 15 minutes. Filter decarburization; wash the three-necked flask with 10 ml of water and filter. The filtrate was combined into a 500 ml three-necked flask and stirred. Next, 350 ml of ethanol was added dropwise, and the mixed solution was added over 2 hours to precipitate a solid. The mixture was stirred at 15 to 20 ° C for 2 hours, filtered, washed once with 20 ml of absolute ethanol, and dried under vacuum at 35 to 40 ° C. 6. g of compound I.
核磁1 HNMR(DMS0- d6, D20 , 500Hz) , 红外 IR (KBr, cm-1), 及 X-射线粉末 衍射数据同例 1。 实施例 4 Nuclear magnetic 1 H NMR (DMS0-d 6 , D 2 0 , 500 Hz), infrared IR (KBr, cm -1 ), and X-ray powder diffraction data were the same as in Example 1. Example 4
在 100ml三颈瓶中加入 40ml水和 5ml乙醇,于 10〜15°C下加入 10g化 合物 II, 搅拌下滴加甲磺酸 2. 5g, 搅拌 20分钟, 加入 0. 5g针用活性碳, 搅拌 15分钟, 过滤脱碳; 10ml水洗涤三颈瓶, 过滤。 合并滤液至 500ml 三颈瓶中, 搅拌下, 滴加丙酮 1000ml, 4小时加完丙酮, 析出固体。 保温 15〜20°C搅拌 2小时, 过滤, 丙酮 20ml洗涤一次, 35〜40°C真空干燥。 得 5. 9g化合物 I。 实施例 5 In a three-necked 100ml flask was added 40ml of water and 5ml of ethanol, 10g of compound II at 10~15 ° C, added dropwise with stirring methanesulfonic acid 2. 5g, stirred for 20 minutes, was added 0. 5 g needle with activated carbon, Stir for 15 minutes, filter for decarburization; wash the three-necked flask with 10 ml of water and filter. The filtrate was combined into a 500 ml three-necked flask. Under stirring, 1000 ml of acetone was added dropwise, and acetone was added over 4 hours to precipitate a solid. The mixture was stirred at 15 to 20 ° C for 2 hours, filtered, and washed with acetone 20 ml once, and dried at 35 to 40 ° C under vacuum. 5. g of compound I was obtained. Example 5
在 100ml三颈瓶中加入 200ml水和 50ml甲醇, 于 55〜60°C下加入 5g 化合物 II,搅拌下滴加甲磺酸 1. lg,搅拌 30分钟,加入 0. 2g针用活性碳, 搅拌 15分钟, 过滤脱碳; 10ml水洗涤三颈瓶, 过滤。 合并滤液至 250ml 三颈瓶中, 室温搅拌, 滴加四氢呋喃 500ml, 2 小时加完四氢呋喃, 析出 固体。 保温 15〜2(TC搅拌 2小时, 过滤, 丙酮 20ml洗涤一次, 35〜40°C 真空干燥, 得 2. lg化合物 I。 实施例 6  2克针用活性碳,搅拌搅拌。 Add a mixture of 0. 2g of the activated carbon, stir, add 0g of the compound II, add 5g of the compound II. 15 minutes, decarburization by filtration; wash the three-necked flask with 10 ml of water and filter. The filtrate was combined into a 250 ml three-necked flask, stirred at room temperature, and 500 ml of tetrahydrofuran was added dropwise thereto, and tetrahydrofuran was added over 2 hours to precipitate a solid. Insulation 15~2 (TC stirring for 2 hours, filtration, acetone 20 ml once, 35~40 ° C vacuum drying, 2. lg compound I. Example 6
