WO2022001847A1 - 四氢异喹啉类衍生物的盐、其制备方法及其医药应用 - Google Patents

四氢异喹啉类衍生物的盐、其制备方法及其医药应用 Download PDF

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WO2022001847A1
WO2022001847A1 PCT/CN2021/102261 CN2021102261W WO2022001847A1 WO 2022001847 A1 WO2022001847 A1 WO 2022001847A1 CN 2021102261 W CN2021102261 W CN 2021102261W WO 2022001847 A1 WO2022001847 A1 WO 2022001847A1
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methoxy
sodium
pharmaceutically acceptable
acceptable salt
alkali metal
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PCT/CN2021/102261
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English (en)
French (fr)
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郭阳辉
孟力陈
黄贤贵
廖伟伟
龚永祥
徐代旺
胡泰山
陈磊
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浙江海正药业股份有限公司
上海昂睿医药技术有限公司
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Priority to EP21832154.5A priority Critical patent/EP4177253A1/en
Priority to US18/012,269 priority patent/US20230242522A1/en
Priority to JP2023521813A priority patent/JP2023531097A/ja
Publication of WO2022001847A1 publication Critical patent/WO2022001847A1/zh

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/472—Non-condensed isoquinolines, e.g. papaverine
    • A61K31/4725—Non-condensed isoquinolines, e.g. papaverine containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/02—Drugs for disorders of the nervous system for peripheral neuropathies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/04—Centrally acting analgesics, e.g. opioids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/07—Optical isomers

Definitions

  • the present invention relates to (S)-2-(6-fluorobenzo[d]oxazol-2-yl)-6-methoxy-5-((5-methoxypyridin-2-yl)methoxy )-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid in a pharmaceutically acceptable salt form, a process for its preparation, a pharmaceutical composition comprising the pharmaceutically acceptable salt and its use as a therapeutic agent, in particular as angiotensin Use of hormone II type 2 receptor antagonists.
  • Neuropathic pain is a chronic pain disease caused by primary injury or dysfunction of the nervous system. Trauma, inflammation, infection, or compression can cause neuropathic pain, such as diabetic neuralgia (DNP), post-herpetic neuralgia (PHN), primary neuropathy, secondary neuropathy, peripheral neuropathy, Neuropathic diseases caused by mechanical nerve injury or biochemical nerve injury, etc.
  • DNP diabetic neuralgia
  • PPN post-herpetic neuralgia
  • primary neuropathy secondary neuropathy
  • peripheral neuropathy mainly include antiepileptic drugs, antidepressants and narcotic analgesics, such as gabapentin, pregabalin, tricyclic antidepressants, etc.
  • Angiotensin II receptor is a G protein-coupled receptor with angiotensin II as a ligand, and it is an important part of the renin-angiotensin system.
  • the major subtypes of angiotensin II receptors include the type 1 receptor (AT 1 R) and the type 2 receptor (AT 2 R).
  • AT 1 R and AT 2 R have only about 30% of the same amino acid sequence, but angiotensin II, as its main ligand, has similar affinity with both.
  • AT 2 R is abundantly expressed in various embryonic tissues and less distributed in adult normal tissues, but its expression is elevated after tissue injury.
  • AT 2 R is related to blood pressure regulation, nerve growth, pain control and myocardial regeneration .
  • Drugs targeting AT 2 R can improve cardiovascular function and relieve neuropathic pain.
  • the compound olodanrigan (EMA401) developed by Spinifex in Australia, is a highly selective AT 2 R antagonist and is currently in Phase II clinical trials. Good therapeutic effect, while Spinifex is also developing AT 2 R antagonist EMA-400.
  • WO 93/23378 discloses the preparation method of Olodanrigan and EMA-400, and its structures are respectively as follows:
  • AT 2 R antagonists At present, a series of patent applications involving AT 2 R antagonists have been published, including WO2016113668, WO2015003223 and WO2013110135, etc.
  • the research and application of AT 2 R antagonists have made certain progress, but they are still far from meeting the needs of clinical medication.
  • the room for improvement is still huge, and it is still necessary to continue the research and development of new AT 2 R antagonists, especially new selective AT 2 R antagonists.
  • (S)-2-(6-Fluorobenzo[d]oxazol-2-yl)-6-methoxy-5-((5-methoxypyridin-2-yl)methoxy)-1 ,2,3,4-tetrahydroisoquinoline-3-carboxylic acid (compound 1m) is an angiotensin II type 2 receptor antagonist, the preparation and biological activity of which are disclosed in WO2019242599.
  • the compound has insufficient water solubility and unsatisfactory pharmacokinetic properties, which greatly limit its application. Therefore, how to improve its solubility and pharmacokinetic properties is still a problem in the prior art.
  • the inventors have unexpectedly found that the salts of this compound, especially the sodium salt thereof, have particularly favorable water solubility and significantly improved pharmacokinetic properties.
  • the present invention provides a (S)-2-(6-fluorobenzo[d]oxazol-2-yl)-6-methoxy-5-((5-methoxypyridine- Pharmaceutically acceptable salt of 2-yl)methoxy)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid (Compound 1m), method for its preparation, and pharmaceutical combination comprising the same and their use as therapeutic agents, especially as angiotensin II type 2 receptor antagonists.
  • the pharmaceutically acceptable salt is (S)-2-(6-fluorobenzo[d]oxazol-2-yl)-6-methoxy-5-((5-methoxypyridine-2- yl)methoxy)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid with an organic or inorganic base as an alkali metal, calcium or magnesium salt; the sodium salt thereof is particularly preferred.
  • the salt-formed form is excellent for the treatment of primary neuropathy, secondary neuropathy, peripheral neuropathy, neuropathy due to mechanical or biochemical nerve damage, post-herpetic neuralgia, diabetic neuralgia or related neuropathies
  • the disease activity and solubility are obviously improved, the animal body has good pharmacokinetic properties, and the toxicity is low, and it is suitable for preparing preparations for treating neurological diseases.
  • the alkali metal salts of the present invention are selected from sodium salts, potassium salts and lithium salts; preferably sodium salts.
