TWI334351B - Substituted 4-phenyl-4-[1h-imidazol-2-yl]-piperidine derivatives for reducing ischaemic damage - Google Patents

Description

1334351 A7 B7 V. INSTRUCTION DESCRIPTION (1) ' Scope of the invention The present invention relates to 4-phenyl-4-[1H-imidazol-2-yl]-hexahydropyridine derivatives for reducing ischemic injury of organs And a pharmaceutical composition comprising the hexahydropyridine derivative and the hexahydrogen. ratio. Derivatives are used for the prevention and treatment of 5 organ ischemic injuries, especially for the reduction of cardiac and cerebral ischemic injuries. BACKGROUND OF THE INVENTION - ~... In the framework of this application, a lack of an enterprise is defined as reducing or losing blood flow to the tissue, and its associated reduction or loss of oxygen supply to the group. In the framework of the present application, ischemic injury is defined as a negative effect associated with an ischemic event, such as ischemic necrosis or infarction, where metabolic events under cellular degeneration and cell death include: energy consumed via ATP Failure, cytotoxicity, glutamate release, calcium pooling, stimulation of 15 membrane phospholipid metamorphism and subsequent free fatty acid aggregation, and free radical production. · Increased demand for compounds that provide protection against ischemia and its associated side effects. The Ministry of Economic Affairs, the Intellectual Property Office, and the Consumer Cooperatives, have recently discovered that some highly specific Deta-2 opioid receptor agents 20 can provide prolonged pharmacology of the myocardium via a method similar to that occurring in pre-ischemic conditions (IPC). Induced ischemic protection (00乂丨11 (135% 3111161 & 丨., in Proceedings of the 11th International Hibernation Symposium 2000, pp. 377-384, Springer-Verlag, Berlin, Germany), pre-ischemic conditions depicting the heart Short-term pre-ischemic conditions make this paper scale applicable to China National Standard (CNS) A4 specification (210 χ 297 mm) Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed 1334351 A7 B7 V. Invention Description (2) Blood causes less damage, which in turn leads to less myocardial infarction and less arrhythmia, the mechanism of which is based on the function of improved mitochondrial ATP-sensitive K-channel (mitoKATP) (open), a It is known that DADLE (D-Ala2-D-Leu5-enkephalin) 5 is shown to cause the same effect on the organ as before the ischemic condition, therefore, pre-ischemic condition and chemical The mode of action of the substance must be viewed as a starting point to activate a more basic or protective cell pattern, as the German opioid receptor is found in most tissues, including the heart and brain tissue, and is expected to exhibit ischemic protection such as cardiac tissue. Compounds also protect brain tissue, as well as, for example, lung, kidney or liver tissue. Currently, ischemia plays an important role in two major medical aspects: cardiac ischemia and cerebral ischemia or stroke. 'Heart ischemia heart surgery has been Associated with controlled episodes of one or more ischemia and reperfusion 15 episodes, in traditional cardiac surgery, the heart stops and is cooled with a cardiac paralysis solution to reduce myocardial oxygen consumption, allowing the heart to have longer periods of ischemia without Too much damage, but this requires maintaining blood circulation through the external circulatory system, which has several major drawbacks: it causes a pronounced inflammatory body reaction that causes microembolism and it completely interferes with the condensation of the blood 20 and the fibrinolytic system, Moreover, during the surgery of the external circulation, microembolism and non-pulsation cycles with vital organs such as the brain and kidney The most moderate perfusion of the small intestine is associated with, for example, increased anaerobic metabolism (increased lactate during the postoperative period), impaired renal function, and mental confusion. Due to the development of local (mechanical) stabilizers, in the past few years to avoid -4- This paper scale applies to China National Standard (CNS) A4 specification (210x297 mm)

Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed 1334351 A7 B7 V. Description of invention (3) The above shortcomings, coronary surgery is not carried out by means of external circulation, but the disadvantage is that the heart maintains normal body temperature and must perform mechanical work while forcing the area Ischemia, at present, more than 30% of coronary surgery in the world is performed without the use of external circulation. For this application, cardioprotective agents are particularly useful, and such agents must be able to exert protective effects on myocardial tissue using basal cell mechanisms. Thus prolonged forced ischemia time. In addition, this application has a potential area of preservation in donor hearts, which is still a problem at present, as acceptable ischemic time is still limited to 4 to 6 hours, and cardiac surgery requires complex reconstruction, except for 10 Before the use of cardiac paralysis stopped, cardioprotective agents are beneficial for long-term surgical cardiac ischemia. In general, the present application has potential fields in methods of ischemic-reperfusion sequence forcing all surgical and percutaneous interventions in any organ, such as transplantation surgery, aneurysm surgery, vascular surgery for obstructive vascular disease, and 15 percutaneous Intervene in the narrow coronary arteries, carotid arteries and peripheral arteries. In particular, the present application has potential areas for patients before anesthesia for any reason, where it is appropriate to reduce blood supply to the organ, such as in stable and restless angina, or via hemodynamic effects of anesthesia. The resulting conditions, such as loss of blood pressure, and the patient who developed the blood clot during the first hour after the heart attack. _· The cerebral ischemic brain is more than any other organ in the body. Its survival and normal function are determined by a fairly stable supply of oxygenated blood. Although it only accounts for 2% of body weight, the brain receives 15% of the heart's blood and consumes 20% of the body. Oxygen used This paper scale applies to China National Standard (CNS) A4 specification (210x297 mm)

1334351 A7 B7 V. Inventive Note (4) In addition, fixed supply of blood is required to provide glucose to the brain, and the primary energy source used by the brain is to produce high energy phosphates such as ATP (see, for example, WO 96/27380 (Interneuron Pharmaceuticals, Inc.)). In the framework of this application, cerebral ischemia is defined as the interruption or reduction of blood flow from the arteries to the brain, usually because blood clots (thrombus) or other substances (embolism) block the arteries, leading to ischemic stroke, according to this article. Definitions, ischemic stroke is a syndrome caused by a variety of causes, such as atherosclerotic cerebrovascular diseases such as hypoperfusion and arterial embolism, penetrating arterial disease, cardiac embolism such as but not limited to atrial fibers 10 tremors, membranous disease and ventricular thrombosis, cryptogenic stroke, and other more common causes of sputum such as thrombin status, anatomy, arteritis, migraine or vascular dysplasia and drug abuse (see for example CarcZ/ovascw/ar 77z /-〇m 厶edited by M-

Verstraete, V. Fuster and E.J. Topol, Second Edition, 15 Lippincot-Raven Publishers, Philadelphia, 1998). Stroke is the third cause of death in the United States, and about 500,000 new cases occur each year. Due to the high incidence of stroke in Asia, the Ministry of Economic Affairs' Intellectual Property Bureau employee consumption cooperatives print strokes, which is the leading cause of death in the world. Ischemic stroke is the most common form of stroke and is associated with approximately 85 percent of all strokes. 20 This application has potential areas in wind prevention in some cases, such as the surgical procedure for the risk of jk deficiency, the reduction of ischemic injury in the case of stroke, the reduction of cerebral infarction after cerebral ischemia, and the lack of treatment. Bloody stroke, especially in acute treatment of stroke after an ischemic event. Background of Previous Techniques-6- This paper scale applies to National Standards (CNS) A4 (210x297 mm) Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperatives Printed 1334351 Α7 Β7 V. Invention Description (ο WO 99/04795 ( Toray Industries Inc.) discloses a tetracyclic pyridine and pyridinium derivative having pharmacological effects of an opioid receptor agent and its use for reducing organ ischemic injury, wherein the disclosed compound and the compound of the present invention are not Structural Correlation. 5 WO 00/37470 (Janssen Pharmaceutica NV) discloses a variety of 4-phenyl-4-[1Η-imidazol-2-yl]-hexahydropyridine derivatives, but does not form part of the invention and serves as The intermediate is used to synthesize a compound resistant to histamine. Inventive Note 10 In the present application, based on a substituted 4-phenyl-4-[1Η-imidazol-2-yl]-hexahydro-β ratio derivative, Revealing a novel compound that has important clinical utility in reducing ischemic injury to organs in drinking milk animals, especially in the heart and brain tissue, particularly in mammals, by inducing cardioprotective effects to prevent complications And with Caused by coronary artery 15-disease. The subject of the present invention is an agent for reducing organ ischemic injury in a mammal, which comprises as an active ingredient a substituted 4-benzene according to formula (I) 4-[1Η-imidazol-2-yl]-hexahydropyridine derivative

This paper scale applies to the Chinese National Standard (CNS) A4 specification (2丨Οχ 297 mm)

1334351 A7 B7 V. INSTRUCTIONS (6) A pharmaceutically acceptable acid or base addition salt thereof, a stereochemically isomeric form thereof, a tautomeric form thereof and an N-oxide form thereof, wherein: A= B is a divalent π-bond group; X is a covalent bond, -CH2- or CH2CH2-; 5 R1 is hydrogen, alkoxy, alkylcarbonyloxy, Ar-oxy, Het-oxy, Ar-carbonyl , Het-carbonyloxy, Ar-alkoxy, Het-alkoxy, alkyl, polydentate, methoxyl, Ar-alkyl, Het-alkyl, Ar, Het, thio Sulfur-based, Ar-thio, Het-thio or NR9R10, wherein R9 and R10 are each independently hydrogen, alkyl, Ar, 1〇Ar-alkyl, Het, Het-alkyl, Ar-. , Het-single base bite · ester alkyl group; or A = B and R1 - forming a substituted semi-aromatic or aromatic carbocyclic or heterocyclic ring Het2 or Het3; R2 is a hydroxyl group, alkoxy group , alkoxycarbonyl, phenoxy, phenyloxyoxyhalyl, cyano, alkyl, polydentate alkyl, alkoxyalkyl, formazanyl, carboxy, carbonyl, azide, amine a carbonyl group, a mono-n-alkylaminocarbonyl group, a phenyl group, a nitro group, an amine group, a mono or a diamined amine Alkylthio; the R3 of the Intellectual Property Office of the Ministry of Economic Affairs printed by the Consumer Cooperatives is R, G, Ar, Ar-alkyl, Ar-diyl, Ar-hetero-Het-alkyl, Het-alkenyl or Het-carbonyl; 20 R4, R5 are each independently hydrogen, alkyl, carboxy 'aminocarbonyl, alkyl ester. Its halogen or hydroxyalkyl group; i P is an integer equal to 0, 1, 2 or 3. In the organization of the present application, the alkyl group is a linear or branched saturated hydrocarbon group having from 丨 to 6 carbon atoms; or a cyclic saturated group containing from 3 to 7 carbon atoms.

Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperative Printed 1334351 A7 B7 V. Inventive Note (7) and Hydrocarbyl (cycloalkyl); or a cyclic saturated hydrocarbon group containing from 3 to 7 carbon atoms to be attached to from 1 to 6 a linear or branched saturated hydrocarbon group of one carbon atom; wherein each carbon atom may be optionally substituted with an amine group, a nitro group, a thio group, a hydroxyl group, an oxy group, a cyano group, a decyl group or a carboxyl group, and a preferred alkyl group is a Base, ethyl 5, propyl, isopropyl, butyl, tert-butyl, cyclopropyl, cyclopentyl, decyl, cyclohexylmethyl and cyclohexylethyl. In the organization of the present application, an alkenyl group is a hospital group having one or more double bonds according to the above definition, and a preferred dilute group is a vinyl group and an acryl group. In the organization of the present application, Ar is selected from the group consisting of a 10 carbocyclic ring comprising a phenyl group and a naphthyl group, each optionally substituted with one or more substituents, each substituent being independently selected from the group consisting of a hydroxyl group, an alkoxy group, Alkylcarbonyloxy, phenoxy, phenylcarbonyloxy, halo, cyano, alkyl, polyhaloalkyl, alkoxyalkyl, mercapto, methylidene, carboxy, alkylcarbonyl, alkyl ester Alkyl, aminocarbonyl, mono or monoamine, phenyl, phenyl, decyl, amine, mono 15 or dialkylamino, thio, alkylthio or S〇2_CH3, preferably Ar is a naphthyl group or a phenyl group, each optionally having a hydroxyl group, a methoxy group, an ethoxy group, a phenoxy group, a trihalomethoxy group, a dentate group, a methyl group, a trifluoromethyl group, a gas methyl group, a carboxyl group. , a methyl ester group, an ethyl ester group, a diethylaminocarbonyl group, a strepyl group, a nitro group, a methylthio group, a trifluoromethoxy group or an SC^-Cu alkyl group. 20 In the organization of the present application, the functional group is selected from a substituent including fluorine, gas V bromine and hydrazine, and the poly-group is a hydrocarbon group saturated with a straight bond or a branched chain of _ carbon atoms or contains from 3 to A cyclic saturated nicotine group of 7 carbon atoms, wherein one or more carbon atoms are replaced by - or a plurality of self-atomic atoms, preferably a desert, fluorine or gas, and a preferred polyterpenoid is trimethylmethyl. . -9- This paper scale applies to China National Standard (CNS) A4; ^7i7〇 297 297 ~ -----

Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperative Printed 1334451 A7 B7 V. INSTRUCTIONS (o In the organization of the present application, Het is selected from heterocyclic groups including Het1, Het2 and Het3, and Het1 is selected from pyrrolidinyl groups. , dioxin, imidazolium, pyrazolyl, hexahydropyridyl, decyl, dioxy, morphine 11 linyl, dithiazide, thiofolfonyl, hexahydropyrryl and tetrahydrofuranyl 5 aliphatic monocyclic heterocyclic group, Het2 is a semi-aromatic monocyclic heterocyclic group selected from the group consisting of 2H-pyrrolyl, pyrrolinyl, imidazolium and pyrazolyl, and Het3 is selected from the group consisting of batch β , 吼α, 咪, 17, 坐, 吱, π, D, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyrimidinyl, pyridinyl, argon Or a tri-negative aromatic monocyclic heterocyclic group; or 10 selected from the group consisting of fluorenyl, 喳哼σ-linyl, fluorenyl, benzoimidazolyl, benzo-α. Aromatic bicyclic heterocyclic groups of the basic, stupid and 嗟β-based, stupid and iso-indolyl, benzofuranyl and benzothienyl groups; each monocyclic and bicyclic heterocyclic group may be optionally Substituted on a carbon and/or hetero atom by a halogen group, a hydroxyl group, a hydroxy group, a decyl group, an Ar, an Ar-hospital group or a ratio of α to a group. 15 A valuable compound is a compound according to formula (I), a pharmaceutically acceptable acid or base addition salt, a stereochemically isomeric form thereof, an tautomeric form thereof, and a quinone-oxide form thereof, wherein Α=Β is selected from the group consisting of C=0, C= N-R6 wherein R6 is hydrogen or cyano, C=S, S=0, S02 and C=CR7R8 wherein R7 and R8 are each independently hydrogen, nitro or alkyl. 20 Other valuable compounds are according to formula ( a compound of I), a pharmaceutically acceptable acid or base addition salt thereof, a stereochemically isomeric form thereof, a tautomeric form thereof, and an N-oxide form thereof, wherein R1 is selected from the group consisting of alkoxy groups , Ar-alkoxy, alkyl, polyalkyl, oxyalkyl, Ar-hospital, Het-alkyl, Ar, hexahydrogen ratio, well base, 吼° base, 嗟-10- This paper scale applies to the Chinese National Standard (CNS) A4 specification (210 X 297 mm)

Ministry of Economic Affairs, Intellectual Property Bureau, Staff and Consumers Co., Ltd. Printed 1334351 A7 B7 V. Description of Invention (9). Azolyl, pyrrolidinyl and NR9RIG wherein R9 and R1G are each independently hydrogen, alkyl, Ar, Ar-alkyl, Hey. Base or burnt base. Other valuable compounds are compounds according to formula (I), pharmaceutically acceptable acid or base addition salts thereof, stereochemically isomeric forms thereof, 5 tautomeric forms thereof and their N-oxide forms Wherein A=B and R1 together form a group selected from the group consisting of Het2 and Het3, and more preferably A=B and R1 are formed to be selected from the group consisting of benzoxazolyl, thiazolyl, benzothiazolyl, and cumin Base and mouth. Set the foundation. Still other valuable compounds are those according to formula (I), their pharmaceutically acceptable acid or base addition salts, their stereochemically isomeric forms, their tautomeric forms and their N-oxidation. a form wherein X is a covalent bond or a -CH2- group, and preferably X is a covalent bond. ' Further valuable compounds are compounds according to formula (I), pharmaceutically acceptable acid or base addition salts thereof, stereochemically isomeric forms thereof, tautomeric forms thereof and their N-oxidation Form of matter wherein R2 is a oxy group or a group. 1 Further valuable compound is a compound according to formula (I), a pharmaceutically acceptable acid or base addition salt thereof, a stereochemically isomeric form thereof, a tautomeric form thereof and an N-oxide thereof Form wherein R3 is selected from the group consisting of phenylalkyl and naphthyl, each independently substituted with at least one substituent selected from the group consisting of a halo group, an alkyl ester group, a hydroxyl group, an alkoxy group, and a dialkylaminocarbonyl group. When R3 is an alkyl group, the preferred alkyl group is a cyclohexylfluorenyl group. Other valuable compounds are compounds according to formula (I), their drugs -11- This paper scale applies to China National Standard (CNS) A4 specification (210x297 mm)

334351 Α7 _ Β7 真, invention description (IG). A scientifically acceptable acid or base addition salt, a stereochemically isomeric form thereof, a mutual wheat isomer form and an N-oxide form thereof, among which 〇〇 or S02, R is a pitoxyoxyalkyl group & or Nr9r1 wherein R and R are each independently hydrogen or Ar; or eight = 8 and 艮1 together form a stupid 5 oxazolyl group, P Is 0, r3 is a benzyl group optionally substituted with a hydroxy group, an alkyl group or an alkyl ester group, and R4 and R5 are each hydrogen. More specifically, the following compounds are the most preferred compounds: phenethyl 4-phenylphenyl hydrazino) _1H-imidazole _2 yl] hexahydropyryl; 1 〇 1-propyl phenyl Styloinyl)-1Η-imidazol-2-yl]-hexahydroindole 0; b-ethyl 4-pyridyl-4-[l-[(4-hydroxyphenyl)methyl]-1H- Imidazolyl-2-yl]-hexahydrogen ratio. 1-ethyl ester-4-phenylphenylethyl)-lH-imidazol-2-yl]-hexahydro 15 . Ratio of α; 4 isopropyl ester-4-phenyl-4-[1-(phenylmethyl)-lH-imidazol-2-yl]-hexahydro. ratio. Butyl; ethyl 4-phenyl]-4-[1-[[4-(methyl)phenyl]methyl]-1Η-imidazole - Ministry of Economic Affairs, Intellectual Property Office, Staff Consumer Cooperative, Printed 2-Base ]- Hexazapine. 20 1-Benzylmercapto-4-phenyl-4-[1-(phenylmethyl)-1Η-flavor. 2-yl]-hexahydropyridine; 1-(methoxyethenyl)-4-phenyl-4-[1-(1-phenylethyl)-1Η-imidazol-2-yl] - hexahydropine bite; 4-[[2-(1-benzoylmethyl-4-phenyl-4-pyridinyl)-1Η-imidazol-1-yl] -12- This paper size applies _ country National Standard (CNS) A4 Specification (210x297 mm) 1334351 A7 B7

Mercapto]-mercaptophenyl phthalate; 4-[[2-[1-(2-benzopyrene<1 yl)-4-phenyl-4-hexahydro). ratio. Stationary)-1^;-imidol 1-yl]methyl]-methylbenzoate; 1-benzylidene-4-phenyl-4-[l-(l-phenylethyl)_ih -imidazol-2-yl] 5 hydropyridine; 1-ethyl ester 4·phenyl-4-[1-[1-[4-(ethyl)phenyl]ethyl]-1H-isoxazole- 2-yl]-hexahydropyridine; and hydrazine, 4-diphenyl_4_[1-(stylmethylmethane.sodium-2-yl)-1-hexahydroindole. An acceptable acid addition salt is a medically active non-toxic acid addition salt form which may be formed by the inclusion of a compound of formula (I), which may be obtained from a suitable form according to formula (I). Acid treatment, such as inorganic acids such as hydrohalic acid, especially hydrogen acid, hydrobromic acid, sulfuric acid, nitric acid and acid; organic acids such as acetic acid, glycolic acid, propionic acid, lactic acid 15, pyruvic acid, oxalic acid, malonic acid, Succinic acid, maleic acid, fumaric acid « , malic acid, tartaric acid, citric acid, mandelic acid, pseudo sulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluic acid, cyclohexyl sulfonic acid, salicylic acid , Aminosalicylic acid and Bamo acid. Ministry of Economic Affairs, Intellectual Property Bureau, Staff Consumer Cooperative, printed with acid The compound according to formula (1) can also be converted into its medically active non-toxic base addition salt form by treatment with an appropriate organic and 20 inorganic treatment. Suitable base salt forms include, for example, ammonium salts, alkali metal and alkaline earth metal salts, especially Mercury, sodium, potassium, magnesium and dance salts, with organic bases such as ethylenedibenzylamine 'N-mercapto-D-reducing glucosamine salt, sea salt of hybamine and with amino acids such as arginine And the salt of lysine. -13- The paper size is applicable to the National Solidarity Standard (CNS) A4 specification (2丨〇X 297 public) 1334351 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed A7 B7 V. Invention Description (Instead, the acid or base addition salt form can be converted to the free form by treatment with a suitable base or acid. The noun addition salts used in the context of the present application also include compounds according to formula (I) and a solvate from which a salt can be formed, such as a hydrate and an alcoholate. The term "stereochemically isomeric form" as used herein, defines that the compound of formula (I) may have all possible differences. Structure form unless otherwise It is mentioned or pointed out that the chemical name of a compound represents a mixture of all possible stereochemically isomeric forms containing all of the non-parent isomers and palmar isomers of the basic molecular structure 10, more specifically, stereo The center may have an R- or S-configuration, and the substituent on the divalent cyclic (partial) saturated group may have a cis- or anti-configuration, and the stereochemically isomeric form of the compound of formula (I) is clearly It is included in the scope of the present invention. According to the CAS-naming convention, when two stereos 15 of the absolute configuration are known, '^ is present in the numerator, and the R or S description is specified (based on Cahn-Ingold-

Prelog order principle) to the minimum number of palm center, reference center, the second two stereo center configuration is to use the relative description [R *, R *] or [R *, S *], where R * is always specific As a reference center and [R*, R*] represents the center of the same pair of palms and [R*, S*] represents the center of different pairs of palms, such as 20, if the smallest number in the molecule has a S configuration And the second center is R, the stereo description will be specific to S-[R*, S*], and if "and" Θ", the highest order substituent on the asymmetric carbon atom in the ring system with the least number of rings The position is any "α" position in the plane determined by the ring system forever, relative to the highest order substituent on the reference atom - 14 - This paper scale applies to the Chinese National Standard (CNS) A4 specification (210x297 mm)

