[go: up one dir, main page]

WO2018130207A1 - 4,4-二苯基哌啶类化合物或其可药用盐、药物组合物及用途 - Google Patents

4,4-二苯基哌啶类化合物或其可药用盐、药物组合物及用途 Download PDF

Info

Publication number
WO2018130207A1
WO2018130207A1 PCT/CN2018/072426 CN2018072426W WO2018130207A1 WO 2018130207 A1 WO2018130207 A1 WO 2018130207A1 CN 2018072426 W CN2018072426 W CN 2018072426W WO 2018130207 A1 WO2018130207 A1 WO 2018130207A1
Authority
WO
WIPO (PCT)
Prior art keywords
group
pharmaceutically acceptable
acceptable salt
pain
compound
Prior art date
Application number
PCT/CN2018/072426
Other languages
English (en)
French (fr)
Inventor
张城
王好山
董国兴
吴宁
张树卓
庄笑梅
庞冲
王娟
杨日芳
郑建全
李锦�
恽榴红
Original Assignee
中国人民解放军军事科学院军事医学研究院
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 中国人民解放军军事科学院军事医学研究院 filed Critical 中国人民解放军军事科学院军事医学研究院
Priority to KR1020197023802A priority Critical patent/KR102507380B1/ko
Priority to JP2019559149A priority patent/JP7068338B2/ja
Priority to US16/477,655 priority patent/US10889575B2/en
Publication of WO2018130207A1 publication Critical patent/WO2018130207A1/zh

Links

Images

Classifications

    • 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
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/06—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
    • C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
    • C07D409/06—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
    • 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
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/451—Non condensed piperidines, e.g. piperocaine having a carbocyclic group directly attached to the heterocyclic ring, e.g. glutethimide, meperidine, loperamide, phencyclidine, piminodine
    • 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
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
    • A61K31/4535—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a heterocyclic ring having sulfur as a ring hetero atom, e.g. pizotifen
    • 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
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
    • A61K31/454—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. pimozide, domperidone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/12—Drugs for disorders of the urinary system of the kidneys
    • 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
    • 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
    • A61P25/06—Antimigraine agents
    • 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
    • A61P25/30—Drugs for disorders of the nervous system for treating abuse or dependence
    • A61P25/32—Alcohol-abuse
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D211/00—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings
    • C07D211/04—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D211/06—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
    • C07D211/08—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms
    • C07D211/10—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms with radicals containing only carbon and hydrogen atoms attached to ring carbon atoms
    • C07D211/14—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms with radicals containing only carbon and hydrogen atoms attached to ring carbon atoms with hydrocarbon or substituted hydrocarbon radicals attached to the ring nitrogen atom
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D221/00—Heterocyclic compounds containing six-membered rings having one nitrogen atom as the only ring hetero atom, not provided for by groups C07D211/00 - C07D219/00
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D241/00—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings
    • C07D241/02—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings
    • C07D241/04—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings having no double bonds between ring members or between ring members and non-ring members
    • 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
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/10—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a carbon chain containing aromatic rings