  • Typical pharmaceutically acceptable salts of the compounds of formula (1m) of the present invention include, but are not limited to:
  • the present invention also relates to a process for the preparation of a pharmaceutically acceptable salt of a compound of formula (1m), said process comprising making (S)-2-(6-fluorobenzo[d]oxazol-2-yl)-6-methoxy Base-5-((5-methoxypyridin-2-yl)methoxy)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid with metal hydroxide, alkali metal carbonate , the step that alkali metal bicarbonate or alkali metal alkoxide react, wherein said metal hydroxide is selected from sodium hydroxide, potassium hydroxide, lithium hydroxide, calcium hydroxide, magnesium hydroxide, preferably sodium hydroxide ; Described alkali metal carbonate is selected from sodium carbonate and potassium carbonate; Described alkali metal bicarbonate is selected from sodium bicarbonate and potassium bicarbonate; Described alkali metal alkoxide is selected from sodium methoxide, sodium ethoxide
  • reaction temperature has a certain influence on different salt-forming reactions, which is also well known to those skilled in the art.
  • the salt-forming reaction of the present invention The temperature is from normal temperature to the boiling point of the solvent used, and those skilled in the art can easily determine the most preferred reaction temperature for the specific salt-forming reaction through conventional technical means in the art.
  • the present invention also provides a pharmaceutical composition, which contains a therapeutically effective dose of a pharmaceutically acceptable salt of the compound of formula (1m), and a pharmaceutically acceptable carrier; Use in the medicament of primary neuropathy, secondary neuropathy, peripheral neuropathy, neuropathy caused by mechanical or biochemical nerve damage, post-herpetic neuralgia, diabetic neuralgia or related neurological diseases.
  • the present invention also provides a pharmaceutically acceptable salt of the compound of formula (1m) or a pharmaceutical composition thereof in the preparation and treatment of primary neuropathy, secondary neuropathy, peripheral neuropathy, neuropathy caused by mechanical nerve injury or biochemical nerve injury, Use in the medicament of postherpetic neuralgia, diabetic neuralgia or related neurological diseases.
  • the present invention also provides the use of the pharmaceutically acceptable salt of the compound of formula (1m) or the pharmaceutical composition thereof in the preparation of angiotensin II type 2 receptor antagonist.
  • the present invention also relates to a treatment for primary neuropathy, secondary neuropathy, peripheral neuropathy, neuropathy caused by mechanical or biochemical nerve damage, post-herpetic neuralgia, diabetic neuralgia or related neurological diseases , comprising administering to a patient in need thereof a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of formula (Im) or a pharmaceutical composition thereof.
  • the present invention also relates to a method of inhibiting the angiotensin II type 2 receptor, the method comprising contacting the angiotensin II type 2 receptor with a pharmaceutically acceptable salt of a compound of formula (1m) or a pharmaceutical composition thereof.
  • the present invention also relates to a pharmaceutically acceptable salt of a compound of formula (1m) or a pharmaceutical composition thereof for use in the treatment of primary neuropathy, secondary neuropathy, peripheral neuropathy, neuropathy caused by mechanical or biochemical nerve damage, Medications for postherpetic neuralgia, diabetic neuralgia, or related neurological disorders.
  • the present invention also relates to a pharmaceutically acceptable salt of the compound of formula (1m) or a pharmaceutical composition thereof for use as a medicament for inhibiting angiotensin II type 2 receptors.
  • “Pharmaceutical composition” means a mixture containing a pharmaceutically acceptable salt or prodrug of a compound described herein with other chemical components, which, in addition to the active pharmaceutical ingredient, also contains other components such as pharmaceutically acceptable carriers and/or excipients Form.
  • the purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitate the absorption of the active ingredient and thereby exert biological activity.
  • the present invention provides the following technical solutions:
  • the metal hydroxide is selected from sodium hydroxide, potassium hydroxide, lithium hydroxide, calcium hydroxide, magnesium hydroxide, preferably sodium hydroxide;
  • the alkali metal carbonate is selected from sodium carbonate and potassium carbonate
  • the alkali metal bicarbonate is selected from sodium bicarbonate and potassium bicarbonate;
  • the alkali metal alkoxide is selected from sodium methoxide, sodium ethoxide, sodium tert-butoxide, potassium methoxide, potassium ethoxide and potassium tert-butoxide, preferably sodium methoxide and sodium ethoxide.
  • the examples provide the preparation and related structural identification data of the compound represented by formula (1m) and its pharmaceutically acceptable salts (eg compound 1). It must be noted that the following examples are used to illustrate the present invention rather than limit it.
  • the 1 H NMR spectrum was measured with a Bruker instrument (400 MHz), and the chemical shifts were expressed in ppm. A tetramethylsilane internal standard (0.00 ppm) was used.
  • Mass spectrum is obtained by LC/MS instrument, and the ionization mode can be ESI or APCI.
  • the thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate, the size of the silica gel plate used for thin layer chromatography (TLC) is 0.15mm ⁇ 0.2mm, and the size of the TLC separation and purification products is 0.4mm ⁇ 0.5mm.
  • the solution in the reaction refers to an aqueous solution.
  • the compound is purified by silica gel column chromatography eluent system and thin layer chromatography, wherein the eluent system is selected from: A: petroleum ether and ethyl acetate system; B: dichloromethane and methanol system; C: dichloromethane Methyl chloride and ethyl acetate system; the volume ratio of the solvent varies according to the polarity of the compound, and can also be adjusted by adding a small amount of acidic or basic reagents, such as acetic acid or triethylamine.
  • the eluent system is selected from: A: petroleum ether and ethyl acetate system; B: dichloromethane and methanol system; C: dichloromethane Methyl chloride and ethyl acetate system; the volume ratio of the solvent varies according to the polarity of the compound, and can also be adjusted by adding a small amount of acidic or basic reagents, such as
  • Test Example 1 Test of the antagonistic activity of the compounds of the present invention to human AT 2 R ligand binding
  • Angiotensin II Type 2 Receptor (AT 2 R) is involved in neuronal differentiation and regeneration, cell proliferation and angiogenesis, and maintenance of bone mass.
  • AT 2 R inhibitors can be used in the treatment of pain and abnormal neuroregenerative diseases, inhibit the proliferation of tumor cells and increase bone mass.
  • a method by AT 2 binding assays, the compounds of this invention antagonize study for the AT 2 R.