The "α,," which is the highest order substituent on the other asymmetric carbon atoms of the system f, is named "厶" in the same side of the plane determined by the ring system, and is in the plane determined by the ring system. On the other side, we named the mountain. We have thought that the substituted mountain atom at the 4-position of the hexahydropyridyl moiety is not a polar atom. Therefore, the compound of formula (1) is via the palm substituent R, R2. R3' R4 or R5 may have only at least one stereocenter in its structure. The tautomeric form of the compound of formula (I) is meant to include the compound of formula (1) wherein, for example, the yl group is converted to a keto group ( a keto-enolyl tautomer). A group oxide form of a compound according to formula (I) is meant to include a compound of formula (1) wherein one or more nitrogen atoms are oxidized to a so-called N-oxide 15 These N-oxides, among which hexahydroanthracene, and the nitrogen of the sulfhydryl moiety and/or the stilbene moiety are oxidized. Ministry of Economic Affairs, Smart Finance, Bureaux X, Consumer Cooperatives 20 The compounds of formula (1) prepared according to the following method can be synthesized into pairs.昱 昱. The structure of the racemic mixture, its root The dissociation methods known in the art separate them from each other, and the ruthenium compound of formula (1) can be converted into the corresponding non-palphalinic salt form by reaction with a suitable palmitic acid. Separation of the conformational salt form, for example via selective or stepwise crystallization and the release of the palmo isomer from it with a base, the separation of the alternative to the isomerized form of the compound of formula (1) involves the use of a liquid layer of the palmitic stationary phase The pure stereochemically isomeric form can also be derived from the appropriate starting material in the form of the corresponding pure stereochemical isomer. The conditions of this paper are applicable to the National Standard (CNS) A4 specification (21〇χ297). Public interest) 1334351 A7 B7 V. Description of the invention (14) The reaction is stereospecific, and it is preferred that if a specific stereoisomer is required, the compound will be synthesized by stereospecific preparation methods. The present invention also includes a derivatized compound (hereinafter commonly referred to as a "precursor") of a pharmacologically active compound according to the present invention, which is decomposed in vivo to obtain a root. The compound of the present invention, the efficacy of the prodrug at the target receptor is usually (but not exclusively) lower than that of the originally decomposed compound, and the prodrug is particularly useful when the chemical or physical properties of the desired compound make it difficult or ineffective for administration, for example, The desired compound may have only poor solubility, which may not be transported through the mucosal epithelium, or it may have unwanted transient plasma half-lives. Other discussion of prodrugs can be found in Stella, VJ ei α/., “Prodrugs” , 1985, ρρ. 112-1.76 and 1985, 29, ρρ· 455-473. Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperative Printed According to the present invention, the pharmacologically active compound pre-drug form is usually root 15 according to formula (I) a pharmaceutically acceptable acid or base addition salt thereof in the form of a stereochemically isomeric form, a tautomeric form thereof and a ruthenium-oxide form thereof, which may be esterified or amided, including The esterified acid group is a group of the formula -C00Rx wherein Rx is alkyl, strepyl, benzyl or one of the following groups:

20

-CH2 amide group includes a formula -CONRyRz group, wherein R>1H, Ci_6 alkyl, strepyl or benzyl and RzS-〇H, H, C, _6 alkyl, phenyl or benzyl-16- paper Zhang scale applies China National Standard (CNS) A4 specification (210x297 mm) 1334351 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed A7 B7 V. Description of invention (15) Amine-containing compounds according to the invention may be used with ketones or aldehydes For example, the enthalpy derivatization forms a Mannich test, which can be hydrolyzed in aqueous solution under the first order kinetics. 5 The preferred organ is the heart, brain, liver, lungs or kidneys, and the mammal is a human. - more specifically, a preferred embodiment of the present invention relates to an agent for reducing ischemic heart damage in a mammal comprising as an active ingredient a compound according to formula (I), a pharmaceutically acceptable acid thereof Or a base plus 10 salts, a stereochemically isomeric form thereof, a tautomeric form thereof and an N-oxide form thereof, preferably a mammal is a human. Conditions associated with ischemic heart damage are, for example, post-coronary ischemic signs, angina pectoris, unstable angina, angina after myocardial infarction, myocardial infarction, acute myocardial infarction, traditional cardiopulmonary bypass (CPB), and isolation Cardiac protection from off-bypass cardiac surgery, non-cardiac surgery in patients with known or coronary artery disease (CAD) risk, and coronary restenosis after PTC A, in the framework of this application, Ischemic injury must be interpreted as any injury to any part of the heart, including the coronary arteries, and includes direct cellular damage and inter-contact damage due to, for example, lack of oxygen. More specifically, a preferred embodiment of the present invention relates to an agent for reducing cerebral ischemic injury in a mammal comprising as an active ingredient a compound according to formula (I), a pharmaceutically acceptable acid thereof or Alkali addition salts, their stereochemically isomeric forms, their tautomeric forms and their N--17- paper scales are applicable to the Chinese National Standard (CNS) A4 specification (210 x 297 mm)

1334351 A7 B7 V. INSTRUCTIONS (16) In the form of an oxide, the preferred mammal is a human. Conditions associated with stroke are, for example, atherosclerotic cerebrovascular diseases such as ash perfusion and arterial embolism, penetrating arterial disease, cardiac embolism such as, but not limited to, atrial fibrillation, membrane disease, and Ventricular thrombosis, cryptogenic stroke, and other more common causes such as thrombin status, anatomy, arteritis, migraine or vasospasm and drug abuse, ischemic injury is interpreted as any damage to any part of the brain, including for example due to Direct cell damage and indirect damage such as increased intracranial pressure in the absence of oxygen. More specifically, a preferred embodiment of the present invention relates to an agent for eliciting a cardioprotective effect in a mammal comprising as an active ingredient a compound according to formula (I), a pharmaceutically acceptable acid thereof Or a base addition salt, a stereochemically isomeric form thereof, a tautomeric form thereof, and an N-vapor form thereof, the cardioprotective effect means that the heart tissue is better for ischemia than the unprotected heart 15 tissue Effect of Protection: Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperative Printed In another specific embodiment, the organ is the lung, liver or kidney. The present invention also relates to a pharmaceutical composition for reducing ischemic injury to a mammalian organ, comprising a pharmaceutically acceptable carrier and an active ingredient as an active ingredient, a compound according to formula (I), which is pharmaceutically acceptable A 20 acid or base addition salt, a stereochemically isomeric form thereof, a tautomeric form thereof, an N-oxide form thereof, and a precursor thereof. The preferred organ is the heart, brain, liver, lung or kidney, and the mammal is a human. More specifically, the preferred embodiment of the present invention relates to a -18-paper scale applicable national standard (CNS) A4 specification (210x297 mm) 1334351 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed A7 B7 V. INSTRUCTION DESCRIPTION (π) A pharmaceutical composition for reducing ischemic injury in a mammalian heart, comprising a pharmaceutically acceptable carrier and an active ingredient as an active ingredient, according to formula (I), pharmaceutically Acceptable acid or base addition salts, stereochemically isomeric forms thereof, tautomeric forms thereof, bismuth-oxide forms 5 and precursors thereof, preferably mammals are human. More specifically, a preferred embodiment of the present invention relates to a pharmaceutical composition for reducing cerebral ischemic injury in a mammal, comprising a pharmaceutically acceptable carrier and an effective medical amount as an active ingredient. (I) a compound, a pharmaceutically acceptable acid or base addition salt thereof, a stereoisomer thereof, a tautomeric form thereof, a ruthenium-oxide form thereof, and a prodrug thereof, preferably The suckling animal is a human. Another preferred embodiment of the present invention relates to a pharmaceutical composition for inducing a cardioprotective effect in a mammal comprising a pharmaceutically acceptable carrier and an effective amount of the active ingredient according to formula (I) a compound, a pharmaceutically acceptable acid or base addition salt thereof, a stereochemically isomeric form thereof, a tautomeric form thereof, a bismuth-oxide form thereof, and a prodrug thereof, preferably a mammal Humanity. Another preferred embodiment of the present invention relates to a pharmaceutical composition in the form of a cardioplegia solution comprising an effective amount of a compound 20 according to formula (I), a pharmaceutically acceptable acid or base addition salt thereof, and a stereo The chemical isomer form, its tautomeric form, its bismuth-oxide form, its prodrug, and a suitable carrier. In another preferred embodiment, the compound according to the present invention may be simultaneously or sequentially applied to an antithrombotic agent and/or an angiogenic growth factor-19-this paper scale to the Chinese National Standard (CNS) A4 specification (210x297 male). PCT)

1334351 A7 B7 V. Description of invention (l8

In combination with anti-thrombotic agents, any agent known to achieve this effect can be selected, such as, but not limited to, glycoprotein Ilb/Ixia inhibitor, thrombus inhibitor, factor Xa inhibitor, tissue factor channel inhibitor, thromboembolic antagonist Agents or low molecular weight heparin, angiogenic growth factors may be selected from blood 5 tube endothelial growth factor (VEGF), for example, as disclosed in GB-2332 373 A (Merck & Co, Inc.), the contents of which are incorporated herein by reference. For example, as disclosed in WO 98/07832 (Ludwig Institute for Cancer research) or WO 98/49300 (Collateral Therapeutics), the above specific examples provide for reducing the risk of acute 10 coronary ischemia syndrome in patients at risk for this syndrome. Advantages, risky patients include those with initial coronary ischemic symptoms and those who are more likely to experience further thrombosis and ischemic tissue damage than those without this syndrome. Coronary artery infarction and administration of the above-mentioned specific embodiment of the pharmaceutical composition before the formation of the final blood clot within 6 hours after the occurrence of the infarction, the above-mentioned The embodiment then provides for the patient to reduce further clot formation while reducing tissue damage and enhancing tissue repair, and is also faced with a thrombus treatment for peripheral arterial occlusion or generally for thrombosis and intracranial vascular occlusion via release of a thrombus. treatment. The invention therefore also relates to a pharmaceutical composition for treating a mammalian 20 animal undergoing an ischemic event, comprising an effective amount of a compound according to the invention and at least one second medical agent, including an antithrombotic agent and/or angiogenicity Growth factors, or their respective pharmaceutically acceptable acid or base addition salts, stereochemically isomeric forms thereof, tautomeric forms thereof, N-oxide forms thereof, and precursors thereof, and suitable carriers, And according to this issue -20- This paper scale applies China National Standard (CNS) A4 specification (210x297 mm) Binding Line Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed 1334351 A7 B7 V. Invention Description (19) Compound Preparation of Ming Use for reducing ischemic heart damage comprising a compound according to the invention and at least one second medical agent, including an antithrombotic agent and/or an angiogenic growth factor. Specifically, a compound according to formula (I), a pharmaceutically acceptable 5-acid or base addition salt thereof, a stereochemically isomeric form thereof, a tautomeric form thereof, an N-oxide form thereof, and a precursor thereof The drug and its constituents also have a high potential for preservation in donor organs. The Ministry of Economic Affairs, the Intellectual Property Office, the employee consumption cooperative, printed according to the present invention, which can be used for reducing ischemic injury in mammals, can be used by itself, but it can be formulated into different medical forms for the purpose of 10, and suitable compositions can be enumerated. It is a composition which is generally used for a systemic drug, and when the pharmaceutical composition of the present invention is prepared, it is an effective amount of a specific compound as an active ingredient, if necessary, in the form of an addition salt, and is closely related to a pharmaceutically acceptable carrier. Mixing, depending on the form of preparation of the drug to be administered, the carrier may be in various forms, and these pharmaceutical compositions are suitably in unit dosage form, especially for parenteral injection or infusion, for example, when preparing a composition. Any conventional pharmaceutical medium may be used. For parenteral compositions, the carrier usually comprises sterile water, at least for the most part, although other ingredients may be included, for example to aid solubility, for example, an injection solution may be prepared in which the carrier contains a saline solution 20, a glucose solution or a mixture of saline and glucose solution, Injectable suspensions can be prepared wherein the liquid carriers, suspending agents and the like, Also included are solid form preparations which are converted, shortly before use, to liquid form of the formulation. Depending on the mode of administration, the pharmaceutical composition preferably contains from 0.05 to 99 weight - 21 - the paper size applies to the national standard (CNS) A4 specification (210x297 mm) 1334351 A7 B7 V. Invention description (20) %, More preferably, it is from 0.1 to 70% by weight of the active ingredient, and from 1 to 99.95% by weight, more preferably from 30 to 99.9% by weight of the pharmaceutically acceptable carrier, all percentages being based on the total composition. The amount to be administered is appropriately selected depending on factors such as symptoms, age, body weight, and the method of administration 5, but in the case of parenteral administration such as injection into an adult, 0.01 mg to 1.0 g of the compound according to the present invention can be administered per day. The compound according to the present invention may be administered from 0.1 mg to 3 g per day, whether administered once or in divided doses, in the case of oral administration or in several divided doses, in the case of oral administration to an adult. 10 The pharmaceutical composition may additionally contain various other ingredients known in the art, such as stabilizers, buffers, emulsifiers, viscosity modifiers, surfactants or preservatives. The pharmaceutical composition is preferably administered intravenously, for example, by infusion (continuous intravenous administration) or bolus administration. In addition, the present invention also relates to an acid or base addition salt, a stereochemically isomeric form thereof, and a tautomeric bridge form thereof, which are printed by the compound of formula (I), its pharmaceutically acceptable Ministry of Commerce, Intellectual Property Office, and the Consumer Cooperative. The use of the N-oxide form and its precursors, and any of the above-described pharmaceutical compositions, for the manufacture of a medicament for reducing organ ischemic injury in a mammal. 20 The preferred organ is the heart, brain, liver, lungs or kidneys, and the mammal is a human. More particularly, preferred embodiments of the invention are directed to compounds of formula (I), pharmaceutically acceptable acid or base addition salts thereof, stereochemically isomeric forms thereof, tautomeric forms thereof, N-oxide form and its -22- This paper scale applies to China National Standard (CNS) A4 specification (2丨0x297 mm) 1334351 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed A7 B7 V. Invention description (21) The prodrug and any of the above pharmaceutical compositions are useful for the manufacture of a medicament for reducing ischemic heart damage in a mammal, preferably a human. More particularly, preferred embodiments of the invention are directed to a compound of formula (I), a pharmaceutically acceptable acid or base addition salt thereof, a stereochemical 5 isomer form thereof, a tautomeric form thereof, The N-oxide form and its prodrug, and any of the above-described pharmaceutical compositions, are useful for the manufacture of a medicament for reducing cerebral ischemic injury in a mammal, preferably a human. More particularly, preferred embodiments of the invention are directed to a compound of formula (I), a pharmaceutically acceptable acid or base addition salt thereof, a stereochemical 10 isomer form thereof, a tautomeric form thereof, The N-oxide form and precursor thereof, and any of the above-described pharmaceutical compositions, are useful for the manufacture of a medicament for eliciting a cardioprotective effect in a mammal, preferably a human. Further, the present invention also includes a compound of the formula (I), a pharmaceutically acceptable acid or base addition salt thereof, a stereochemically isomeric form thereof, a tautomeric 15-form thereof, an N-oxide form thereof, and A prodrug and a pharmaceutical composition thereof, for use in the manufacture of a medicament for treating a mammal undergoing an ischemic event, comprising administering an effective amount of a compound according to the invention and at least one second agent, including an antithrombotic agent and/or a blood vessel a growth factor, or a pharmaceutically acceptable acid or base addition salt thereof, a pharmaceutically isomeric form thereof, a tautomeric form thereof, an N-oxide form thereof, and a prodrug thereof, and a suitable Carrier. The invention also relates to a method for reducing organ, especially cardiac and/or cerebral ischemic injury, in mammals, particularly humans, or a method for inducing a cardioprotective effect in a mammal, the steps comprising - This paper size applies to the National Standard (CNS) A4 specification (2丨0 x 297 mm)