Definitions

  • the invention belongs to the field of medicinal chemical industry and relates to a 4,4-diphenylpiperidine compound or a pharmaceutically acceptable salt thereof, a pharmaceutical composition comprising the compound or a pharmaceutically acceptable salt thereof and use thereof.
  • N-type calcium channel belongs to a subtype of voltage-gated calcium channel (VDCC). It is composed of ⁇ 1 ⁇ subunit and has high voltage activation and rapid inactivation. It is mainly distributed in nerve tissue and can be used by w-conotoxin GVIA. (wCgTx) blockade has been identified as a clinically relevant novel drug target.
  • N-type calcium channel blockers have good treatment for stroke and cerebral ischemia, analgesia, especially neuropathic pain, reduction of alcohol cravings and treatment of alcoholism [1], and kidney protection [2, 3, 4] . Application prospects.
  • N-type calcium channel is an important part of pain production and pain conduction. Because the blocker for N-type calcium channel directly acts on N-type calcium channel, it does not involve second messenger or G protein. Therefore, it is not easy to produce addiction.
  • the highly selective N-type calcium channel blocker ⁇ -conotoxin MVIIA It was approved by the US FDA in December 2004 and its clinical practice shows that N-type calcium channel blockers have been proven to be a new target for the treatment of pain and have good application prospects.
  • the blockade of N-type calcium channels reduces the release of norepinephrine and renin from the sympathetic nerve terminals, reduces the resistance to the arteries and the arteries, and reduces the pressure of the glomerulus [2] .
  • ACE angiotensin-converting enzyme
  • Ang II angiotensin II
  • PLC protein kinase C
  • OS oxidative stress
  • DN diabetic nephropathy
  • the N-type calcium channel is located at the sympathetic nerve endings, and the sympathetic nerves excite the N-type calcium channel calcium influx.
  • the release of norepinephrine causes the renal arteries and the small arteries to contract, and stimulates the release of renin from the paracellular cells.
  • renal hemodynamics especially glomerular hyperperfusion pressure, plays a key role in renal function damage.
  • Tomoyuki [3] reported that in the rat model of hypertensive nephropathy, L/N calcium channel blocker cilnidipine significantly reduced plasma renin and norepinephrine levels, as well as goals and goals.
  • Arterial pressure reduces microalbuminuria. It may reduce the amount of urinary albumin to protect the kidney by inhibiting the N-type calcium channel.
  • IR insulin resistance
  • hyperinsulinemia can excite the hypothalamic sympathetic nerve center, and also affect renal hemodynamics, directly acting on the glomerular arteries, dilating into the small arterioles, aggravating glomerular hyperfiltration, Hyperperfusion state promotes glomerular sclerosis by causing indirect mechanisms such as hypertension, hyperlipidemia, and hyperuricemia.
  • Hyperinsulinemia aggravates the occurrence of glomerular hypertrophy by stimulating cytokines such as insulin-like growth factors.
  • cytokines such as insulin-like growth factors.
  • the early pathological manifestations of type 2 diabetic nephropathy are a slight widening of the mesangial matrix and a slight thickening of the glomerular basement membrane (GBM).
  • GBM glomerular basement membrane
  • N-type calcium channel blockers [5] .
  • the 4-piperidinyl aniline small molecule compound disclosed in WO99/43658 exhibits significant analgesic activity as an orally selectable N-type calcium channel blocker; Teodori et al [6] , Knutsen et al [7] ] , Yamamoto et al [8,9] , Tyagarajan et al [10] have also made some progress; small molecule N-type calcium channel blocker NMED-160 has entered phase II clinical studies, and related compounds have also been studied. Better results [11,12] .
  • N-type calcium channel blockers currently available have defects: on the one hand, these compounds have insufficient biological activity and channel selectivity; on the other hand, the pharmacokinetic properties of these compounds are poor. Sufficient pharmacological activity can be produced by administration in a clinically non-simple and easy route such as side ventricle administration. Therefore, there is still a need to develop a novel N-type calcium channel blocker.
  • the inventors have conducted research and creative work to obtain a class of 4,4-diphenylpiperidine compounds or pharmaceutically acceptable salts thereof.
  • the present inventors have found that such a compound or a pharmaceutically acceptable salt thereof exhibits remarkable N-type calcium channel blocking activity and has good pharmacokinetic properties, and can effectively prevent or alleviate pain, and has new prevention.
  • And/or treatment of pain especially neuropathic pain
  • stroke cerebral ischemia
  • alcohol addiction alcohol addiction
  • alcoholism kidney disease
  • addictive conditions caused by analgesic drugs or tolerant conditions caused by analgesic drugs The potential of a drug or pharmaceutically active ingredient.
  • a first aspect of the invention relates to a compound of formula I or a pharmaceutically acceptable salt thereof:
  • g is selected from the group consisting of C 1-8 alkylene, substituted C 1-8 alkylene, carbonyl, and C 1-8 alkylene;
  • R is selected from a C 5-20 aryl group, a substituted C 5-20 aryl group, a C 4-20 heterocyclic group, and a substituted C 4-20 heterocyclic group;
  • substitution is independently substituted by one or more substituents selected from the group consisting of halogen, C 1-8 alkyl, C 1-8 alkoxy, C 3-8 cycloalkyl, cyano, Nitro, fluorenyl, methylthio, ethylthio, trifluoromethyl, amino, amido, mono C 1-8 alkylamino, di C 1-8 alkylamino, C 1-8 alkylsulfonyl, Hydroxy, aryloxy, heteroaryloxy and heterocyclyloxy.
  • substituents selected from the group consisting of halogen, C 1-8 alkyl, C 1-8 alkoxy, C 3-8 cycloalkyl, cyano, Nitro, fluorenyl, methylthio, ethylthio, trifluoromethyl, amino, amido, mono C 1-8 alkylamino, di C 1-8 alkylamino, C 1-8 alkylsulfonyl, Hydroxy, aryloxy, heteroary
  • pharmaceutically acceptable salts of the compounds of formula I include inorganic or organic acid salts thereof including, but not limited to, hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, hydrogen sulfate.
  • Salt phosphate, hydrogen phosphate, acetate, glycolate, propionate, butyrate, oxalate, adipate, alginate, lactate, citrate, tartrate, Succinate, maleate, fumarate, picrate, aspartate, gluconate, benzoate, methanesulfonate, ethanesulfonate, besylate, Tosylate, pamoate, pyruvate, glycolate, malonate, trifluoroacetate, malate, salicylate, p-aminosalicylate, pamoate and Ascorbate, etc.; for example, the hydrochloride salt of a compound of formula I.
  • the compound of any of the first aspects of the invention may also be a prodrug or a form that releases the active ingredient after metabolic changes in the body.
  • the selection and preparation of suitable prodrug derivatives are well known to those skilled in the art.
  • g is selected from the group consisting of C 1-8 alkylene, carbonyl, and C 1-8 alkylene; R is selected from C 5-20 aryl, substituted C 5-20 aryl, and C 4-20 heterocyclic group.
  • the substituent is selected from the group consisting of C 1-8 alkyl, C 1-8 alkoxy, C 3-8 cycloalkyl, decyl, methylthio, ethylthio, Amino group, mono C 1-8 alkylamino group, di C 1-8 alkylamino group and hydroxyl group.
  • the halogen is fluorine, chlorine, bromine or iodine.
  • g is selected from the group consisting of C 1-8 alkylene, carbonyl and C 1-8 alkylene; R is selected from C 6-12 aryl, substituted C 6-12 aryl and C 4-12 aromatic heterocyclic group.
  • the substituent is selected from the group consisting of C 1-8 alkoxy, methylthio, ethylthio, amino, mono C 1-8 alkylamino, and di C 1-8 alkane. Amino group.
  • the C 4-12 aromatic heterocyclic group is a C 4-5 aromatic heterocyclic group or a C 6-12 benzoheterocyclyl group.
  • g is selected from the group consisting of methylene, carbonyl, benzoyl, acetylidene, 1,3-propionyl, 1,2-propionyl, 1,4-butyryl And 1, 3-butyryl, 1,2-butyryl;
  • R is selected from the group consisting of phenyl, substituted phenyl, thienyl and benzoxazolinone (for example, 2-benzoxazolinone).
  • the substituent is selected from the group consisting of methoxy and dimethylamino.
  • the compound or a pharmaceutically acceptable salt thereof is selected from the group consisting of the following compounds (1) to (6) and pharmaceutically acceptable salts thereof:
  • a second aspect of the invention relates to a process for the preparation of a compound according to any one of the first aspects of the invention, or a pharmaceutically acceptable salt thereof.
  • the compounds of formula I of the present invention can be prepared using synthetic routes as shown below, and in addition, reference can be made to the detailed description in the examples of the invention. It can also be converted to its pharmaceutically acceptable salt by reaction with an acid, as needed.
  • N-benzyl-4-piperidone is used as a raw material
  • benzene is used as a raw material and a reaction solvent to react at room temperature in the presence of trifluoromethanesulfonic acid to obtain 4,4-diphenyl-1-benzylpiperidine.
  • 4,4-diphenyl-1-benzylpiperidine (formula II) is reacted with ethyl chloroformate and then hydrolyzed to give 4,4-diphenylpiperidine (formula IV); , 4-diphenylpiperidine (formula IV) in a dry acetone, with a commercial halogenated or aldehyde under alkaline conditions such as triethylamine, stirred at room temperature for 2 to 24 hours for halogenation or amine reduction The reaction is carried out to give the target compound of formula I.
  • the method for producing a compound according to any one of the first aspects of the present invention, or a pharmaceutically acceptable salt thereof comprises the steps of:
  • R or g in the above formula I is as defined in the compound of any one of the first aspects of the invention or a pharmaceutically acceptable salt thereof.
  • a third aspect of the invention relates to a pharmaceutical composition
  • a pharmaceutical composition comprising a compound according to any one of the first aspects of the invention, or a pharmaceutically acceptable salt thereof; optionally, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier and/or Or accessories.
  • the compounds of the invention may be administered in their own form or in the form of a pharmaceutical composition.
  • the compound may be admixed with one or more pharmaceutically acceptable carriers, excipients or diluents.
  • the pharmaceutical compositions of the present invention are usually formulated in a conventional manner using one or more physiologically acceptable carriers and/or excipients which facilitate the processing of the active compounds into preparations which may be employed in a pharmaceutical form, suitable formulations Depending on the route of administration chosen, it can be prepared according to common knowledge well known in the art.
  • Pharmaceutically acceptable carriers or excipients that can be used in the pharmaceutical compositions of the present invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum proteins, buffer substances such as phosphates, glycerol, Sorbic acid, potassium sorbate, a partial glyceride mixture of saturated vegetable fatty acids, water, salt or electrolyte, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salt, colloidal silica, Magnesium trisilicate, polyvinylpyrrolidone, cellulosic material, polyethylene glycol, sodium carboxymethylcellulose, polyacrylate, beeswax, polyethylene-polyoxypropylene block polymer, lanolin.
  • ion exchangers alumina, aluminum stearate, lecithin
  • serum proteins such as human serum proteins
  • buffer substances such as phosphates
  • the pharmaceutical composition of the compound of formula I of the present invention, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof may be administered enterally or parentally, such as orally, muscle, subcutaneously, nasally, orally. Mucosa, skin, peritoneum or rectum.
  • Formulations such as tablets, capsules, pills, aerosols, pills, powders, solutions, suspensions, emulsions, granules, liposomes, transdermal agents, buccal tablets, suppositories, lyophilized powders Wait. It can also be prepared as a sustained release preparation, a controlled release preparation, and various microparticle delivery systems.
  • a fourth aspect of the invention relates to a method of blocking or inhibiting a N-type calcium ion channel in vivo or in vitro, comprising administering to a subject in need thereof an effective amount of a compound according to any one of the first aspects of the invention or A pharmaceutically acceptable salt or a step of administering to a subject in need thereof an effective amount of the pharmaceutical composition of the third aspect of the invention.
  • the fifth aspect of the invention relates to the compound of any one of the first aspect of the invention, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of the third aspect of the invention, for the preparation of an N-type calcium channel blocker or inhibitor use.
  • a sixth aspect of the present invention relates to a compound according to any one of the first aspects of the present invention, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to the third aspect of the present invention, which is useful for the preparation or prevention of pain, stroke, cerebral ischemia, alcohol formation Use in addiction, alcoholism, kidney disease, an addictive disorder caused by an analgesic drug, or a drug caused by an analgesic drug; for example, the pain is postoperative pain, migraine, visceral pain or Neuropathic pain; for example, the renal disease is acute renal failure, chronic renal failure, or renal insufficiency.
  • a seventh aspect of the invention relates to the prevention or treatment of pain, stroke, cerebral ischemia, alcohol addiction, alcoholism, kidney disease, an addictive disorder caused by an analgesic drug or a tolerable disorder caused by an analgesic drug
  • a method comprising the step of administering to a subject in need thereof an effective amount of a compound according to any one of the first aspects of the invention, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to the third aspect of the invention; for example, the pain It is post-operative pain, migraine, visceral pain or neuropathic pain; for example, the kidney disease is acute renal failure, chronic renal failure or renal insufficiency.
  • the compound of the present invention or a pharmaceutically acceptable salt thereof can be administered alone or in combination with other compounds of the present invention or a pharmaceutically acceptable salt thereof, and/or in combination with other known therapeutic agents.
  • the specific dosage and method of use of the compound of the present invention or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of the present invention for different patients depends on various factors including the age, weight, sex, natural health status, nutritional status, and compound of the patient.
  • the intensity of the activity, the time of administration, the rate of metabolism, the severity of the condition, and the subjective judgment of the treating physician depends on various factors including the age, weight, sex, natural health status, nutritional status, and compound of the patient.
  • the intensity of the activity, the time of administration, the rate of metabolism, the severity of the condition, and the subjective judgment of the treating physician depends on various factors including the age, weight, sex, natural health status, nutritional status, and compound of the patient.
  • the intensity of the activity, the time of administration, the rate of metabolism, the severity of the condition, and the subjective judgment of the treating physician depends on various factors including the age, weight, sex, natural health status, nutritional status, and compound of the patient.
  • the unit dosage form will generally contain from 0.1% to 99% by weight of active substance, more typically from 5% to 75% by weight of active substance.
  • unit dosage forms can contain from 1 mg to 1 g of the compound, from 10 mg to 500 mg of the compound, from 50 mg to 400 mg of the compound, or from 100 mg to 200 mg of the compound.
  • the eighth aspect of the invention relates to the compound of any one of the first aspect of the invention, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to the third aspect of the invention for blocking or inhibiting the N-type calcium ion channel.
  • the ninth aspect of the invention relates to a compound of any one of the first aspect of the invention, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to the third aspect of the invention for use in the prevention or treatment of pain, stroke, or cerebral ischemia , alcohol addiction, alcoholism, kidney disease, an addictive disorder caused by an analgesic drug or a tolerable disorder caused by an analgesic drug; for example, the pain is postoperative pain, migraine, visceral pain or neuropathy Pain; for example, the kidney disease is acute renal failure, chronic renal failure, or renal insufficiency.
  • halogen refers to a Group VIIA element, including fluorine (F), chlorine (Cl), bromine (Br), iodine (I), and ruthenium (At).
  • C 1-8 alkyl refers to a straight or branched alkyl group having from 1 to 8 carbon atoms.
  • a linear or branched alkyl group having 1 to 6 carbon atoms for example, a linear or branched alkyl group having 1 to 4 carbon atoms, such as a methyl group, an ethyl group, a propyl group, an isopropyl group, Butyl, isobutyl, sec-butyl, tert-butyl, pentyl, 2-ethyl-butyl, hexyl, heptyl and octyl.
  • C 1-8 alkylene refers to a straight or branched alkyl group having from 1 to 8 carbon atoms in the form of two hydrogen atoms, for example, C 1-6 alkylene, C 1-4 Alkylene group, methylene group, 1,2-ethylene group, ethylene group, isopropylidene group, 1,3-propylene group and the like.
  • carbonyl refers to a divalent group in which two atoms of carbon and oxygen are linked by a double bond.
  • C 1-8 alkylene refers to a group which is left after the C 1-8 alkyl fatty acid has been removed from the hydroxyl group and which also eliminates one hydrogen atom in form.
  • C 1-8 alkoxy means "C 1-8 alkyl-O-" wherein C 1-8 alkyl is as defined above.
  • C 3-8 cycloalkyl means a cyclic alkyl group having 3 to 8 carbon atoms.
