CN1068566A - 2-(4-hydroxy piperidine subbase)-1-alkanol as antiischemic agents - Google Patents

Abstract

A series of 2-(4-hydroxypiperidino)-1-alkanol derivatives, Can be used as a drug to treat traumatic brain and spinal cord injuries and neurodegenerative diseases Diseases include senile dementia in mammals, especially humans.

Description

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The present invention relates to neuroprotective (anti-ischemic excitatory amino acid receptor blockade) 2-(4-hydroxypiperidino)-1-alkanol derivatives defined by the following formula (I); Acceptable salts; involving the use of these compounds in the treatment of stroke and trauma of the brain and spinal cord, and of neuronal degeneration in mammals, especially humans, including (but not limited to) senile dementias such as Alzheimer's, Humtington's and Parkinson's diseases method; also relates to some of its intermediates.

Benphenamine (A) is a racemic so-called dl-erythro compound having the following relative stereochemical formula:

It is marketed as an antihypertensive agent, and many analogues are also available as antihypertensive agents. See Carron et al., US Patent 3,509,164; Carron et al., Drug Res., Vol. 21 pp 1992-1999 (1971). More recently, benzphenamine has been found to have antiischemic and excitatory aminophenol receptor blocking activity, see Cotti et al., J.Pharm.Exp.Therap., Vol. 247, pp 1211-21 (1988); Carter et al., loc.cit., pp1222-32 (1988).

See also French patent 2546166, published January 17, 1990, and EPO publication EP-A1-351282. One goal is to find highly potent compounds with neuroprotective activity and low or insignificant hypotensive effects. This goal is substantially achieved in the present invention.

U.S. Patent 3,294,804 reports: certain 1-phenyl-3-(4-aryl-4-acetoxy-piperidino)-1-propanols can be used as analgesics; Japanese publication 53-02,474 ( CA89:146938W; Derwent Abs.14858A) and 53-59,675 (CA89:146938W; Derwent Abs.48671A) reported: 1-[4-(amino- and hydroxy-alkyl)phenyl]-2-(4 -Hydroxy-4-tolylpiperidino)-1-alkanols and alkanones with analgesic, antihypertensive, psychotropic or antiinfective activity; EP398,578-A and Der90-350,327/47. Report: 2-piperidino-1-alkanol derivatives have anti-ischemic activity.

The present invention is dedicated to compounds of the following formula (I) and pharmaceutically acceptable acid addition salts thereof,

Wherein R ₁ , R ₂ and R ₃ are respectively selected from hydrogen, an alkyl group containing 1 to 6 carbon atoms, and a substituted phenyl group, wherein the substituent on the substituted phenyl group is selected from hydroxyl, containing 1 to 4 Alkyl, chlorine, bromine, fluorine, trifluoromethyl, amino, nitro and alkoxy groups containing 1 to 4 carbon atoms; or _R1 and _R2 together form methylene, ethylene, Propylene or butylene;

m is 0 to 2;

m is 1 or 2;

X and Y are respectively selected from hydrogen, chlorine, bromine, fluorine, trifluoromethyl, alkoxy containing 1 to 4 carbon atoms, alkyl containing 1 to 4 carbon atoms, hydroxyl, amino, nitro and substituted phenoxy, wherein the substituents on the substituted phenyl are selected from the group consisting of hydrogen, hydroxyl, alkyl containing 1 to 4 carbon atoms, chlorine, bromine, fluorine, trifluoromethyl, nitro, amino and An alkoxy group of 1 to 4 carbon atoms; M and Q are respectively selected from hydrogen, hydroxyl, amino, chlorine, bromine, fluorine, trifluoromethyl, nitro, an alkyl group containing 1 to 4 carbon atoms, containing 1 Alkoxy with 4 to 4 carbon atoms, N,N-dialkylamino with 1 to 4 carbon atoms in each alkyl group, N-alkylamino with 1 to 4 carbon atoms, NHCOR ₄ , NHCOOR _{5 and NHSO2R6} ;

In the formula, R is selected from _hydrogen , alkyl containing 1 to 6 carbon atoms, phenyl and substituted phenyl, wherein the substituent on the substituted phenyl is selected from hydroxyl, chlorine, bromine, fluorine, trifluoro Methyl, amino, nitro, alkyl with 1 to 4 carbon atoms and alkoxy with 1 to 4 carbon atoms;

Wherein R ₅ and R ₆ are selected from alkyl, phenyl and substituted phenyl containing 1 to 6 carbon atoms respectively, wherein the substituent on the substituted phenyl is selected from hydroxyl, chlorine, bromine, fluorine, tri Fluoromethyl, amino, nitro, alkyl with 1 to 4 carbon atoms and alkoxy with 1 to 4 carbon atoms;

Or M and Q together form a divalent group Z, wherein Z is selected from:

In the formula, _R7 and _R8 are respectively selected from hydrogen and methyl.

"Pharmaceutically acceptable acid addition salt" refers to (but not limited to) such as hydrochloride, hydrobromide, hydroiodide, nitrate, hydrogen sulfate, dihydrogen phosphate, methanesulfonate , maleate and succinate. Said salts are generally prepared by reacting the free base form of compound (I) with an appropriate acid (generally one molar equivalent) in a solvent. Those salts which do not precipitate directly are generally isolated by evaporation of the solvent and/or addition of a non-solvent followed by filtration.

，并且化合物在丙醇链上的1位和2位具有

或赤式相对立体化学结构，即A preferred group of compounds of the invention are those wherein M and Q form a free radical Z, R _{and R} are hydrogen and _R is methyl, wherein Z is

, and the compound has the 1 and 2 positions on the propanol chain

or erythro relative stereochemical structure, namely

，并且化合物在丙醇链上的1位和2位具有

或苏式相对立体化学结构，即Another group of preferred compounds of the invention are those wherein M and Q form a free radical Z, R _{and R} are hydrogen and _R is methyl, wherein Z is

, and the compound has the 1 and 2 positions on the propanol chain

or threo relative stereochemistry, ie

The present invention is also directed to a pharmaceutical composition comprising a compound of formula I of the present invention; and a method of treating central nervous diseases in mammals, especially humans, which comprises administering to said mammal a neuroprotective effective amount of formula (I) compound. The compositions and methods are particularly useful in the treatment of brain and spinal cord trauma, stroke, Alzheimer's disease, Parkinson's disease, Huntington's disease and related central nervous system disorders.

