A new nalmefene dimer and its preparation method and use

Abstract

The present invention relates to a novel nalmefene dimer compound or its salt having a structure shown in formula (I), as well as a preparation method and application of the compound. The compound has a strong inhibitory effect on prostate cancer cell lines LNCaP and 22Rv1, and can be used to prepare anti-prostate cancer drugs. #imgabs0#

Description

Technical Field

[0001] The present invention relates to a compound and a preparation method and use thereof, in particular to a new nalmefene dimer compound and a preparation method and use thereof, belonging to the field of pharmaceutical chemistry. Background Art

[0002] Nalmefene hydrochloride was synthesized in 1975. It is another new pure opioid receptor antagonist synthesized after naloxone (NAL) and naltrexone (NTX). It can bind to opioid receptors μ, κ, and δ, and its binding effect with μ receptors is the strongest. Nalmefene hydrochloride is a water-soluble derivative of naltrexone. Its chemical structure of the 6th methylene group gives it the characteristics of long action time, multiple routes of administration, high bioavailability, few side effects, stronger physiological activity, and easier penetration of biological membranes. It has different degrees of effect on maintaining the normal functions of the respiratory, circulatory, digestive, endocrine and nervous systems. It has been used in the antagonism of respiratory depression caused by narcotic analgesics, the treatment of heart failure and shock, the treatment of alcohol poisoning and addiction, and weight loss. At the same time, nalmefene hydrochloride also has a certain antagonistic effect on poisoning caused by some non-opioid drugs, such as reversing respiratory depression and coma caused by excessive alcohol.

[0003] The structural formula of nalmefene is as follows:

[0004]

[0005] The structure of nalmefene is similar to that of morphine, and it has a phenolic hydroxyl group, which is easily oxidized and has a certain degree of increase in related substances. So far, there are no reports on the separation, structural confirmation and use research of the new nalmefene dimer compound of the present invention. Summary of the invention

[0006] The object of the present invention is to provide a new nalmefene dimer compound having a structure shown in formula (I):

[0007]

[0008] Its chemical name is 17,17'-bis(cyclopropylmethyl)-4,5α:4',5'α-diepoxy-6,6'-bis(methylenedimorphinan-3,3'-methylenedioxy-14,14'-diol.

[0009] Another object of the present invention is to provide a method for preparing the novel nalmefene dimer compound, wherein nalmefene is reacted with dihalomethane in a reaction solvent under base catalysis to obtain the novel nalmefene dimer compound represented by formula (I).

[0010] Furthermore, the reaction solvent is at least one of N,N-dimethylformamide and tetrahydrofuran; the base catalyst used for the base catalysis is at least one selected from potassium tert-butoxide, sodium methoxide, sodium ethoxide and sodium propoxide; the dihalomethane is at least one selected from diiodomethane, dibromomethane and dichloromethane.

[0011] The molar feed ratio of nalmefene to the base catalyst is 1:0.5-4.0, and the molar feed ratio of nalmefene to dihalomethane is 1:0.5-100.

[0012] Furthermore, the molar feed ratio of nalmefene to dihalomethane is 1:1.0-10.

[0013] Furthermore, the reaction temperature of the reaction is controlled at 0°C to 70°C.

[0014] Furthermore, the reaction temperature of the reaction is controlled at 20°C to 50°C.

[0015] In one embodiment, the preparation method further comprises the step of separation and purification using high performance preparative liquid chromatography gradient elution, wherein the chromatographic conditions are: using a carbon 18 bonded silica gel chromatographic column; mobile phase A is 0.03% ammonia solution, mobile phase B is acetonitrile, gradient elution; and the detection wavelength is 210±10nm.

[0016] The novel nalmefene dimer compound of the present invention can be used as a reference substance for controlling the quality of nalmefene free base or its pharmaceutically acceptable salt and its preparations, and for detecting the shelf life of nalmefene free base or its pharmaceutically acceptable salt and its preparations. Therefore, another object of the present invention is to provide the use of the novel nalmefene dimer compound in the preparation of a reference substance for a nalmefene drug or its preparations.