  • Examples of the C 3-6 cycloalkyl group, the C 3-5 cycloalkyl group, and the C 3-8 cycloalkyl group include a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, a cycloheptyl group, a cyclooctyl group, and the like. .
  • C 5-20 aryl refers to an aromatic ring (including fused rings) containing from 5 to 20 carbon atoms which formally eliminates one hydrogen atom, such as a C 6-20 aryl group, a C 6-18 aryl group, C 6-12 aryl.
  • the aryl group specifically include a methoxy group, a phenyl group, a naphthyl group, an anthracenyl group, an anthracenyl group and the like.
  • aryloxy means "aryl-O-" wherein aryl is as defined above for the C5-20 aryl.
  • C 4-20 heterocyclic group means a heterocyclic group containing 4 to 20 atoms (1-3 atoms selected from the group consisting of hetero atoms such as oxygen, nitrogen and sulfur), and is classified into an aliphatic heterocyclic group and an aromatic heterocyclic group.
  • Two types. Examples of the heterocyclic group include a benzoheterocyclyl group, a tetrahydrofuranyl group, a pyrrolidinyl group, a piperidinyl group, a morpholinyl group, a thiazolidinyl group, a thiazolinethione group, a thiothiazolyl group, a benzothiazolyl group, a thiophene group.
  • Base thiadiazolyl, benzoxazolin-2-one, and the like.
  • heterocyclyloxy means “heterocyclyl-O-" wherein heterocyclyl is as defined above for the C 4-20 heterocyclyl.
  • heteroaryl refers to an aromatic heterocyclic ring wherein one to three atoms are selected from the group consisting of heteroatoms oxygen, nitrogen and sulfur, and the other atoms are carbon atoms.
  • heteroaryl groups include C 5-20 heteroaryl, C 5-12 heteroaryl, pyrrolyl, pyridyl, imidazolyl, furyl, pyranyl, thienyl, pyrimidinyl, pyrazinyl, pyridazine Base, fluorenyl, quinolyl, pyridopyridyl, oxazolyl and the like.
  • heteroaryloxy means “heteroaryl-O-" wherein heteroaryl is as defined above for the C5-20 heteroaryl.
  • polysubstituted means substituted by a plurality of substituents such as disubstituted, trisubstituted, tetrasubstituted and the like.
  • administering includes all means of directly or indirectly releasing a compound/pharmaceutical composition to the intended site of action.
  • amido refers to a group in which the hydroxy group of the carboxylic acid is substituted with an amino group (or an amine group) and formally eliminates one hydrogen atom.
  • C 1-8 alkylsulfonyl refers to a sulfonyl group substituted with a C 1-8 alkyl group, wherein the C 1-8 alkyl group is as defined above for a C 1-8 alkyl group.
  • mercapto refers to the abbreviation of sulfhydryl (-SH), also known as “thiol”.
  • thiol A monovalent atomic group composed of two elements of hydrogen and sulfur.
  • Thiol is one of the essential groups of certain enzyme proteins.
  • C 1-8 alkylamino means that the hydrogen atom on the amino group is substituted by a C 1-8 alkyl group, wherein the C 1-8 alkyl group is as defined above for the C 1-8 alkyl group.
  • Single C 1-8 alkylamino means that one hydrogen atom on the amino group is substituted by a C 1-8 alkyl group.
  • Di-C 1-8 alkylamino means that two hydrogen atoms on the amino group are substituted by a C 1-8 alkyl group.
  • C 4-12 aromatic heterocyclic group means having 4 to 12 carbon atoms, the ring system is relatively stable, and the ring including the hetero atom is a planar type, and 4n + 2 ⁇ electrons in the ring are in a ring-closed state.
  • the group of the conjugate system is, for example, a benzoxazolinone group, a pyridyl group in a C 4-5 aromatic heterocyclic group, a thienyl group, a furyl group or the like.
  • C 6-12 benzoheterocyclyl refers to a group containing 6 to 12 carbon atoms and a benzene ring fused to a monoheterocyclic ring, such as a benzoxazolinone group.
  • blocking refers to complete (all) blocking or partial blocking, such as a blocking rate of 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, and 70% or more. , 80% or more or 100% blocking.
  • Blocking agent refers to the drug used to achieve this blockade.
  • the compound of the present invention, a pharmaceutically acceptable salt thereof and a pharmaceutical composition comprising the same have a significant specific blocking or inhibitory effect on the N-type calcium ion channel.
  • the compound of the present invention, a pharmaceutically acceptable salt thereof and a pharmaceutical composition comprising the same have good pharmacokinetic properties and are effective for relieving pain.
  • a compound of the present invention, a pharmaceutically acceptable salt thereof, and a pharmaceutical composition comprising the same can prevent or treat post-operative pain, migraine, visceral pain, neuropathic pain, stroke, cerebral ischemia, alcohol addiction, alcoholism, Acute renal failure, chronic renal failure, renal insufficiency, an addictive disorder caused by an analgesic drug, or a tolerable disorder caused by an analgesic drug.
  • Figure 1 is a concentration of the compound (1) in the plasma of 6 rats in Test Example 4 of the present invention
  • Figure 2 is a graph showing the concentration of the compound (1) in rat plasma after intravenous administration and oral administration in Test Example 4 of the present invention.
  • Test Example 1 Analgesic effect of the compound of the present invention
  • mice were weighed and labeled, and each group consisted of 10 (male and female) divided into 7 groups, including 5 experimental groups (using the hydrochloride of compound (4) and compounds (1), (2), ( 3), (5)), positive control group (using NMED-160) and negative control group (using physiological saline).
  • the positive control group used NMED-160 is a selective small molecule N-type calcium channel blocker, has entered the phase II clinical trial, can be synthesized by reference [11], the structural formula is as follows:
  • mice were intragastrically administered (30 mg/kg). After 40 minutes, 0.6% (v/v) acetic acid 0.4 mL was intraperitoneally injected. After 5 minutes, the number of mouse writhings in the subsequent 15 minutes was recorded. The inhibition rate of the drug on the acetic acid writhing in mice was calculated, and the analgesic effect of the drug was evaluated. The results are shown in Table 2.
  • A represents the number of writhings in the negative control group
  • B represents the number of writhings of the experimental group or the positive control group.
  • the compound of the present invention and its pharmaceutically acceptable salt have an obvious analgesic effect; wherein the analgesic effects of the compounds (1) to (2) are comparable to those of NMED-160.
  • Test Example 2 N-type calcium channel current suppressing activity of the compound of the present invention
  • N-type calcium channel cDNAs plasmid ⁇ 1B (GeneBank accession no. AF055477)/ ⁇ 2 ⁇ (AF286488)/ ⁇ 1b (L06110), rabbit ⁇ 1C (X15539), human ⁇ 1A (NM000068), rat ⁇ 1E (NM 009782) And rat HERG (U04270).
  • E. coli competent cells containing ⁇ 1B , ⁇ 2 ⁇ , ⁇ 1b plasmid were placed in LB solution (100 mL) containing ampicillin (50 ⁇ g/mL) at 37 ° C, 200 rpm. Shake for 12-17 hours and reserve the next day.
  • Alkaline proteases are capable of inactivating nucleases and other proteins released during bacterial lysis that affect the quality of the isolated plasmid. Then, 350 ⁇ L of the neutralizing solution was added to the centrifuge tube and rapidly inverted 4 times to thoroughly mix, and the centrifuge tube was centrifuged at a maximum speed (14,000 g) for 10 minutes at room temperature.
  • the surgical instruments were immersed in 75% (w/w) ethanol for 30 minutes and taken out to dry.
  • the 5th thread was sterilized in boiling water for 10 minutes.
  • the claws were buried in crushed ice for about 40 minutes to anesthetize them.
  • Remove the anesthetized paw place the abdomen up on the tiled crushed ice, and bury its head and limbs with crushed ice.
  • Disinfect the lower abdomen skin with an alcohol cotton ball then pick it up with a needle (middle or left in the center) and use a pair of ophthalmic scissors to cut a small opening of about 1 cm. Cut the muscle layer in the same way (be careful not to damage any internal organs, and cut the muscle layer to see the oocytes).
  • the leaflets of 1 cm 3 size were taken out with tweezers and scissors, and placed in a prepared culture dish containing OR-2 (containing penicillin), and the muscle layer and the skin layer were respectively sutured.
  • the oocytes are transferred to a sterile glass tube and washed repeatedly with an OR-2 solution until the residual blood is washed.
  • the collagenase solution was added and shaken for about 1 hour, and the new collagenase solution was replaced and the shaking was continued for about 1 hour (when most of the isolated or single cells were seen).
  • Plasmids ⁇ 1B , ⁇ 2 ⁇ , ⁇ 1b were injected into Xenopus oocytes at a concentration of 1:1:1, and the total volume of the three plasmids injected per cell was 46 nL.
  • the injected cells were placed in ND-96 medium, and the expressed current was recorded after standing at a temperature of 18 ° C for 48 hours.
  • the perfusion administration method has a flow rate of 3 mL/min and a drug concentration of 10 ⁇ M to be tested.
  • the cells were clamped at -100 mV, stepped at 10 mV, depolarized to +60 mV, and the current was recorded.
  • Table 3 shows the results of inhibition of N-type calcium channel currents (10 ⁇ M)
  • Test Example 3 Compounds of the invention inhibit P/Q-type calcium channels, Herg channels, sodium channels, and potassium channels Activity
  • the plasmid DNA was digested with the above reaction system, and incubated at 37 ° C for 1 hour.
  • the reactants were added in the following order of the reaction system:
  • rNTPs 25 mM ATP, CTP, UT 7.5 ⁇ L each + 3 mM GTP 0.6 ⁇ L + denuclease water 6.9 ⁇ L;
  • Linear DNA template (5-10 ⁇ g total) + denuclease water 32.5 ⁇ L;
  • Ribom7 hat analog, 40 mM, 7.5 ⁇ L;
  • the needle is punctured on the clean thin tissue paper to make the diameter of the needle tip thick, and polished on the polishing instrument, so that the needle tip is smooth and flat.
  • the diameter of the tip is preferably 6-10 ⁇ m.
  • the mineral oil Light mineral oil
  • the adjusted injection concentration was approximately 2 ng/nL mixed (1:1:1). 1 ⁇ L of the mixed cRNA was carefully dropped onto a clean parafilm, and the cRNA was inhaled into the injection needle using a microinjector.
  • the puncture cells are not too deep, and it is advisable that the tip of the needle just passes through the cell membrane, and 46.5 nl of cRNA is injected into the oocyte, and the cells will slightly swell after the injection. After waiting for 30 s, pull out the needle and observe if the cells have spilled contents. If there is any, give up. In addition, if the tip of the needle is easily pierced into the cell, there is no breakthrough, indicating that the cell is not strained, and the cell state is not good, meaning that the cell may not be able to clamp to the desired potential, and the cell should be discarded.
  • the injected oocytes were placed in ND-96 solution, and after 48 hours of incubation in a biochemical incubator (18 ° C) (replacement of ND-96 liquid daily), the expressed P/Q-type calcium channels were recorded separately. Current and Herg channel current.
  • Extracellular fluid BaCl 2 5, N-methyl-D-glucamine 50, KCl 5, HEPES 5, adjusted to pH 7.4 with methanesulfonic acid.
  • the intracellular fluid was 3 mM KCl.
  • Preparation of BAPTA 10 mM Hepes dissolved BAPTA and the pH was adjusted to 7.2 with CsOH.
  • Newborn Wistar rats were taken and sterilized with 75% (w/w) ethanol.
  • the brain was taken under aseptic conditions, and the hippocampus was peeled off and placed in an anatomical solution of an ice bath.
  • the hippocampus was cut into 1-2 mm 3 tissue blocks, digested with 0.25% trypsin at 37 ° C for 30 minutes, then the digested tissue pieces were transferred to the planting solution to terminate digestion, and the appropriate caliber in the planting solution.
  • the pipette (the tip diameter of 2 mm) was blown to the cells, and uniformly dispersed to prepare a cell suspension, and a small amount of the suspension was taken to count the cells in the trypan blue dye solution.
  • planting solution inoculate the cells at a density of 1 ⁇ 10 5 /mL in a 35 mm culture dish coated with polylysine, place in a 10 ° carbon dioxide incubator at 36 ° C overnight, and change culture after 24 hours.
  • Base replace the planting solution with 2 mL of stock solution.
  • the medium was changed once every three days, and the cultured cells were used for patch clamp experiments for 12-15 days.
  • an appropriate amount of cytarabine was added to the medium on the third day of culture (6 ⁇ L of cytarabine stock solution was added to each dish to a final concentration of 3 ⁇ g/mL).
  • the recording fluid used for whole-cell recording is mainly an intra-electrode fluid and an extracellular fluid.
  • the liquid content (mM) of the recording electrode was recorded: KCl 140, HEPES 10, EGTA 10, and the pH was adjusted to 7.2-7.4.
  • the composition of the extracellular fluid NaCl 140, KCl 5, MgCl 2 1, HEPES 10, Glucose 10, CaCl 2 3, adjusted to pH 7.2-7.4 with NaOH. If the experiment requires blocking sodium current, tetrodotoxin (TTX) 1 ⁇ M can be added to the extracellular fluid. If the experiment requires blocking the potassium current, and only the CsCl can not completely block the potassium current, tetraethylammonium (TEA) or 4-aminopyridine (4-AP) can be added to the electrode liquid and the extracellular fluid.
  • TX tetrodotoxin
  • the compound of the present invention is capable of specifically inhibiting an N-type calcium ion channel.
  • Test Example 4 Rapid evaluation of pharmacokinetics in rats
  • Compound (1) is dissolved in 25% (w/v) aqueous solution of hydroxypropyl- ⁇ -cyclodextrin;
  • Megestrol acetate internal standard, purchased from the Central Inspection Office;
  • Hydroxypropyl- ⁇ -cyclodextrin purchased from Xi'an Deli Biochemical Co., Ltd., configured as a 25% (w/v) aqueous solution of hydroxypropyl- ⁇ -cyclodextrin;
  • Acetonitrile, methanol chromatographically pure, purchased from Fisher Company;
  • LC-MS/MS Liquid chromatography-mass spectrometry
  • API 5000 triple quadrupole cascade LC-MS/MS
  • LDZ5-2 centrifuge (Beijing Medical Centrifuge Factory);
  • MicroCL21R high speed refrigerated centrifuge (Thermo Company, USA);
  • JJ-1 precision booster electric mixer (Changzhou Guohua Electric Co., Ltd.);
  • QC sample The mother solution of compound (1) (1 mg/mL) was diluted stepwise into three standard solutions of low, medium and high concentrations. Take 0.05mL of blank rat plasma, add 0.005mL of standard solution of low, medium and high concentration and 0.3mL of acetonitrile containing endogenous progesterone (10ng/mL).
  • Mobile phase composition mobile phase A was water (containing 0.1% formic acid), mobile phase B was acetonitrile (containing 10 mM ammonium acetate and 0.1% formic acid), and the gradient elution procedure is shown in Table 5.
  • the ionization mode is electrospray (ESI) condition, positive ion detection;
  • the m/z detected by the compound (1) and the internal standard were 427.168 and 385.2, respectively.
  • the selective ion (SRM) method detects the fragment ion of m/z 427.168 as m/z 250.1 and CE as 18; the fragment ion of internal standard m/z 385.2 is m/z 267.3 and CE is 25.
  • the measured peak area was plotted against the concentration of the compound (1), and the linear range was 1 ng/mL-2000 ng/mL, and the lowest detection line was 1 ng/mL, and the linearity was good.
  • the precision of the concentration of the quality control sample and the accuracy between the true concentration and the true concentration are within ⁇ 15%, indicating that the detection method is accurate and reliable.
  • the results of 1#-3# were averaged, and the results of 4#-6# were averaged to obtain the relationship between the average concentration and time of compound (1) in iv-administered and po-administered rats, as shown in Fig. 2.
  • the Winnolin pharmacokinetic software was used to analyze the measured data and calculate the main pharmacokinetic parameters, including the area under the curve (AUC), clearance rate (CL), apparent volume of distribution (V), and end. Elimination half-life (t 1/2 ), mean residence time (MRT), and area under the curve (AUC) for oral administration, terminal elimination half-life (t1/2), mean residence time (MRT), peak concentration (C max ), peak time (T max ) and biological strength (F), the obtained pharmacokinetic parameters are shown in Table 6.
  • the compound as an analgesic drug has good pharmacokinetic properties in rats, and further research can be carried out, including the distribution of target sites of brain tissue under steady state conditions.
  • Compound (1) was formulated into a solution having a concentration of 4 mg/mL with a 25% (w/v) aqueous solution of hydroxypropyl- ⁇ cyclodextrin.
  • Three adult SD rats were randomly numbered, and 0.4 mL of the compound (1) solution was intravenously administered, respectively, and then the compound (1) solution (0.4 mL/h) was slowly instilled for two hours.
  • 0.1 mL of blood was collected before administration and 30 minutes, 60 minutes and 2 hours after administration, placed in an anticoagulant tube, and the plasma was separated by centrifugation for 15 minutes (3500 rpm), and 0.05 mL of the plasma sample was accurately taken, and the refrigerator was frozen at -30 ° C. Save the test.
  • the filter paper was blotted dry and stored in a freezer at -30 °C for testing.
  • Rat brain tissue and spinal cord samples were weighed and added to normal saline at 1:4 (v/v) to prepare homogenate.
  • the compound (1) easily enters the brain tissue and the spinal cord through the blood-brain barrier, and is 3-4 times in the brain tissue and the spinal cord, and is an analgesic drug as a central target. This distribution is beneficial to the efficacy of the drug.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Epidemiology (AREA)
  • Biomedical Technology (AREA)
  • Neurosurgery (AREA)
  • Neurology (AREA)
  • Pain & Pain Management (AREA)
  • Addiction (AREA)
  • Urology & Nephrology (AREA)
  • Cardiology (AREA)
  • Vascular Medicine (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Psychiatry (AREA)
  • Rheumatology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Hydrogenated Pyridines (AREA)
  • Plural Heterocyclic Compounds (AREA)