1334351 A7 B7 V. INSTRUCTIONS 22 10 15 The amount of a compound according to formula (1), a pharmaceutically acceptable acid or test addition salt thereof, a stereochemically isomeric form thereof, a tautomeric form thereof, The N-oxide form and its prodrug and any of the above pharmaceutical compositions are administered to the mammal in need of such treatment. Further, the present invention also relates to a compound of the formula (1), a pharmaceutically acceptable acid or base addition salt thereof, a stereochemically isomeric form thereof, a tautomeric form thereof, an oxide form thereof, and a precursor thereof, and any of the above Pharmaceutical Compositions The use of pharmaceutical agents for the prevention and/or treatment of cardiac or cerebral ischemic events in a mammal. The compounds according to the invention can generally be prepared via a series of steps, each step being known to those skilled in the art, in particular, a compound according to formula (Ζ-α) can be via the formula (1) according to the reaction scheme (1) The reaction of the intermediate is carried out in a suitable reaction solvent such as toluene in the presence of a suitable base such as triethylamine. In the reaction scheme, all variables are as defined in formula (I) And wi and the portion to which it is attached is equal to R1; the W1 example is an alkyl group, Ar^Het, and the wi〇c(=〇)c丨 example is a gas formate. ” i counts the Ministry of Economic Affairs, the Intellectual Property Bureau, the employee consumption cooperative, printed

Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperative Printed 1334351 A7 B7 V. Description of Invention (23) According to formulas (Ia), (Ib), (Ic), (Id), (Ie), (If), (Ig) and The compound of (Ig) can also be prepared according to the reaction of any of the reactions shown in the reaction scheme (2) via the intermediate of the formula (III), in which all the variables are as defined in the formula (1) and W1 is attached thereto. The part is equal to R1; 5 W1 instances are alkyl, Ar or Het. The reaction (a) is carried out in a suitable solvent such as dichloroethane and using BOC20, and the reaction is conveniently carried out under reflux for several hours. The reaction (b) is carried out in a suitable solvent such as THF, and the reaction is conveniently carried out at room temperature for 1 to several hours. The reaction (c) is carried out in a suitable solvent such as di-methane in the presence of a suitable base such as Et3N at room temperature for 1 hour. Reaction (d) is carried out in a suitable solvent such as THF or DMF without the need for a base and at room temperature for several hours. The reaction (e) is carried out in refluxing acetone or in a suitable assay such as DMF in the presence of potassium carbonate 15 and can conveniently be carried out at 80 °C. The reaction (f) is carried out in a suitable solvent such as dichloromethane in the presence of a suitable base such as triethylamine at room temperature for about 30 to 120 minutes. The reaction (g) is carried out in a suitable solvent such as acetonitrile under reflux for 24 hours. 20 Reaction (h) is carried out according to R1 under different conditions, for example, when R 2 CF; the reaction is carried out in dioxane in the presence of triethylamine at -78 ° C for 1 hour, for R 1 = NH 2 , the reaction It is at the reflux temperature in two. The reaction was carried out for 12 hours in the hexane. For Ri = CH3, the reaction was carried out in dioxethane at room temperature in the presence of triethylamine for 3 hours. -25- This paper size applies to national standard (CNS) A4 specification (210x297 mm)

1334351 A7 B7 V. INSTRUCTION DESCRIPTION (24) The reaction (1) is carried out in a suitable solvent such as isopropyl alcohol at a reflux temperature for 12 to 36 hours. The reaction (j) is carried out in a suitable solvent such as acetonitrile at reflux temperature for 24 hours. έ Ordering Line Printed by the Intellectual Property Office of the Ministry of Economic Affairs. 6 2 This paper scale applies to the Chinese National Standard (CNS) A4 specification (210x297 mm) 1334351 A7 B7 V. Invention Description (25

Figure 2 0 0 everyone

〇 0—W w'n=c=b (b) (c) (d) (e) (g) (h)

B = 0. S o W '-(la) B -W '-NQH (Lb) (HjC)Si—N=C=0 Λ (1*c) 0 h2n*^q dd) h2n X—Cl h2n X (>-e) R1—U—Cl 0 person (lc) ax?1 r6, n person (ΜΪ R〆W2 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative print

Q

W2=r\ Cl, nh2 Het-Hal Hal = CI,BM.FR \ R〆, Q dg) Het-Q (fh) R^R8w.XKQ 0-i) -27- This paper scale applies to Chinese national standards ( CNS) A4 size (210x297 mm) 1334351 A7 B7 V. Description of the invention (26) A compound according to formula (Ic) can also be prepared by reacting an intermediate of formula (IV) with a halide, in which all The variables are as defined for formula (I) and the reaction is carried out with a base such as NaH (60% in mineral oil) and in a reaction inert solvent such as DMF or THF.

(IV) (iv) 10 Starting materials and intermediate compounds according to formula (II), (III) and (IV) are commercially available compounds or may be prepared according to conventional reaction methods generally known in the art. The intermediate of the formula (II) can be produced according to the following reaction scheme (4), wherein all of the variables of the formula 15 are as defined in the formula (I). , Ministry of Economic Affairs, Intellectual Property Bureau, employee consumption cooperation, du printing, this paper scale applies to China National Standard (CNS) A4 specification (210x297 mm) W4351

Five', invention description (27 (Rs) p schema 4

Nh3 (a)

, R3 SOCI2, "0 h2n

10 (6) (R2)p (R2)p t (d) Order 15

'QVC R5 R4 Line Economy Zou Wisdom Property Officer w Consumer Co., Ltd. Printing (II) Reaction Scheme 4 includes the step (a) of the hydrazine-based gas of the type shown with a substituted primary amine such as sulfhydryl in a suitable base such as Et3N In the presence and reaction in a suitable reaction inert solvent such as di-methane, the reaction is conveniently carried out at room temperature 'in the next step (b), the adduct obtained in the step is refluxed with soc丨2' and then The resulting product is reacted with an appropriately substituted 2,2-dimethoxyethylamine in a reaction inert solvent such as DMF, for example at room temperature (step c), in step (d), step (c) The compound is obtained only by (: ring-29- this paper scale applies Chinese National Standard (CNS) A4 specification (210x297 mm)' ~ 1334351 A7 B7 V. Invention description (μ) to obtain substituted microphone. The intermediate compound of the formula (10) can be produced from a compound according to the formula (IV) by selective reduction of the (four) part (4) moiety of the compound according to the following reaction:

(III) Ministry of Economic Affairs, Intellectual Property Office, Employees' Consumption Cooperative Printing System The intermediate compound of formula (IV) can be prepared by hydrogenating a compound according to formula (I-c) according to the following reaction:

The reaction is carried out in the presence of a suitable base such as WK〇H in a suitable reaction inert solvent such as 2-propanol and under reflux. 15 20 Let all the variables be as defined in formula (1), and the reaction is carried out in a sterol in the presence of a catalyst such as Pd/C (10%) and at a moderate temperature. Obviously in the foregoing and the following reactions, the reaction product can be separated from the reaction medium, and if necessary, according to the general -30- paper scale of the art, the Chinese National Standard (CNS) A4 specification ϋ^Τ297 public 1334351 Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed A7 B7 V. INSTRUCTIONS (29) Known methods for further purification, such as extraction, crystallization and chromatography, the presence of more than one reaction product of the palm isomer form It is also apparent that it can be separated from its mixture via known techniques, in particular preparative chromatography, such as preparative HPLC. The following examples illustrate the invention without limiting it. In the experimental part of the compound, the absolute stereochemical configuration of the stereoscopic carbon atom was not determined experimentally. In these cases, the first separated stereochemical isomer form is called ''A' and the second is called '' B" without further indicating the stereochemistry of the real 10, but the "A" and "B" isomer forms can be used by those skilled in the art using methods known in the art such as X- The light diffraction method is clearly identified, and the separation method is described in detail below. · Hereinafter, "DMF" is defined as ruthenium, Ν-dimethyl decylamine, "TMF" is defined as tetrahydrofuran and ''DIPE' is defined as diisopropyl" ether. 15 Α. Preparation of intermediate compounds. Example Α1 · 1-Mercapto-4-phenyl-4-pyropyridine carbon helium (0.49 mol) was added portionwise to benzylamine (0.49 mol) at room temperature And Ν, a mixture of hydrazine-diethylethylamine (1.223 mol) in CH2C12 (2500 ml), the mixture was stirred at room temperature for 1 hour, K2C03 (150 g) and H20 were added, and the mixture was stirred and The layers were separated, the aqueous layer was extracted with CH.sub.2 C.sub.2, and the combined organic layer was dried (MgS04), filtered and evaporated to yield 144 g (95%) of 1-methyl-4-phenyl-N-( Stylosyl)-4-hexahydropyridiniumamine (Intermediate 1). -31- This paper size is applicable to China National Standard (CNS) A4 specification (210 x 297 mm)

Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperative Printed 1334451 A7 87 V. INSTRUCTIONS (3〇) EXAMPLE A2 A mixture of Intermediate 1 (0.47 mol) in S0C12 (750 ml) was stirred and refluxed for 1 hour to evaporate the solvent. Adding toluene twice and re-evaporating, yield: 190 g (100%) of N-[gas (1-methyl-4-phenyl-4-hexahydro-5 pyridyl) fluorenylene] alum Salt (intermediate 2). Example A3 A mixture of Intermediate 2 (0.47 mol) in DMF (750 mL) was cooled in an ice bath, and 2,2-dimethoxyethylamine (0.54 mol) dissolved in DMF was added dropwise. Stir at room temperature overnight, evaporate the solvent, 10 yield: 210 g (100%) of N-(2,2-dimethoxyethyl)-1-methyl-4-phenyl-indole--曱 ))-4-hexahydropyridine decylamine dihydrochloride (Intermediate 3). Example A4 A mixture of Intermediate 3 (0.47 mol) in 6 equivalents of HCl (1500 mL) was stirred to a turbid solution, then washed with CH.sub.2Cl.sub.2 (900 mL), stirred at 80 ° C for 1 hour, cooled, and used. The NaOH 50% solution was basified and extracted with CH2C12, the organic layer was separated, dried (MgSO4), filtered and evaporated, and the residue was crystallised from CH3CN, filtered and dried, yield: 38.3 g (25%) 1-Methyl-4-phenyl-4-[1-(phenyl 20 decyl)-1 Η-imidazol-2-yl]hexahydropyridine (Intermediate 4). - Example A5 A mixture of compound 1 (0.089 mol) in methanol (250 ml) was hydrogenated at 50 ° C with Pd/C 10% (3 g) as a catalyst. After hydrogen (1 eq.) was consumed, it was touched. The medium is filtered and the filtrate is evaporated to make the residue from the -32- paper scale applicable to the Chinese National Standard (CNS) A4 specification (210x297 mm).