[0017] Another object of the present invention is to provide a pharmaceutical composition, which contains nalmefene or a pharmaceutically acceptable salt thereof and a compound of the structure represented by formula (I) in an amount not higher than 0.5% by weight, i.e., the novel nalmefene dimer compound of the present invention:

[0018]

[0019] The pharmaceutically acceptable salt of nalmefene is selected from any one of maleate, methanesulfonate, sulfonate, sulfate, hydrochloride, hydrobromide, phosphate, nitrate, p-toluenesulfonate, tartrate, fumarate, acetate, formate, benzoate, cinnamate, succinate, malonate, and citrate, or a combination thereof.

[0020] Furthermore, the content of nalmefene or a pharmaceutically acceptable salt thereof in the pharmaceutical composition is not less than 90%, preferably not less than 95%, and more preferably not less than 98%.

[0021] Furthermore, the content of the compound represented by the structure of formula (I) is not higher than 0.4%, preferably not higher than 0.3%, and more preferably not higher than 0.2%.

[0022] Furthermore, the weight ratio between nalmefene or a pharmaceutically acceptable salt thereof and the compound of formula (I) is not less than 200:1, preferably not less than 300:1, and more preferably not less than 500:1.

[0023] In one embodiment, the pharmaceutical composition contains not less than 98% of nalmefene or a pharmaceutically acceptable salt thereof, not more than 0.20% of the new nalmefene dimer compound represented by formula (I), not more than 0.5% of naltrexone hydrochloride, not more than 0.5% of dinalmefene, and the remainder is other related substances.

[0024] The preparation method of the above-mentioned pharmaceutical composition is as follows: under nitrogen or inert gas protection conditions, naltrexone and methyl triphenylphosphonium bromide are subjected to Wittig reaction under the catalysis of potassium tert-butoxide to obtain nalmefene, and nalmefene is further reacted with an acid to prepare a pharmaceutically acceptable salt.

[0025] The present invention also provides a pharmaceutical preparation, which comprises the above pharmaceutical composition and an optional pharmaceutically acceptable carrier.

[0026] Furthermore, the present invention provides a pharmaceutical preparation, wherein the weight ratio between nalmefene or a pharmaceutically acceptable salt thereof and the compound of formula (I) is not less than 200:1, preferably not less than 300:1, and more preferably not less than 500:1.

[0027] The pharmaceutical preparation of the present invention can be various dosage forms well known in the art. The dosage form suitable for the present invention is selected from oral preparations, external preparations or injections, preferably oral preparations or injections, and more preferably injections. The injection is selected from injections (injections), infusions, lyophilized powder injections or sterile packaging preparations, preferably injections. The pharmaceutical preparation of the present invention can be prepared by formulation technology means well known in the art.

[0028] The pharmaceutically acceptable carrier of the present invention is a commonly used excipient or adjuvant for preparing the preparation well known in the art. The commonly used excipients or adjuvants for the injection include, but are not limited to: antioxidants, such as phenols such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), nordihydroguaiaretic acid (NDGA), sulfur-containing compounds such as thiodipropionic acid, sulfites, bisulfites, dithiocarbamates, sodium sulfite, sodium bisulfite, sodium pyrosulfite, sodium thiosulfate; organic acids / alcohols / esters such as ascorbic acid, citric acid, malic acid, sorbitol, glycerol, propylene glycol, ascorbyl palmitate, esters such as hydroquinone, hydroxycoumarin, vitamin E, amines such as ethanolamine, soybean lecithin, cephalin, plant lecithin or animal lecithin, inorganic acid or its salt, phosphoric acid or its salt, phosphorous acid or its salt; osmotic pressure regulators such as sodium chloride, glucose Glucose, potassium chloride, magnesium chloride, calcium chloride, sorbitol, mannitol, etc., preferably sodium chloride or glucose; antibacterial agents, such as 0.5% phenol, 0.3% cresol, 0.5% chlorobutanol; pH adjusters, such as hydrochloric acid, tartaric acid, citric acid, potassium hydroxide, sodium hydroxide, sodium dihydrogen phosphate, disodium hydrogen phosphate, acetic acid, sodium acetate, lactic acid, citric acid, sodium citrate, sodium bicarbonate, sodium carbonate, any one or a combination thereof; emulsifiers, such as polysorbate-80, sorbitan monophosphate, Pluronic F-68, lecithin, soy lecithin; solubilizers, such as Tween-80, bile, glycerol, propylene glycol, lecithin, polyoxyethylene castor oil, etc.; fillers or excipients, such as lactose, mannitol, sorbitol, dextran, etc.