Abstract

本发明属于医药化工领域，公开了4,4-二苯基哌啶类化合物或其可药用盐、药物组合物及用途。具体地，本发明公开了式I所示的化合物或其可药用盐，还公开了包含该化合物或其可药用盐的药物组合物。本发明的化合物或其可药用盐、药用组合物具有明显的N型钙离子通道阻断活性，并具有良好的药代动力学性质，能够有效地缓解疼痛，具有作为新的预防或治疗疼痛、中风、脑缺血、酒精成瘾、酒精中毒、肾病、由镇痛药物引起的成瘾性病症或由镇痛药物引起的耐受性病症的药物的潜力。

Description

4,4-二苯基哌啶类化合物或其可药用盐、药物组合物及用途 技术领域
本发明属于医药化工领域，涉及4,4-二苯基哌啶类化合物或其可药用盐、包含该化合物或其可药用盐的药物组合物及用途。
背景技术
N型钙离子通道属于电压门控钙通道(VDCC)的一个亚型，由α1β亚单位构成，具有高电压激活、较快速失活的特点，主要分布在神经组织中，可被w-conotoxin GVIA(wCgTx)阻滞，已被确证为具有临床相关性的新型药物作用靶标。
N型钙离子通道阻断剂在治疗中风与脑缺血、镇痛尤其是神经源性疼痛、减少酒精嗜求和治疗酒精中毒 [1]以及肾保护 [2,3,4]等方面具有良好的应用前景。
研究结果表明，N型钙离子通道是疼痛产生和痛觉传导过程中的一个重要环节，由于针对N型钙离子通道的阻断剂直接作用于N型钙离子通道，不涉及第二信使或G蛋白，因此不易产生成瘾性。目前，高选择性的N型钙离子通道阻断剂ω-芋螺毒素(ω-conotoxin)MVIIA
Figure PCTCN2018072426-appb-000001
已于2004年12月被美国FDA批准进入市场销售，其临床实践表明，N型钙离子通道阻断剂已被证实为治疗疼痛的一个新型靶标，具有良好的应用前景。
目前的研究表明，N型钙通道的阻断与治疗肾病有关：
一方面，N型钙离子通道的阻断减少了交感神经末梢去甲肾上腺素、肾素的释放，降低了入球动脉及出球动脉的阻力，减少了肾小球的压力 [2]。糖尿病状态下，肾脏内肾素血管紧张素系统被不适当激活，血管紧张素转换酶(ACE)活性增高，血管紧张素Ⅱ(AngⅡ)表达增多，直接参与了肾脏的进行性损害；它不仅通过影响全身及肾脏局部的血流动力，造成肾小球内的高压，还与糖尿病肾病(DN)发病机制中的蛋白激酶C(PKC)学说、氧化应激(OS)学说、细胞因子学说及遗传分子学说紧密相关。肾内有交感神经和副交感神经分布，肾内交感神经大部分是血管运动性的，使肾血管收缩；N型钙离子通道位于交感神经末梢，交感神经兴奋后N型钙离子通道钙离子内流，去甲肾上腺素释放，使肾脏的入球小动脉及出球小动脉均收缩，并刺激球旁细胞释放肾素。而且在众多的糖尿病肾病发展因素中，肾血液动力学，特别是肾小球高灌注压在肾功能损伤中起关键作用。2005年Tomoyuki [3] 等报道在高血压肾病大鼠模型观察，L/N型钙离子通道阻断剂西尼地平明显的减少了血浆肾素及去甲肾上腺素水平，以及入球及出球动脉的压力，降低了微量尿白蛋白。其可能通过抑制N型钙通道，来实现降低微量尿白蛋白来保护肾脏。
另一方面，N型钙通道的阻断增加了对胰岛素的敏感性，降低了胰岛素抵抗 [4]。胰岛素抵抗(IR)引起糖尿病肾脏病变的机制可能是刺激细胞外基质集聚，升高各种炎症性细胞因子，增加钠的储留，造成对肾脏的损伤。由于IR产生血胰岛素增高，高胰岛素血症可兴奋下丘脑交感神经中枢，还会影响肾脏血流动力学，直接作用于肾小球动脉，扩张入球小动脉，加重肾小球高滤过、高灌注状态，通过引起高血压、高血脂、高血尿酸等间接机制，促使肾小球硬化，高胰岛素血症通过刺激细胞因子，如胰岛素样生长因子来加重肾小球肥大的发生。2型糖尿病肾病的早期病理表现为系膜基质轻度增宽及肾小球基底膜(GBM)轻度增厚正说明这一点。有专家提出IR不单是糖尿病发生根源，更是糖尿病患者肾脏病变形成的基础。1999年，Ishikawa [4]等在高血压肾病模型中观察N型钙离子通道阻断剂西尼地平降低了胰岛素抵抗，而L型钙通道阻断剂阿洛地平没有此作用。这些进一步证实了N型钙通道可能作为治疗肾病潜在的新治疗靶点。
目前，针对N型钙离子通道阻断剂已发展出多个系列的小分子化合物 [5]。其中，WO99/43658公开的4-哌啶基苯胺类小分子化合物作为可口服的选择性的N型钙离子通道阻断剂表现出明显的镇痛活性；Teodori等 [6]、Knutsen等 [7]、Yamamoto等 [8,9]、Tyagarajan等 [10]的研究也取得了一定进展；小分子N型钙离子通道阻断剂NMED-160已经进入II期临床研究，其相关化合物的研究也得到了较好结果 [11,12]。
但是，目前研究得到的N型钙离子通道阻断剂仍存在缺陷：一方面，这些化合物在生物活性及通道选择性方面存在不足；另一方面，这些化合物的药代动力学性质较差，需通过侧脑室给药等临床上非简单易行的途径给药才能产生足够的药理活性。因此，目前尚需开发新型的N型钙离子通道阻断剂。
发明内容
本发明人经过研究和创造性劳动，得到了一类4,4-二苯基哌啶类化合物或其可药用盐。本发明人发现，该类化合物或其可药用盐表现出明显的N型钙离子通道阻断活性，并具有良好的药代动力学性质，能够有效地防治或缓解疼痛，具有作为新的预防和/或治疗疼痛(尤其是神经源性疼痛)、中风、脑缺血、酒精成瘾、酒精中毒、肾病、由镇痛药物引起的成瘾性病症或由镇痛药物引起的耐受性病症的药物或药物活性成分的潜力。
本发明第一方面涉及式Ⅰ所示的化合物或其可药用盐：
Figure PCTCN2018072426-appb-000002
其中：
g选自C 1-8亚烷基、取代的C 1-8亚烷基、羰基和C 1-8亚酰基；
R选自C 5-20芳基、取代的C 5-20芳基、C 4-20杂环基和取代的C 4-20杂环基；
所述取代独立地为被一个或多个取代基所取代，所述取代基选自卤素、C 1-8烷基、C 1-8烷氧基、C 3-8环烷基、氰基、硝基、巯基、甲硫基、乙硫基、三氟甲基、氨基、酰胺基、单C 1-8烷基氨基、二C 1-8烷基氨基、C 1-8烷基磺酰基、羟基、芳氧基、杂芳氧基和杂环基氧基。
本发明中，式I化合物的可药用盐包括其无机酸盐或有机酸盐，其包括但不限于：盐酸盐、氢溴酸盐、氢碘酸盐、硝酸盐、硫酸盐、硫酸氢盐、磷酸盐、磷酸氢盐、乙酸盐、羟乙酸盐、丙酸盐、丁酸盐、草酸盐、己二酸盐、藻酸盐、乳酸盐、柠檬酸盐、酒石酸盐、琥珀酸盐、马来酸盐、富马酸盐、苦味酸盐、天冬氨酸盐、葡糖酸盐、苯甲酸盐、甲磺酸盐、乙磺酸盐、苯磺酸盐、对甲苯磺酸盐、双羟萘酸盐、丙酮酸盐、乙醇酸盐、丙二酸盐、三氟乙酸盐、苹果酸盐、水杨酸盐、对氨基水杨酸盐、扑酸盐和抗坏血酸盐等；例如为式I化合物的盐酸盐。
本发明第一方面任一所述化合物还可以是前药或可在体内代谢变化后释放出所述活性成分的形式。选择和制备适当的前药衍生物是本领域技术人员的公知技术。
本发明第一方面的一个实施方式中，g选自C 1-8亚烷基、羰基和C 1-8亚酰基；R选自C 5-20芳基、取代的C 5-20芳基和C 4-20杂环基。
本发明第一方面的一个实施方式中，所述取代基选自C 1-8烷基、C 1-8烷氧基、C 3-8环烷基、巯基、甲硫基、乙硫基、氨基、单C 1-8烷基氨基、二C 1-8烷基氨基和羟基。
本发明第一方面的一个实施方式中，所述卤素为氟、氯、溴或碘。
本发明第一方面的一个实施方式中，g选自C 1-8亚烷基、羰基和C 1-8亚酰基；R 选自C 6-12芳基、取代的C 6-12芳基和C 4-12芳杂环基。
本发明第一方面的一个实施方式中，所述取代基选自C 1-8烷氧基、甲硫基、乙硫基、氨基、单C 1-8烷基氨基和二C 1-8烷基氨基。
本发明第一方面的一个实施方式中，所述C 4-12芳杂环基为C 4-5芳杂环基或C 6-12苯并杂环基。
本发明第一方面的一个实施方式中，g选自亚甲基、羰基、亚甲酰基、亚乙酰基、1,3-亚丙酰基、1,2-亚丙酰基、1,4-亚丁酰基、1,3-亚丁酰基、1,2-亚丁酰基；R选自苯基、取代苯基、噻吩基和苯并噁唑啉酮基(例如2-苯并噁唑啉酮基)。
本发明第一方面的一个实施方式中，所述取代基选自甲氧基和二甲基氨基。
本发明第一方面的一个实施方式中，所述的化合物或其可药用盐选自如下(1)-(6)化合物及其可药用盐：
(1)6-(3-(4,4-二苯基哌啶)-丙酰基)苯并噁唑啉-2-酮；
(2)6-(3-(4,4-二苯基哌啶)-乙酰基)苯并噁唑啉-2-酮；
(3)1-(4-二甲氨基苄基)-4,4-二苯基哌啶；
(4)1-(3,4,5-三甲氧基苄基)-4,4-二苯基哌啶；
(5)1-(2-甲基噻吩基)-4,4-二苯基哌啶；
(6)4,4-二苯基-1-苄基哌啶。
上述化合物(1)-(6)的名称和结构式如下面的表1所示。
表1化合物(1)-(6)的名称和结构式
Figure PCTCN2018072426-appb-000003
Figure PCTCN2018072426-appb-000004
本发明第二方面涉及本发明第一方面任一所述的化合物或其可药用盐的制备方法。
可以采用如下所示的合成路线制备本发明的式I化合物，另外，也可参见本发明实施例中的详细描述。根据需要，还可以与酸反应转化为其可药用盐。
首先，以N-苄基-4哌啶酮为原料，以苯为原料和反应溶剂，在三氟甲磺酸存在下室温反应得到4,4-二苯基-1-苄基哌啶(式Ⅱ)；而后，4,4-二苯基-1-苄基哌啶(式Ⅱ)与氯甲酸乙酯反应，然后碱水解，得到4,4-二苯基哌啶(式Ⅳ)；4,4-二苯基哌啶(式Ⅳ)在干燥的丙酮中，与商品化的卤代物或醛在三乙胺等碱性条件下，室温下搅拌2～24小时进行卤代反应或还原胺化反应，得到式Ⅰ目标化合物。
Figure PCTCN2018072426-appb-000005
Figure PCTCN2018072426-appb-000006
本发明第二方面的一个实施方案中，本发明第一方面任一所述的化合物或其可药用盐的制备方法，包括下述步骤：
(1)N-苄基-4哌啶酮与苯在三氟甲磺酸存在下反应，生成式Ⅱ所示的化合物，
Figure PCTCN2018072426-appb-000007
(2)式Ⅱ化合物与氯甲酸乙酯反应，生成式Ⅲ所示的化合物，
Figure PCTCN2018072426-appb-000008
(3)式Ⅲ化合物在碱条件下水解，生成式Ⅳ所示的化合物，
Figure PCTCN2018072426-appb-000009
(4)式Ⅳ化合物与卤代物或醛反应，生成式Ⅰ所示的化合物，
Figure PCTCN2018072426-appb-000010
上述式Ⅰ中的R或g的定义如本发明第一方面任一的化合物或其可药用盐所述。
本发明第三方面涉及一种药物组合物，包含本发明第一方面任一所述的化合物或其可药用盐；可选地，所述药物组合物还包含药学上可接受的载体和/或辅料。
本发明的化合物可以本身形式给药，或者以药物组合物的形式给药。本发明的药物组合物中，化合物可以是与一种或多种药学上可接受的载体、赋形剂或稀释剂相混合的。本发明的药物组合物通常是按常规方式配制的，使用一种或多种生理学上可接受的载体和/或辅料，它们有利于将活性化合物加工成可以在药学上使用的制剂，适当的制剂取决于所选择的给药途径，可以按照本领域熟知的常识进行制备。
可用于本发明药物组合物的药用载体或赋形剂包括但不限于：离子交换剂、氧化铝、硬脂酸铝、卵磷脂、血清蛋白如人血清蛋白、缓冲物质如磷酸盐、甘油、山梨酸、山梨酸钾、饱和植物脂肪酸的部分甘油酯混合物、水、盐或电解质，如硫酸鱼精蛋白、磷酸氢二钠、磷酸氢钾、氯化钠、锌盐、胶态二氧化硅、三硅酸镁、聚乙烯吡咯烷酮、纤维素物质、聚乙二醇、羧甲基纤维素钠、聚丙烯酸酯、蜂蜡、聚乙烯-聚氧丙烯嵌段聚合物、羊毛脂。
本发明的式I化合物或其可药用盐或者含有式I化合物或其可药用盐的药物组合物的给药途径可为肠道或非肠道，如口服、肌肉、皮下、鼻腔、口腔粘膜、皮肤、腹膜或直肠等。给药剂型例如片剂、胶囊、滴丸、气雾剂、丸剂、粉剂、溶液剂、混悬剂、乳剂、颗粒剂、脂质体、透皮剂、口含片、栓剂、冻干粉针剂等。还可以制备成缓释制剂、控释制剂及各种微粒给药系统。
本发明第四方面涉及一种在体内或体外阻断或抑制N型钙离子通道的方法，包括给予有需求的受试者以有效量的本发明第一方面任一所述的化合物或其可药用盐或者给予有需求的受试者以有效量的本发明第三方面所述的药物组合物的步骤。
本发明第五方面涉及本发明第一方面任一所述的化合物或其可药用盐或者本发明第三方面所述的药物组合物在制备N型钙离子通道阻断剂或抑制剂中的用途。
本发明第六方面涉及本发明第一方面任一所述的化合物或其可药用盐或者本发明第三方面所述的药物组合物在制备预防或治疗疼痛、中风、脑缺血、酒精成瘾、酒精中毒、肾病、由镇痛药物引起的成瘾性病症或由镇痛药物引起的耐受性病症的药物中的用途；例如，所述疼痛为手术后疼痛、偏头痛、内脏痛或神经性疼痛；例如，所述肾病为急性肾衰竭、慢性肾衰竭或肾机能不全。
本发明第七方面涉及一种预防或治疗疼痛、中风、脑缺血、酒精成瘾、酒精中毒、肾病、由镇痛药物引起的成瘾性病症或由镇痛药物引起的耐受性病症的方法，包括给予有需求的受试者有效量的本发明第一方面任一所述的化合物或其可药用盐或者本发明第三方面所述的药物组合物的步骤；例如，所述疼痛为手术后疼痛、偏头痛、内脏痛或神经性疼痛；例如，所述肾病为急性肾衰竭、慢性肾衰竭或肾机能不全。
本发明的化合物或其可药用盐可以单独给药或与其它的本发明的化合物或其可药用盐联合给药，和/或与其它已知治疗剂联合给药。
本发明的化合物或其可药用盐或者本发明的药物组合物针对不同患者的特定使用剂量和使用方法决定于诸多因素，包括患者的年龄，体重，性别，自然健康状况，营养状况，化合物的活性强度，服用时间，代谢速率，病症的严重程度以及诊治医师的主观判断。
单位剂型通常含有0.1重量％至99重量％的活性物质，更通常为5重量％至75重量％的活性物质。举例来说，单位剂型可以含有1mg至1g化合物、10mg至500mg化合物、50mg至400mg化合物或者100mg至200mg化合物。
本发明第八方面涉及本发明第一方面中任一所述的化合物或其可药用盐或者本发明第三方面所述的药物组合物，其用于阻断或抑制N型钙离子通道。
本发明第九方面涉及本发明第一方面中任一所述的化合物或其可药用盐或者本发明第三方面所述的药物组合物，其用于预防或治疗疼痛、中风、脑缺血、酒精成瘾、 酒精中毒、肾病、由镇痛药物引起的成瘾性病症或由镇痛药物引起的耐受性病症；例如，所述疼痛为手术后疼痛、偏头痛、内脏痛或神经性疼痛；例如，所述肾病为急性肾衰竭、慢性肾衰竭或肾机能不全。
在本发明中，如无特别说明：
术语“卤素”是指ⅦA族元素，包括氟(F)、氯(Cl)、溴(Br)、碘(I)和砹(At)。
术语“C 1-8烷基”是指具有1-8个碳原子的直链或支链烷基。例如为具有1-6个碳原子的直链或支链烷基，例如为具有1-4个碳原子的直链或支链烷基，例如甲基、乙基、丙基、异丙基、丁基、异丁基、仲丁基、叔丁基、戊基、2-乙基-丁基、己基、庚基和辛基。
术语“C 1-8亚烷基”是指具有1-8个碳原子形式上消除了两个氢原子的直链或支链烷基，例如为C 1-6亚烷基、C 1-4亚烷基、亚甲基、1,2-亚乙基、亚乙基、亚异丙基、1,3-亚丙基等。
术语“羰基”是指碳和氧两种原子通过双键连接而成的二价基团。
术语“C 1-8亚酰基”是指C 1-8烷基脂肪酸去掉羟基后剩下的并且在形式上还消除了一个氢原子的基团。例如C 2-8亚酰基、C 2-6亚酰基、C 1-6亚酰基、亚甲酰基、亚乙酰基、1,3-亚丙酰基、1,2-亚丙酰基、1,4-亚丁酰基、1,3-亚丁酰基、1,2-亚丁酰基等。
术语“C 1-8烷氧基”表示“C 1-8烷基-O-”，其中C 1-8烷基如上述定义。
术语“C 3-8环烷基”表示具有3-8个碳原子的环状烷基。例如C 3-6环烷基、C 3-5环烷基，C 3-8环烷基的例子包括环丙基、环丁基、环戊基、环己基、环庚基、环辛基等。
术语“C 5-20芳基”是指包含5-20个碳原子的形式上消除了一个氢原子的芳香环(包括稠环)，例如C 6-20芳基、C 6-18芳基、C 6-12芳基。芳基的例子具体包括茂基、苯基、萘基、蒽基、芴基等。
术语“芳氧基”表示“芳基-O-”，其中芳基如上述C 5-20芳基定义。
术语“C 4-20杂环基”是指包含4-20个原子(1-3个原子选自杂原子氧、氮和硫)的杂环基，分为脂杂环基和芳杂环基两类。杂环基的例子包括苯并杂环基、四氢呋喃基、吡咯烷基、哌啶基、吗啉基等、噻唑烷基、噻唑啉硫酮基、硫代噻唑啉基、苯并噻唑基、噻吩基、噻二唑基、苯并噁唑啉-2-酮基等。
术语“杂环基氧基”表示“杂环基-O-”，其中杂环基如上述C 4-20杂环基的定义。
术语“杂芳基”是指芳香杂环，其中1-3个原子选自杂原子氧、氮和硫，其它原子 为碳原子。杂芳基的例子包括C 5-20杂芳基、C 5-12杂芳基、吡咯基、吡啶基、咪唑基、呋喃基、吡喃基、噻吩基、嘧啶基、吡嗪基、哒嗪基、吲哚基、喹啉基、吡啶并吡啶基、咔唑基等。