  • Test compounds (compound 1m and compounds 1-4) were dissolved in dimethyl sulfoxide and prepared as 10 mM stock solutions, respectively, according to standard procedures.
  • Tag-lite angiotensin receptor red agonist prepare 8600nM stock solution, and store it at -80°C after packaging;
  • test compound is diluted 5 times, with a total of 10 concentration gradients
  • step (3) Transfer the diluted compounds in step (2) to a working plate (3657, Corning) at 160nL/well, 200g, room temperature, 1 minute;
  • Tb-labeled-AT 1 R cells were used instead of Tb-labeled-AT 2 R to test the IC 50 of the antagonistic activity of the compounds of the present invention against AT 1 R .
  • Antagonistic activity of the test compounds of the present invention the AT 2 R IC 50 values in the table below.
  • the compounds of the present invention are highly selective antagonistic action of AT 2 R pair.
  • the LC/MS/MS method was used to determine the compounds of the present invention by intravenous injection or gavage in the rats, and the drug concentrations in the plasma at different times were measured to study the pharmacokinetics of the compounds of the present invention in rats. dynamic characteristics.
  • SD male rats Six healthy adult Sprague Dawley (SD) male rats were purchased from Weitong Lihua Laboratory Animal Technology Co., Ltd.
  • Compounds 2-4 were weighed, dissolved in an appropriate amount of .5% sodium carboxymethylcellulose (CMC-Na), vortexed for 1 minute until the compounds were completely suspended, and the final concentration was 6 mg/mL.
  • CMC-Na sodium carboxymethylcellulose
  • the sodium salt of the compound of the present invention (Compound 1) and its free acid (Compound 1m)
  • the plasma concentration and the area under the curve are significantly improved, and the plasma concentration is increased by more than 50 times, and the area under the curve is increased by 14%.
  • the sodium salts of the present invention have significant pharmacokinetic properties relative to other salts (such as lithium, potassium, calcium, magnesium salts), with both plasma concentrations and area under the curve Significant improvement, with unexpected technical effects.

Abstract

本发明涉及(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸的可药用盐、其制备方法、包含该可药用盐的药物组合物以及其作为治疗剂特别是作为血管紧张素Ⅱ2型受体拮抗剂的用途。

Description

四氢异喹啉类衍生物的盐、其制备方法及其医药应用
本申请要求于2020年7月1日提交到中国专利局的发明名称为“四氢异喹啉类衍生物的盐、其制备方法及其医药应用”的中国专利申请202010625224.7的优先权，其内容通过引用以整体并入本文。
技术领域
本发明涉及(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸的可药用盐形式、其制备方法、包含该可药用盐的药物组合物以及其作为治疗剂特别是作为血管紧张素Ⅱ2型受体拮抗剂的用途。
背景技术
神经性疼痛是由神经系统出现原发性损伤或功能障碍引起的慢性疼痛疾病，按病变的部位可分为外周神经性疼痛和中枢神经性疼痛。外伤、炎症、感染或压迫等原因均可引起神经性疼痛，例如糖尿病性神经痛(DNP)、带状疱疹后遗神经痛(PHN)、原发性神经病、继发性神经病、周围神经病、由机械性神经损伤或生化神经损伤引起的神经病疾病等。目前临床上用于治疗神经性疼痛的药物主要有抗癫痫药物、抗抑郁药物和麻醉性镇痛药，如加巴喷丁、普瑞巴林、三环类抗抑郁药等。但这些药物没有针对性，治疗效果非常有限，而且有严重的副作用，包括认知改变、镇静作用、恶心以及耐受性和依赖性，远未满足临床用药需求。因此，需要研究神经性疼痛的发病机制、找到药物作用明确的靶点，开发新型的能有效治疗神经性疼痛且不良反应少的药物。