1334351 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed A7 B7 V. Invention description (3ι) CH3CN crystallized, filtered and dried, and yield: 23.89 g (90%) of 4-phenyl-4-(1Η-imidazole Ethyl-2-yl)-1-hexahydropyridinecarboxylate (Intermediate 5). Example A6 5 A mixture of Intermediate 5 (0.026 mol) and KOH (0.26 mol) in 2-propanol (150 mL) was stirred and refluxed for 10 hr. The solvent was evaporated and the residue dissolved in H20 and mixture Extracted with CH2C12, the organic layer was separated, dried, filtered and evaporated and evaporated. Intermediate 6). 10 Example A7 was reacted under N2 pressure, and a mixture of Intermediate 5 (0.0033 mol) in DMF (5 mL) and THF (5 mL) was added dropwise to 60% NaH (0.004 m) in mineral oil. The solution was stirred with THF (10 mL) at room temperature and the mixture was stirred at room temperature for 1 hour, then a solution of 4-(15-ethoxyethoxy)phenylhydranol (0.004 mol) in THF was added dropwise. The resulting reaction mixture was extracted with CH.sub.2Cl.sub.sub.sub.sub.sub.sub.sub.sub.sub.sub.sub.sub.ssssssssssssssssssssssssssssssssssssssssssssssssssssssss NH3) 95/5), the pure stream washing fraction was collected and solvent 20 was evaporated. Yield: 1.33 g of 4-phenyl-4-[l-((4-methylcarboxy)phenylmethyl)-1Η-imidazole Ethyl-2-yl]-1-hexahydropyridinecarboxylate (Intermediate 7). B. Preparation of final compound Example B1 Intermediate 4 (0.05 mol) and N,N-diethylethylamine (0.15 mol) -33- This paper scale applies to China National Standard (CNS) A4 specification (210x297 PCT)

1334351 A7 B7 V. INSTRUCTIONS (32) A mixture of toluene (750 ml) was stirred at 100 ° C, ethyl phthalate (0.2 5 mol) was added dropwise and the reaction mixture was stirred and refluxed for 1 hour and then cooled. Pour the mixture into K2C03 aqueous solution (35 g K2C03), separate the liquid layer, extract the aqueous layer with CH2C12, dry the separated organic layer (MgS04) 5, filter and evaporate the solvent, and leave the residue on glass filter Filtration via silica gel (flow wash: CH 2 Cl 2 / C 2 H 5 OH 98/2), the desired fractions were collected and the solvent was evaporated, the residue crystallised from CH3CN, filtered and dried, yield: 16.7 g (86%) of 4-phenyl 4-[1-(Styryl)-indolyl-imidazol-2-yl]-1-hexahydropyridinecarboxylate (Compound 1). 10 Example B2 Preparation of Compound 2 〇

Ministry of Economic Affairs, Intellectual Property Bureau, Staff Consumer Cooperative, printed with arachnid (0.0023 mol) added to Intermediate 6 (0.0019 mol·) and Ν, Ν-diethylethylamine (0.0024 mol) in CH2C12 (15 (ml) and stirred at room temperature, the reaction mixture was stirred at room temperature for 30 minutes, water was added, the layers were separated, the aqueous layer was extracted with CH2C12, and the combined 20 layers were dried (Na2S04) and filtered. The solvent was evaporated and the residue was purified on EtOAc EtOAc (EtOAc:EtOAc:EtOAc: Crystallized from n-hexane, filtered and dried, yield: 0.42 g (yield: 52%) Compound 2. -34- This paper scale applies to China National Standard (CNS) A4 specification (210x297 mm) Printed by the Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperative 1334351 A7 V. Description of Invention (33) Example B3 Preparation of Compound 3

Reaction under N2 pressure, a solution of intermediate 5 (0.0054 mol) in DMF (10 mL) and THF (10 mL) was added dropwise to THF (30 mL) NaH. After stirring at room temperature for 1 hour, 10 then decyl 4-(bromohydrazinyl)benzoate (0.00624 mol) in THF (5 mL) was added dropwise and the reaction mixture was stirred at 60 ° C for 3 h, then water was added. The mixture was extracted with CH.sub.2Cl.sub.sub.sub.sub.sub.sub.sub.sub.sub.sub.ssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssss /2), 15 parts of the desired fractions were taken and the solvent was evaporated, the residue was crystallized from DIPE, filtered and dried, yield: 1.7 g (70%) of compound 3, melting point: 149.1 °C. Example B4 Preparation of Compound 4

-35- This paper size applies to China National Standard (CNS) A4 specification (210 X 297 mm)

1334351 Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperative Printed A7 B7 V. Description of Invention (34) Mix a mixture of Intermediate 6 (0.0059 mol) and (0.0059 mol) in CH3CN (70 mL) for 24 hours. The solvent was evaporated, water was added, and the mixture was evaporated, evaporated, evaporated, evaporated, evaporated, 0.33 g of compound 4, m.p. 84.2. Example B5 Preparation of Compound 5 10

The mixture of the intermediate 4 (0.0001 mol) in 6 eq. of HCl (22.8 mL) was stirred and refluxed for 4 hr. The mixture was basified and then extracted with CH.sub.2 C.sub.2. • The solvent was evaporated, the residue was crystallized from DIPE, filtered and dried, yield: 0.24 g (62%) of Compound 5. Example B6 20 Preparation of Compound 6 %

-36- This paper size is applicable to China National Standard (CNS) A4 specification (210x297 mm)

1334351 Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperative Printed A7 B7 V. Description of Invention (35) Add isocyanato stupid (0.0094 mol) dropwise to intermediate 6 in THF (50 ml) (0.0094 mol) The reaction mixture was stirred at room temperature for 30 minutes, water was added and the mixture was extracted with CH.sub.2 C.sub.2, and the organic layer was dried (Na.sub.2SO.sub.4). Dry, yield: 2.7 g (68%) of compound 6. Example B7 Preparation of Compound 7

The isocyanato group (0.0007 mol) was added to Intermediate 6 (0.0007 mol) in THF (10 mL) and the reaction mixture was stirred at room temperature for 3 hr. The separated organic layer was dried (Na.sub.2SO.sub.sub.sub.sub.sub.sub.sub.sub.sub.sub.sub.sub.sub.sub.sub.sub.sub. Yield: 0.2 g (66%) of compound 7 ° 20 Example B8 ... a) Preparation of compound 8

This paper size applies to the Chinese National Standard (CNS) A4 specification (210x297 mm)

1334351 Ministry of Economic Affairs, Intellectual Property Bureau, Consumer Cooperative, Printed A7 B7 V. Description of Invention (% will be compound 3 (0.002 mol) and LiOH (0. 02 mol) in thf (u liter) and Η 2 〇 (π ml) The mixture was stirred at room temperature for 24 hours, and the mixture was adjusted to pH 6 and extracted with CH.sub.2, and the organic layer was separated, dried, filtered and evaporated, and the residue was washed with CH2C12, yield: 0-72 g (83%) Compound 8, melting point 251.6. 〇 b) Preparation of compound 9

10 Under the pressure of N2, transfer NaH60% (0.000642 mol) in DMF (2 house liter) Loose solution at room temperature, add intermediate 6 (0-000642 mol) dropwise in DMF (8 ml) The solution was stirred at room temperature for 1 hour, CHJ (0.001284 moles) was added and the reaction mixture was stirred at 60 ° C for 2 hours in a Parr pressure vessel. The solvent was evaporated and the residue was passed on a silica gel. Purification by high pressure liquid chromatography (flow washing: Ci^Clz/CHsOH 98/2), the desired fractions were collected, and the solvent was evaporated, yield: 0.14 g (49%) of Compound 9. c) Preparation of compound 10

Add the IMo concentration of THF to LiAlH4 (0.000444 mol) -38- This paper size is applicable to China National Standard (CNS) A4 specification (210 X 297 mm)

1334351 Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperative Printed A7 B7 V. Description of Invention (37) Add to Intermediate 7 (0.000404 mol) in THF (5 ml) and stir at 0 ° C, the reaction mixture is at 0 After stirring for 30 minutes, a mixture of 10% aq. EtOAc and EtOAc EtOAc (EtOAc)EtOAc. : CH2C12/CH30H 96/4), the desired fractions were collected and evaporated, and the residue was crystallised from CH3OH/H2, filtered and dried, yield: 0.020 g (10%) of Compound 10. d) Preparation of compound 11

LiOH (0.001423 mol) was added dropwise to a solution of the intermediate 7 15 (0.0006469 mol) in dioxane / H20 1 / 1 (6 mL), and the resulting suspension was stirred at room temperature for 18 hr. The residue was taken up in EtOAc (EtOAc m.) The organic layer was separated, washed with brine, dried over NaH~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Example B9 Adding LiOH (0.018 mol) to Intermediate 7 (0.0018 mol) at -39- This paper scale applies to China National Standard (CNS) A4 specification (210x297 mm)

1334351 A7 _____ —___B7 _____ V. Illustrative (π) Mixture of THF (10 ml) and 屮〇〇〇ml) The reaction mixture was stirred at room temperature for 3 hours, water was added, and CH2C12 was added to extract the mixture. The layers were separated, dried (Na.sub.2.sub.4), filtered, and evaporated. The residue was washed with methanol and <RTI ID=0.0> Ethyl phenyl fluorenyl)-1 hydrazine-sodium aceto-2-yl]-1-hexahydropyridinecarboxylate (Compound 12). Prepare the following compounds listed in Tables 1-5: - (All melting points (mp) are in .c) Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperatives Printed on this paper scale Applicable to China National Standard (CNS) A4 specification (210x297 mm) ) 1334351 A7 B7 V. INSTRUCTIONS (39) TABLE 1: 〇

Ministry of Economic Affairs Intellectual Property Bureau Employees Consumption Cooperatives Printed Compound No. Experiment No. R1— RJ—Physical Properties'· 110 B2 —Η cr 13 B1 , 〇〆·· cr 14 cr m.p_=137 1 B1 A/ CT 12 " B9 "" /v7_—' jcr - Ϊ5 "...B3~ '' Tv7,. XJ 4 Ϊ6... B3">ν··ΓFxr"_ mp=l 17 \1 B3~ " A/ 贤 - mp=127 18 B3 A0:.·. Real: mp =125 8 __ B6 八. ··: oyCr-"· OH mp=252 -41- This paper size is applicable to China National Standard (CNS) A4 specification (210x297 mm) 1334351 A7 B7 V. Invention description (40)