[0029] The injection is selected from injection solution, sterile powder for injection and concentrated solution for injection, and can be used for intramuscular injection, intravenous injection, intravenous drip, etc. The specifications of the injection of the present invention are selected from 1mL, 2mL, 5mL, 10mL, 20mL, 50mL, 100mL, 200mL, 250mL or 500mL.

[0030] The pharmaceutical preparation of the present invention can be a unit preparation, calculated as nalmefene, containing 0.01 mg to 50 mg of the pharmaceutical composition of the present invention as an essential active ingredient, preferably 0.01 mg to 30 mg, more preferably 0.0125 mg, 0.04 mg, 0.08 mg, 0.1 mg, 0.16 mg, 0.2 mg, 0.25 mg, 0.32 mg, 0.4 mg, 0.5 mg, 0.75 mg, 1 mg, 1.25 mg, 1.5 mg, 1.75 mg, 2 mg, 2.5 mg, 5 mg, 10 mg.

[0031] In particular, the present invention also provides a method for preparing nalmefene hydrochloride injection, which is prepared by mixing the above-mentioned pharmaceutical composition with a pharmaceutically acceptable injection carrier.

[0032] Furthermore, the pharmaceutically acceptable injection carrier is selected from any one of antioxidants, antibacterial agents, pH regulators, or a combination thereof.

[0033] Further, the antioxidant is selected from phenols such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), nordihydroguaiaretic acid (NDGA), sulfur-containing compounds such as dithiopropionic acid, sulfites, bisulfites, dithiocarbamates, sodium sulfite, sodium bisulfite, sodium metabisulfite, sodium thiosulfate; organic acids / alcohols / esters such as ascorbic acid, citric acid, malic acid, sorbitol, glycerol, propylene glycol, ascorbyl palmitate, esters such as hydroquinone, hydroxycoumarin, vitamin E, amines such as ethanolamine, soybean lecithin, cephalin, plant lecithin or animal lecithin, inorganic acids or their salts, phosphoric acid or its salts, phosphorous acid or its salts.

[0034] Furthermore, the pH adjuster is selected from any one of hydrochloric acid, tartaric acid, citric acid, potassium hydroxide, sodium hydroxide, sodium dihydrogen phosphate, disodium hydrogen phosphate, acetic acid, sodium acetate, lactic acid, citric acid, sodium citrate, sodium bicarbonate, and sodium carbonate, or a combination thereof.

[0035] Furthermore, the antibacterial agent is selected from any one of 0.5% phenol, 0.3% cresol, 0.5% chlorobutanol or a combination thereof.

[0036] The present invention also provides the use of the above-mentioned pharmaceutical composition and pharmaceutical preparation in the preparation of opioid receptor antagonist drugs.

[0037] The inventors were pleasantly surprised to find in their research that the new nalmefene dimer compound of the present invention has strong anti-prostate cancer activity. Therefore, another object of the present invention is to provide a use of a new nalmefene dimer compound or a composition thereof in the preparation of an anti-prostate cancer drug, and to provide a novel anti-prostate cancer drug composition, wherein the composition contains an effective amount of the new nalmefene dimer compound.

[0038] The structure of the new nalmefene dimer compound represented by formula (I) of the present invention is confirmed by nuclear magnetic resonance spectrum and mass spectrometry, and is analyzed as follows (the data is derived from the test results of the new nalmefene dimer compound sample prepared in Example 1 of the present invention):

[0039] 1. The mass spectrometry data are shown in Table 1 below:

[0040] Table 1 Mass spectral data of the novel nalmefene dimer compound of the present invention

[0041]

[0042] Mass spectrometry (MS) showed that the product was [1 / 2M+H] +The mass-to-nuclear ratio of the peak is 346, and the deviation between the measured value of the mass spectrometry signal and the theoretical value is very small, which is consistent with the molecular formula of the new nalmefene dimer compound (C 43 H 50 N 2 O 6 ) is consistent.