血管紧张素Ⅱ受体是以血管紧张素Ⅱ作为配体的G蛋白偶联受体，它是肾素-血管紧张素系统的重要组成部分。血管紧张素Ⅱ受体主要亚型包括1型受体(AT 1R)和2型受体(AT 2R)。AT 1R和AT 2R仅有约30％的氨基酸序列相同，但血管紧张素Ⅱ作为其主要配体，与二者有相近的亲和力。
AT 2R在各种胚胎组织中大量表达，在成年正常组织中分布较少，但是在组织损伤后，其表达升高。AT 2R与血压调控、神经生长、疼痛控制和心肌再生相关，靶向AT 2R的药物可以改善心血管功能、缓解神经性疼痛等。由澳大利亚Spinifex公司开发的化合物olodanrigan(EMA401)是一种高选择性AT 2R拮抗剂，目前处于临床二期，该候选药物对糖尿病性神经痛、带状疱疹后遗神经痛等神经性疼痛具有良好的治疗效果，同时Spinifex公司也在开发AT 2R拮抗剂EMA-400。WO 93/23378公开了Olodanrigan和EMA-400的制备方法，其结构分别如下：
Figure PCTCN2021102261-appb-000001
目前已经公开了一系列的涉及AT 2R拮抗剂专利申请，其中包括WO2016113668、WO2015003223和WO2013110135等，AT 2R拮抗剂的研究和应用已取得一定的进展，但是仍远未能满足临床用药需求，提高的空间仍然巨大，仍有必要继续研究和开发新的AT 2R拮抗剂，尤其是新的选择性AT 2R拮抗剂。
(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸(化合物1m)是一种血管紧张素Ⅱ2型受体拮抗剂，其制备和生物活性公开于WO2019242599中。然而，该化合物的水溶解性不够好，药代动力学性质也不理想，这些大大限制了其应用。因此，如何改善其溶解性和药代动力学性质，仍然是现有技术中面临的一个问题。
发明内容
本发明人意外发现，该化合物的盐特别是其钠盐具有特别有利的水溶性和显著改善的药代动力学性质。
基于上述发现，本发明提供了一种(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸(化合物1m)的可药用盐、其制备方法、及包含该可药用盐的药物组合物以及其作为治疗剂特别是作为血管紧张素Ⅱ2型受体拮抗剂的用途。其中所述可药用盐为(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸与有机碱或无机碱形成的碱金属盐、钙盐或镁盐；尤其优选其钠盐。
该成盐形式具有优异的治疗原发性神经病、继发性神经病、周围神经病、由机械性神经损伤或生化神经损伤引起的神经病、带状疱疹后遗神经痛、糖尿病性神经痛或相关神经性疾病的活性，溶解度明显改善，在动物体内具有良好的药代动力学性质，毒性低，适用于制备治疗神经性疾病的制剂。
Figure PCTCN2021102261-appb-000002
本发明所述的碱金属盐选自钠盐、钾盐和锂盐；优选为钠盐。
本发明式(1m)化合物的典型的可药用盐包括，但不限于：
Figure PCTCN2021102261-appb-000003
本发明还涉及式(1m)化合物的可药用盐的制备方法，所述方法包括使(S)-2-(6-氟苯并[d] 噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸与金属氢氧化物、碱金属碳酸盐、碱金属碳酸氢盐或碱金属醇盐进行反应的步骤，其中所述金属氢氧化物选自氢氧化钠、氢氧化钾、氢氧化锂、氢氧化钙、氢氧化镁，优选为氢氧化钠；所述碱金属碳酸盐选自碳酸钠和碳酸钾；所述碱金属碳酸氢盐选自碳酸氢钠和碳酸氢钾；所述碱金属醇盐选自甲醇钠、乙醇钠、叔丁醇钠、甲醇钾、乙醇钾和叔丁醇钾，优选为甲醇钠或乙醇钠。
通常上述制备过程可以在冷却、常温或者加热条件下进行，值得注意的是反应温度的选择对不同的成盐反应有一定的影响，这也是本领域技术人员技术所熟知的，本发明成盐反应温度为常温至所用溶剂的沸点，本领域技术人员通过本领域常规的技术手段就能容易地确定具体成盐反应的最优选反应温度。
本发明还提供了一种药用组合物，其含有治疗有效剂量的式(1m)化合物的可药用盐，及药学上可以接受的载体；且本发明还提供了该药物组合物在制备治疗原发性神经病、继发性神经病、周围神经病、由机械性神经损伤或生化神经损伤引起的神经病、带状疱疹后遗神经痛、糖尿病性神经痛或相关神经性疾病的药物中的用途。
本发明还提供了式(1m)化合物的可药用盐或其药物组合物在在制备治疗原发性神经病、继发性神经病、周围神经病、由机械性神经损伤或生化神经损伤引起的神经病、带状疱疹后遗神经痛、糖尿病性神经痛或相关神经性疾病的药物中的用途。
本发明还提供了式(1m)化合物的可药用盐或其药物组合物在制备血管紧张素Ⅱ2型受体拮抗剂中的用途。
本发明还涉及一种治疗原发性神经病、继发性神经病、周围神经病、由机械性神经损伤或生化神经损伤引起的神经病、带状疱疹后遗神经痛、糖尿病性神经痛或相关神经性疾病的方法，所述方法包括给予需要治疗的患者有效治疗量的式(1m)化合物的可药用盐或其药物组合物。
本发明还涉及一种抑制血管紧张素Ⅱ2型受体的方法，所述方法包括使血管紧张素Ⅱ2型受体与式(1m)化合物的可药用盐或其药物组合物接触。
本发明还涉及式(1m)化合物的可药用盐或其药物组合物，其用作治疗原发性神经病、继发性神经病、周围神经病、由机械性神经损伤或生化神经损伤引起的神经病、带状疱疹后遗神经痛、糖尿病性神经痛或相关神经性疾病的药物。
本发明还涉及式(1m)化合物的可药用盐或其药物组合物，其用作抑制血管紧张素Ⅱ2型受体的药物。
发明的详细说明
除非有相反陈述，否则说明书和权利要求中的术语具有下述含义。
“药物组合物”表示含有本文所述化合物的可药用盐或前体药物与其他化学组分的混合物，除了活性药物成分外，其还包含其他组分例如可药用的载体和/或赋形剂。药物组合物的目的是促进对生物体的给药，利于活性成分的吸收并进而发挥生物活性。
本发明化合物的合成方法
为了实现本发明的目的，本发明提供了如下技术方案：
(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸碱加成盐可采用如下方法制备：
使(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸在与水混溶的有机溶剂中与金属氢氧化物、碱金属碳酸盐、碱金属碳酸氢盐或碱金属醇盐进行反应，其中：
所述金属氢氧化物选自氢氧化钠、氢氧化钾、氢氧化锂、氢氧化钙、氢氧化镁，优选为氢氧化钠；
所述碱金属碳酸盐选自碳酸钠和碳酸钾；
所述碱金属碳酸氢盐选自碳酸氢钠和碳酸氢钾；