术语“杂芳氧基”表示“杂芳基-O-”，其中杂芳基如上述C 5-20杂芳基的定义。
术语“多取代”表示被多个取代基所取代，例如为二取代、三取代、四取代等。
术语“给药”包括所有直接或间接释放化合物/药物组合物到预期作用部位的手段。
术语“酰胺基”是指羧酸的羟基被氨基(或胺基)取代并且形式上消除了一个氢原子的基团。
术语“C 1-8烷基磺酰基”是指含C 1-8烷基取代的磺酰基，其中C 1-8烷基如上述C 1-8烷基的定义。
术语“巯基”是指硫氢基(-SH)的简称,又称“硫醇基”。由氢和硫两种元素组成的一价原子团。巯基是某些酶蛋白的必需基团之一。
术语“C 1-8烷基氨基”是指氨基上的氢原子被C 1-8烷基所取代，其中C 1-8烷基如上述C 1-8烷基的定义。“单C 1-8烷基氨基”是指氨基上的一个氢原子被C 1-8烷基所取代。“二C 1-8烷基氨基”是指氨基上的两个氢原子被C 1-8烷基所取代。
术语“C 4-12芳杂环基”是指包含4-12个碳原子，环系较稳定，包括杂原子在内的环是平面型，环内有4n+2个π电子处于环闭共轭体系的基团，例如苯并噁唑啉酮基、C 4-5芳杂环基中的吡啶基、噻吩基、呋喃基等。
术语“C 6-12苯并杂环基”是指包含6-12个碳原子，苯环与单杂环稠和形成的基团，例如苯并噁唑啉酮基。
术语“阻断”是指完全(全部)阻断或部分阻断，例如阻断率为10％以上、20％以上、30％以上、40％以上、50％以上、60％以上、70％以上、80％以上或100％的阻断。“阻断剂”指实现这一阻断所使用的药物。
本发明取得的有益效果：
1、本发明化合物、其可药用盐及包含该化合物的药物组合物对N型钙离子通道有明显的特异性阻断或抑制作用。
2、本发明化合物、其可药用盐及包含该化合物的药物组合物具有良好的药代动力学性质，能够有效地缓解疼痛。
3、本发明化合物、其可药用盐及包含该化合物的药物组合物可以预防或治疗手术 后疼痛、偏头痛、内脏痛、神经性疼痛、中风、脑缺血、酒精成瘾、酒精中毒、急性肾衰竭、慢性肾衰竭、肾机能不全、由镇痛药物引起的成瘾性病症或由镇痛药物引起的耐受性病症。
附图说明
为了使本发明的内容更容易被清楚的理解，下面根据本发明的具体实施例并结合附图，对本发明作进一步详细的说明，其中：
图1为本发明测试例4中6只大鼠血浆中的化合物(1)浓度；
图2为本发明测试例4中静脉注射和口服给药后大鼠血浆中的化合物(1)浓度。
具体实施方式
实施例1：6-(3-(4,4-二苯基哌啶)-丙酰基)苯并噁唑啉-2-酮(化合物(1))的合 成
Figure PCTCN2018072426-appb-000011
称取N-苄基-4-哌啶酮1.89g(0.01mol)溶于20mL干燥的苯中，搅拌下加入三氟甲磺酸20mL，室温下搅拌反应4小时，将反应液倒入冰水中，加入40％的氢氧化钠水溶液调pH值至10-12，用30mL二氯甲烷进行提取，共提取3次，合并后的提取液用饱和氯化钠溶液30mL和水30mL各洗涤1次，加入适量无水硫酸钠干燥过夜，滤除干燥剂，减压蒸除溶剂，得白色固体(4,4-二苯基-1-苄基哌啶，化合物(6))3.24g，产率99％。
将上面制备的3.24g 4,4-二苯基-1-苄基哌啶(化合物(6))溶于100mL甲苯中，加入20mL氯甲酸乙酯，加热回流反应8小时，减压蒸除溶剂，残留物中加入100mL蒸馏水和8mL40％的氢氧化钠水溶液，加热回流反应24小时。减压蒸除溶剂，加入100mL蒸馏水，用40mL二氯甲烷进行提取，共提取3次，合并后的提取液用饱和氯化钠溶液30mL和水30mL各洗涤1次，加入适量无水硫酸钠干燥过夜，滤除干燥剂，减压蒸除溶剂，得白色固体1.81g，为4,4-二苯基哌啶，m.p.148℃-150℃，产率76.0％。 1H-NMR(CDCl 3，ppm)δ：7.30-7.34(m，4H)，7.20-7.23(m，6H)，3.21(t，4H，J＝5.32Hz)，2.68(t，4H，J＝5.32Hz)。
取上面制备的4,4-二苯基哌啶0.47g(0.002mol)和6-(3-氯丙酰基)苯并噁唑啉-2-酮0.45g(0.002mol)，溶于30mL丙酮中，加入0.22g(0.002mol)三乙胺，于室温下搅拌反应24小时，抽滤，固体用乙醚和水洗涤，干燥后得白色固体(化合物(1))0.52g，m.p.236℃(分解)，产率61.0％。 1H-NMR(DMSO-d 6，ppm)δ：7.79-7.81(m，2H)，7.10-7.34(m，11H)，3.12(t，2H，J＝7.00Hz)，2.41-2.58(m，10H)。MS[M+H] +：427.3。
实施例2：6-(3-(4,4-二苯基哌啶)-乙酰基)苯并噁唑啉-2-酮(化合物(2))的合 成
Figure PCTCN2018072426-appb-000012
取0.47g(0.002mol)的4,4-二苯基哌啶(参照实施例1方法制备)和0.45g(0.002mol)6-(2-氯乙酰基)苯并噁唑啉-2-酮，溶于30mL丙酮中，加入0.22g(0.002mol)三乙胺，于室温下搅拌反应24小时，抽滤，固体用乙醚和水洗涤，干燥后得白色固体(化合物(2))0.44g，产率52.0％。 1H-NMR(DMSO-d 6，ppm)δ：7.79-7.81(m，2H)，7.10-7.34(m，11H)，3.76(s，2H)，2.40-2.53(m，8H)。MS[M+H] +：425.2。
实施例3：1-(4-二甲氨基苄基)-4,4-二苯基哌啶(化合物(3))的合成
Figure PCTCN2018072426-appb-000013
称取0.47g(0.002mol)的4,4-二苯基哌啶(参照实施例1方法制备)和0.30g(0.002mol)4-二甲氨基苯甲醛，溶于25mL干燥的二氯甲烷中，室温下搅拌反应1小时，冰水浴冷却下加入0.53g(0.0025mol)三乙酰氧基硼氢化钠，于冰水浴中继续搅拌反应30分钟，升至室温，继续搅拌反应8小时。加入20mL二氯甲烷，用饱和碳酸氢钠水溶液、饱和氯化钠水溶液和水各30mL洗涤。分出二氯甲烷层，加入适量无水硫酸钠干燥过夜。滤除干燥剂，减压蒸除溶剂，硅胶柱分离得白色固体(化合物(3))0.49g，m.p.128℃-130℃，产率66.1％。 1H-NMR(CDCl 3，ppm)δ：7.12-7.29(m，12H)，6.68(d，2H，J＝8.68Hz)，3.34(s，2H)，2.48-2.51(brs，8H)。MS[M+H] +：371.0。
实施例4：1-(3,4,5-三甲氧基苄基)-4,4-二苯基哌啶盐酸盐(化合物(4)盐酸盐) 的合成
Figure PCTCN2018072426-appb-000014
称取0.47g(0.002mol)4,4-二苯基哌啶(参照实施例1方法制备)和0.19g(0.002mol)3,4,5-三甲氧基苯甲醛，参照实施例3的方法进行合成，得淡黄色油状物(化合物(4))，氯化氢的乙醚溶液成盐得白色固体(化合物(4)盐酸盐)0.46g，m.p.235℃-237℃，产率55.1％。 1H-NMR(CDCl 3，ppm)δ：12.51(br，1H)，7.44-7.48(t，2H，J＝7.56Hz)，7.03-7.46(m，10H)，3.91(s，6H)，3.85(s，3H)，3.45(t，2H)，2.13(t，2H)，2.71(t，4H)。MS[M+H] +：418.6。
实施例5：1-(2-甲基噻吩基)-4,4-二苯基哌啶(化合物(5))的合成
Figure PCTCN2018072426-appb-000015
称取0.47g(0.002mol)4,4-二苯基哌啶(参照实施例1方法制备)和0.22g(0.002mol)2-噻吩甲醛，参照实施例3的方法进行合成，得到白色固体(化合物(5))0.56g，产率84.1％。 1H-NMR(DMSO-d 6，ppm)δ：6.88-7.41(m，11H)，3.54(s，2H)，2.41(m，8H)。MS[M+H] +：334.4。
测试例1：本发明化合物的镇痛作用
1.实验目的：测定化合物(4)盐酸盐及化合物(1)、(2)、(3)、(5)在小鼠醋酸扭体模型上的镇痛活性。
2.实验材料：昆明种小鼠(18-22g)，雌雄各半，由军事医学科学院实验动物中心提供。
3.实验方法及结果：
将小鼠称重、标记，每组10只(雌雄各半)，分为7组，包括5个实验组(分别使用化合物(4)的盐酸盐及化合物(1)、(2)、(3)、(5))、阳性对照组(使用NMED-160)及阴性对照组(使用生理盐水)。其中，阳性对照组使用的NMED-160是一种选择性的小分子N型钙离子通道阻断剂，已进入II期临床试验，可以参照参考文献[11]合成，结构式如下：
Figure PCTCN2018072426-appb-000016
对小鼠灌胃给药(30mg/kg)，40分钟后，腹腔注射0.6％(v/v)醋酸0.4mL，5分钟后开始记录随后的15分钟内小鼠扭体的次数，按下式计算药物对小鼠醋酸扭体的抑制率，评价药物的镇痛效果，结果见表2。
Figure PCTCN2018072426-appb-000017
其中：
A代表阴性对照组的扭体次数；
B代表实验组或阳性对照组的扭体次数。
表2镇痛作用结果
药物 扭体抑制率(％) 每组动物数量(只)
NMED-160 51.75 10
化合物(1) 58.77 10
化合物(2) 54.66 10
化合物(3) 23.17 10
化合物(4)盐酸盐 39.68 10
化合物(5) 46.87 10
由表2可知，本发明的化合物、其可药用盐具有明显的镇痛作用；其中，化合物 (1)-(2)的镇痛作用与NMED-160相当。
测试例2：本发明化合物的N-型钙离子通道电流抑制活性
1.实验目的：测定本发明化合物(4)盐酸盐及化合物(1)、(2)、(3)、(5)对爪蟾卵母细胞瞬时表达的N型钙离子通道(α 1B/β 1b/α2 δ)的电流抑制作用。
2.实验材料：
非洲爪蟾卵母细胞。
N型钙通道cDNAs质粒α 1B(GeneBank accession no.AF055477)/α 2δ(AF286488)/β 1b(L06110)，兔α 1C(X15539)，人α 1A(NM000068)，大鼠α 1E(NM 009782)和大鼠HERG(U04270)。
3.实验方法：
(1)钙通道不同亚基cDNA扩增：
将含α 1B，α 2δ，β 1b质粒(cDNAs质粒)的大肠杆菌感受态细胞分别置于LB溶液(100mL)，含氨苄青霉素(50μg/mL)，以37℃，200转/分的转速振摇12-17小时，次日备用。
(2)提取质粒：
取4mL上述大肠杆菌的培养物用台式离心机(12000g)离心5分钟，弃取上清液，并将试管倒置于纸巾上吸去剩余的培养液。向上述离心管加入250μL细胞悬浮液，旋涡震荡或吹打以充分悬浮细胞，将悬浮的细胞转移至5mL的消毒离心管中。加入250μL细胞裂解液并颠倒4次，以充分混合。孵育时间大约1-5分钟。再加入10μL碱性蛋白酶溶液并颠倒离心管4次以充分混合，于室温孵育5分钟。碱性蛋白酶能够灭活核酸酶以及其他细菌裂解过程中释放出的能够影响分离质粒质量的蛋白。然后向离心管加入350μL中和液并迅速颠倒4次以充分混合，于室温将离心管以最大速度(14000g)离心10分钟。
将上述离心后澄清的裂解液大约850μL转移到准备好的离心柱中，不要搅动或将任何白色沉淀与上清液一同转移。用离心机以最大速度于室温离心1分钟，从收集管上取下离心管并弃去收集管中的液体，将离心柱重新插入到收集管上。加入750μL先前用95％(W/W)的乙醇稀释过的柱清洗液。用离心机以最大速度于室温离心1分钟，从收集管上取下离心管并弃去收集管中的液体，将离心柱重新插入到收集管上。加入250μL柱清洗，重复清洗一次。用离心机以最大速度于室温离心2分钟。
将离心柱转移至一个灭菌的1.5mL离心管中，操作时需小心，不要将清洗液与离心柱一同转移。加入100μL去核酸酶的水，以洗脱质粒DNA(可等2分钟，便于完全溶解)，于室温用离心机以最大速度离心1分钟。洗脱后将离心柱取出，并弃之。合并离心液于-20℃条件下保存备用，并测其浓度。
(3)非洲爪蟾卵母细胞的分离和培养：
将手术器械在75％(W/W)的乙醇中浸泡30分钟，取出晾干。5号丝线在沸水中消毒10分钟。
将爪蟾掩埋于碎冰中约40分钟，使其麻醉。取出已麻醉的爪蟾，腹部向上置于平铺的碎冰上，并用碎冰掩埋其头部和四肢。用酒精棉球消毒下腹部皮肤，然后用针头挑起(正中偏左或右)，用眼科剪刀剪开1cm左右的小口。用同样的方法剪开肌肉层(注意不要损伤任何内脏器官，剪开肌肉层后即可看到卵母细胞)。用镊子和剪刀取出1cm 3大小的小叶，放入预先准备好的含有OR-2(含青霉素)的培养液皿中，分别缝合肌肉层和皮肤层。
将卵母细胞转移到无菌的玻璃管中重复用OR-2溶液清洗，至洗净残留的血液为止。加入胶原酶溶液振摇消化约1小时，更换新的胶原酶溶液继续振摇约1小时(此时可看到大多数已分离或单个细胞)。
除去消化液，用OR-2溶液清洗5-6次，转入盛有ND-96溶液的培养皿中挑选V期成熟的细胞置于ND-96溶液中，于1℃条件下用生化培养箱保存备用，每天换液一次。
(4)注入钙通道质粒：
将质粒α 1B，α 2δ，β 1b按照浓度1:1:1的比例注入爪蟾卵母细胞，每个细胞约注入的三种质粒的总体积为46nL。注入后的细胞置于ND-96培养液中，温度18℃放置培养48小时后记录表达的电流。
(5)电流记录：
灌流给药方法，流速为3mL/min，待测药物浓度为10μM。采用双电极电压钳技术，将细胞钳制在-100mV，以10mV为步阶，去极化至+60mV，记录电流。
N-型钙离子通道电流抑制实验结果见表3。
电流抑制率(％)＝100×(给药后电流幅度-给药前电流幅度)/给药前电流幅度
表3化合物抑制N-型钙离子通道电流实验结果(10μM)
Figure PCTCN2018072426-appb-000018
以上结果表明，本发明的化合物、其可药用盐对N-型钙离子通道具有较强的抑制作用。
测试例3：本发明化合物对P/Q-型钙通道、Herg通道、钠通道以及钾通道电流抑 制活性
(一)P/Q-型及Herg通道DNA的酶切线性化
1.P/Q-型钙通道各亚基(α 2δ、β 1b、α 1B、α 1E、α 1A、α 1C)和Herg通道DNA的来源同测试例2，按下列体系分别进行酶切：
Figure PCTCN2018072426-appb-000019
Figure PCTCN2018072426-appb-000020
质粒DNA按上述反应体系进行酶切，37℃下温育反应1小时。
2.DNA的纯化和去RNA酶:
a.在已酶切的质粒DNA样品中加入0.1倍体积的10×蛋白酶K缓冲液、0.1倍体积的5％SDS溶液、20mg/mL的蛋白酶K(终浓度为100μg/mL)，37℃下温育反应1小时；
b.将上述反应物转移至干净的1.5mLeppendorf管中，加入等体积的Tris平衡酚：氯仿：异戊醇(25：24：1)，振荡1分钟，室温下(20－25℃)以12000rpm的速度离心2分钟(若有机相和无机相不能充分分开，延长时间重新离心)；
c.小心将上层水相转移至另一干净的1.5mL eppendorf管中，弃去两相间的界面层和有机相；
d.加入等体积的氯仿：异戊醇(24：1)，室温下(20－25℃)以12000rpm的速 度离心2分钟；
e.小心将上层水相转移至另一干净的1.5mL eppendorf管中，加入0.1倍体积的3M醋酸钠(pH值为5.2)和2.5倍体积的95％(W/W)乙醇，振荡混匀，放置于-20℃中过夜(12－16小时)；
f.低温(<4℃)下以12000rpm的速度离心40分钟；
g.小心移出上清液，注意不要搅动沉淀，用吸头吸尽附于管壁的所有液滴；
h.在上一步所得沉淀中加入半管70％(W/W)乙醇，振荡混匀，低温(<4℃)下以12000rpm的速度离心2分钟；
i.小心移出上清液，重复上一步；
j.室温下(20－25℃)下将1.5mLeppendorf管敞口放置，直至残留液体挥干；
k.加入适当体积的去核酶水(Nuclease-free water)，充分漂洗管壁，混匀，使模板DNA完全溶解，测其浓度值后，保存于-20℃备用。
3.钙离子通道各亚基及HERG通道cDNA体外转录成cRNA [13,14]：
按照下列反应体系顺序加入各反应物：
α 1A、β 1b、α 2δ、α 1E、α 1A、α 1C亚基及Herg通道亚基的线性化模板；
T7Transcription 5×Buffer 20μL；
rNTPs(25mM ATP，CTP，UT各7.5μL+3mM GTP 0.6μL+去核酶水6.9μL)；