R

L

'NIR Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed

This paper scale applies to China National Standard (CNS) A4 specification (210x297 mm) 1334351 A7 B7 V. Invention description (4i) 0

Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperatives Printed Compound No. Experiment No. R丨... R,-- Physical Properties 27 B1 σ' mp=80 28 "B1 σ'. CT mp=215 29 B2 ,〇/···, · CT·· mp= l 11 30 B3 \〇, oyCT""3Ϊ" B3 Υ cy... 32 B1 0^..' cr 4 33 B2 ch3— cr. mp= 183 '—34... B2 ch3ch2— Cr mp=133 —孖一— "'B2 isopropyl... cr mp= 107 36'—""132D>- cr. mp^l 11 B2 third butyl cr. t mp=165 -43 -

This paper scale applies to China National Standard (CNS) A4 specification (210x297 mm) 1334351 A7 B7 V. Description of invention (42

0II

Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperatives Printed Compound Numbers Serial Number R1... Physical Properties. 2 B2 0·.··· cr·· mp==123 38 B3 〇-' v 39 B3 0···.· ... 40 ...B3... σ—— o ""......'41............ —σ—— (fV.·' 4 42 a...B2 — F3V mp=151 cf3 43 B2 γ —ζΓ: mp=79 ,. 44 B2 Λ cr-· mp =149 —.4?—— cr·· cr 1·· . — j -44- This paper scale applies to the Chinese National Standard (CNS) A4 specification. (210x297 mm) 1334351 A7 B7 V. Description of invention (43 0 Ministry of Economic Affairs Intellectual Property Office Staff Consumption Cooperative Printed Compound No. Experiment No. R1... κΛ- Physical te quality 46 Έ2 nh2— σ····· mp=208 47 ► A B2 /V Η CT mp= 144 48. B2 ζΓ:... 49 B2 yLy σ' 50 B2 /νγ CT:... 51 B2 σ' 6 B6 σ-· « 52 B3 σ'- 〇, 53 . B3 σ' 54. B3 —σ····· * ^ — ____ -45-

This paper size applies to the Chinese National Standard (CNS) A4 specification (210 x 297 mm) 1334351 A7 B7

Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperatives Printed Compound No. Experiment No. R1... R Rule - Physical Properties 55 B3 σκ'- 0^'" 56 A...B3 : '57 Β3 — "ΧΓ· 58 Β3 σ- xr· ·· 59 Β3 σ- σ··' "60 Β3 σ"- α1··· 4 61 Β3 σν oy 0 _ 62 Β3 χτ·. ·. —......63 Β3 pr' 64 ''βΤ " —— C?:. ζΤ·· -46-

This paper scale applies to China National Standard (CNS) A4 specification (210x297 mm) 1334351 A7 B7 V. Invention description (45) 0

Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperatives Printed Compound No. Experiment No. R1... R3... Physical Properties 65 B2 α:: CT· 66 B2 α:; —ζΤ— 67" ~ B2 α: cr: 68— _ B2 Sf. _ 〇τ:· 7 B7 Οφ 0— 〇r-'·· 69 B2 "Xr"- 〇τ· « 7 B2 Ί· ζΤ. 70 '一B2 严1 Η 0τΝ··__ CT 71 B2 cf: - 72 B2 χτ"·· cr -47- This paper size is applicable to China National Standard (CNS) A4 specification (2丨0x297 mm) 1334351 V. Invention description (46) 0

Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperatives Printed Compound No. Experiment No. R丨... RJ...]Physical Properties 73 B2 CT.· ...74 B2 • σ' - 10 B6 cr.— - - ........ ... 75 B2 〇χΗ σ'·.. U ' 76 B2 0τκ·" cr 4 77 B2 F3C-o —cr _ 11 B6 .......... —ζΓ_:— » - · 78 B2 丨 cr - cr -48- This paper scale applies to China National Standard (CNS) A4 specification (210x297 mm) 1334351 A7 B7 V. Invention description (47 0IIR1—^ Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing compound No. Experiment number R1... R\- Physical properties 79 B2, square.... cr. —9 B6 σ1· ' ^cr::— ............. 80 B2 furnace; Cr:.· 81 B2 〇/.·· cr 113 B2 ΗΝ*· C0 cr 82 B2 r^N··- cr « 83 B2 W·. cr mp=74 84 B2 cf 85 B2 S—^ h... .. cr. mp= 165 86 B2 σ"· cr·. -49-

This paper scale applies to China National Standard (CNS) A4 specification (210x297 mm) 1334351 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing A7 B7 V. Invention description (48) 0

Table 2 :

Compound No. Experiment No. Ra... R, -- R2... Position of R2 Physical Data 4 * 88 89 90 114 115 B3 B3 B3 B3 B3 0_·.·· σ " σ Η Η Η ;σ; Η /0,, — -F —FCC a . -50- This paper scale applies to National Standard (CNS) A4 (117x297 mm)

1334351 A7 B7 V. Description of invention (49

Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperatives Printed Compound No. 骀 Α = Β R1... Physical Data 5 Β 5 ΟΝΗ H w 91 Β 5 C=NH 4 Β 4 C=N-CN H .· mp =84 92 Β4 C= N-CN 93 Β4 C=CN〇2 σ'·,. 95 Β2 c=s .................~H........... ...— w· mp=172 96 Β2 c=s (X:- 94 Β2 S〇2 —ch3 mp=167 97 Β2 S〇2 —nh2 mp=212 Γιϊ — Β2 S〇2 ...CF3 mp =104 98 Β2 S〇2 a"- -51- This paper scale applies to China National Standard (CNS) A4 specification (210 X 297 mm) 1334351 A7 B7 V. Description of invention (50) Table 4 ·

Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperatives Printed Compound No. Experiment No. Z (A=B and R1 - R) R?•... Physical Data 99 B3 ΟΛ 0 100 B3 101 B3 〇U, α1.·, 4 102 B3 W :... « 103 B2 0;' σ'- mp=204 104 B2 1 CT···· mp=181 105 B2 σ.···· . mp =190 106 B2 σ·,'· mp=107 - 52- This paper size applies to the national standard (CNS) A4 specification (210x297 mm) 1334351 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed A7 B7 V. Invention description (52) PS, Sultana M., Takemori AE: Naltrindole, a highly selective and potent non-peptide δ opioid receptor antagonist. Eur. J. Pharmacol. 146, 185-186, 1988) U69593 and Nobenaprofen (nor-BNI) in Kappa opioid receptor 5 as a reference agent and antagonist, for opioid receptors, morphine as a reference agent and naloxone as a reference antagonist 儿 " children, Mansour A., Akil H., Medzihradsky F., Traynor JR,

Woods JH: Stimulation of guanosine-5'-0-(3-[35S]thio)triphosphate binding by endogenous opioids 10 acting at a cloned Mu receptor. J. Pharmacol. Exp. Ther. 286, 282-288, 1998) and (Smart D·, Hirst RA, Hirota K·, Grandy DK, Lambert DG: The effects of recombinant rat μ-opioid receptor activation in CHO cells on phospholipase C, [Ca2+]I and adenylyl cylase. Br. J. 15 Pharmacol. 120, 1165-1171, 1997) 〇. C. 1. Materials and Methods 'Cell culture coils will be permanently transfected with Cba cells or opaque receptor CHO cells supplemented with 10% heat-deactivated burdock serum and containing 100 IU/ Dulbecco's Modified Eagle's Medium (DMEM)/Nutrition with milliliter penicillin G, 100 micrograms/ml streptomycin sulfate, 110 micrograms/ml pyruvate and 300 microliters/ml L-glutamate antibiotic solution The mixture was cultured in Ham F12 (ratio 1:1) and will be permanently transfected with C6 glioma cells of the opiate receptor in DMEM medium supplemented with the above 10% heat-activated burdock serum and antibiotic solution-54- paper scale Applicable to national standards of t countries (CNS> A4 specifications (2丨Οχ 29 7 mm)

Ministry of Economic Affairs, Intellectual Property Bureau, Staff Consumer Cooperatives, Printing 1334351 A7 B7 V. Inventions (53) Medium training. Membrane Preparation The membrane was prepared as a total particulate matter component. All cell lines were grown to 90% confluence in a 145 mm Petri dish and 5 mM molar sodium butyrate was added 24 hours prior to collection. The medium was removed and the cells were removed. Wash with ice-cold phosphate buffered saline (PBS w/o Ca2+ and Mg2+), scrape from the culture dish to 50 mM Tris-HCl buffer pH 7.4, and centrifuge (16,000 RPM at 4 ° C) After 10 minutes) collection, the cell pellet was resuspended in hypotonic 5 mM Tris-HCl buffer pH 7.4 and homogenized again with an Ultra 10 Turrax homogenizer, and the homogeneous solution was centrifuged at 18000 RPM at 4 °C. After 20 minutes, the final pellet was resuspended in 50 mM Tris-HCl buffer pH 7.4 and aliquoted at -7 (TC, using the Biorad Protein Assay (Bradford) using Burdock Serum Albumin (BSA) MM.. d ropzW •seAw/iz've 15 method for the quantification of microgram quantities qf protein utilizing the principle of protein-dye binding.

Analytical Biochem. 72: 248-254, 1976) 〇 匕 Radioligand Binding A preliminary radioligand binding assay was performed to show the optimal test conditions for these opioids in their corresponding mammalian cell membranes. Competitive inhibition of compound [3H]DPDPE at different single-line concentration compounds with a radioligand concentration of 2 nanomolar (Kd = 1.7 nanomolar) and ranging from PIC50 to at least three orders, for gram Competitive binding of Ba and 缪 receptors, [3H]U69539 (Kd=0.4 nano Mo-55- This paper scale applies to 'Home Standard (CNS) M Specification (2) G η97

1334351 A7 B7 V. INSTRUCTIONS 〇4) Ear concentration) and [3H]DAMGO (Kd=0.6 nanomolar concentration) were used at a density of 1 nanomolar, respectively, and the membrane was thawed and released on ice. In a 50 mM Tris-HCl buffer pH 7.4, for the opiate receptor, the culture buffer was supplemented with 2 mmol concentration MgCl2, 1 mM molar EGTA and 0.1% BSA, non-specific Sexual association is defined for the Dessert, Keba and Oscar receptors in the presence of 1 micromolar Natto, Spiradoline and dextromoramide, found in 25 . (: The competitive binding test method for the most suitable three receptor subtypes for 1 hour was cultured. The test method was carried out in 10 rows in the final volume of 500 μl, and was quickly passed under reduced pressure using a Filtermate 196 (Packard).