所述碱金属醇盐选自甲醇钠、乙醇钠、叔丁醇钠、甲醇钾、乙醇钾和叔丁醇钾，优选为甲醇钠和乙醇钠。
具体实施方式
以下实施例用于进一步描述本发明，但这些实施例并非限制着本发明的范围和精神实质。
实施例
实施例给出了式(1m)所表示的化合物及其可药用盐(例如化合物1)的制备及相关结构鉴定数据。必须说明，下述实施例是用于说明本发明而不是对本发明的限制。 1H NMR图谱是用Bruker仪器(400MHz)测定而得，化学位移用ppm表示。使用四甲基硅烷内标准(0.00ppm)。 1H NMR的表示方法：s＝单峰，d＝双重峰，t＝三重峰，m＝多重峰，br＝变宽的，dd＝双重峰的双重峰，dt＝三重峰的双重峰。若提供偶合常数时，其单位为Hz。
质谱是用LC/MS仪测定得到，离子化方式可为ESI或APCI。
薄层层析硅胶板使用烟台黄海HSGF254或青岛GF254硅胶板，薄层色谱法(TLC)使用的硅 胶板采用的规格是0.15mm～0.2mm，薄层层析分离纯化产品采用的规格是0.4mm～0.5mm。
柱层析一般使用烟台黄海硅胶200～300目硅胶为载体。
在下列实施例中，除非另有指明，所有温度为摄氏温度，除非另有指明，各种起始原料和试剂来自市售或者是根据已知的方法合成，市售原料和试剂均不经进一步纯化直接使用，除非另有指明，市售厂家包括但不限于Aldrich Chemical Company，ABCR GmbH&Co.KG，Acros Organics，广赞化工科技有限公司和景颜化工科技有限公司等。
CDCl 3：氘代氯仿。
DMSO-d 6：氘代二甲基亚砜。
实施例中如无特殊说明，则反应中的溶液是指水溶液。
对化合物进行纯化，采用硅胶柱层析洗脱剂体系和薄层色谱法，其中洗脱剂体系选自：A：石油醚和乙酸乙酯体系；B：二氯甲烷和甲醇体系；C：二氯甲烷和乙酸乙酯体系；其中溶剂的体积比根据化合物的极性不同而不同，也可以加入少量的酸性或碱性试剂进行调节，如醋酸或三乙胺等。
实施例1
(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸钠
Figure PCTCN2021102261-appb-000004
Figure PCTCN2021102261-appb-000005
第一步
(Z)-3-(2-(苄氧基)-3-甲氧基苯基)-2-((叔丁氧基羰基)氨基)丙烯酸甲酯
将(±)-BOC-A-膦酰基甘氨酸三甲酯1b(9.8g,33mmol)和四甲基胍(4.0g,34.4mmol)溶于100mL四氢呋喃中，将反应液降至0℃，加入2-(苄氧基)-3-甲氧基苯甲醛1a(7.0g,28.7mmol)的四氢呋喃溶液(5mL)，室温下反应过夜。反应结束后，减压浓缩，加入乙酸乙酯(40mL)溶解残留物，依次以10％柠檬酸溶液(30mL)和饱和食盐水(30mL)洗涤，以无水硫酸钠干燥，过滤，减压浓缩，得到的残留物用硅胶柱层析法(洗脱剂：A体系)纯化，得到(Z)-3-(2-(苄氧基)-3-甲氧基苯基)-2-((叔丁氧基羰基)氨基)丙烯酸甲酯1c(9.5g,白色固体)，产率：80％
MS m/z(ESI)：314.0[M-100]
第二步
(S)-3-(2-(苄氧基)-3-甲氧基苯基)-2-((叔丁氧基羰基)氨基)丙酸甲酯
将(Z)-3-(2-(苄氧基)-3-甲氧基苯基)-2-((叔丁氧基羰基)氨基)丙烯酸甲酯1c(5.0g,12.0mmol)和(R)-N-二苯基膦-N-甲基-(S)-2-(二苯基膦)二茂铁基乙胺(90mg,0.06mmol)和二(1,5-环辛二烯)四氟硼酸铑(I)(100mg,0.024mmol)溶于50mL甲醇中，置换氢气三次，并插氢气球，室温下反应过夜。反应结束后，过滤，将滤液减压浓缩，得到的残留物用硅胶柱层析法(洗脱剂：A体系)纯化，得到(S)-3-(2-(苄氧基)-3-甲氧基苯基)-2-((叔丁氧基羰基)氨基)丙酸甲酯1d(3.2g,无色油状)，产率：64％。
MS m/z(ESI)：316.0[M-100]
1H NMR(400MHz,DMSO-d 6)δ7.48-7.32(m,5H),7.23(d,J＝8.0Hz,1H),6.98-6.96(m,2H),6.78(dd,J＝6.4,2.0Hz,1H),4.96(q,J＝10.4Hz,2H),4.20(td,J＝8.8,5.2Hz,1H),3.82(s,3H),3.55(s,3H),3.05(dd,J＝13.4,5.0Hz,1H),2.71(dd,J＝13.2,10.0Hz,1H),1.30(s,9H).
第三步
(S)-2-氨基-3-(2-(苄氧基)-3-甲氧基苯基)丙酸甲酯盐酸盐
将(S)-3-(2-(苄氧基)-3-甲氧基苯基)-2-((叔丁氧基羰基)氨基)丙酸甲酯1d(3.2g,7.7mmol)溶于10mL 1,4-二氧六环，加入氯化氢的1,4-二氧六环溶液(9.6mL,38.5mmol,4M)，室温下反应2小时。反应结束后，减压浓缩，得到(S)-2-氨基-3-(2-(苄氧基)-3-甲氧基苯基)丙酸甲酯盐酸盐1e(2.7g,白色固体)，产率：100％。
MS m/z(ESI)：316.0[M+1]
1H NMR(400MHz,DMSO-d 6)δ8.62(s,3H),7.47-7.32(m,5H),7.03-7.02(m,2H),6.81–6.78(m,1H),4.94(q,J＝11.2Hz,2H),4.04(t,J＝7.2Hz,1H),3.83(s,3H),3.50(s,2H),3.05(d,J＝7.2Hz,2H).
第四步
(S)-5-(苄氧基)-6-甲氧基-1,2,3,4-四氢异喹啉-3-甲酸甲酯盐酸盐
将(S)-2-氨基-3-(2-(苄氧基)-3-甲氧基苯基)丙酸甲酯盐酸盐1e(1.3g,3.7mmol)溶于2N稀盐酸(26mL)中，置换氩气三次，室温下搅拌30分钟，依次加入甲醛水溶液(2.8mL,37mmol,37wt.％)和四氢呋喃(5mL)，再置换氩气3次，室温下反应过夜。反应结束后，在反应液中加入乙腈，减压浓缩，重复多次，得到(S)-5-(苄氧基)-6-甲氧基-1,2,3,4-四氢异喹啉-3-甲酸甲酯盐酸盐1f(400mg,白色固体)，产率：30％。
MS m/z(ESI)：328.0[M+1]
1H NMR(400MHz,DMSO-d 6)δ10.12(s,2H),7.45–7.33(m,5H),7.05(d,J＝8.8Hz,1H),7.00(d,J＝8.4Hz,1H),4.96(d,J＝2.0Hz,2H),4.41(dd,J＝10.8,5.2Hz,1H),4.22(q,J＝15.6Hz,2H),3.82(s,3H),3.77(s,3H),3.21(dd,J＝17.2,5.2Hz,1H),2.92(dd,J＝17.6,11.2Hz,1H).