线性DNA模板(共5-10μg)+去核酶水32.5μL；
Ribom7帽子类似物，40mM，7.5μL；
MixE，10μL。
a.从恒温箱中取出反应物，加入RQ1Rnase-Free Dnase(按每μg模板DNA加1u酶的比例)，37℃下温育反应15分钟；
b.加入等体积的水平衡酚：氯仿：异戊醇(25：24：1)，振荡1分钟，室温下(20－25℃)以12000rpm的速度离心2分钟(若有机相和无机相不能充分分开，延长时间重新离心)；
c.小心将上层水相转移至另一干净的1.5mLeppendorf管中并加入等体积的氯仿：异戊醇(24：1)，室温下(20－25℃)以12000rpm的速度离心2分钟；
d.小心将上清液转移至另一干净的1.5mLeppendorf管中，加入0.1倍体积的3M醋酸钠(pH值为5.2)和2.5倍体积的95％(W/W)乙醇，振荡混匀，放置于-20℃ 中过夜(12－16小时)；
e.低温(<4℃)下以12000rpm的速度离心40分钟；
f.小心移出上清液，注意不要搅动沉淀，用吸头吸尽附于管壁的所有液滴；
g.在上一步所得沉淀中加入半管70％(W/W)乙醇；
h.小心移出上清液，重复上一步；
i.室温下(20－25℃)下将1.5mLeppendorf管敞口放置，直至残留液体挥干，加入适当体积的去核酶水(nuclease-free water)，充分漂洗管壁，混匀，使mRNA完全溶解，保存于-70℃备用。
4.非洲爪蟾卵母细胞的注射：
将专用注射针用两步法电极拉制仪拉制好后，在干净的薄面巾纸上刺一下，使针尖的口径变粗，并在抛光仪上抛光，使得针尖光滑平整。针尖的直径在6－10μm较为合适。安装注射用针之前，先在针内注射矿物油(Light mineral oil)以润滑管壁。根据原始各亚基cRNA的浓度，调整注射浓度大约为2ng/nL分别混合(1:1:1)。取1μL混合好的cRNA小心滴在干净的parafilm上，用微量注射仪将cRNA吸入到注射用针中。(注意：防止RNA酶的污染；防止空气被吸入到注射用针中，这两点均会影响cRNA在卵母细胞中的表达)。将挑选好的健康的Ⅴ、Ⅵ期成熟卵母细胞置入底部粘有网格(防止细胞在平皿中滑动)的培养皿中。仔细调节三维微操纵仪，使针尖接触细胞表面。若细胞状态良好，针尖接触细胞时会感到一定的阻力，同时可看到细胞膜上因张力形成的皱褶。穿刺细胞不易过深，以针尖刚刚穿过细胞膜为宜，将46.5nl cRNA注射入卵母细胞，注射后细胞会轻微鼓胀。等待30s之后，将注射针拔出，观察细胞是否有内容物溢出，如果有应放弃之。另外，若针尖极易刺入细胞时无突破感，表明细胞无张力，细胞状态不佳，意味着此细胞可能无法钳制到所需电位，也应放弃此细胞。将注射好的卵母细胞置于ND-96溶液中，在生化培养箱(18℃)培养48小时后(需每天更换ND-96液体)，即可分别记录表达的P/Q-型钙通道电流及Herg通道电流。
5.记录爪蟾表达的钙通道细胞外液及细胞内液：
细胞外液(mM)：BaCl 2 5，N-methyl-D-glucamine 50，KCl 5，HEPES 5，用methanesulfonic acid调pH值至7.4。
细胞内液为3mM的KCl。BAPTA的配制10mM Hepes溶解BAPTA，用CsOH调节pH值至7.2。
(二)原代培养的海马神经元
1.海马神经元培养：
取当天新生Wistar大鼠，用75％(W/W)乙醇消毒。在无菌条件下取脑，剥离出海马置于冰浴的解剖液中。将海马剪成1－2mm 3的组织块，用含0.25％胰蛋白酶的解剖液在37℃下消化30分钟，然后将消化好的组织块移入种植液中终止消化，在种植液中用适当口径(尖端直径2mm)的滴管吹打细胞，使之均匀分散，制成细胞悬液，取少量悬液以台盼蓝染液记数细胞。加入适量种植液，将细胞按1×10 5/mL的密度接种在事先涂有多聚赖氨酸的35mm培养皿中，置于36℃10％的二氧化碳培养箱中过夜，24小时后更换培养基，将种植液换为2mL饲养液。以后每3天半量换液一次，培养细胞在12－15天期间用于膜片钳实验。为抑制非神经元过度增殖，在培养的第3天向培养基中加入适量阿糖胞苷(每皿中加阿糖胞苷储备液6μL，终浓度为3μg/mL)。
2.记录海马神经元电流的电极内液和细胞外液：
全细胞记录所用的记录液主要是电极内液和细胞外液。
记录电极内液成份(mM)：KCl 140，HEPES 10，EGTA 10，调pH值为7.2－7.4。
细胞外液的成分(mM)：NaCl 140，KCl 5，MgCl 2 1，HEPES 10，Glucose 10，CaCl 2 3，用NaOH调pH值为7.2－7.4。如果实验需要阻断钠电流，可在细胞外液中加入河豚毒素(TTX)1μM。如果实验需要阻断钾电流，而只用CsCl又不能完全阻断钾电流时，可在电极内液和细胞外液中加入四乙基铵(TEA)或4-氨基吡啶(4-AP)。
测定结果如表4所示。
表4P/Q-型钙通道、Herg通道、钠通道和钾通道电流抑制结果
Figure PCTCN2018072426-appb-000021
“-”表示化合物对此通道电流无作用。
实验结果表明，本发明化合物、其可药用盐对P/Q-型钙通道、Herg通道、TTX敏感钠通道和电压门控钾通道均无阻断或抑制作用。
因此，本发明的化合物、其可药用盐能够特异性地抑制N-型钙离子通道。
测试例4：大鼠体内药代动力学快速评价
1.材料和仪器：
化合物(1)溶于25％(w/v)的羟丙基-β-环糊精水溶液中；
甲地孕酮：内标物，购自中检所；
羟丙基-β-环糊精：购自西安德立生物化工有限公司，配置成25％(w/v)的羟丙基-β-环糊精水溶液；
乙腈、甲醇：色谱纯，购自Fisher公司；
纯净水；
成年SD大鼠，体重180-220g，雄性，购自军事医学科学院动物中心，动物合格证SCXK-(军)-2012-0004；
液相色谱－质谱联用仪(LC-MS/MS)，三重四极杆串接LC-MS/MS(API 5000)；
LDZ5-2型离心机(北京医用离心机厂)；
MicroCL21R高速冷冻离心机(美国Thermo公司)；
JJ-1型精密增力电动搅拌器(常州国华电器有限公司)；
XW-80A型漩涡混合器(上海青浦沪西是仪器厂)；
Sartorius BS110S电子分析天平(北京赛多利斯天平有限公司)，Libror EB-330D电子分析天平(日本，岛津)。
2.标准曲线绘制：
用乙腈配制化合物(1)的母液1mg/mL，逐级稀释成系列浓度的标准溶液；取多份空白大鼠血浆0.05mL，分别加入系列浓度的标准溶液0.005mL以及0.3mL含内标物甲地孕酮(10ng/mL)的乙腈，化合物(1)的终浓度分别为1ng/mL、2ng/mL、5ng/mL、20ng/mL、100ng/mL、200ng/mL、500ng/mL、1000ng/mL、2000ng/mL，在漩涡混合器上充分涡旋1分钟，离心10分钟(14000xg)，取上清液约0.15mL于进样小瓶内，进样5μL应用LC-MS/MS检测分析。
质控样品：用化合物(1)的母液(1mg/mL)，逐级稀释成低、中、高三个浓度的标 准溶液。取多份空白大鼠血浆0.05mL，分别加入低、中、高三个浓度的标准溶液0.005mL以及0.3mL含内标物甲地孕酮(10ng/mL)的乙腈，化合物(1)的终浓度分别为2ng/mL，50ng/mL，1600ng/mL，作为化合物(1)的质控样品(每个浓度重复3个样品，n＝3)，在漩涡混合器上充分涡旋1分钟，离心10分钟(14000xg)，取上清液约0.15mL于进样小瓶内，进样5μL应用LC-MS/MS检测分析。
液相色谱操作条件：
色谱柱：Kromasil100-5C18，Dim 50×2.1mm，(Sweden)；
流动相组成：流动相A为水(含0.1％甲酸)，流动相B为乙腈(含10mM乙酸铵和0.1％甲酸)，梯度洗脱程序见表5。
表5流动相梯度洗脱程序
时间(分钟) 流速(mL/分钟) 流动相A(％) 流动相B(％)
0.5 0.6 90 10
2 0.6 5 95
2.5 0.6 5 95
2.6 0.6 90 10
4 0.6 90 10
MS/MS定量分析条件：
离子化方式为电喷雾(ESI)条件，正离子检测；
化合物(1)和内标物检测的m/z分别为427.168和385.2。选择性离子(SRM)方式检测m/z 427.168的碎片离子为m/z 250.1，CE为18；内标物m/z 385.2的碎片离子为m/z 267.3，CE为25。
将测定得到的峰面积相对于化合物(1)浓度绘制标准曲线，线性范围：1ng/mL-2000ng/mL，最低检测线1ng/mL，线性良好。质控样品检测浓度的精密度以及与真实浓度之间的准确度均在±15％以内，说明检测方法准确可靠。
3.大鼠体内PK快速评价：
6只成年SD大鼠，随机编号，分两组分别给予静注(i.v.，1.85mg/kg)和口服(p.o.，11mg/kg)的化合物(1)，于给药前与给药后2分钟、5分钟、15分钟、30分钟、60分钟、2小时、4小时、7小时和24小时采血0.1mL，置于抗凝管中，离心15分钟(3500rpm)分离血浆，准确取血浆0.05mL，置-30℃冰箱冻存备测。
向大鼠血浆样品0.05mL加入0.3mL含内标物甲地孕酮的乙腈(100ng/mL)和空白乙腈0.005mL，在漩涡混合器上充分涡旋1分钟，离心10分钟(14000xg)，取上清约0.15mL于进样小瓶内，进样5μL，参照上面2.的方法用LC-MS/MS检测分析，结合上面2.得到的标准曲线计算得到各时间点6只大鼠体内化合物(1)的浓度，如图1所示，其中，1#-3#为i.v.给药，4#-6#为p.o.给药。将1#-3#的结果取平均值，4#-6#的结果取平均值，得到i.v.给药和p.o.给药大鼠体内化合物(1)平均浓度和时间的关系曲线，如图2所示，应用Winnolin药代动力学软件对所测数据进行分析，计算主要药代动力学参数，包括静脉注射的曲线下面积(AUC)、清除率(CL)、表观分布容积(V)、末端消除半衰期(t 1/2)、平均驻留时间(MRT)以及口服给药的曲线下面积(AUC)、末端消除半衰期(t1/2)、平均驻留时间(MRT)、峰浓度(C max)、达峰时间(T max)和生物力度(F)，得到的药代参数如表6所示。
表6大鼠静注(1.85mg/kg)和口服(11mg/kg)化合物(1)后的药代参数(n＝3)
Figure PCTCN2018072426-appb-000022
大鼠体内药代动力学研究结果表明，化合物(1)经静脉注射给药后，在体内的平均清除率为2.38L/h/kg，略低于大鼠的肝血流量(3.3L/h/kg) [15]，提示肝脏代谢对化合物(1)的体内清除作用约70％；稳态表观分布容积V为12.39L/kg，远远大于大鼠血浆的容积(0.03L/kg) [15]及大鼠体内总水量(0.6L/kg) [15]，提示大部分药物主要分布到血管外组织中，在靶组织中暴露的几率也较大；口服给药后平均达峰时间为1-2h，提示药物肠道吸收良好，口服给药后体内药物暴露量相比，平均生物利用度为44.7％，代谢性质良好。综 上，该化合物作为镇痛药物，大鼠体内的药代动力学性质良好，可进一步进行相关研究，包括药物在稳态状态下脑组织靶部位的分布等。
测试例5：中枢神经系统分布评价
1.材料和仪器:同测试例4。
2.标准曲线绘制：
参照测试例4中2.的方法进行；
另外，将空白大鼠血浆替换为空白大鼠脑匀浆，绘制关于大鼠脑匀浆的化合物(1)标准曲线。
3.大鼠血浆及脑匀浆中化合物(1)的中枢神经系统分布评价：
化合物(1)用25％(w/v)的羟丙基-β环糊精水溶液配制成浓度为4mg/mL的溶液。成年SD大鼠3只，随机编号，分别先给与静推0.4mL的化合物(1)溶液，后缓慢滴注化合物(1)溶液(0.4mL/h)，连续两个小时。分别于给药前与给药后30分钟、60分钟和2小时采血0.1mL，置于抗凝管中，离心15分钟(3500rpm)分离血浆，准确取血浆样品0.05mL，置-30℃冰箱冻存备测。
2小时采血后股动脉放血致死后，取全脑及脊髓，用生理盐水冲洗干净后，滤纸吸干水分，置-30℃冰箱冻存备测。
向大鼠血浆样品0.05mL加入0.3mL含内标物甲地孕酮的乙腈(100ng/mL)和空白乙腈0.005mL，在漩涡混合器上充分涡旋1分钟，离心10分钟(14000xg)，取上清液约0.15mL于进样小瓶内，进样5μL，参照测试例4的方法用LC-MS/MS检测分析，结合上面2.得到的标准曲线计算得到大鼠血浆样品中化合物(1)浓度。
大鼠脑组织及脊髓样品称重后按照1:4(v/v)加入生理盐水制备匀浆，取匀浆样品0.05mL加入0.3mL含内标物甲地孕酮的乙腈(100ng/mL)和空白乙腈0.005mL，在漩涡混合器上充分涡旋1分钟，离心10分钟(14000xg)，取上清约0.15mL于进样小瓶内，进样5μL，参照测试例4的方法用LC-MS/MS检测分析，结合上面2.得到的标准曲线计算得到大鼠脑组织及脊髓中的化合物(1)浓度。
表7大鼠给药2h得到的血浆、大鼠脑组织、大鼠脊髓中的化合物(1)浓度(n＝3).
Figure PCTCN2018072426-appb-000023
Figure PCTCN2018072426-appb-000024
结果见表3。结果可见，化合物(1)在血浆中暴露后，容易透过血脑屏障进入脑组织和脊髓，在脑组织和脊髓中的含量是血浆中的3-4倍，作为中枢靶标的镇痛药物，这种分布特点有利于药效的发挥。
显然，上述实施例仅仅是为清楚地说明所作的举例，而并非对实施方式的限定。对于所属领域的普通技术人员来说，在上述说明的基础上还可以做出其它不同形式的变化或变动。这里无需也无法对所有的实施方式予以穷举。而由此所引伸出的显而易见的变化或变动仍处于本发明创造的保护范围之中。
参考文献：
[1]Newton PM,Messing RO.Channels,2009,3(2):77-81.
[2]Tomoyuki KONDA,Azusa ENOMOTO,Akira TAKAHARA,and hiroshi YAMAMOTO，Effects of L/N-Type Calcium Channel Antagonist,Cilnidipine on Progressive Renal Injuries in Dahl Salt-Sensitive Rats.Biol.Pharm.Bull.2006,29(5):933—937.
[3]TomoyukiKonda,Azusa Enomoto,etal.The N-and L-Type Calcium Channel Blocker Cilnidipine Suppresses Renal Injury in Dahl Rats Fed a high-Sucrose Diet.an Experimental Model of Metabolic Syndrome Nephron Physiol,2005,101:1–13.
[4]Ishikawa T,Nobukata h,etal.Effects of Ca 2+channel blocker cilnidipine in comparison with that of amLodipine and nifedipine on insulin resistance in Dahl S rats.Jpn Pharmacol Ther.1999,27,53-60.
[5]Yamamoto T,Takahara A.Current Topics in Med Chem.,2009,9:377-395.
[6]Teodori.J Med Chem,2004,47:6070-6081.
[7]Knutsen.Bioorg Med Chem Lett,2007,17(3):662-667.
[8]Yamamoto.Bioorg Med Chem,2006,14:5333-5339.
[9]Yamamoto.Bioorg Med Chem Lett,2008,18(17):4813-4816.
[10]Tyagarajan.Bioorg Med Chem Lett,2011,21(17):869-873.
[11]Zamponi GW,etal.Bioorg Med Chem Lett,2009,19:6467-6472.
[12]Pajouheshh,etal.Bioorg Med Chem Lett,2010,20:1378-1383.
[13]候云德.分子克隆实验指南(第二版)，科学出版社，2002.
[14]来茂德.医学分子生物学，人民卫生出版社，1999.
[15]Davis B,Morris T.Pharma Res,1993,10,1093-1095.