The reaction was terminated by filtration of UniFilterTM_96, GF/Btm, and the amount of bound radioactivity was measured on a filter paper unit after drying on a filter paper and adding a scintillator (Microscint-O; Packard) via liquid scintillation counting. C.3. "35SlGTPr S. Combine 15 Use the modified Lazareno method fLazare like 51..· Measwrew/wi of agonist-stimulated[35S]GTP ^ S binding to cell . Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative prints membranes. Meth. Molec. Biol. 106, 231-243, 1999), the determination of [35S]GTP7 S binding to G-protein, in the initial [35S] GTP rs binding assay, the optimization of the test conditions led to the selection of the following buffer 2 Sputum: 20 millimolar Hepes and 100 millimolar NaCl, containing 3 micromolar GDP and 1 millimolar concentration 1 : 邑 (: 12 for 缪 opioid receptor CHO membrane, containing 10 micromoles Concentration of GDP and 1 millimolar concentration of MgCl^ in the opiate receptor C6 neuroglioma cell membrane, and 1 〇 micromolar concentration of GDP and 0.3 millimolar concentration of MgCl2 for the Kappa opioid receptor CHO membrane, test mixing - 56- This paper scale applies to China National Standard (CNS) A4 specification (2丨0x 297 mm) 1334351 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed A7 B7 V. Invention description (55) The compound contains 10 μg membrane protein , additionally adding 10 μg/ml soap to the diluted membrane as a cleaning agent to maximize [35S]GTPr S penetrating membrane. For the test activity, 175 μl of diluted membrane in the above buffer with 25 μl of 25 μl buffer and 25 μl of different concentrations of compound 5 in total volume is 225 μm Before raising the culture together, for the antagonistic activity, 25 μl of buffer was added to the reference agent to stimulate the basal value. For all three cell lines, DPDPE, U69593 and morphine at a concentration of 300 nanomolar were used for Corresponding receptor subtype, after incubation at 37 ° C for 20 minutes, 25 μl of [35S]GTPr S was added to a final concentration of 0.25 10 nanomolar and the test mixture was incubated at 37 °C. At 0 min, the bound and free-state [35S]GTPr S was separated by filtration through UniFilterTM-96, GF/BTM under reduced pressure using a Filtermate 196 (Packard), dried on a filter paper by liquid flash counting and scintillation was added (Microscint) -O; Packard) Determines the amount of bound radiation 15 activity on the filter paper unit. « The base [35s]GTPr s binding is measured in the absence of compound, and the stimulation via the agent is calculated as a percentage increase above the substrate value, using

The GraphPad Prism program analyzed the S-shaped agent concentration response curve and the inhibition reference effect of the 20-agent-stimulated [35S]GTP τ S binding antagonist inhibition curve by non-linear regression analysis in [35s]GTP 7 S binding. , C.4. Results All compounds according to the present invention showed a pICso value of at least 6 for the opiate receptor and a pIC5 〇 value for the 缪 or gram receptor of 6 or lower. The compounds listed in Table 6 show that for the opiate receptors between 7 and 8 -57- This paper scale applies the Chinese National Standard (cns) a4 specification (2丨〇 297 297 public)

1334351 Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperative Printed A7 B7 V. Inventive Note (56) pICso value and pIC5() value for 缪 or keba receptor 6 or lower, the compounds listed in Table 7 show Its opiate receptor is higher than 8 between ρΙ (:5 及 value and pIC5() value for 6 or lower of Pan or Keba receptor, and its selectivity for opioid receptors beyond opioid receptors is up to 6〇〇. MJ-pair, the pic5 of the opiate receptor agent test (^ compound number pIC50 value 1 Γ /IJ >=Π <-^4 〇*\4 compound number pICso value 43 7.9 22 7.3 17 7.9 87 7.3 30 7.9 45 7.3 105 7.9 51 7.3 78 7.9 4 7.3 101 7.8 55 7.3 28 7.8 71 7.3 11 7.8 99 7.3 29 7.8 34 7.2 * 67 7.8 72 7.2 ' 7 7.7 81 7.2 9 7.7 64 7.2 52 7.7 18 7.2 103 7.7 42 7.2 26 7.7 10 7.2 27 7.7 33 7.1 15 7.6 37 7.1 69 7.6 80 7.1 -58- tl This paper scale applies to the Chinese National Standard (CNS) A4 specification (2丨〇X 297 mm) 1334351 A7 B7 V. Invention Description (57) 50 7.6 90 7.1 32 7.6 56 7.1 93 7.5 47 7.1 65 7.5 43 7.1 84 7.5 48 7.1 66 7.5 79 7.0 75 7.4 111 7.0 13 7.4 7 7.0 76 7.4 68 7.0 96 7.4 95 7.0 94 7.4 92 7.0 70 7.4 49 7.0 36 7.3 74 7.0 Table 7: Agent Receptor Binding (pIC5G) and Signal Transmission Binding (pIC5Q) Test Ministry of Economics Intellectual Property The result of the printing of the employee consumption cooperatives of the bureau, nd: no test compounding * learning agent receptor binding agent by the object (pIC5 〇) body binding number which 50 value) De 缪 巴 巴 巴 巴 德 德 德 德Agent 3 0 Know. 〇, 8.8 6 nd 7.3 5 -59-

This paper scale applies to the national standard (CNS) A4 specification (2丨Ox 297 mm) Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed 1334351 A7 B7 V. Invention description (58) 38 0 8.7 6 ndndnd 20 a person 8.6 8.6 6 nd 7 5 102 • 8.5 6 ndndnd 25 . People again QC ) 3⁄4 8.4 6 nd 6.9 5 2 (/〇c ) 3⁄4 8.3 6 nd 6.8 5 « 41 c^qc Λ 3 ) 3⁄4 8.3 6 ndndnd: 98 (X °<- c μ? jV) 8.2 5.6 5.8 6.1 5 19 A person QI? 0 8.2 6 nd 6.5 5 -60- This paper size applies to the Chinese National Standard (CNS) A4 specification (210 X 297 mm)

1334351 A7 B7 V. INSTRUCTIONS (59) 24 0 persons c C 8.2 6 nd 6.9 5 1 0 C 8.1 5 6.3 nd 5 31 k/ C 8.1 6 ndndnd 12 0 c 8.0 6 nd 7 5 D. Clinical trial: heart deficiency Blood D.1. Rat-Langendorff model test for the compound of the Department of Economics, Intellectual Property Office, Staff Consumer Cooperative, printed test compounds 1, 24 and 25. 5 Mode Rapid dissection of the rat heart, connected to the cannula via the aorta and without perfusion of blood-free physiological buffer under 80 mm Hg perfusion pressure (modified Krebs-Henseleit buffer: NaCl 118 mmol/L, KC117 millimoles / liter, MgS041.2 millimoles / liter, KH2P041.2 millimoles / liter 10, CaCl2 1.8 millimoles / liter, NaHC03 23 millimoles / liter, glucose 11 millimoles / liter) Dissolve oxygen in physiological buffer, the heart beats 350 times per minute with the atrium, and will fill the balloon as needed (adjust the preload) -61- This paper scale applies to China National Standard (CNS) A4 specification (2丨Ox297 MM)

1334351 A7 B7 V. INSTRUCTIONS (60) Into the left ventricle, measure the diastolic pressure and the resulting pressure and the maximum rate of pressure generated as a measure of contractility (change in muscle contractility) and the minimum rate of pressure reduction as a measure of relaxation The advantage of this model is that the heart is isolated and studied under control without other organ interactions, 5 and the method is reasonably fast and inexpensive. The disadvantage is that there is no blood perfusion, only dissolved oxygen, so there is a larger crown. The arterial flow rate significantly dilates the coronary system, and there is no closed system and small animal model with no metabolism and no interaction with other organs, which is quite far from clinical practice. Protocol 10 The duration of the ischemic study was 20 minutes, and the measurement after the ischemic (reperfusion) study duration lasted for 40 minutes. The measurement before the ischemic study duration lasted for 30 minutes and was performed after stabilization, before ischemia. The situation group measured the following during these 30 minutes: 10 min baseline, 5 min ischemia, 5 min reperfusion, 5 min ischemia and 5 min reperfusion, in the compound according to the invention (4 μg / 5 0 0 15 ml buffer solution) In the group of 15 minutes, 15 minutes after the baseline, so everything is time-matched. Results The Ministry of Economic Affairs, the Intellectual Property Office, the employee consumption cooperative, printed the isolated heart, and recovered the contractile function after treatment with the compound according to the present invention, at least as good as the pre-ischemic group, and it must be noted that the drug according to the present invention has a negative influence on the heart. The effect of muscle contractility, which is confirmed by an increase in the final diastolic blood pressure of the left ventricle and a decrease in the developed pressure and dP/dUax, which may be related to solvent cyclodextrin because the same effect was observed in 2 experiments. The same concentration of cyclodextrin was used, however, compared with the control group, the functional recovery after the ischemic study time period was not -62- This paper scale was applied to the Chinese National Standard (CNS) A4 specification (210x297 mm) 1334351 Ministry of Economics intellectual property Bureau staff consumption cooperatives printed A7 B7 V. Invention description (61) is better, this shows that before ischemia, it is not the effect of negatively affecting muscle contractility but heart protection. Statistical analysis showed significant changes in developmental stress, final diastolic blood pressure, dP/dtmax, and 5 dP/dtmin between the control group and the group treated with the compound according to the present invention, pO.OOOOOl (see Figures 1 and 2). . The number of ventricular fibrillation that occurred during reperfusion with the group treated with the compound according to the invention was significantly reduced compared to the control group and the pre-ischemic group, and the group treated with the compound according to the invention was compared with the other groups. Less onset of ventricular fibrillation (Table 8). 10 Table 8: Results of the rat-Langendorff experiment-1C: ischemic contracture (mmHg) _vfib: number of ventricular fibrillation during reperfusion · -EDP: degree of normalization of EDP during reperfusion: Ο: restored to Baseline EDP; +.·. mmHg = mmHg above baseline EDP 15 -Recup: % recovery of pressure developed after reperfusion, 4 conjugate number voles No. 1C vfib EDP Recup · XI 10 0 0 91% 24 X2 5 1 (short) +5 85% X3 7 0 +7 87% Y1 7 2 (short) + 10 75% 25 Y2 5 0 +6 79% Y3 7 1 (short) +6 85 % Z1 4 0 0 92% -63- This paper size is applicable to China National Standard (CNS) A4 specification (210x297 mm)

1334351 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed A7 B7 V. Invention description (62) 1 Z2 6 0 +7 85% Z3 8 1 (short) +8 82% Control group C 21 4 +26 48% D.2 In order to deal with the weaknesses of the previous model, the sheep mode uses a complex in-vivo mode in which the heart effect is well controlled. ~ Tested compound ...' 5 Test compound 1. The model was anesthetized with ketamine and isoflurane. The venous cannula of the two vena cava and the arterial cannula of the carotid artery were used to connect the external circulation. The extra cannula with a flow meter in the right ventricle provided the flow velocity of the coronary artery, except for measuring the heartbeat. In addition to arterial blood pressure, the immediate left ventricular pressure and the left ventricular volume are measured via a delivery catheter. A lateral tube is also attached to the pulmonary artery on the arterial intubation of the artificial heart. This path allows the left ventricle to be adjusted in a perfect control mode. Load, in order to obtain measurements of left ventricular contractility and load before and after the ischemic study time course. 15 The advantage of this mode is that it is a large animal model, using the external circulation, in the metabolically active mode, there may be other non-cardiac effects, in a well-controlled heart condition, where the heart is perfused with blood, the technique of this model The nature of difficulty can be seen as a disadvantage. Experimental Protocol 20 The duration of the ischemic study was 30 minutes while maintaining the heart at room temperature and completely unloaded via the external circulation and ventilating ventricle. The control group and other groups were time-64-this paper scale applicable to the National Standard (CNS) A4 specification ( 2丨0x297 mm)

Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed 1334351 A7 B7 V. Invention Description (63) Anastomosis, the pre-ischemic condition group performed 3 times 5 minutes before the situation was ischemia and there was a 5 minute reperfusion interval, and the three groups were in the absence of implementation. The exosomes were started 30 minutes before the blood study time, and the exosomes were stopped 40 minutes after the ischemic study. In five technically successful experiments, the compound 1 was administered 15 minutes before the start of the ischemic study. 78 mg / 20 ml), 10 ml of solution in one experiment and 100 ml in the other four, only the four will be carried out here, in the other groups, 7 experiments have been carried out, Before the pressure-volume ratio of the left ventricle, the system is converted to a right heart shunt mode in which the venous motion is divided into circulation through the vena cava and blood is returned to the sheep via the pulmonary 10 artery, which is during baseline, pre-ischemic conditions, or medication compound 1 After the ischemic study time course and the ischemic study time course, 40, 70 and 100 minutes were completed. Results Traditional parameters 15 There was no significant difference in mean blood pressure between the groups. Compound 1 did not cause significant changes in blood pressure. It is necessary to understand that Compound 1 is a circulating drug in the external body, which significantly affects blood pressure. No significant difference was observed in left arterial pressure, although the left arterial pressure in the control group was slightly higher compared to the other two groups. 20 The same applies to heart flow after reperfusion, with a higher propensity for heart flow in the pre-ischemic group and in the group treated with Compound 1. Pressure-volume relationship The pressure-volume relationship of the left ventricle was studied in the control group, the pre-ischemic group, and the group treated with Compound 1. -65- This paper size is applicable to the National Standard (CNS) A4 specification (210x297 mm)