The present invention is also committed to the intermediate compound of the following formula

Wherein, _R2 and _R3 are respectively selected from hydrogen, an alkyl group containing 1 to 6 carbon atoms, and a substituted phenyl group, wherein the substituents on the substituted phenyl group are selected from hydroxyl, containing 1 to 4 carbon atoms Alkyl, chlorine, bromine, fluorine, trifluoromethyl, amino, nitro and alkoxy groups of 1 to 4 carbon atoms;

m is 0 to 2;

m is 1 or 2;

X and Y are respectively selected from hydrogen, chlorine, bromine, fluorine, trifluoromethyl, alkoxy containing 1 to 4 carbon atoms, alkyl containing 1 to 4 carbon atoms, hydroxyl, amino, nitro and substituted phenoxy, wherein the substituents on the substituted phenyl are selected from the group consisting of hydrogen, hydroxyl, alkyl containing 1 to 4 carbon atoms, chlorine, bromine, fluorine, trifluoromethyl, nitro, amino and Alkoxy groups with 1 to 4 carbon atoms; M and Q are selected from hydrogen, hydroxyl, amino, chlorine, bromine, fluorine, trifluoromethyl, nitro, alkyl groups with 1 to 4 carbon atoms, 1 Alkoxy with 4 to 4 carbon atoms, N,N-dialkylamino with 1 to 4 carbon atoms in each alkyl group, N-alkylamino with 1 to 4 carbon atoms, NHCOR ₄ , NHCOOR _{5 and NHSO2R6} ;

In the formula, R is selected from _hydrogen , alkyl containing 1 to 6 carbon atoms, phenyl and substituted phenyl, wherein the substituent on the substituted phenyl is selected from hydroxyl, chlorine, bromine, fluorine, trifluoro Methyl, amino, nitro, alkyl with 1 to 4 carbon atoms and alkoxy with 1 to 4 carbon atoms;

Wherein R ₅ and R ₆ are selected from alkyl, phenyl and substituted phenyl containing 1 to 6 carbon atoms respectively, wherein the substituent on the substituted phenyl is selected from hydroxyl, chlorine, bromine, fluorine, tri Fluoromethyl, amino, nitro, alkyl with 1 to 4 carbon atoms and alkoxy with 1 to 4 carbon atoms;

Or M and Q together form a divalent group Z, wherein Z is selected from:

In the formula, _R7 and _R8 are respectively selected from hydrogen and methyl.

Depending on the specific values of R ₁ , R ₂ , and R ₃ , compounds of formula (I) may have one or two asymmetric centers and thus exist in various isomeric forms. All such isomers are within the scope of the present invention. The individual isomers can be separated by classical methods well known to those skilled in the art.

The compounds of formula (I) of the invention, as defined above, are generally readily prepared by reacting the chloro compound (II) with piperidine (III), followed by reduction of the resulting ketone to the alcohol, as detailed below.

Usually the -OH and -NH ₂ substituents in the precursor ketones are initially prepared in protected form, eg -OA ₁ , or -NHA ₂ in compounds of formula (IV). A ₁ and A ₂ are as defined below. The above-mentioned protected ketones are usually prepared by reacting an appropriately substituted 2-halo-1-alkanone (II) with an appropriately substituted piperidino derivative (III).

For example:

The reaction of compound (II) with compound (III) is usually carried out under typical nucleophilic substitution conditions. Substantially equal molar equivalents may be employed when the availability of the two reactants is approximately equal. Although when one reactant is more available, it is usually best to use an excess of one reactant so that the bimolecular reaction is completed in a shorter time. The reaction is usually carried out in a reaction-inert solvent such as ethanol in the presence of at least 1 molar equivalent of the base, its piperidine derivative if readily available, but often a tertiary amine at least as strong as the nucleophilic piperidine. conduct. If necessary, the reaction can be carried out by adding up to 1 molar equivalent or more of iodine salt (eg: NaI, KI). The temperature is not critical, but is generally high so that the reaction is complete in a short time, but not so high as to cause excessive decomposition. A temperature range of 50-120°C is generally satisfactory. The temperature is conveniently taken to be the reflux temperature of the reaction mixture.

As used above and elsewhere herein, the term "reaction inert solvent" refers to any solvent that does not interact with starting materials, reagents, intermediates or products in a manner that would adversely affect the yield of the desired product.

If necessary, the ketone intermediate (IV) bearing a protected form of the OH or NH group (OA ₁ or NHA ₂ ) can be deprotected in a conventional manner at this step.

For example, when A is triisopropylsilyl or tert-butyldimethylsilyl, it can be obtained by mixing tetrabutylammonium fluoride (usually, substantially 2 molar equivalents) in a random solvent (such as tetrahydrofuran) ) method to remove the protecting group conveniently. When A is benzyl or A is benzyloxycarbonyl, the protecting group is usually removed by conventional hydrogenolysis in the presence of a noble metal compound in a reaction-inert solvent. For example using 10% Pd/C as catalyst, preferably at low pressure (eg: 1-10atm) and low temperature (eg: 20-75°C) in a reaction inert solvent like methanol.

Typically, the ketone intermediate (IV) is converted to the corresponding alcohol by one of two conventional reduction procedures to selectively produce the threo or erythro compounds of formula (I).

Earlier and elsewhere in this paper, "Soviet style" or Refers to the relative stereochemical formula of the 1- and 2-positions on the propanol chain, namely:

是指在丙醇链上的1-和2-位的相对立体化学式，即：while "red" or

Refers to the relative stereochemical formula of the 1- and 2-positions on the propanol chain, namely:

In order to obtain the desired erythro compound of formula (I), the corresponding ketone intermediate (IV) can be reduced with potassium borohydride, usually in excess (for example, more than 5 molar equivalents), in the presence of glacial acetic acid, in a protic solvent such as In ethanol, usually at 15-20°C.