Claims (13)

  1. 式Ⅰ所示的化合物或其可药用盐：
    Figure PCTCN2018072426-appb-100001
    其中：
    g选自C 1-8亚烷基、取代的C 1-8亚烷基、羰基和C 1-8亚酰基；
    R选自C 5-20芳基、取代的C 5-20芳基、C 4-20杂环基和取代的C 4-20杂环基；
    所述取代独立地为被一个或多个取代基所取代，所述取代基选自卤素、C 1-8烷基、C 1-8烷氧基、C 3-8环烷基、氰基、硝基、巯基、甲硫基、乙硫基、三氟甲基、氨基、酰胺基、单C 1-8烷基氨基、二C 1-8烷基氨基、C 1-8烷基磺酰基、羟基、芳氧基、杂芳氧基和杂环基氧基。
  2. 根据权利要求1所述的化合物或其可药用盐，其中，
    g选自C 1-8亚烷基、羰基和C 1-8亚酰基；
    R选自C 5-20芳基、取代的C 5-20芳基和C 4-20杂环基；
    所述取代基选自C 1-8烷基、C 1-8烷氧基、C 3-8环烷基、巯基、甲硫基、乙硫基、氨基、单C 1-8烷基氨基、二C 1-8烷基氨基和羟基。
  3. 根据权利要求1所述的化合物或其可药用盐，其中，
    g选自C 1-8亚烷基、羰基和C 1-8亚酰基；
    R选自C 6-12芳基、取代的C 6-12芳基和C 4-12芳杂环基；
    所述取代基选自C 1-8烷氧基、甲硫基、乙硫基、氨基、单C 1-8烷基氨基和二C 1-8烷基氨基。
  4. 根据权利要求3所述的化合物或其可药用盐，其中，所述C 4-12芳杂环基为C 4-5 芳杂环基或C 6-12苯并杂环基。
  5. 根据权利要求1所述的化合物或其可药用盐，其中，
    g选自亚甲基、羰基、亚甲酰基、亚乙酰基、1,3-亚丙酰基、1,2-亚丙酰基、1,4-亚丁酰基、1,3-亚丁酰基和1,2-亚丁酰基；
    R选自苯基、取代苯基、噻吩基和苯并噁唑啉酮基；
    所述取代基选自甲氧基和二甲基氨基。
  6. 根据权利要求1所述的化合物或其可药用盐，其选自如下的化合物及其可药用盐：
    6-(3-(4,4-二苯基哌啶)-丙酰基)苯并噁唑啉-2-酮；
    6-(3-(4,4-二苯基哌啶)-乙酰基)苯并噁唑啉-2-酮；
    1-(4-二甲氨基苄基)-4,4-二苯基哌啶；
    1-(3,4,5-三甲氧基苄基)-4,4-二苯基哌啶；
    1-(2-甲基噻吩基)-4,4-二苯基哌啶；
    4,4-二苯基-1-苄基哌啶。
  7. 一种药物组合物，包含权利要求1至6中任一所述的化合物或其可药用盐；
    可选地，还包含药学上可接受的辅料。
  8. 一种在体内或体外阻断或抑制N型钙离子通道的方法，包括给予有需求的受试者以有效量的权利要求1至6中任一所述的化合物或其可药用盐或者给予有需求的受试者以有效量的权利要求7所述的药物组合物的步骤。
  9. 权利要求1至6中任一所述的化合物或其可药用盐或者权利要求7所述的药物组合物在制备N型钙离子通道阻断剂或抑制剂中的用途。
  10. 权利要求1至6中任一所述的化合物或其可药用盐或者权利要求7所述的药物组合物在制备预防或治疗疼痛、中风、脑缺血、酒精成瘾、酒精中毒、肾病、由镇痛药物引起的成瘾性病症或由镇痛药物引起的耐受性病症的药物中的用途；例如，所 述疼痛为手术后疼痛、偏头痛、内脏痛或神经性疼痛；例如，所述肾病为急性肾衰竭、慢性肾衰竭或肾机能不全。
  11. 一种预防或治疗疼痛、中风、脑缺血、酒精成瘾、酒精中毒、肾病、由镇痛药物引起的成瘾性病症或由镇痛药物引起的耐受性病症的方法，包括给予有需求的受试者以有效量的权利要求1至6中任一所述的化合物或其可药用盐或者权利要求7所述的药物组合物的步骤；例如，所述疼痛为手术后疼痛、偏头痛、内脏痛或神经性疼痛；例如，所述肾病为急性肾衰竭、慢性肾衰竭或肾机能不全。
  12. 权利要求1至6中任一所述的化合物或其可药用盐或者权利要求7所述的药物组合物，其用于阻断或抑制N型钙离子通道。
  13. 权利要求1至6中任一所述的化合物或其可药用盐或者权利要求7所述的药物组合物，其用于预防或治疗疼痛、中风、脑缺血、酒精成瘾、酒精中毒、肾病、由镇痛药物引起的成瘾性病症或由镇痛药物引起的耐受性病症；例如，所述疼痛为手术后疼痛、偏头痛、内脏痛或神经性疼痛；例如，所述肾病为急性肾衰竭、慢性肾衰竭或肾机能不全。
PCT/CN2018/072426 2017-01-13 2018-01-12 4,4-二苯基哌啶类化合物或其可药用盐、药物组合物及用途 WO2018130207A1 (zh)