[0043] 2. H NMR and C NMR

[0044]

[0045] The hydrogen spectrum data are shown in Table 2 below:

[0046] Table 2 H NMR spectrum data of the novel nalmefene dimer compound of the present invention

[0047]

[0048]

[0049] Note: s-singlet; d-doublet; m-multiplet; br-broad peak

[0050] The carbon spectrum data are shown in Table 3 below:

[0051] Table 3 C NMR spectrum data of the novel nalmefene dimer compound of the present invention

[0052] Serial number Sample δ(ppm) Attribution 1 3.833 C-19, C-19' 2 3.993 C-20, C-20' 3 9.379 C-18, C-18' 4 22.900 C-11, C-11' 5 28.207 C-14, C-14' 6 30.413 C-15, C-15' 7 32.281 C-13, C-13' 8 44.408 C-12, C-12' 9 48.477 C-8, C-8' 10 59.246 C-17, C-17' 11 62.599 C-10, C-10' 12 70.843 C-9, C-9' 13 89.624 C-7, C-7' 14 93.673 C-22 15 111.603 C-21, C-21' 16 118.622 C-3, C-3' 17 118.871 C-2, C-2' 18 127.428 C-16, C-16' 19 131.585 C-4, C-4' 20 140.065 C-5, C-5' 21 145.329 C-1, C-1' 22 145.679 C-6, C-6'

[0053] NMR spectrum analysis:

[0054] According to the analysis of nuclear magnetic resonance hydrogen spectrum signal, the test sample 1 H-NMR test solvent is CDCl 3 After deducting the solvent peak and the water peak, there are 48 H in total, and 2 active hydrogens are not included in the peak, which is the same as the number of inactive H in the new nalmefene dimer molecular structure.

[0055] According to the analysis of the carbon NMR spectrum signal, the 13 The C-NMR spectrum has 22 groups of carbon peaks. The structure is a symmetrical structure with the ether bond (C-22) as the center. Therefore, except for the central carbon atom (C-22), each of the other groups of carbon peaks has 2 carbon atoms, totaling 43 carbon atoms, which is the same as the C number of the new nalmefene dimer.

[0056] In summary, the NMR data have reasonable attribution and are consistent with the structural formula of the new nalmefene dimer.

[0057] The novel nalmefene dimer compound represented by formula (I) of the present invention has been found through activity studies to have a significant proliferation inhibitory effect on castration-resistant prostate cancer cell lines LNCaP and 22Rv1, indicating that it has an anti-prostate cancer effect.

[0058] This activity study used JQ-1 as a positive control compound. JQ-1 is an effective reversible BET bromodomain inhibitor that has an inhibitory effect on the proliferation activity of various tumor cells. According to literature reports, JQ-1 has a significant proliferation inhibitory effect on castration-resistant prostate cancer cell lines LNCaP and 22Rv1. BRIEF DESCRIPTION OF THE DRAWINGS

[0059] Figure 1 HPLC chart of the compound represented by the structure of formula (Ⅰ);

[0060] Figure 2 The compound represented by the structure of formula (I) 1 H-NMR spectrum;

[0061] Figure 3 The compound represented by the structure of formula (I) 13 C-NMR spectrum;

[0062] Figure 4 It is the mass spectrum of the structural compound represented by formula (I). DETAILED DESCRIPTION

[0063] The present invention will be further described below in conjunction with test examples and embodiments, which will enable professionals and technicians in the field to more fully understand the present invention, but will not limit the present invention in any way.