In order to obtain the desired threo compound of formula (I), the corresponding ketone intermediate (IV) can be conveniently reduced with sodium borohydride, usually in excess (for example, more than 5 molar equivalents), in a protic solvent such as ethanol, usually in Carry out at 15-20°C. The resulting mixture is subjected to silica gel column chromatography to obtain the threo compound of the formula (I).

Any protecting groups remaining after reduction of the ketone are subsequently removed by standard procedures as described above.

The starting materials and reagents used to synthesize the compounds of the present invention are readily available, either commercially, prepared according to literature procedures, or prepared by the preparations exemplified below.

The compound of formula (I) of the present invention has selective neuroprotective activity, which is based on its anti-ischemic activity and ability to block excitatory amino acid receptors, while having low or no hypotensive activity. The anti-ischemic activity of the compounds of the present invention can be determined according to one or more of the methods described in detail in Gotti et al. and Carter et al. above, or in an analogous manner.

The ability of the compounds of the invention to block excitatory amino acids was demonstrated by rescuing neuronal cultures of rats exposed to the excitotoxic amino acid glutamate. The following is a typical approach. Part I: Cell Separation:

Embryos at 17 days of gestation were removed from the mice and placed in Tyrode's solution. Brains were then removed and placed in fresh Tyrode's solution. Remove the rhomboid and thalamus with an iris knife. Then, divide the forebrain into two hemispheres. The meninges are then gently removed. The hippocampus appears as a black folded area inside the dermal margin. The hippocampus was carefully dissected from the remaining tissue and placed in the opposite corner of the dish. When all dissections were completed, the hippocampal tissue stored in the other horn was minced into 1mm pieces. The fragments were removed with a Pasteur pipette and placed into sterile tubes. Gently aspirate Tyrode's solution and add calcium-magnesium-free Tyrode's solution. The tissue was washed three times with Tyrode's solution without calcium and magnesium. The final wash was incubated at 37 °C for 15 min. The buffer was again removed and replaced with 11 M of fresh Tyrode's solution without calcium and magnesium. Then add 0.1% trypsin sodium (100μl, 10mg/ml stock sterile solution), and incubate the tube at 37°C for 1 hour. After the trypsin sodium incubation, wash the tissue with serum containing medium to stop the trypsin sodium effect. Tissue was resuspended in 1 ml of fresh medium and triturated with a fine-bore Pasteur pipette. Cells were then counted with a hemocytometer. Cells were then seeded in 96 wells of Falcon Primeria tissue culture dishes with 75,000 cells per well in Complete medium. The complete medium consisted of minimal essential medium (MEM) with Earle's salts, 10% fetal calf serum, 10% horse serum, L-glutamic acid (2mM), penicillin-lianmycin (100U per mL), and glucose ( A 100-fold stock solution containing 27.8 g per 100 ml was prepared so that the final concentration was 21 mM. On the third day the dish was inoculated with fresh culture. After two more days, cytosine arabinoside was removed and replaced by a maintenance culture (Complete medium minus fetal calf serum). Discs were then inoculated twice a week and three weeks after dissection, the discs were used for glutamate toxicity assays to ensure normal development of nerves in culture.

Part II: Glutamate treatment and post-addition of glutamate.

After three weeks of culture, the medium was removed from the cells and washed three times with chlorine-free adjusted saline solution (CSS-Cl). CSS-Cl contains 69mM Na ₂ SO ₄ , 2.67mM K ₂ SO ₄ , 0.33mM NaHPO ₄ , 0.44mM KH ₂ PO ₄ , 1mM NaHCO ₃ , 1mM MgSO ₄ , 10mM HEPES (N-2-hydroxyethylpiperazine -N'-2-ethylsulfonic acid), 22.2 mM glucose and 71 mM sucrose, pH=7.4. After washing, 1 to 3 mM glutamate in CSS-Cl buffer was added, and appropriate control wells contained buffer without glutamate. Plates were incubated at 37°C for 15 minutes. After glutamate incubation, wash twice with serum-free medium. Prepare the appropriate concentration of the test drug in serum-free medium and add it to the corresponding pool of the microtiter plate (100 μl per pool). Negative control pools received serum-free medium without drug. Several glutamate-treated pools were also given drug-free serum-free medium as a positive control. Plates were incubated overnight at 37°C and viability was assessed the next day with LDH (lactate dehydrogenase) and MTT (methylthiotetrazolium).

Part III: Evaluation of cell viability:

Take out 100μl medium from each plate, transfer to a clean plate, and measure the amount of LDH released. Add 100 μl MTT solution to each pool. MTT solution was prepared by adding 10 μl of MMT stock solution (5 mg/ml in PBS, phosphate-buffered saline) per 100 μl of serum-free medium. Plates were incubated at 37°C for 4 to 6 hours. Then, 100 μl of acid-alcohol solution (0.08 N HCl in isopropanol) was added to each well, and the wells were stirred vigorously to dissolve the pink crystals. Control pools should contain media containing MTT and acid alcohol but no cells. Then, read with a microplate reader, using a dual-wavelength test filter, 570nm, and a parametric filter, 630nm. The disk must be read within 1 hour.

Then, the removed medium was assessed for LDH. An equal volume of the withdrawn sample was added to the LDH reaction mixture. At this point the appropriate pools were mixed to obtain a 500 μl sample. For each sample, the mixture was reconstituted by reconstituting 480 μl 0.1 M sodium phosphate, pH 7.5, 10 μl sodium pyruvate (66 mM) and 10 μl reduced NADH (5 mg NADH per vial, in 440 μl 0.1 N NaOH, each 10 μl for each sample). The sample is quickly added to the reaction mixture in the small cup, and the disappearance of the absorption peak at 340nm is measured with a Beckmean DU-8 spectrophotometer.

Undesired hypotensive activity is also determined by known methods, eg also by Carron et al. above.