在 100ml三颈瓶中加入 15ml水和 25ml四氢呋喃, 于 25〜30°C下加入 10g化合物 II, 搅拌下滴加甲磺酸 2. 3g, 搅拌 60分钟, 加入 0. 5g针用活 性碳, 搅拌 15分钟, 过滤脱碳; 10ml水洗涤三颈瓶, 过滤。 合并滤液至 100ml三颈瓶中, 搅拌下, 滴加丙酮 10ml, 10分钟加完丙酮, 析出固体。 保温 15〜20°C搅拌 2小时, 过滤, 丙酮 10ml洗涤一次, 35〜40°C真空干 燥, 得 2. 5g化合物 I。 实施例 7 5克针用活性碳。 In a 100ml three-necked flask with the addition of 15ml of water and 25ml of tetrahydrofuran, at 25~30 ° C, 10 g of the compound II, stirring with the addition of methane sulfonic acid 2. 3g, stirring for 60 minutes, adding 0. 5 g needle with activated carbon Stir for 15 minutes, filter for decarburization; wash the three-necked flask with 10 ml of water and filter. The filtrate was combined into a 100 ml three-necked flask, and 10 ml of acetone was added dropwise with stirring, and acetone was added for 10 minutes to precipitate a solid. 5克化合物1。 I. Ig. Example 7
在反应罐中加入 100L水, 于 10〜15°C下加入 10Kg化合物 Π, 搅拌下 滴加甲磺酸 2.8Kg, 搅拌 1小时, 加入 0.5Kg针用活性碳, 搅拌 20分钟, 过滤脱碳; 滤液经 0.22 μηι超滤膜过滤, 溶液放入冻干机中冻千, 得到化 合物 I的冻干粉末 8.4Kg。 Add 100L of water to the reaction tank, add 10Kg of compound hydrazine at 10~15 °C, stir Add 2.8Kg of methanesulfonic acid dropwise, stir for 1 hour, add 0.5K g of needle with activated carbon, stir for 20 minutes, filter and decarburize; filter the filtrate through 0.22 μηι ultrafiltration membrane, and put the solution into the freeze dryer to freeze the compound to obtain the compound. I lyophilized powder of 8.4Kg.
实施例 8 注射剂的制备  Example 8 Preparation of Injection
取实施例 1所得的 25.6g化合物 I加到 15〜20°C的 0.9%氯化钠溶液 5L中, 搅拌溶解后, 加入 0.3g针用活性碳, 搅拌 10分钟, 过滤脱碳; 用 0.45μηι滤膜过滤, 再用 0.22 μιη滤膜过滤, 加 0.9%氯化钠溶液至总量为 10L, 测定含量, 检査澄明度, 灌装到 100ml输液瓶中, 压塞轧铝盖, 121 °C灭菌 20min。 实施例 9 注射剂的制备  25.6 g of the compound I obtained in Example 1 was added to 5 L of a 0.9% sodium chloride solution at 15 to 20 ° C, and after stirring and dissolved, 0.3 g of a needle was added with activated carbon, stirred for 10 minutes, and decarburized by filtration; using 0.45 μηι Filter the membrane, filter with 0.22 μηη filter, add 0.9% sodium chloride solution to the total amount of 10L, measure the content, check the clarity, fill into the 100ml infusion bottle, press the rolled aluminum cover, 121 °C Sterilize for 20 min. Example 9 Preparation of Injection
取实施例 1所得的 12.8g化合物 I加到 15〜20°C的 5%葡萄糖溶液 2L 中,搅拌溶解后,加入 0.3g针用活性碳,搅拌 10分钟,过滤脱碳;用 0.45 m滤膜过滤, 再用 0.22 μηι滤膜过滤, 加 5°/。葡萄糖溶液至总量为 10L, 测定含量, 检查澄明度, 灌装到 100ml 输液瓶中, 压塞轧铝盖, 121。C灭 菌 20min。 实施例 10 注射剂的制备  12.8 g of the compound I obtained in Example 1 was added to 2 L of a 5% glucose solution at 15 to 20 ° C, stirred and dissolved, and 0.3 g of a needle was added with activated carbon, stirred for 10 minutes, and decarburized by filtration; a filter of 0.45 m was used. Filter and filter with 0.22 μηι filter, add 5 ° /. Glucose solution to a total of 10L, determine the content, check the clarity, fill the 100ml infusion bottle, press the aluminum cap, 121. C kill bacteria for 20min. Example 10 Preparation of Injection