Priority Applications (3)

Application Number Priority Date Filing Date Title
KR1020197023802A KR102507380B1 (ko) 2017-01-13 2018-01-12 4,4-디페닐피페리딘 화합물 또는 이의 약학적으로 허용 가능한 염, 약학 조성물 및 이의 용도
JP2019559149A JP7068338B2 (ja) 2017-01-13 2018-01-12 ４，４－ジフェニルピペリジン化合物又はその医薬的に許容される塩、医薬組成物及びそれらの使用。
US16/477,655 US10889575B2 (en) 2017-01-13 2018-01-12 4,4-diphenylpiperidine compounds or pharmaceutically acceptable salts thereof, pharmaceutical compositions and uses thereof

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201710023076.XA CN108299411B (zh) 2017-01-13 2017-01-13 4,4-二苯基哌啶类化合物或其可药用盐、药物组合物及用途
CN201710023076.X 2017-01-13

Publications (1)

Publication Number Publication Date
WO2018130207A1 true WO2018130207A1 (zh) 2018-07-19

Family

ID=62840085

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2018/072426 WO2018130207A1 (zh) 2017-01-13 2018-01-12 4,4-二苯基哌啶类化合物或其可药用盐、药物组合物及用途

Country Status (5)

Country Link
US (1) US10889575B2 (zh)
JP (1) JP7068338B2 (zh)
KR (1) KR102507380B1 (zh)
CN (1) CN108299411B (zh)
WO (1) WO2018130207A1 (zh)

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4016280A (en) * 1969-07-17 1977-04-05 Byk Gulden Lomberg Chemische Fabrik Gmbh 4,4-Diarylpiperidine compositions and use
WO2001095856A2 (en) * 2000-06-15 2001-12-20 Chaconne Nsi Co., Ltd. Urea derivative useful as an anti-cancer agent and process for preparing same
CN1361775A (zh) * 1999-07-21 2002-07-31 阿斯特拉曾尼卡英国有限公司 新的二苯基哌啶衍生物
US20040204404A1 (en) * 2002-09-30 2004-10-14 Robert Zelle Human N-type calcium channel blockers
WO2008031227A1 (en) * 2006-09-14 2008-03-20 Neuromed Pharmaceuticals Ltd. Diaryl piperidine compounds as calcium channel blockers
WO2008043183A1 (en) * 2006-10-13 2008-04-17 Neuromed Pharmaceuticals Ltd. Cyclopropyl-piperazine compounds as calcium channel blockers
CN102239146A (zh) * 2008-10-02 2011-11-09 雅培制药有限公司 作为钙通道阻断剂的新化合物

Family Cites Families (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE1966199A1 (de) * 1969-07-17 1971-11-04 Promonta Chem Fab 4,4-Diarylpiperidine
US4022786A (en) * 1969-07-17 1977-05-10 Byk Gulden Lomberg Chemische Fabrik Gmbh 4,4-Diarylpiperidines and process of making the same
IL90858A (en) * 1988-07-07 1994-08-26 Rhone Poulenc Sante History (Aza) Naftalensultam, their preparation and preparations containing them
DE19756235A1 (de) * 1997-12-17 1999-07-01 Klinge Co Chem Pharm Fab Neue piperidinylsubstituierte Pyridylalkan- alken- und -alkincarbonsäureamide
SI2208727T1 (sl) * 2002-09-19 2012-11-30 Lilly Co Eli Diaril etri kot antagonisti opioidnega receptorja
JP2008540395A (ja) 2005-05-03 2008-11-20 ファイザー・インク アミドレソルシノール化合物
JP2009532381A (ja) * 2006-03-31 2009-09-10 アストラゼネカ アクチボラグ 二環式ベンズイミダゾール化合物、および代謝型グルタミン酸受容体増強剤としての該化合物の使用
US9132740B2 (en) * 2009-06-29 2015-09-15 Ford Global Technologies, Llc Automotive exterior lamp assembly with charge port
ES2804545T3 (es) * 2013-03-11 2021-02-08 Univ Temple Nuevos moduladores de la actividad del receptor de 5-hidroxitriptamina 7 y su método de uso
WO2016183150A1 (en) * 2015-05-12 2016-11-17 Temple University-Of The Commonwealth System Of Higher Education Novel sigma-2 receptor binders and their method of use

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4016280A (en) * 1969-07-17 1977-04-05 Byk Gulden Lomberg Chemische Fabrik Gmbh 4,4-Diarylpiperidine compositions and use
CN1361775A (zh) * 1999-07-21 2002-07-31 阿斯特拉曾尼卡英国有限公司 新的二苯基哌啶衍生物
WO2001095856A2 (en) * 2000-06-15 2001-12-20 Chaconne Nsi Co., Ltd. Urea derivative useful as an anti-cancer agent and process for preparing same
US20040204404A1 (en) * 2002-09-30 2004-10-14 Robert Zelle Human N-type calcium channel blockers
WO2008031227A1 (en) * 2006-09-14 2008-03-20 Neuromed Pharmaceuticals Ltd. Diaryl piperidine compounds as calcium channel blockers
WO2008043183A1 (en) * 2006-10-13 2008-04-17 Neuromed Pharmaceuticals Ltd. Cyclopropyl-piperazine compounds as calcium channel blockers
CN102239146A (zh) * 2008-10-02 2011-11-09 雅培制药有限公司 作为钙通道阻断剂的新化合物

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
DATABASE C AS 24 October 2004 (2004-10-24), retrieved from STN Database accession no. 768332-29-2 *
DATABASE CAS 13 June 2004 (2004-06-13), retrieved from STN Database accession no. 692717-79-6 *
DOULGAS A. KLUMPP: "Dicationic Intermediates Involving Protonated Amides: Dual Modes of Reactivity Including the Acylation of Arenes", ORGANIC LETTERS, vol. 6, no. 11, 29 April 2004 (2004-04-29), pages 1789 - 1792, XP055507909, ISSN: 1523-7060 *

Also Published As

Publication number Publication date
CN108299411B (zh) 2021-02-05
US20190359604A1 (en) 2019-11-28
KR102507380B1 (ko) 2023-03-07
US10889575B2 (en) 2021-01-12
JP7068338B2 (ja) 2022-05-16
CN108299411A (zh) 2018-07-20
KR20190103385A (ko) 2019-09-04
JP2020505449A (ja) 2020-02-20

Similar Documents

Publication Publication Date Title
CN105814067B (zh) 作为钠通道调节剂的吡啶酮酰胺的前药
JP5161774B2 (ja) Ａｍｐｋ活性化物質としてのチエノピリドン誘導体の使用およびこれを含む医薬組成物
TWI304062B (en) N-(pyridin-2-yl)-sulfonamide derivatives
TWI380818B (zh) Ｇ蛋白偶聯型受體（ｇｐｃｒ）激動劑
AU2014267974B2 (en) Cycloalkyl acid derivative, preparation method thereof, and pharmaceutical application thereof
JP2017002085A (ja) スピロ−オキシインドール化合物のエナンチオマーおよび治療剤としてのその使用
KR101009554B1 (ko) Ｐｄｅ７ 억제제로서 스피로시클릭 퀴나졸린 유도체
BG104432A (bg) Терапевтично използване на хинолинови производни
JPH07119222B2 (ja) Ｒ（＋）―テラゾシン
TW200410675A (en) An NF-κB activation inhibitor
JP2001511767A (ja) 糖尿病薬
KR20220078644A (ko) 미토콘드리아 질환 치료를 위한 바티퀴논의 퀴논-, 히드로퀴논- 및 나프토퀴논-유사체
TWI552750B (zh) 作為ｋｃｎｑ２／３鉀離子通道調節劑之經取代之２－氧基－及2－硫基－二氫喹啉－３－羧醯胺
US9751847B2 (en) Methods and compositions related to neuroactive thiazoline compounds
WO2018130207A1 (zh) 4,4-二苯基哌啶类化合物或其可药用盐、药物组合物及用途
WO2010040315A1 (zh) 1-丁基-2-羟基芳烷哌嗪衍生物及作为抗抑郁剂的应用
CN101454317A (zh) 作为一氧化氮合酶二聚化抑制剂的咪唑衍生物
JP6595011B2 (ja) 新規ホスホジエステラーゼ５阻害剤とその使用
TW201736366A (zh) 吲嗪類衍生物、組合物及使用方法
CN101321738A (zh) 螺环衍生物
CN112457265A (zh) 四氮唑类衍生物、制备、含其的药物组合物及其应用
TW200303744A (en) Water soluble phenylglycine derivatives
CN114478359B (zh) 氨基甲酸酯类trpv1拮抗/faah抑制双靶点药物及其制备方法和应用
TW201125860A (en) Prodrugs of [4-[4-(5-aminomethyl-2-fluoro-phenyl)-piperidin-1-yl]-(1H-pyrrolo-pyridin-yl)-methanones and synthesis thereof
US9737517B2 (en) Pharmaceutical use of hexahydro-dibenzo[a,g]quinolizine compounds

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 18738843

Country of ref document: EP

Kind code of ref document: A1

ENP Entry into the national phase

Ref document number: 2019559149

Country of ref document: JP

Kind code of ref document: A

NENP Non-entry into the national phase

Ref country code: DE

ENP Entry into the national phase

Ref document number: 20197023802

Country of ref document: KR

Kind code of ref document: A

122 Ep: pct application non-entry in european phase

Ref document number: 18738843

Country of ref document: EP

Kind code of ref document: A1