The selective neuroprotective anti-ischemic activity and excitatory amino acid blocking activity make the compound of the present invention useful for treating brain and spinal cord trauma, degenerative CNS (central nervous system) diseases such as: stroke, Alzheimer's disease, Parkmson's disease and Humtington's disease , without obvious superficial or immediate excessive reduction in blood pressure. When using neuroprotective amounts of the compound of formula (I) of the present invention to treat the above-mentioned diseases in humans, the dosage is generally 0.02 to 10 mg/kg/day (1-500 mg/day for a person with a general weight of 50 kg), in a single dose or in divided doses. dose, regardless of the route of administration. Of course, depending on the particular compound and the nature of the particular patient's condition, the clinician may employ doses outside the above ranges. Oral administration is generally preferred. However, parenteral (i.m, i.v.) or topical administration is the preferred route of administration if the patient cannot swallow it or buccal absorption would reduce efficacy.

The compound of the present invention is usually administered in the form of a pharmaceutical composition comprising at least one compound of formula (I) and a pharmaceutically acceptable carrier or diluent in a ratio of 1:20 to 20:1 usually Adopt conventional method to prepare above-mentioned composition with solid or liquid carrier, to adapt to desired administration method: for oral administration, make tablet, hard or soft gelatin capsule, suspension, granule, powder etc.; For enteric administration, it is made into injection solution or suspension, etc.; for topical administration, it is made into solution, lotion, ointment, ointment, etc.

The invention is illustrated, but not limited, by the following examples.

All non-aqueous reactions were performed under anhydrous, oxygen-free nitrogen to generally maximize yields. All solvents/diluents were dried according to standard published methods or purchased in pre-dried form. All reactions were stirred mechanically or electromagnetically. NMR spectra were recorded at 300 MHz and are expressed in ppm downfield from trimethylsilane. NMR solvent was _CDCl3 unless otherwise stated, and IR spectra are expressed in microns and generally indicated only stronger signals.

Example 1

(±)-3,4-Dihydro-6-(1-hydroxy-2-(1-(4-hydroxy-4-phenoxymethyl)piperidinyl)ethyl)quinoline-2-(1H )-ketone

300mg (1.23mmol) 4-hydroxy-4-(phenoxymethyl)piperidine hydrochloride, 409mg (1.84mmol) 6-(2-chloroacetyl)-3,4-dihydroquinoline-2 A mixture of (1H)-ketone and 0.514ml (0.373g, 3.7mmol) of triethylamine in 25ml of acetonitrile was stirred overnight at 60°C. Then, the solvent was removed in vacuo, the residue was partitioned between water and ethyl acetate, and the organic layer was washed with water and brine. The ethyl acetate layer was dried with brine and magnesium sulfate, and the solvent was evaporated to give 3,4-dihydro-6-(1-oxo-2-(1-(4-hydrogen-4-phenoxymethyl)piperidinyl ) ethyl) quinolin-2-(1H)-one brown solid, which can be directly used in the subsequent reduction step without further purification.

The above ketone was dissolved in 25ml of absolute ethanol, and 500mg (13.1mmol) of NaBH was added in batches over 20 minutes. The reaction mixture was stirred at room temperature for 4 hours, then the solvent was removed and the residue was partitioned between water and ethyl acetate. After drying, the ethyl acetate was removed in vacuo and the residue was chromatographed on silica gel to give 73 mg (15%) of the product, m.p. 186-188°C

NMR ( _CD3OD ) δ1.70-2.10 (4H, m), 2.52-3.07 (10H, m), 3.33 (2H, s), 3.83 (2H, s), 6.82-7.38 (8H, m).

Example 2

(±)-5-(1-Hydroxy-2-(1-(4-hydroxy-4-phenoxymethyl)piperidinyl)ethyl)benzimidazolin-2-one

According to the method of Example 1, from 4-hydroxy-4-(phenoxymethyl)piperidine hydrochloride (1.23mmol), 5-(2-chloroacetyl)-2-hydroxybenzimidazole (1.84mmol ) and triethylamine (3.7mmol) in acetonitrile (25ml) to give the title compound. The resulting ketone was stirred together with sodium borohydride (13.1 mmol) in absolute ethanol, and the desired compound was obtained after silica gel chromatography with a yield of 35%, m.p.232-235°C

Theoretical value C ₂₁ H ₂₅ N ₃ O ₄ ·H ₂ O: C, 62.81; H, 6.77; N, 10.46. Found value: C, 62.98; H, 6.54; N, 10.32.

Example 3

(±)-5-(1-Hydroxy-2-(1-(4-hydroxy-4-phenoxymethyl)piperidinyl)ethyl)-2-oxindole

According to the method of Example 1, from 4-hydroxy-4-(phenoxymethyl)piperidine hydrochloride (1.23mmol), 5-(2-chloroacetyl)oxindole (1.84mmol) and triethyl A solution of the amine (3.7mmol) in acetonitrile (25ml) gave the title compound. The resulting ketone was stirred with sodium borohydride (13.1mmol) in absolute ethanol and the desired compound was obtained after silica gel chromatography. Yield 40%, m.p. 171-174°C.

Example 4

(±)-erythro-5-(1-hydroxy-2-(1-(4-hydroxy-4-phenoxymethyl)piperidinyl)propyl)benzimidazolin-2-one

933 mg (2.36 mmol) of (±)-1-(5-(2-hydroxybenzimidazolyl)-2-(1-(4-hydroxy-4-phenoxymethyl)piperidinyl)propan-1 - A solution of the ketone in 10 ml of glacial acetic acid and 50 ml of absolute ethanol was treated in batches with 944 mg (17.48 mmol) of potassium borohydride at 15-20° C., and the resulting solution was stirred overnight at room temperature. The reaction mixture was evaporated to dryness, leaving The material was dissolved in a minimum amount of water. The pH of the solution was adjusted to 7-8 with solid NaHCO, and a solid precipitated. The material was insoluble in chloroform and substantially insoluble in ethyl acetate. The whole was evaporated to dryness again, and the crystals The residue was dissolved in ethanol and filtered to remove the salt. The ethanol was evaporated and the residue was dissolved in isopropanol and treated with HCl gas in ether to precipitate an amorphous salt which was isolated by filtration and dried under nitrogen The material was dissolved in a hot mixture of ethyl acetate and methanol, clarified with decolorizing charcoal, and the methanol was then boiled off. After cooling, the product was obtained as colorless crystals, 410 mg (40%)

mp254-255°C. IR (KBr) 5.90 μm; NMR (CD ₃ OD) δ1.22 (3H, d, J=7), 1.95-2.09 (2H, m), 2.15-2.30 (2H, m), 3.42 -3.76 (4H, m), 3.91 (2H, s), 5.47 (1H, s), 6.92-7.35 (8H, m).