[0064] Example 1 Preparation of the novel nalmefene dimer compound of the present invention

[0065] Nalmefene (3.39 g, 10 mmol), potassium tert-butoxide (0.56 g, 5 mmol), N,N-dimethylformamide (50 ml) and diiodomethane (1.34 g, 5 mmol) were mixed and stirred, and heated at 70°C for 8 hours. The reaction solution was added to 200 ml of water, extracted with 100 ml of ethyl acetate * 3 times, and the ethyl acetate phase was combined; the ethyl acetate phase was washed with 100 ml of saturated brine; the ethyl acetate phase was separated, and the ethyl acetate was removed by reduced pressure distillation. The residue was the crude product. The sample was separated by preparative liquid phase (chromatographic column was carbon 18 bonded silica gel column, mobile phase A: 0.03% aqueous solution; B: acetonitrile, gradient elution, 10% to 40% B, 60 minutes; flow rate 80 ml / min) to obtain 0.35 g of product, HPLC purity 97.7%.

[0066] Example 2 Preparation of the novel nalmefene dimer compound of the present invention

[0067] Nalmefene (3.39 g, 10 mmol), sodium methoxide (2.16 g, 40 mmol), N,N-dimethylformamide (50 ml) and dichloromethane (84.93 g, 1000 mmol) were mixed and stirred, and heated at 40°C for 24 hours. The reaction solution was added to 200 ml of water, extracted with 100 ml of dichloromethane 3 times, and the dichloromethane phase was combined; the dichloromethane phase was washed with 100 ml of saturated salt water; the dichloromethane phase was separated, and dichloromethane was removed by reduced pressure distillation. The residue was the crude product. The sample was separated by preparative liquid phase (chromatographic column was carbon 18 bonded silica gel column, mobile phase A: 0.03% aqueous solution; B: acetonitrile, gradient elution, 10% to 40% B, 60 minutes; flow rate 80 ml / min) to obtain 0.22 g of product, HPLC purity 96.4%.

[0068] Example 3 Preparation of the novel nalmefene dimer compound of the present invention

[0069] Nalmefene (3.39 g, 10 mmol), sodium ethoxide (1.36 g, 20 mmol), tetrahydrofuran (50 ml) and dibromomethane (8.69 g, 50 mmol) were mixed and stirred, and reacted at 0°C for 24 hours. The reaction solution was added to 200 ml of water, extracted with 100 ml of ethyl acetate * 3 times, and the ethyl acetate phase was combined; the ethyl acetate phase was washed with 100 ml of saturated brine; the ethyl acetate phase was separated, and the ethyl acetate was removed by vacuum distillation. The residue was the crude product. The sample was separated by preparative liquid phase (chromatographic column was carbon 18 bonded silica gel column, mobile phase A: 0.03% aqueous solution; B: acetonitrile, gradient elution, 10% to 40% B, 60 minutes; flow rate 80 ml / min) to obtain 0.32 g of product, HPLC purity 96.0%.

[0070] Example 4 Preparation of the novel nalmefene dimer compound of the present invention

[0071] Nalmefene (3.39 g, 10 mmol), sodium ethoxide (1.36 g, 20 mmol), tetrahydrofuran (50 ml) and dibromomethane (3.48 g, 20 mmol) were mixed and stirred, and reacted at 70°C for 12 hours. The reaction solution was concentrated to dryness under reduced pressure, and the residue was dissolved in 100 ml of ethyl acetate, and the ethyl acetate solution was washed with 100 ml of saturated brine; the ethyl acetate phase was separated, and the ethyl acetate was removed by vacuum distillation. The residue was the crude product, and the sample was separated by preparative liquid phase (chromatographic column was carbon 18 bonded silica gel column, mobile phase A: 0.03% aqueous solution; B: acetonitrile, gradient elution, 10% to 40% B, 60 minutes; flow rate 80 ml / min) to obtain 0.41 g of product, HPLC purity 98.0%.

[0072] Example 5 Preparation of the novel nalmefene dimer compound of the present invention

[0073] Nalmefene (3.39 g, 10 mmol), sodium propoxide (0.82 g, 10 mmol), tetrahydrofuran (50 ml) and dichloromethane (4.25 g, 50 mmol) were mixed and stirred, and reacted at 40°C for 24 hours. The reaction solution was concentrated to dryness under reduced pressure, and the residue was dissolved in 100 ml of ethyl acetate, and the ethyl acetate solution was washed with 100 ml of saturated brine; the ethyl acetate phase was separated, and the ethyl acetate was removed by reduced pressure distillation. The residue was the crude product, and the sample was separated by preparative liquid phase (chromatographic column was carbon 18 bonded silica gel column, mobile phase A: 0.03% aqueous solution; B: acetonitrile, gradient elution, 10% to 40% B, 60 minutes; flow rate 80 ml / min) to obtain 0.21 g of product, HPLC purity 98.3%.