取实施例 1所得的 128g化合物 I加到 15〜20°C的注射用水 500ml中, 搅拌溶解后, 加入 5g针用活性碳, 搅拌 10分钟, 过滤脱碳; 0.45μπτ滤 膜过滤, 再用 0.22 μπι滤膜过滤, 加注射用水至总量为 lOOOOml, 测定含 量, 检查澄明度, 充氮灌封到 10ml的安锫瓶中, 121Ό灭菌 20rain。 实施例 11 粉针剂的制备  The compound I obtained in Example 1 was added to 500 ml of water for injection at 15 to 20 ° C, stirred and dissolved, and then 5 g of the needle was added with activated carbon, stirred for 10 minutes, and decarburized by filtration; 0.45 μπτ filter was filtered, and 0.22 was further used. Filter through μπι filter, add water for injection to a total of lOOOOml, determine the content, check the clarity, fill with nitrogen in a 10ml ampoule, and sterilize for 20rain at 121Ό. Example 11 Preparation of powder injection
取实施例 2所得的化合物 I的无菌粉在无菌条件下分装成 0.128g/瓶、 0.192g/瓶、 0.256g/瓶、 0.320g/瓶、 0.384g/瓶、 0.512g/瓶或 0.640g/ 瓶, 得到化合物 I的无菌粉末注射制剂。 实施例 12 注射剂的制备  The sterile powder of Compound I obtained in Example 2 was dispensed under sterile conditions into 0.128 g/bottle, 0.192 g/bottle, 0.256 g/bottle, 0.320 g/bottle, 0.384 g/bottle, 0.512 g/bottle or 0.640 g / bottle, a sterile powder injection preparation of Compound I was obtained. Example 12 Preparation of Injection
取实施例 7所得的化合物 I的无菌粉在无菌条件下分装成 0.128g/瓶、 0.192g/瓶、 0.256g/瓶、 0.320g/瓶、 0.384g/瓶、 0.512g/瓶或 0.640g/ 瓶, 得到化合物 I的无菌冻干粉末注射制剂。 实施例 13 片剂的制备 The sterile powder of Compound I obtained in Example 7 was dispensed under sterile conditions into 0.128 g/bottle, 0.192 g/bottle, 0.256 g /bottle, 0.320 g /bottle, 0.384 g /bottle, 0.512 g/bottle or 0.640 g / bottle, a sterile lyophilized powder injection preparation of Compound I was obtained. Example 13 Preparation of tablets
处方(每千片用量): 化合物 I 128g、 乳糖 1. 9g、 10%PVP溶液 10ml、 硬脂 酸镁 0. 52g。  Prescription (per 1,000 tablets): Compound I 128 g, lactose 1. 9 g, 10% PVP solution 10 ml, magnesium stearate 0. 52 g.
制备: 取实施例 1所得的化合物 I与乳糖在流化床制粒机中吹风处理 10分钟, 使其混匀, 再喷入 10%PVP溶液, 风速约 120米 /小时, 喷入速度 约为 2〜3ml/分, 60°C热气流千燥约 15分钟,加入硬脂酸镁混匀后,压片, 包衣即得。 实施例 14 胶囊剂的制备  Preparation: The compound I obtained in Example 1 and lactose were air-dried in a fluidized bed granulator for 10 minutes, mixed, and then sprayed into a 10% PVP solution at a wind speed of about 120 m/hr. 2~3ml/min, 60°C hot air flow for about 15 minutes, add magnesium stearate to mix, press, and coat. Example 14 Preparation of Capsules
处方(一 立处方量): 化合物 I 128g、微晶纤维素 40g、羧甲淀¾ ^内 50g、硬 脂藤 3.4g。 Prescription (a prescription amount of a): Compound I 128g, microcrystalline cellulose 40g, starch carboxymethylcellulose within ¾ ^ 50 g, stearyl vine 3.4g.
制备: 取处方中的原辅料, 充分混合均匀后, 填充至 4 号空胶囊中, 即得。 实施例 15 颗粒剂的制备  Preparation: Take the original and auxiliary materials in the prescription, mix well and fill them into the No. 4 empty capsules. Example 15 Preparation of granules
处方: 化合物 I 128g、 糊精 120g、 蔗糖糖 280g。  Prescription: Compound I 128g, dextrin 120g, sucrose sugar 280g.