Example 5

(±)-threo-5-(1-hydroxy-2-(1-(4-hydroxy-4-phenoxymethyl)piperidinyl)propyl)benzimidazolin-2-one

Add a total of 700 mg (18.4 mmol) of sodium borohydride in portions to 325 mg (0.82 mmol) of (±)-1-(5-(2-hydroxybenzimidazolyl)-2-(1-(4-hydroxy -4-phenoxymethyl)piperidinyl)propan-1-one in absolute ethanol (20ml) suspension, the reaction mixture was stirred overnight at room temperature.Then the solvent was evaporated and the residual foam was dissolved in ethyl acetate and In water, the aqueous layer was extracted with ethyl acetate. The ethyl acetate extracts were combined, dried and evaporated, and the residual foam was subjected to silica gel column chromatography, eluting with 1:1 ethanol/ethyl acetate, to obtain a white solid product.

mp>250°C. NMR (acetone-d ₆ ) δ0.79 (3H, d, J=7), 1.71-1.88 (2H, m), 11.90-2.08 (2H, m), 2.48-2.88 (4H, m ), 3.01 (1H, t, J=7), 3.88 (2H, s), 4.26 (1H, d, J=7), 6.86-7.32 (8H, m); theoretical value

C ₂₂ H ₂₇ N ₃ O ₄ .1.5 H ₂ O: C/62.24; H, 7.12; N, 9.89. Found C, 61.72; H, 6.73; N, 9.03.

Example 6

(±)-erythro-3,4-dihydro-6-(1-hydroxy-2-(1-(4-hydroxy-4-phenoxymethyl)piperidinyl)propyl)quinoline-2 (1H)-ketone

At 15-20°C, 7.13g (17.5mmol) of (±)-1-(6-(1,2,3,4-tetrahydro-2-oxoquinolinyl))-2-(1- (4-Hydroxy-4-phenoxymethyl)piperidinyl)propan-1-one was dissolved in 135 ml of absolute ethanol and 70 ml of glacial acetic acid and treated in batches with 6.22 g (115 mmol) of KBH ₄ at 15–20 °C After that, it was then allowed to warm to room temperature for 30 min, the reaction mixture was evaporated to dryness, and the residue was dissolved in ice water and basified with solid NaHCO ₃ . The precipitated solid was separated by filtration, washed with water, and air-dried to give 3.66 g of crystalline free base, mp 192-196°C. The filtrate was extracted with ethyl acetate and the combined organic phases were dried over brine and _MgSO4 and evaporated to give another 786 mg of product (62% overall yield). A 510 mg sample of this material was dissolved in ethyl acetate and treated with ethereal HCl gas to give 475 mg of the crystalline hydrochloride salt.

mp214-216°C (dec).IR (KBr) μm; NMR (CD ₃ OD) δ1.15 (3H, d, J=7), 1.86-2.04 (2H, m), 3.52-3.66 (2H, m) , 3.69-3.80 (1H, m), 3.86 (2H, s), 5.34 (1H, s), 6.81-6.96 (4H, m), 7.17-7.28 (4H, m).

(±)-threo-3,4-dihydro-6-(1-hydroxy-2-(1-(4-hydroxy-4-phenoxymethyl)piperidinyl)propyl)quinoline-2 (1H)ketone

Add a total of 1.50 g (39.5 mmol) of NaBH in portions to 700 mg (1.71 mmol) of (±)-1-(5-(2-hydroxybenzimidazolyl))-2-(1-(4-hydroxy -4-phenoxymethyl)piperidin)propan-1-one in absolute ethanol (50ml) suspension, stirred overnight at room temperature. Then, the solvent was evaporated, the residual foam was dissolved in ethyl acetate and water, and the aqueous layer was extracted with acetic acid. The organic phases were combined, dried and evaporated, and the residual foam was chromatographed on a silica gel column eluting with 1:1 ethanol/ethyl acetate to give the product as a white solid, m.p.192-196°C. A small amount of erythro compound is formed in this reduction, which can be separated by column.

NMR (CD ₃ OD) δ-0.82 (3H, d, J=7), 1.72-2.06 (4H, m), 2.50-2.82 (6H, m), 2.88-3.02 (2H, t, J=7), 3.02 (1H, t, J=7), 3.84 (2H, s), 4.28 (1H, d, J-7), 6.80-7.34 (8H, m);

Theoretical for _{C22H27N3O4.1.5H2O :} C, _65.88 ; H, _7.60 ; _N , 6.40.

Found values: C, 65.74; H, 7.09; N, 6.31.

Example 8

(±) Erythro-5-(1-hydroxy-2-(1-(4-hydroxy-4-phenoxymethyl)piperidinyl)propyl)-2-oxindole

0.5g (2.05mmol) 4-hydroxy-4-phenoxymethyl) piperidine hydrochloride, 0.5g (2.25mmol) 5-(2-chloropropionyl)-2-oxindole and 1ml (0.725 g, 7.18 mmol) a mixture of triethylamine and 20 ml of acetonitrile was refluxed for 24 hours. The solvent was removed in vacuo, the residue was partitioned between dry ethyl acetate and water, the acetic acid layer was washed with water and brine, dried over magnesium sulfate, and concentrated to give ketone, 537 mg (66%) of brown foam, which was directly used in the next reaction without further purification.