[0074] Example 6 Preparation of the novel nalmefene dimer compound of the present invention

[0075] Nalmefene (3.39 g, 10 mmol), sodium methoxide (1.08 g, 20 mmol), tetrahydrofuran (50 ml) and diiodomethane (5.36 g, 20 mmol) were mixed and stirred, and reacted at 50°C for 12 hours. The reaction solution was concentrated to dryness under reduced pressure, and the residue was dissolved in 100 ml of ethyl acetate, and the ethyl acetate solution was washed with 100 ml of saturated brine; the ethyl acetate phase was separated, and the ethyl acetate was removed by vacuum distillation. The residue was the crude product, and the sample was separated by preparative liquid phase (chromatographic column was carbon 18 bonded silica gel column, mobile phase A: 0.03% aqueous solution; B: acetonitrile, gradient elution, 10% to 40% B, 60 minutes; flow rate 80 ml / min) to obtain 0.31 g of product, HPLC purity 99.3%.

[0076] Example 7 Preparation of the novel nalmefene dimer compound of the present invention

[0077] Nalmefene (3.39 g, 10 mmol), sodium ethoxide (2.72 g, 40 mmol), N,N-dimethylformamide (50 ml) and dibromomethane (17.4 g, 100 mmol) were mixed and stirred, and reacted at 20°C for 24 hours. The reaction solution was added to 200 ml of water, extracted with 100 ml of dichloromethane * 3 times, and the dichloromethane phase was combined; the dichloromethane phase was washed with 100 ml of saturated salt water; the dichloromethane phase was separated, and dichloromethane was removed by reduced pressure distillation. The residue was the crude product. The sample was separated by preparative liquid phase (chromatographic column was carbon 18 bonded silica gel column, mobile phase A: 0.03% aqueous solution; B: acetonitrile, gradient elution, 10% to 40% B, 60 minutes; flow rate 80 ml / min) to obtain 0.26 g of product, HPLC purity 97.4%.

[0078] Example 8 Study on the activity of the novel nalmefene dimer compound of the present invention

[0079] Experimental methods:

[0080] LNCaP cells (from ATCC) were cultured in RPMI-1640 medium containing 10% fetal bovine serum and double antibody (1X), and 22Rv1 cells (from ATCC) were cultured in RPMI-1640 medium containing 15% fetal bovine serum and double antibody (1X). When the cells reached 85% confluency, they were inoculated into 96-well plates, with 1500 cells per well. T 0 Plate, incubate at 37°C, 5% CO 2 Incubate overnight in an incubator.

[0081] Test T before adding medication on the second day 0 Plate cell reading. The starting concentration of positive compounds was 10 μM, and the starting concentration of test compounds was 100 μM. Both were diluted 3-fold in 9 gradients. Incubate at 37°C, 5% CO 2 After 72 hours of incubation in the incubator, 100 μL CellTiter-Glo was added to each well. TM (Promega, Cat. G7573) detection reagent, mix for 2 minutes, incubate at room temperature for 10 minutes, and measure the chemiluminescence reading using a microplate reader (BMG, PHERAstar FSX).

[0082] According to the following formula (1), the inhibition rate of each concentration of the compound was calculated, and the concentration GI of the compound when the proliferation rate was 50% was calculated using the origin9.2 software and the DoseResp function. 50 Value. Where T 0 is the chemiluminescence reading before administration, T 72给药 T is the chemiluminescence reading after 72 hours of incubation with the compound. 72溶媒 The chemiluminescence reading is for the vehicle control after 72 hours of incubation.