制备: 取实施例 1 所得的化合物 I和蔗糖粉碎成细粉, 混匀, 以湿法 制粒, 置 60Ό下, 充分干燥, 分装, 即得。 实施例 16 滴眼剂的制备  Preparation: The compound I obtained in Example 1 and sucrose were pulverized into fine powder, mixed, and granulated by a wet method, placed under 60 Torr, sufficiently dried, and dispensed. Example 16 Preparation of Eye Drops
处方: 化合物 I 3.84g、 氯化钠 0.9g、 硼酸缓冲溶液适量、 苯乙醇 3g、 7 , 共 1000 ml c  Prescription: Compound I 3.84g, sodium chloride 0.9g, boric acid buffer solution, phenylethyl alcohol 3g, 7 , a total of 1000 ml c
制备: 取实施例 1所得的化合物 I和氯化钠加到 600ml蒸馏水中, 搅 拌下完全溶解后, 加入硼酸缓冲溶液调节溶液 pH为 6. 5; 另外取苯乙醇, 加沸蒸馏水 200ml 使之溶解; 将上述两溶液混匀, 加蒸馏水至总量为 1000ml, 搅匀, 用 0. 22 μ πι微孔滤膜过滤, 灌装, 严封, 以 10CTC灭菌 30 分钟流通蒸汽灭菌, 分装, 即得。 实施例 17 滴耳液的制备  Preparation: The compound I obtained in Example 1 and sodium chloride was added to 600 ml of distilled water, and dissolved completely after stirring, and the pH of the solution was adjusted to be 6.5 by adding a boric acid buffer solution. Further, phenylethyl alcohol was added, and 200 ml of boiling distilled water was added to dissolve it. Mix the above two solutions, add distilled water to a total amount of 1000ml, stir well, filter with 0.22 μππ microporous membrane, fill, seal tightly, sterilize with 10CTC for 30 minutes, steam sterilization, dispense , that is. Example 17 Preparation of ear drops
处 方 : 化 合 物 I 12.8g 、 乙 醇 300 ml 、 甘 油 300 ml、水 ¾¾、共 1000 ml  Where: Compound I 12.8g, ethanol 300 ml, glycerin 300 ml, water 3⁄43⁄4, total 1000 ml
制备: 取实施例 1所得的 12. 8g化合物 I溶解于注射用水 200ml中, 加入乙醇 300ml和甘油 2.00ml, 再加入注射用水至总量为 1000ml, 搅匀, 用 0. 22 μ微孔滤膜过滤, 灌装, 即得。 实施例 18 栓剂的制备 Preparation: 12. 8g of Compound I obtained in Example 1 was dissolved in 200ml of water for injection, Add 300 ml of ethanol and 2.00 ml of glycerin, add water for injection to a total amount of 1000 ml, stir well, filter with 0.22 μm microfiltration membrane, and fill. Example 18 Preparation of suppository
处方: 化合物 1 12. 8g、 可可豆脂 200g, 共制成 100粒。  Prescription: Compound 1 12. 8g, cocoa butter 200g, 100 capsules.
制备: 将可可豆脂 200g, 在水浴上加热熔化(温度控制在 5CTC以下), 取实施例 1所得的化合物 I细粉' 12. 8g (过 100 目筛) 加入熔化的可可豆 脂中, 边加边搅拌, 使药物均匀分散在基质中, 然后趁热倾入冷却并涂有 润滑剂的栓模中, 待凝固后刮平, 启模, 包装, 即得。 实施例 12 外用皮肤制剂的制备  Preparation: 200 g of cocoa butter was heated and melted on a water bath (temperature controlled below 5 CTC), and the fine powder of Compound I obtained in Example 1 was taken. 12.8 g (over 100 mesh sieve) was added to the melted cocoa butter. Stirring is carried out to uniformly disperse the drug in the matrix, and then poured into a bolt which is cooled and coated with a lubricant, which is to be solidified, then flattened, opened, and packaged. Example 12 Preparation of topical skin preparation
处方: 化合物 I 128g、 硬脂酸 85g、 白凡士林 170g、 单硬脂酸甘油酯 43g、 十二垸基硫酸钠 2g、 甘油 100g、 三乙醇胺 4g、 X寸羟基苯甲酸乙酯 lg、 水适量, 共 1000g。  Prescription: Compound I 128g, stearic acid 85g, white petrolatum 170g, glyceryl monostearate 43g, sodium decyl sulfate 2g, glycerol 100g, triethanolamine 4g, X-hydroxy hydroxybenzoate lg, water amount, A total of 1000g.