A solution of the above ketone (500 mg, 1.26 mmol) in ethanol (20 mL) was treated in portions with 1.0 g (26.3 mmol) of _NaBH4 and the resulting mixture was stirred at room temperature for 24 hours. The solvent was removed in vacuo and the residue was partitioned between ethyl acetate and water. The ethyl acetate layer was washed with water, dried over magnesium sulfate and brine, then evaporated to dryness. The residue was chromatographed on silica gel eluting with a mixture of ethyl acetate and increasing ethanol to give 121 mg (24%) of the pure fraction of the threo product

mp204-207°C. NMR (DMSO- _d6 ) δ0.70 (3H, d, J=7), 1.58-1.92 (4H, m), 2.40-2.65 (4H, m), 2.86 (1H, m), 3.32-3.40 (2H, m), 3.79 (2H, s), 4.20 (1H, d, J=7), 6.70-7.35 (8H, m), 10.34 (1H, s).

Example 9

(±)-1-(6-(1,2,3,4-tetrahydro-2-oxoquinolinyl))-2-(1-(4-hydrogen-4-phenoxymethyl)piperene pyridyl)propan-1-one

8.30 g (34.06 mmol) of 4-hydroxy-4-phenoxymethylpiperidine hydrochloride and 8.09 g (34.04 mmol) of 6-(2-chloro- 1-Propionyl)-1,2,3,4-tetrahydroquinolin-2(1H)-one was suspended in acetonitrile (100ml), and the mixture was heated to reflux for 3 hours and then stirred at room temperature overnight.

The reaction mixture was poured into water and extracted 3 times with ethyl acetate, the organic phases were combined, dried over brine solution and magnesium sulfate, evaporated to a foam, dissolved in ethyl acetate and cooled to give a brown solid which was found to be the starting chloroketone , discard it. The filtrate was evaporated, the residue was dissolved in ethyl acetate and diethyl ether was added to facilitate crystallization. The product was filtered and washed with ether to give 8.84 g (63.6%) of product as a cream solid, m.p. 137-139°C. Analytical samples were crystallized from hot ethyl acetate.

NMR

(CD ₃ OD) δ1.28 (3H, d, J=7), 1.60-1.92 (4H, m), 2.52-2.84 (6H, m), 3.00 (2H, t, J=7), 3.75 (2H , s), 4.22 (1H, q, J=7), 6.82-7.00 (4H, m), 7.16 (2H, m), 7.82-7.98 (2H, m);

Theoretical C ₂₄ H ₂₈ N ₂ O ₄ : C, 70.56; H, 6.91; N, 6.86. Found C, 70.16; H, 6.78; N, 6.76.

Example 10

(±)-1-(5-(2-Hydroxybenzimidazolyl))-2-(1-(4-hydroxy-4-phenoxymethyl)piperidinyl)propan-1-one

2.43 g (10.0 mmol) of 4-hydroxy-4-phenoxymethylpiperidine hydrochloride and 2.25 g (10.0 mmol) of 5-(2-chloro- 1-Propionyl)-2-hydroxybenzimidazole was suspended in acetonitrile (40ml). The reaction mixture was heated to reflux for 90 minutes and allowed to stand at room temperature over the weekend.

The reaction mixture was then poured into a mixture of water and ethyl acetate and the suspended solid was separated by filtration and found to be pure product weighing 1.15 g after drying. The filtrate was adjusted to pH=7.0, extracted several times with ethyl acetate, and after drying with brine solution and magnesium sulfate, a colorless solid was precipitated from hot ethyl acetate/methanol to give another 560 mg of product (total yield 43%)

m.p.230-235℃（dec.）.NMR

(CD ₃ OD/DMSO-d ₆ ) δ1.29 (2H, d, J=7), 1.60-1.92 (4H, m), 2.54-2.84 (4H, m), 3.77 (2H, s), 4.26 ( 1H, q, J=7), 6.86-7.10 (6H, m), 7.75-7.92 (2H, m).

Example 11

(±)-1-(5-(2-oxindolyl))-2-(1-(4-hydroxy-4-phenoxymethyl)piperidin)propan-1-one

According to the method of preparation 10, from 4-hydroxy-4-phenoxymethylpiperidine hydrochloride (10.0mmol), 5-(2-chloropropionyl)-2-oxindole (10mmol) and triethylamine (35mmol) in acetonitrile (50ml) to give the title compound. The title compound was isolated by crystallization from hot ethyl acetate/methanol as an amorphous foam in 66.4% yield.

NMR ( _CDCl3 ) δ1.28 (3H, d, J=7), 1.58-1.78 (4H, m), 2.40-2.84 (4H, m), 3.54 (2H, s), 3.76 (2H, s), 4.09 (1H, q, J = 7), 6.78-6.96 (3H, m), 7.14-7.26 (2H, m), 7.84-8.05 (3H, m), 9.52 (1H, broad s), 9.64 (1H, broads).

preparation 1

3,4-Dihydroquinolin-2-(1H)-one

A slurry of 50.0 g (0.259 mol) of o-nitrocinnamic acid in ethanol (500 ml) was treated with 5 scoops of Raney nickel and hydrogenated in a Parr shaker with an initial pressure of 50 psi. In the morning the pressure was increased again to 50 Psi and the reaction was allowed to proceed for an additional 5 hours. The reaction mixture was filtered to remove the catalyst, then passed through a bed of silica gel washed with a mixture of ethyl acetate and ethanol to remove traces of nickel salts. Evaporation of the filtrate afforded the desired product in 57% yield.