[0083] Growth%=(T 72给药 -T 0 ) / (T 72溶媒 -T 0 )×100 Formula (1)

[0084] The experimental results are shown in Table 4 below:

[0085] Table 4: Proliferation inhibition GI of LNCaP and 22Rv1 cells by JQ-1 and new nalmefene dimer compounds 50 value

[0086]

[0087] The above results show that the new nalmefene dimer compound has an inhibitory effect on the proliferation of prostate cancer cells LNCaP and 22Rv1, GI50 The value was 5 to 10 μM, showing good inhibitory activity.

[0088] Example 6 Preparation of Nalmefene (Composition 1) and Nalmefene Hydrochloride (Composition 2) of the Present Invention

[0089] Under nitrogen protection, 32.43 g of methyltriphenylphosphonium bromide, 10.19 g of potassium tert-butoxide, and 200 mL of N,N-dimethylformamide were added to a 1 L reaction bottle in sequence, and stirred at 25°C for 0.5 h. 10.00 g of naltrexone was dissolved in 30 mL of N,N-dimethylformamide and added dropwise to the reaction solution, and stirred at 25°C for 1 h.

[0090] After the reaction, add 150mL of 20% ammonium chloride solution and then add 200mL of dichloromethane; let stand for separation, extract the aqueous phase with 200mL and 100mL of dichloromethane in turn, combine the organic phases; extract with 100mL*2 times of 2mol / L HCl solution, separate the water layer, and then wash the aqueous phase with 100mL*2 times of petroleum ether; after separation, adjust the pH of the aqueous phase to 8-9 with ammonia water, filter, and wash the filter cake with water until neutral. The filter cake is vacuum dried at 40°C for 20h to obtain 6.42g of nalmefene with a purity of 98.67%.

[0091] 5.00 g of nalmefene was added to 100 mL of acetone, stirred to dissolve, concentrated hydrochloric acid was added dropwise to adjust the pH to 2-3, a white solid was precipitated, filtered, and the filter cake was vacuum dried at 40° C. for 8 h to obtain 5.12 g of nalmefene hydrochloride.

[0092] Example 7 Preparation of Nalmefene Mesylate (Composition 3) of the Present Invention

[0093] 5.00 g of nalmefene was added to 100 mL of dichloromethane, stirred to dissolve, and methanesulfonic acid was added dropwise to adjust the pH to 2-3. A white solid was precipitated, which was filtered and the filter cake was dried in vacuum at 40° C. for 8 h to obtain 5.32 g of nalmefene mesylate.

[0094] Example 8 Preparation of Nalmefene Phosphate (Composition 4) of the Present Invention

[0095] 5.00 g of nalmefene was added to 100 mL of acetone, stirred to dissolve, phosphoric acid was added dropwise to adjust the pH to 2-3, a white solid was precipitated, filtered, and the filter cake was vacuum dried at 40° C. for 8 h to obtain 5.22 g of nalmefene phosphate.

[0096] Example 9 Stability Study of the Pharmaceutical Composition of the Present Invention

[0097] Take an appropriate amount of the samples of nalmefene (composition 1) and nalmefene hydrochloride (composition 2) prepared in Example 6 and commercially available nalmefene and nalmefene hydrochloride samples, put them in a weighing bottle with a thickness of about 5 mm, open them and place them under high temperature (60° C.), high humidity (25° C., relative humidity (90±5)%), and light (intensity of (4500±500) lx) environment for 10 days, take samples on the 5th and 10th days, observe that their appearance and color do not change (white powder), and HPLC determines the related substances. Compared with the samples before (0d) under each environment, the results are shown in Table 5 below:

[0098] Table 5 Stability test results of the pharmaceutical composition of the present invention and existing products (%)

[0099]

[0100] Chromatographic conditions: octadecylsilane bonded silica gel as filler (Kromasil C18, 4.6 mm × 250 mm, 5 μm or chromatographic column of equivalent performance); acetonitrile-phosphate buffer (7.8 g of sodium dihydrogen phosphate, 2 ml of triethylamine, add water to 1000 ml, adjust the pH value to 4.2 with 85% phosphoric acid) (20:80) as mobile phase; detection wavelength is 210 nm; column temperature is 30°C.