第五步
(S)-5-(苄氧基)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-1,2,3,4-四氢异喹啉-3-甲酸甲酯
将(S)-5-(苄氧基)-6-甲氧基-1,2,3,4-四氢异喹啉-3-甲酸甲酯盐酸盐1f(80mg,0.22mmol)、2-氯-6-氟苯并[d]噁唑1g(37mg,0.22mmol)和三乙胺(91μL，0.66mmol)溶于2mL四氢呋喃中，50～60℃下反应5小时。反应结束后，冷却至室温，减压浓缩，得到的残留物用薄层色谱法(展开剂：A体系)纯化，得到(S)-5-(苄氧基)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-1,2,3,4-四氢异喹啉-3-甲酸甲酯1h(60mg)，产率：59％。
MS m/z(ESI)：462.9[M+1]
1H NMR(400MHz,CDCl 3)δ7.49-7.29(m,6H),7.06(dd,J＝7.8,2.2Hz,1H),6.96-6.86(m,3H),5.19(dd,J＝6.4,2.4Hz,1H),5.05(d,J＝10.8Hz,1H),4.95(d,J＝11.2Hz,1H),4.90(d,J＝15.6Hz,1H),4.76(d,J＝15.2Hz,1H),3.89(s,3H),3.66-3.61(m,4H),2.94(dd,J＝16.4,6.4Hz,1H).
第六步
(S)-2-(6-氟苯并[d]噁唑-2-基)-5-羟基-6-甲氧基-1,2,3,4-四氢异喹啉-3-甲酸甲酯
将(S)-5-(苄氧基)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-1,2,3,4-四氢异喹啉-3-甲酸甲酯1h(600mg,1.3mmol)和10％钯碳(300mg,50％w)溶于10mL甲醇中，插氢气球，置换氢气4次，室温下反应过夜。反应结束后，用硅藻土过滤反应液，依次以乙酸乙酯和甲醇(V:V＝1:1)的混合溶剂(100mL×3)、二氯甲烷(100mL×3)洗涤硅藻土，将滤液减压浓缩，得到粗品(S)-2-(6-氟苯并[d]噁唑-2-基)-5-羟基-6-甲氧基-1,2,3,4-四氢异喹啉-3-甲酸甲酯1i(500mg)，产率：100％。
MS m/z(ESI)：372.9[M+1]
1H NMR(400MHz,DMSO-d 6)δ8.86(br,1H),7.50(dd,J＝8.8,2.4Hz,1H),7.35(dd,J＝8.6,5.4Hz,1H),7.10-7.04(m,1H),6.88(d,J＝8.4Hz,1H),6.74(d,J＝8.4Hz,1H),5.25(dd,J＝6.4,2.4Hz,1H),4.81(d,J＝15.2Hz,1H),4.63(d,J＝15.6Hz,1H),3.79(s,3H),3.59(s,3H),3.46(dd,J＝16.2,1.8Hz,1H),3.04(dd,J＝16.8,6.4Hz,1H).
第七步
(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸甲酯
将2-(氯甲基)-5-甲氧基吡啶盐酸盐1j(113mg,0.72mmol)、碳酸钾(149mg,1.08mmol)和(S)-2-(6-氟苯并[d]噁唑-2-基)-5-羟基-6-甲氧基-1,2,3,4-四氢异喹啉-3-甲酸甲酯1i(100mg,0.27mmol)依次溶于6mLN,N-二甲基甲酰胺中，70℃下反应6小时。反应结束后，冷却至室温，加入100mL乙酸乙酯和50mL水，分液，收集有机相，水相以乙酸乙酯(50mL×2)萃取，合并有机相，以饱和盐水(50mL×2)洗涤，以无水硫酸钠干燥，过滤，减压浓缩，得到粗品(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸甲酯1k(133mg)，产率：100％。粗品可进一步通过柱层析(洗脱剂：B体系)进一步分离纯化用于表征。
MS m/z(ESI)：493.9[M+1]
1H NMR(400MHz,CDCl 3)δ8.30(d,J＝2.4Hz,1H),7.59(d,J＝8.8Hz,1H),7.32-7.29(m,2H),7.07(dd,J＝7.8,2.2Hz,1H),6.96-6.86(m,3H),5.21(dd,J＝6.4,2.4Hz,1H),5.12(d,J＝12.0 Hz,1H),5.05(d,J＝11.6Hz,1H),4.91(d,J＝15.6Hz,1H),4.77(d,J＝15.6Hz,1H),3.89(s,3H),3.87(s,3H),3.67(dd,J＝16.4,2.4Hz,1H),3.63(s,3H),3.02(dd,J＝16.4,6.4Hz,1H).
第八步
(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸
将(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸甲酯1k(133mg,0.27mmol)溶于4mL四氢呋喃中，加入3mL氯化钙(481.74mg,4.34mmol)的异丙醇和水(V:V＝2:1)的混合溶液，再加入3mL氢氧化钠溶液(56mg,1.4mmol)，室温下反应过夜。反应结束后，加入80mL乙酸乙酯和100mL水，以1M稀盐酸调节反应液酸碱度至pH＝5～6，分液，收集有机相，水相以乙酸乙酯(50mL×2)萃取，合并有机相，有机相以饱和盐水(50mL×2)洗涤，以无水硫酸钠干燥，过滤，减压浓缩，得到的残留物用制备柱分离纯化，得到(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸1m(15mg)，产率：12％。
MS m/z(ESI)：479.9[M+1]
1H NMR(400MHz,DMSO-d 6)δ8.30(d,J＝2.8Hz,1H),7.58(d,J＝8.4Hz,1H),7.52-7.48(m,2H),7.34(dd,J＝8.6,5.0Hz,1H),7.09-7.01(m,3H),5.07(dd,J＝6.2,2.6Hz,1H),5.00(d,J＝11.2Hz,1H),4.93(d,J＝11.2Hz,1H),4.79(d,J＝15.6Hz,1H),4.66(d,J＝15.2Hz,1H),3.87(s,3H),3.83(s,3H),3.51(dd,J＝16.0,2.4Hz,1H),3.00(dd,J＝16.2,6.6Hz,1H).