NMR

(DMSO-d ₆ ) δ2.45 (2H, t, J=7), 2.87 (2H, t, J=7), 6.87 (2H, d of d, J=7, 7), 7.12 (2H, d of d, J=7, 10), 10.08 (1H, s).mp165-166℃.

preparation 2

6-(2-Chloropropionyl)-3,4-dihydroquinolin-2-(1H)-one

72.5g (0.544mol) _AlCl3 in _CS2 (800ml) suspension was stirred in dry nitrogen atmosphere, 14.1ml (20.0g, 0.177mol) 2-chloropropionyl chloride and 20.0g (0.136ml) 3,4 were added successively -Dihydroquinolin-2(1H)-one, the reaction mixture was refluxed, phase separation occurred. The reaction was quenched by pouring onto ice with vigorous stirring. The resulting pale yellow precipitate was isolated by filtration, washed with water, and dried overnight over P ₂ O ₅ to obtain 27.7 g (91%) of the desired product. mp236.5-238°C.

preparation 3

5-(2-Chloropropionyl)-2-hydroxybenzimidazole

The title compound was prepared as in Preparation 2 from a _CS₂ (800 mol) solution of 2-hydroxybenzimidazole (0.136 mol), aluminum chloride (0.544 mol) and 2-chloropropionyl chloride (0.177 mol). The title compound was collected by filtration in 92% yield.

mp245°dec. Theoretical C ₁₀ H ₉ ClN ₂ O ₂ : C, 53.47; H, 4.04; N, 12.47. Found C, 54.41; H, 4.07; N, 13.25.

preparation 4

5-(2-Chloropropionyl)-2-oxindole

The title compound was prepared as in Preparation 2 from a solution of 2-oxindole (0.136 mol), aluminum chloride (0.544 mol) and 2-chloropropionyl chloride in CS ₂ (800 ml). The title compound was isolated by filtration. Yield 91%. mp157-158°C.

Preparation 5

6-(2-Chloroacetyl)-3,4-dihydroquinolin-2(1H)-one

From 3,4-dihydroquinolin-2(1H)-one (0.136 mol), aluminum chloride (0.544 mol) and 2-chloroacetyl chloride (0.177 mol) in _CS2 (800 ml) as in Preparation 2 The title compound was obtained from solution, and the title compound was isolated by filtration with a yield of 50%. mp215-216°C.

Preparation 6

5-(2-Chloroacetyl)-2-hydroxybenzimidazole

In a manner analogous to Preparation 2, the title compound was prepared from a solution of 2-hydroxybenzimidazole (0.136 mol), aluminum chloride (0.544 mol) and 2-chloroacetyl chloride (0.177 mol) in CS ₂ (800 ml). The title compound was isolated by filtration. Quantitative yield mp 273-275°C (dec.).

Preparation 7

5-(2-Chloroacetyl)-2-oxindole

The title compound was obtained from a solution of 2-oxindole (0.136 mol), aluminum chloride (0.544 mol) and 2-chloroacetyl chloride (0.177 mol) in _CS2 (800 ml) as in Preparation 2, and the title compound was isolated by filtration compound. Yield 90%, mp236.5-239℃

Preparation 8

Under a nitrogen atmosphere, oil-free sodium hydride (2.16g, 0.09M) was added to dry dimethylsulfoxide (250ml) and heated to 60-65°C until a uniform black solution formed, about 1 hour. Then, 19.83 g (0.09M) trimethyliodosulfonic acid was added (slightly exothermic) and the mixture was stirred until a brown solution formed, about 30 minutes. Then, a solution of 13.40 g (67.3 mM) of N-t-butoxycarbonyl-4-piperidone in dimethyl sulfoxide (50 ml) was stirred at room temperature for 1 hour. The reaction mixture was poured into 1L of cold water and the whole was extracted with hexane (4 x 100ml). The combined hexane extracts were backwashed with 50 mL of water and saline solution, dried over magnesium sulfate, and evaporated to obtain 11.75 g of white crystalline product 6-tert-butoxycarbonyl-1-oxa-6-azaspiro[2.5]octane (78% Yield).

Then extract the aqueous layer (3×50ml) with hexane to obtain another 650mg product, the total yield is 82.5%

m.p.57.5-59.5℃; IR (KBr) 5.90μm; NMRδ1.32-1.48 (2H, m), 1.42 (9H, s), 1.74-1.80 (2H, m), 2.65 (2H, s), 3.31-3.43 (2H, m), 3.61-3.72 (2H, m); theoretical value

_C11H19NO3 : C, 61.94; H _, 8.98; N _, 6.57. Found: C, 62.05; H, 9.09; K, 6.58.

A solution of 10.37g (0.11M) of phenol in dry dimethylsulfoxide (100ml) was treated in portions with 1.99g (82.8mmol) of oil-free sodium hydride in a cold water bath maintained at 20-25°C. The reaction mixture was stirred at room temperature for 45 minutes to give a gray suspension. 11.75 g (55.2 mmol) of 6-tert-butoxycarbonyl-1-oxa-6-azaspiro[2.5]octane dissolved in 65 mL of dimethyl sulfoxide was added dropwise, then, the reaction mixture was heated to 55-60 °C 7 hours, then stirred overnight at room temperature.

Then, the reaction mixture was poured into 1 L of cold water and extracted 4 times with ether. The ether extracts were combined, washed back with 10% NaOH and brine, dried over magnesium sulfate, and evaporated to give the desired product N-tert-butoxycarbonyl-4-hydroxyl-4-phenoxymethylpiperidine, weighing 17.01g (100% ), oily substance.

IR (Film) 5.91, 2.95 μM; NMR (CDCl ₃ ) δ 1.46 (9H, s); 1.53-1.80 (4H, m), 3.13-3.30 (2H, m), 3.80 (2H, s), 3.80- 3.98 (2H, m), 6.84-6.99 (2H, m), 7.22-7.44 (3H, m);

Theoretical value for C ₁₇ H ₂₅ NO ₄ : C, 66.42; H, 8.20; N, 4.56. Found value: C, 65.72; H, 8.21; N, 4.77.

A solution of 17.0 g (0.055M) of N-tert-butoxycarbonyl-4-hydroxy-4-phenoxymethylpiperidine in methanol (150 ml) was saturated with HCl gas and the mixture was cooled and treated with HCl gas. Step by step and repeat again. After crystals formed, the reaction mixture was treated with 500 ml of anhydrous ether and stirred at room temperature overnight.