[0101] Table 5 above shows that compared with the commercially available products, the content of the new nalmefene dimer compound in Composition 1 and Composition 2 of the present invention is lower, and the growth of the related substance A and total impurities under high temperature, high humidity and light conditions is much less than that of the commercially available products, indicating that controlling the content of the new nalmefene dimer compound of the present invention is beneficial to controlling the content of the related substance A and total impurities in the composition. Studies have shown that when the content of the new nalmefene dimer compound exceeds 0.5%, the total impurities increase faster in the stability study.

[0102] Example 10 Preparation of Nalmefene Hydrochloride Injection of the Present Invention

[0103] prescription:

[0104]

[0105] Preparation process:

[0106] Weigh the prescribed amount of glycerol, dissolve it completely with 70% of the prescribed amount of water for injection cooled to room temperature, add the prescribed amount of nalmefene hydrochloride (composition 2) and stir to dissolve (concentrated), add 0.05% (W / V) of the concentrated volume of medicinal activated carbon and stir evenly, let it stand for adsorption for 10 minutes, filter and remove carbon; add water for injection to 95% of the full amount, stir evenly, adjust the pH to 3.8 with 0.1 mol / L hydrochloric acid solution, add water for injection to the full amount, stir evenly; take samples to determine their properties, pH value, content and bacterial endotoxins, filter finely, fill with nitrogen, fill, seal, and sterilize at 121°C for 12 to 15 minutes. According to high performance liquid chromatography, the mass ratio between nalmefene hydrochloride and the compound of formula (I) in the nalmefene hydrochloride injection is 610:1.

[0107] Example 11 Preparation of Nalmefene Hydrochloride Injection of the Present Invention

[0108] prescription:

[0109]

[0110] Preparation process:

[0111] Weigh the prescribed amount of mannitol, dissolve it completely with 70% of the prescribed amount of water for injection cooled to room temperature, add the prescribed amount of nalmefene hydrochloride (composition 2) and stir to dissolve (concentrated), add 0.05% (W / V) of the concentrated volume of medicinal activated carbon and stir evenly, let it stand for adsorption for 10 minutes, filter and remove carbon; add water for injection to 95% of the full amount, stir evenly, adjust the pH to 3.8 with 0.1 mol / L hydrochloric acid solution, add water for injection to the full amount, stir evenly; take samples to determine their properties, pH value, content and bacterial endotoxins, filter finely, fill with nitrogen, fill, seal, and sterilize at 121°C for 12 to 15 minutes. According to high performance liquid chromatography, the mass ratio between nalmefene hydrochloride and the compound of formula (I) in the nalmefene hydrochloride injection is 1050:1.

Claims

1. A nalmefene dimer compound, characterized in that: It has the structure shown in formula (I):

2. A method for preparing the nalmefene dimer compound according to claim 1, characterized in that: It is prepared by reacting nalmefene with dihalomethane in a reaction solvent under base catalysis; The reaction solvent is selected from at least one of N,N-dimethylformamide and tetrahydrofuran; the base catalyst used for the base catalysis is selected from at least one of potassium tert-butoxide, sodium methoxide, sodium ethoxide and sodium propoxide; the dihalomethane is selected from at least one of diiodomethane, dibromomethane and dichloromethane; The molar feed ratio of nalmefene to the base catalyst is 1:0.5-4.0, and the molar feed ratio of nalmefene to the dihalomethane is 1:1.0-10; The separation and purification was carried out by high performance preparative liquid chromatography gradient elution, and the chromatographic conditions were as follows: a carbon 18 bonded silica gel chromatographic column was used; mobile phase A was 0.03% ammonia solution, mobile phase B was acetonitrile, gradient elution; and the detection wavelength was 210±10nm.

3. The preparation method according to claim 2, characterized in that: The reaction temperature of the reaction is controlled at 0°C to 70°C.

4. The preparation method according to claim 3, characterized in that: The reaction temperature of the reaction is controlled at 20°C to 50°C.

5. Use of the nalmefene dimer compound according to claim 1 in the preparation of anti-prostate cancer drugs.

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