1334351 A7 B7 V. Description of the description (64) Regarding the load-related parameters, study the pre-loadable stroke work (PRSW, msw), the results are shown in Figure 3, PRSW points out, stroke work, and the final diastolic volume The ratio of stroke work is the effective mechanical work performed by the heart to expel the ash. When the lower final diastolic volume (due to the shorter sarcomere length) 5 completes the same external mechanical work, the p R s W is larger and its contraction The definition of the state 〇 ' After reperfusion at 7 〇 and 100 minutes and approaching the baseline value, compared with the control group, the pre-ischemic group and the group treated with Compound 1 had significantly better PRSW, therefore, ischemia The pre-existing group and the drug-carrying compound caused a better recovery of the systolic state after 30 minutes of ischemia at room temperature during the period of no ventricular load.滔(τ) is a measure of ventricular relaxation. The smaller the 滔 value, the faster the ventricular relaxation occurs. This measurement is important because the ventricular diastolic function is known to be more sensitive to ischemia than systolic function. After 70 and 100 minutes and 15 minutes, the sputum value of the group treated with Compound 1 was significantly smaller than that of the control group, and the group before ischemia was significantly lower than the control group 100 minutes after reperfusion. Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing parameters SW / PVA is the measurement of the efficacy of its use of cardiac mechanical work, A wind is external mechanical work and PVA is the mechanical energy produced by the heart as a stroke and achieve pressure increase in a specific volume The required potential energy, so far, this PVA (20 pressure-volume-area) is the most relevant mechanical parameter related to oxygen consumption. The greater the proportion of % SW/PVA, the more ventricular work is used as external work to discharge blood, and the lack of In the pre-blood group and the group treated with Compound 1, the control group had a significantly smaller SW/PVA ratio after 4, 70, and 100 minutes of the end of the ischemic study (Fig. 5). -66- This paper size applies to China National Standard (CNS) A4 specification (2丨〇 X 297 public Chu) A7

Ministry of Economic Affairs Intellectual Property Bureau Η Consumer Cooperative Printed 1^34351 The contractile effect is the consumption of Ο 2 converted to PVA or mechanical energy. The measurement was performed in the pre-ischemic group (44.5%) and the compound 1 (46.7%). Asked about the comparison (32.8%, p<〇.〇〇1). Another advantage of the compounds according to the invention is that the group treated with Compound 1 has a softer heart feel compared to the control group and the ischemic condition group. In summary, during the external circulation, 30 minutes of normal temperature myocardial ischemia 'The compound according to the invention is at least as effective as the pre-ischemic condition of the conventional _ as a cardioprotective mechanism, which enhances contraction and diastolic function, including consumption 〇 2 for The development of PVA and PVA into a foreign power SW is also more effective. The pharmacological protection of the compound according to the present invention is superior to that of the pre-ischemic condition because it provides the same efficacy as the pre-ischemic condition, including less risk ( The aorta cross-clamps, gently loses the sensation) and is less time consuming, and compared to the DADLE concentrations used in similar experiments, the compounds according to the invention are administered at a concentration that is at least 10 times lower.

15 to experiment: brain deficiency A <· The reduction of cerebral ischemia or cerebral protection according to the compound of the present invention can be used in a large voles using a transient forebrain ischemia model, in which, for example, sutures are used to close the middle cerebral artery The infarct size was measured after (MCA), which is disclosed in WO 96/27380, and in order to assess the functional node after ischemia, various behavioral tests can be performed according to the disclosure of WO 96/27380. -67- This paper size is applicable to China National Standard (CNS) A4 specification (210x297 mm)

Claims (1)

1334351 ab ψ B8 C8 D8 VI. Patent application scope 年 曰 ( 更 ( ( 专利 专利 专利 专利 专利 专利 专利 专利 专利 专利 专利 123 123 123 123 123 123 123 123 123 123 123 123 123 123 123 123 123 123 123 123 123 123 123 123 123 (II) Amended Claims in Chinese - Enel. (II) (Submitted on April 20, 2010) (Submitted on April 20, 2010) 1. A pharmaceutical composition used to reduce ischemic injury in organs in mammals , which contains as an active ingredient according to the compound of the formula (1) Rf B II -Α· Ri R (I) a pharmaceutically acceptable acid or base addition salt thereof, a stereochemically isomeric form thereof, a tautomeric form thereof and an N-oxide form thereof, wherein: A = B is selected from C = 0, C=N-R6 (wherein R6 is hydrogen or cyano), C=S, S02 and OCR7 R8, wherein R7 and R8 are each independently hydrogen and nitro; X is a covalent bond or -CH2-; The Intellectual Property Office employee consumption cooperative printed R1 is hydrogen, alkoxy, Ar-alkoxy, alkyl, polyalkyl, alkoxyalkyl, Ar-alkyl, Ar or NR9R1Q, of which R9 and RIG are independent The ground is hydrogen, alkyl, Ar, Ar-alkyl, hexahydro. Or a benzoxazolyl group, a thiazolyl group, a benzothiazolyl group, a benzocarbazole group, or a benzoxazolyl group. Sit-base and pyrimidine 11 base; -68 - This paper scale applies to China National Standard (CNS) A4 specification (210 X 297 mm) 1334351 A8 B8 C8 D8 VI. Patent application scope R2 is alkoxy or halogen; R3 is Alkyl, Ar, Ar-alkyl, Ar-alkenyl, pyridyl or isoxazolyl 1 R4, R5 are each independently hydrogen; p is an integer equal to 0 or 1; alkyl is from 1 to 6 a linear or branched saturated hydrocarbon group of a carbon atom; or a cyclic saturated hydrocarbon group (cycloalkyl group) having from 3 to 7 carbon atoms; or a cyclic saturated hydrocarbon group having from 3 to 7 carbon atoms a linear or branched saturated hydrocarbon group of 1 to 6 carbon atoms; wherein each carbon atom may be optionally substituted with an oxy group; the alkenyl group is an alkyl group having one or more double bonds; and Ar is selected from the group consisting of a phenyl group and a carbocyclic carbon ring, each optionally substituted with one or more substituents, each substituent being independently selected from the group consisting of an alkoxy group, a polyhaloalkoxy group, a halogen group, a alkyl group, a polypyrene group, a thiol group , a hydrazine group, a di-alkali group, a phenyl group, a nitro group or a SO2-CH3 group; the halogen group is selected from the group consisting of fluorine, gas and bromine; The property bureau employee consumption cooperative prints a polyhaloalkyl group which is a linear or branched saturated hydrocarbon group having from 1 to 6 carbon atoms or a cyclic saturated hydrocarbon group having from 3 to 7 carbon atoms, wherein one or more carbonaceous charcoal The atom is replaced by one or more halogen atoms. 2. The pharmaceutical composition according to claim 1, wherein R1 is selected from the group consisting of alkoxy groups, Ar-alkoxy groups, alkyl groups, polyhalogenated groups, alkoxyalkyl groups, Ar-alkyl groups. , Ar, or NR9R1G wherein R9 and R1Q are each independently hydrogen, alkyl, Ar, Ar-alkyl or alkyl ester. -69 - This paper size applies to China National Standard (CNS) A4 specification (210x297 mm) Patent application scope 3. According to the scope of the patent application, the composition of the drug is characterized by the fact that x is a covalent bond* 4. The pharmaceutical composition according to item 丨 or item 2 of the patent application is characterized by r3 疋k self-contained The benzyl group and the naphthyl group are each independently substituted with at least one substituent selected from the group consisting of a benzyl group, an alkyl ester group, a hydroxyl group, an alkoxy group, and a dialkylaminocarbonyl group. 5. The pharmaceutical composition according to claim 1, wherein A=B is C-0 or S〇2, R1 is alkoxy, alkoxyalkyl, Ai^nr9rio wherein R and R are each independently Hydrogen or Ar; or a = b and Ri together form a benzoheptyl group, p is hydrazine, R3 is a benzyl group optionally substituted with an alkyl or alkyl ester group, and R4 and R5 are each hydrogen. 6. The pharmaceutical composition according to claim 1, wherein the compound is selected from the group consisting of 1-ethyl ester-4-phenyl-4.[1-(phenylmethyl)·1Η-imidazole-2- ]]-hexahydropyridine; 1-propyl ester-4-phenyl·4·[1-(phenylmethylimidazol-2-yl)-hexahydropyridine; Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 1 -ethyl ester-4-phenyl-4-[1-[(4-pyridylphenyl)methyl]-1 oxime-imidazol-2-yl]-hexahydro D ratio bite; 1-ethyl ester group-4 -phenyl-4-[l-(l-phenylethyl)-glucizin-2-yl]-hexahydropyridine; 1-isopropyl ester-4-phenyl-4-[1-(benzene Methyl)·ιη-imidazol-2-yl]-hexahydropyridine; 1-ethyl ester-4-phenyl-4-[1-[[4-(decyl)phenyl]methyl]- 1Η-imidazole--70 - This paper scale is applicable to China National Standard (CNS) A4 specification (210x297 public hair) χ33435ΐ 6. Patent application scope 2-base]-hexachloro-n-specific bite; 1-phenylhydrazin-4- Phenylphenyl fluorenyl)-1 Η-imidazol-2-yl]-hexahydro σ-bipyridine; 1-(methoxyethenyl)-4-phenyl-4-[1-(1-phenylethyl) -1Η-imidazol-2-yl]-hexahydro-β ratio bite; 4-[[2-(1-benzylidinyl-4-phenyl-4-hexahydropyridyl)-1Η-imidazole-1 -base] Methyl]-methyl benzoate; 4-[[2-[1-(2-benzoxazolyl)>4-phenyl-4-olipopyridine)-1Η-imidazole-1-yl Methyl]•methylbenzoate; 1-benzylidene-4-ylphenyl-4-[1-(1·phenylethyl)-1Η-imidazol-2-yl]-hexahydro 0 ratio. 1-ethyl ester group _4·phenyl-4-[1-[1-[4-(ethyl)phenyl]ethyl]-1 Η-mi. Sodium-2-yl]-hexahydropyridin; and Ν,4-diphenylphenylmethyl)-1Η-imidazol-2-yl]-1-hexahydropyridine. 7. The pharmaceutical composition according to claim 1 or 2 of the patent application, characterized in that the organ is a heart, brain, liver, lung or kidney and the mammal is a human being. The species is used to induce a cardioprotective effect in a mammal. Medicinal composition comprising as an active ingredient a pharmaceutically acceptable acid or base addition salt of the compound according to formula (I), a stereochemically isomeric form thereof, its tautomeric form and its oxime-oxidation Form of matter. Pharmacy 9. Pharmaceutical composition according to the scope of the patent application, which additionally contains acceptable carrier 3 71 - This paper scale applies to China National Standard (CNS) A4 specification (21〇χ297 mm) 1334351 A8 B8 C8 D8 VI. Patent Application Range 10. The pharmaceutical composition according to claim 9 of the patent application, characterized in that the organ is a heart, a brain, a liver, a lung or a kidney and the mammal is a human. A pharmaceutical composition for inducing a cardioprotective effect in a mammal, comprising a pharmaceutically acceptable carrier and, as an active ingredient, a compound according to formula (I), a pharmaceutically acceptable acid or base thereof Salts, their stereochemically isomeric forms, their tautomeric forms, and their N-oxide forms. 12. A pharmaceutical composition in the form of a cardiac paralysis solution, comprising an effective amount of a compound according to formula (I), a pharmaceutically acceptable acid or base addition salt thereof, a stereochemically isomeric form thereof, and a mutual variation thereof The conformational form, its N-oxide form and a suitable carrier. 13. A pharmaceutical composition for treating a mammal undergoing an ischemic event, comprising an effective amount of a compound according to formula (1) and at least one second medical agent, including an antithrombotic agent and/or an angiogenic growth factor, or Each of the pharmaceutically acceptable acid or base addition salts, stereochemically isomeric forms thereof, tautomeric forms thereof, N-oxide forms thereof, and suitable carriers. Printed by the Ministry of Economic Affairs, Intellectual Property Office, Staff Consumer Cooperatives 14. The pharmaceutical composition according to item 1 of the patent application is characterized in that the mammal is a human. -72 - This paper size is applicable to China National Standard (CNS) A4 specification (210x297 mm)