The product was filtered, washed with anhydrous ether in a dry ammonia atmosphere to obtain 10.85 g (80.6%) crystals,

mp202-204°C. IR (KBr) 3.06, 3.14, 3.44, 3.57, 3.56, 6.33, 8.06 μm; NMR (D ₂ O) δ 2.00 (4H, broad s), 3.34 (4H, broad s), 4.00 ( 2H, s), 6.98-7.09 (3H, m), 7.30-7.43 (2H, m). Theoretical value C ₁₂ H ₁₇ NO ₂ .HCl: C, 59.13; H, 7.44; N, 5.75. Measured value: C , 58.98; H, 7.11; N, 5.65.

Claims (10)

1. The preparation method of the compound of the following formula and its pharmaceutically acceptable salt:

Wherein R ₁ , R ₂ and R ₃ are respectively selected from hydrogen, alkyl containing 1 to 6 carbon atoms and substituted phenyl, wherein the substituent on the substituted phenyl is selected from hydroxyl, containing 1 to 4 Alkyl with carbon atoms, chlorine, bromine, fluorine, trifluoromethyl, amino, nitro and alkoxy with 1 to 4 carbon atoms; or R and _R together form methylene, ethylene, _propylene or butylene; m is 0 to 2; m is 1 or 2; X and Y are respectively selected from hydrogen, chlorine, bromine, fluorine, trifluoromethyl, alkoxy containing 1 to 4 carbon atoms, alkyl containing 1 to 4 carbon atoms, hydroxyl, amino, nitro and substituted phenoxy, wherein the substituents on the substituted phenyl are selected from the group consisting of hydrogen, hydroxyl, alkyl containing 1 to 4 carbon atoms, chlorine, bromine, fluorine, trifluoromethyl, nitro, amino and An alkoxy group of 1 to 4 carbon atoms; M and Q are respectively selected from hydrogen, hydroxyl, amino, chlorine, bromine, fluorine, trifluoromethyl, nitro, an alkyl group containing 1 to 4 carbon atoms, containing 1 Alkoxy with 4 to 4 carbon atoms, N,N-dialkylamino with 1 to 4 carbon atoms in each alkyl group, N-alkylamino with 1 to 4 carbon atoms, NHCOR ₄ , NHCOOR _{5 and NHSO2R6} ; In the formula, R is selected from _hydrogen , alkyl containing 1 to 6 carbon atoms, phenyl and substituted phenyl, wherein the substituent on the substituted phenyl is selected from hydroxyl, chlorine, bromine, fluorine, trifluoro Methyl, amino, nitro, alkyl with 1 to 4 carbon atoms and alkoxy with 1 to 4 carbon atoms; Wherein R ₅ and R ₆ are selected from alkyl, phenyl and substituted phenyl containing 1 to 6 carbon atoms respectively, wherein the substituent on the substituted phenyl is selected from hydroxyl, chlorine, bromine, fluorine, tri Fluoromethyl, amino, nitro, alkyl with 1 to 4 carbon atoms and alkoxy with 1 to 4 carbon atoms; Or M and Q together form a divalent group Z, wherein Z is selected from:

In the formula, _R7 and _R8 are respectively selected from hydrogen and methyl. 2. The method of claim 1, wherein in the compound of formula (I) _R2 is hydrogen; _R3 is hydrogen or methyl; M and Q form a group Z, wherein Z is selected from: 3. The method of claim 2, wherein n is 1; m is 0; _R1 is hydrogen; _R3 is hydrogen; and X and Y are each hydrogen. 4, the method for claim 2, prepare following formula compound

wherein n is 1; m is 0; _R1 is hydrogen; X and Y are each hydrogen. 5, the method for claim 2, prepare following formula compound

wherein n is 1; m is 0; _R1 is hydrogen; X and Y are each hydrogen. 6. The method of claim 4, wherein Z in the compound is

7. The method of claim 4, wherein Z in the compound is 8. The method of claim 4, wherein Z in the compound is

9. The method of claim 5, wherein Z in the compound is

10. The preparation method of the compound of the following formula:

Wherein, R ₂ and R ₃ are selected from hydrogen, alkyl and substituted phenyl containing 1 to 6 carbon atoms, wherein the substituent on the substituted phenyl is selected from hydroxyl, containing 1 to 4 carbon atoms Alkyl, chlorine, bromine, fluorine, trifluoromethyl, amino, nitro and alkoxy groups of 1 to 4 carbon atoms; m is 0 to 2; m is 1 or 2; X and Y are respectively selected from hydrogen, chlorine, bromine, fluorine, trifluoromethyl, alkoxy containing 1 to 4 carbon atoms, alkyl containing 1 to 4 carbon atoms, hydroxyl, amino, nitro and substituted phenoxy, wherein the substituents on the substituted phenyl are selected from the group consisting of hydrogen, hydroxyl, alkyl containing 1 to 4 carbon atoms, chlorine, bromine, fluorine, trifluoromethyl, nitro, amino and Alkoxy groups with 1 to 4 carbon atoms; M and Q are selected from hydrogen, hydroxyl, amino, chlorine, bromine, fluorine, trifluoromethyl, nitro, alkyl groups with 1 to 4 carbon atoms, 1 Alkoxy with 4 to 4 carbon atoms, N,N-dialkylamino with 1 to 4 carbon atoms in each alkyl group, N-alkylamino with 1 to 4 carbon atoms, NHCOR ₄ , NHCOOR _{5 and NHSO2R6} ; In the formula, R is selected from _hydrogen , alkyl containing 1 to 6 carbon atoms, phenyl and substituted phenyl, wherein the substituent on the substituted phenyl is selected from hydroxyl, chlorine, bromine, fluorine, trifluoro Methyl, amino, nitro, alkyl with 1 to 4 carbon atoms and alkoxy with 1 to 4 carbon atoms; Wherein R ₅ and R ₆ are selected from alkyl, phenyl and substituted phenyl containing 1 to 6 carbon atoms respectively, wherein the substituent on the substituted phenyl is selected from hydroxyl, chlorine, bromine, fluorine, tri Fluoromethyl, amino, nitro, alkyl with 1 to 4 carbon atoms and alkoxy with 1 to 4 carbon atoms; Or M and Q together form a divalent group Z, wherein Z is selected from:

In the formula, _R7 and _R8 are respectively selected from hydrogen and methyl.