制备: 取实施例 1所得的 128g化合物 I溶解于 300ml水中, 再取十二 烷基硫酸钠、 甘油、 三乙醇胺、 对羟基苯甲酸乙脂及化合物 1的水溶液, 置水浴上加热至全部溶解, 并使温度在 70°C左右; 另取硬脂酸、 白凡士林 及单硬脂酸甘油酯, 加热至全部熔化, 待温度降至 70°C左右, 在不断搅拌 下缓缓加入上项溶液, 并不断搅拌至乳化完全, 冷凝, 即得。 工业应用性 本发明提供了一种喹诺酮类抗感染的药物,具有结构确定,性质稳定, 抗菌疗效确切, 改善了活性药物化合物 (Π ) 的水溶性, 可以方便地制备 成各种适宜的剂型用于临床。 血管剌激, 肌肉剌激试验表明本发明化合物 适用于肌肉注射和静脉注射, 扩大临床应用范围, 增加了抗感染药物制剂 新品种。 本发明通过制备化合物(II ) 的甲磺酸盐, 降低了活性物质(II ) 的毒性, 增加了临床用药安全性。此外本发明的制备生产工艺简便、合理, 生产成本低, 具有工业适用性。  Preparation: 128 g of Compound I obtained in Example 1 was dissolved in 300 ml of water, and then an aqueous solution of sodium lauryl sulfate, glycerin, triethanolamine, ethyl p-hydroxybenzoate and Compound 1 was placed, and heated to a total dissolution in a water bath. And the temperature is about 70 ° C; another take stearic acid, white petrolatum and glyceryl monostearate, heated to full melting, until the temperature drops to about 70 ° C, slowly add the above solution with constant stirring, And continue to stir until the emulsification is complete, condensation, that is. Industrial Applicability The present invention provides a quinolone anti-infective drug which has a structure-determined, stable property, and has an antibacterial effect, and improves the water solubility of the active drug compound (Π), and can be conveniently prepared into various suitable dosage forms. In the clinic. Vaso-irritation, muscle stimulation tests indicate that the compounds of the present invention are suitable for intramuscular and intravenous injections, expand the range of clinical applications, and increase new varieties of anti-infective pharmaceutical preparations. The preparation of the mesylate salt of the compound (II) by the invention reduces the toxicity of the active substance (II) and increases the safety of clinical use. In addition, the preparation and production process of the invention is simple and reasonable, the production cost is low, and the industrial applicability is obtained.

Claims

权利要求 Rights request
1、 一种具有结构式 ( I ) 所示结构的抗感染化合物, 该化合物的化 学名称是 6-氟- 1-甲基- 4-氧代- 7-(1-哌嗪基 )-4Η-[1, 3]硫氮杂环丁烷并 [3, 2- a]喹啉 -3-羧酸甲磺酸盐:  An anti-infective compound having the structure of the formula (I), the chemical name of which is 6-fluoro-1-methyl-4-oxo-7-(1-piperazinyl)-4Η-[ 1, 3]thiazetino[3,2-a]quinoline-3-carboxylic acid mesylate:
其特征在于, 该化合物的结晶性粉末用 Cu-K a 射线测量得到的 X-劑 线粉末衍射图中的 2 Θ、 d-面间距和相对强度具有如下数值:  It is characterized in that the 2 Θ, d-plane pitch and relative intensity in the X-ray line powder diffraction pattern of the crystalline powder of the compound measured by Cu-K a ray have the following values:
2 Θ d 1/1。  2 Θ d 1/1.