第九步
(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸钠
将(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸1m(0.40g，0.84mmol)加入6mL乙酸乙酯中，冰水浴搅拌条件下加入NaOH/乙醇溶液(0.05g/mL，0.668mL)，使pH至7～8。将溶液浓缩至干，得到(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸钠1(0.42g)，产率100％。
MS m/z(ESI)：480.2[M+1]
1H NMR(400MHz,DMSO-d 6)δ8.26(s,1H),7.62(d,J＝8.5Hz,1H),7.46(d,J＝8.6Hz,1H),7.38(d,J＝8.4Hz,1H),7.24(dd,J＝8.2,4.9Hz,1H),7.01-6.90(m,3H),4.93(s,2H),4.71(s,2H),4.66(d,J＝5.5Hz,1H),3.84(s,3H),3.79(s,3H),3.67(d,J＝16.1Hz,1H),2.78(dd,J＝ 16.1,6.4Hz,1H).
实施例2
(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸锂
Figure PCTCN2021102261-appb-000006
将(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸1m(0.20g，0.42mmol)加入至6mL乙酸乙酯中，冰水浴搅拌条件下加入LiOH/乙醇溶液(0.2M，2.1mL)，使pH至7～8。将溶液浓缩至干，得到(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸锂2(0.205g)，产率100％。
MS m/z(ESI)：480.2[M+1]
实施例3
(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸钾
Figure PCTCN2021102261-appb-000007
将(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸1m(0.20g，0.42mmol)加入至6mL乙酸乙酯中，冰水浴搅拌条件下加入KOH/乙醇溶液(0.2M，2.1mL)，使pH至7～8。将溶液浓缩至干，得到(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸钾3(0.215g)，产率100％。
MS m/z(ESI)：480.2[M+1]
实施例4
(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸钙
Figure PCTCN2021102261-appb-000008
将(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸1m(0.20g，0.42mmol)加入至20mL四氢呋喃中，冰水浴搅拌条件下加入氢氧化钙/水溶液(0.01M，21mL)，使pH至7～8。将溶液浓缩并冷冻干燥，得到(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸钙4(0.208g)，产率100％。
MS m/z(ESI)：480.2[M+1]
测试例：
溶解度实验
按照常规溶解度测定方法，测试本发明化合物在以下四种不同系统中的溶解度：磷酸盐缓冲液PBS(pH7.4)，甲醇，0.1％HCl及水，结果如下表所示：
Figure PCTCN2021102261-appb-000009
结论：本发明化合物的钠盐(化合物1)与其游离酸(化合物1m)相比，溶解度明显改善。
由上表结果可见，与化合物1m相比，钠盐化合物1在水中溶解度提高了200倍以上，这一结果是出乎意料的。
生物学评价
测试例1本发明化合物对人源AT 2R配体结合拮抗活性的测试
Angiotensin II Type 2 Receptor(AT 2R)参与神经元分化与再生、细胞增殖与血管发生以及骨量的维持。AT 2R抑制剂可以用于疼痛和异常神经再生性疾病的治疗，抑制肿瘤细胞的增殖以及增加骨量。以下方法通过AT 2配体结合试验，研究本发明化合物对于AT 2R的拮抗程度。
1、试剂与耗材
Figure PCTCN2021102261-appb-000010
Figure PCTCN2021102261-appb-000011
2、试剂配制
(1)10mM人源血管紧张素II(Angiotensin II human)：将10mg Angiotensin II human(纯度99.09％)溶解于0.947mL去离子水，分装后于-80℃保存；
(2)化合物储液的制备
根据标准的方法，将测试化合物(化合物1m和化合物1～4)溶于二甲基亚砜，分别制备成10mM的储液。
(3)Tag-lite angiotensin receptor red agonist:配制8600nM储液，分装后于-80℃保存；
(4)1X Tag-Lite Buffer(TLB):将5X TLB用去离子水稀释至1X。
3、实验步骤
(1)配制适量的1X TLB，混匀待用；
(2)测试化合物进行5倍稀释，共10个浓度梯度；
(3)将步骤(2)中稀释好的化合物各按160nL/孔转移至工作板中(3657,Corning)，200g，室温，1分钟；
(4)将40μl 1X TLB加至上述工作板中，室温条件下200g离心1分钟，于振荡器振荡15分钟混匀后，室温条件下200g离心1分钟待用(化合物的工作浓度为4X)；
(5)用1X TLB将Tag-lite angiotensin receptor red agonist(8600nM储液)稀释至12nM待用；
(6)取5mL 1X TLB于15mL离心管中；
(7)于37℃水浴中将1支Tb-labeled-AT 2R细胞冻融，直至冰全部融化(1～2分钟)；
(8)迅速将冻融的细胞转移至步骤(6)中的1X TLB中，轻柔混匀后，于室温下1200g离心5分钟；
(9)轻柔的将上清液吸出，用1mL 1X TLB将细胞重悬混匀后，再加入1.7mL 1X TLB混匀后至于室温待用；
(10)加入10μL细胞至所有试验孔中，室温下200g离心3秒；加入5μL步骤(4)中的化合物工作液4X至相应的孔中；加入5μL步骤(5)中稀释的4X Tag-lite angiotensin receptor red agonist至所有试验孔中。
(11)将反应板于室温下200g离心1分钟，室温25℃静置1小时后，室温下200g离心1分钟，利用Envision HTRF酶标仪收集数据，利用非线性拟合公式计算IC 50。
(12)同理，采用基本相同的方法，不同的是使用Tb-labeled-AT 1R细胞代替Tb-labeled-AT 2R，测试本发明化合物对于AT 1R的拮抗活性的IC 50。
4、实验结果
本发明化合物对AT 2R拮抗活性测试的IC 50值见下表。
化合物编号 IC 50(nM)/AT 2R IC 50(μM)/AT 1R
化合物1m 10 >10
化合物1 6 >10
结论：(1)本发明化合物的钠盐(化合物1)和其游离酸(化合物1m)相比，对于AT 2R具有显著拮抗活性；
(2)本发明化合物的钠盐(化合物1)和其游离酸(化合物1m)，对于AT 1R拮抗的IC 50值>10μM，对于AT 1R无拮抗活性；