5.520 15.99±0.2 0. 84-1. 00  5.520 15.99±0.2 0. 84-1. 00
10.320 8. 56±0. 2 0. 65-0. 78  10.320 8. 56±0. 2 0. 65-0. 78
11.100 7. 96±0. 2 0. 83-0. 96  11.100 7. 96±0. 2 0. 83-0. 96
14.040 6. 30±0. 2 0. 42-0. 64  14.040 6. 30±0. 2 0. 42-0. 64
14.420 6. 13±0. 2 0. 24-0. 45  14.420 6. 13±0. 2 0. 24-0. 45
16.340 5. 42 + 0. 2 0. 25-0. 46  16.340 5. 42 + 0. 2 0. 25-0. 46
16.880 5. 24 ±0. 2 0. 27-0. 51  16.880 5. 24 ±0. 2 0. 27-0. 51
17.780 4. 98 ±0. 2 0. 29-0. 49  17.780 4. 98 ±0. 2 0. 29-0. 49
18.740 4. 73 + 0. 1 0. 46-0. 72  18.740 4. 73 + 0. 1 0. 46-0. 72
19.820 4. 47 + 0. 1 0. 39-0. 58  19.820 4. 47 + 0. 1 0. 39-0. 58
22.720 3. 91±0. 1 0. 35-0. 63  22.720 3. 91±0. 1 0. 35-0. 63
26.140 3. 40 ±0. 1 0. 52-1. 00  26.140 3. 40 ±0. 1 0. 52-1. 00
26.800 3. 32 ±0. 1 0. 45-1. 00  26.800 3. 32 ±0. 1 0. 45-1. 00
2、 一种制备如权利要求 1所述的抗感染化合物的方法, 其特征在于 用水或含有机溶剂体积百分比为 5〜50%的水作溶剂, 于 0〜60°C加入 6 - 氟- 1-甲基- 4-氧代- 7-(l-哌嗪基 )- 4H- [1, 3] 硫氮杂环丁烷并 [3,2-a] 喹啉- 3-羧酸 (II ) 和甲磺酸, 所用溶剂量为化合物 (II) 重量的 5〜50 倍, 搅拌溶解后, 加入化合物 (Π) 重量的 1〜100倍的有机溶剂, 即析 出化合物 ( I ) , 化合物 (II) 和甲磺酸的投料摩尔比为 1: 0.8〜1.5, 其中所说的有机溶剂是甲醇、 乙醇、 异丙醇、 丙酮、 四氢呋喃中的任意〜 2. A method for preparing an anti-infective compound according to claim 1, characterized in that water or a solvent containing 5 to 50% by volume of organic solvent is used as a solvent, and 6-60 ° C is added to 6 - Fluoro-1-methyl-4-oxo-7-(l-piperazinyl)-4H- [1,3]thiazepine[3,2-a]quinoline-3-carboxylic acid (II) and methanesulfonic acid, the amount of the solvent used is 5 to 50 times the weight of the compound (II), and after stirring and dissolved, the organic solvent is added in an amount of 1 to 100 times by weight of the compound (Π), thereby precipitating the compound (I), the compound The molar ratio of (II) to methanesulfonic acid is 1: 0.8~1.5, wherein the organic solvent is any of methanol, ethanol, isopropanol, acetone, tetrahydrofuran~
3、 一种抗感染药物组合物, 该组合物含有权利要求 1所述的抗感染 类化合物作为有效成分, 并含有常规药用载体。 An anti-infective pharmaceutical composition comprising the anti-infective compound of claim 1 as an active ingredient and comprising a conventional pharmaceutical carrier.
4、 根据权利要求 3所说的抗感染药物组合物, 其特征在于, 该组合 物是一种胃肠道给药的片剂、 胶囊剂或颗粒剂。 The anti-infective pharmaceutical composition according to claim 3, which is a tablet, capsule or granule for gastrointestinal administration.
5、 根据权利要求 3所说的抗感染药物组合物, 其特征在于, 该组合 物是非胃肠道给药的注射剂、 眼用制剂、 耳用制剂、 妇科用制剂或皮肤外 用制剂。 The anti-infective pharmaceutical composition according to claim 3, which is a parenteral injection, an ophthalmic preparation, an otic preparation, a gynecological preparation or an external preparation for skin.
PCT/CN2007/000179 2006-01-18 2007-01-18 Anti-infective compound, preparation method thereof and use thereof WO2007082471A1 (en)

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