因此，本发明化合物对AT 2R的拮抗作用具有高度选择性。
药代动力学测试
1、实验目的
以SD大鼠为受试动物，采用LC/MS/MS法测定大鼠静脉注射或灌胃给予本发明化合物，测定其不同时刻血浆中的药物浓度，研究本发明化合物在大鼠体内的药代动力学特征。
2、实验方案
2.1、实验药品与动物
化合物1m及化合物1～4
健康成年Sprague Dawley(SD)雄性大鼠6只，购自维通利华实验动物技术有限公司。
2.2药物配制与给药
(1)药物配置：
称取76.32mg化合物1m，溶于12.72mL 0.5％羧甲基纤维素钠(CMC-Na)中，涡旋1分钟至化合物完全悬浮，最终配置浓度为6mg/mL；
称取76.61mg化合物1，溶于12.768mL 0.5％羧甲基纤维素钠(CMC-Na)中，涡旋1分钟至化合物完全悬浮，最终配置浓度为6mg/mL。
称取化合物2～4，溶于适量的.5％羧甲基纤维素钠(CMC-Na)中，涡旋1分钟至化合物完全悬浮，最终配置浓度为6mg/mL。
(2)给药
健康成年SD大鼠6只，雌雄各半，平均分成2组；每组3只。禁食过夜后灌胃给药(po)给予本发明化合物1m或化合物1～4，给药剂量均为60mg/kg。
2.3样品采集
于给药前和给药后0.25小时、0.5小时、1小时、2小时、4小时、6小时、8小时和24小时经颈静脉采约0.25mL血液，肝素钠抗凝。血液样本采集后置于冰上，离心分离血浆(离心条件：7000转/分钟，5分钟)。收集的血浆分析前存放于-70℃以下。
2.4样品前处理
取100μL血浆样品，向其中加入300μL甲醇(包含内标工作液，氯雷他定800ng/mL)，涡旋5分钟，10000转/分钟离心10分钟，取1μL混合液至LC-MS/MS进样分析。
3、药代动力学参数结果
本发明的盐化合物和阳性对照化合物(化合物1m)的药代动力学参数如下表所示。
Figure PCTCN2021102261-appb-000012
Figure PCTCN2021102261-appb-000013
结论：本发明化合物的钠盐(化合物1)与其游离酸(化合物1m)相比，血药浓度和曲线下面积均有明显改善，其中血药浓度提高了50倍以上，曲线下面积提高了14倍以上；令人惊讶的是，本发明的钠盐相对于其他盐(例如锂盐、钾盐、钙盐、镁盐)具有显著的药代动力学性质，血药浓度和曲线下面积均有明显提升，具有意料不到的技术效果。

Claims (10)

  1. 一种(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸的可药用盐，其为(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸与有机碱或无机碱形成的碱金属盐、钙盐或镁盐。
  2. 根据权利要求1所述的可药用盐，其中所述碱金属盐选自钠盐、锂盐和钾盐，优选为钠盐。
  3. 根据权利要求1-2任一项所述的可药用盐，其选自：
    Figure PCTCN2021102261-appb-100001
  4. 一种制备如权利要求1-3任一项所述的可药用盐的方法，该方法包括使(S)-2-(6-氟苯并[d]噁唑-2-基)-6-甲氧基-5-((5-甲氧基吡啶-2-基)甲氧基)-1,2,3,4-四氢异喹啉-3-甲酸与金属氢氧化物、碱金属碳酸盐、碱金属碳酸氢盐或碱金属醇盐进行反应的步骤。
  5. 根据权利要求4所述的方法，其中
    所述金属氢氧化物选自氢氧化钠、氢氧化钾、氢氧化锂、氢氧化钙和氢氧化镁，优选为氢氧化钠；所述碱金属碳酸盐选自碳酸钠和碳酸钾；所述碱金属碳酸氢盐选自碳酸氢钠和碳酸氢钾；所述碱金属醇盐选自甲醇钠、乙醇钠、叔丁醇钠、甲醇钾、乙醇钾和叔丁醇钾，优选为甲醇钠或乙醇钠。
  6. 一种药物组合物，其含有治疗有效剂量的根据权利要求1-3任一项所述的可药用盐以及药学上可以接受的载体。
  7. 根据权利要求1-3任一项所述的可药用盐或根据权利要求6所述的药物组合物在制备用于治疗或预防原发性神经病、继发性神经病、周围神经病、由机械性神经损伤或生化神经损伤引起的神经病、带状疱疹后遗神经痛、糖尿病性神经痛或相关神经性疾病的药物中的用途。
  8. 根据权利要求1-3任一项所述的可药用盐或根据权利要求6所述的药物组合物在制备血管紧张素Ⅱ2型受体拮抗剂中的用途。
  9. 根据权利要求1-3任一项所述的可药用盐或根据权利要求6所述的药物组合物，其用作治疗原发性神经病、继发性神经病、周围神经病、由机械性神经损伤或生化神经损伤引起的神经病、带状疱疹后遗神经痛、糖尿病性神经痛或相关神经性疾病的药物。
  10. 根据权利要求1-3任一项所述的可药用盐或根据权利要求6所述的药物组合物，其用作抑制血管紧张素Ⅱ2型受体的药物。
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WO1993023378A1 (en) 1992-05-19 1993-11-25 Warner-Lambert Company Substituted 1,2,3,4-tetrahydroisoquinolines with angiotensin ii receptor antagonist properties
WO2013110135A1 (en) 2012-01-25 2013-08-01 Spinifex Pharmaceuticals Pty Ltd Heterocyclic compounds and methods for their use
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WO2016113668A1 (en) 2015-01-13 2016-07-21 Novartis Ag Pyrrolidine derivatives as angiotensin ii type 2 antagonists
WO2019242599A1 (zh) 2018-06-19 2019-12-26 浙江海正药业股份有限公司 四氢异喹啉类衍生物、其制备方法及其用途

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WO1993023378A1 (en) 1992-05-19 1993-11-25 Warner-Lambert Company Substituted 1,2,3,4-tetrahydroisoquinolines with angiotensin ii receptor antagonist properties
WO2013110135A1 (en) 2012-01-25 2013-08-01 Spinifex Pharmaceuticals Pty Ltd Heterocyclic compounds and methods for their use
WO2015003223A1 (en) 2013-07-08 2015-01-15 Spinifex Pharmaceuticals Pty Ltd Heterocyclic compounds and methods of their use
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WO2019242599A1 (zh) 2018-06-19 2019-12-26 浙江海正药业股份有限公司 四氢异喹啉类衍生物、其制备方法及其用途

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