CN108003164A - Benzodiazepine * class compounds - Google Patents

Abstract

The present invention relates to a compound represented by formula (I) and its preparation method, as well as its application in the preparation of antithrombotic drugs or cardiovascular and cerebrovascular drugs.

Description

Benzodiazepine* compounds

technical field

The present invention relates to a short 3-benzodiazepine Compounds and preparation methods thereof, and the application of compounds represented by formula (I) as antithrombotic drugs in cardiovascular and cerebrovascular pharmaceutical composition preparations.

Background technique

Benzodiazepines are widely used clinically for anxiolytic, sedative and hypnotic effects. As the first water-soluble benzodiazepine derivative, midazolam has been widely used in clinical sedation, hypnosis, analgesia, antiepileptic, anxiolytic and general anesthesia. When midazolam is injected into the human body as an anesthetic, it can be oxidized to α-hydroxy midazolam by cytochrome P450 isoenzyme. However, the oxide still has pharmacological activity, so the anesthesia takes a long time and the recovery is slow. Therefore, the development of new water-soluble derivatives of benzodiazepines with fast anesthesia induction time and short maintenance time has always been paid attention to by medicinal chemists.

The chemical name of the compound of formula (II) is 3-[(4S)-8-bromo-1-methyl-6-(2-pyridyl)-4H-imidazol[1,2-a][1,4]benzene Methyl diazepine-4-yl]propionate,

Wherein, R is hydrogen, methyl, ethyl, isopropyl.

Patent EP1183243 reports that such compounds are short-acting central nervous system (CNS, Central Nervous System) inhibitors, which include sedative hypnotic, anxiolytic, muscle relaxation and anticonvulsant effects. It can be used for intravenous administration in the following clinical treatment regimens: such as preoperative sedation, anxiolytic and amnestic use during surgery; conscious sedation during short-term diagnostic, surgical or endoscopic procedures; when administering other anesthetics and analgesics Before and/or concurrently, as a component for induction and maintenance of general anesthesia; ICU sedation, etc. The compound metabolizes rapidly, does not rely on cellular P450 enzyme metabolism, and can be metabolized by various organs, and its metabolites have very low activity, which reduces the interaction between drugs, and also provides a great possibility for the use of patients with impaired metabolic organ function. possible.

However, the compound shown in formula (II) is extremely unstable. In the test of the influencing factors of forced degradation, it is easy to produce degraded impurities. The content of these degraded impurities is very small, which requires long-term separation and enrichment for structural identification. So far , have not yet seen the relevant reports on the separation, structure confirmation and application research of these degradation impurities.

Contents of the invention

When we carried out stability research on formula (II), we found that the compound was very unstable and could produce more degradation impurities. Therefore, the compound shown in formula (II) is salified with benzenesulfonic acid or p-toluenesulfonic acid to increase its stability, and it is found that the stability of its benzenesulfonate or p-toluenesulfonate is greatly enhanced than its base, but Degradation impurities were still produced during the stability studies of forced degradation, but none of these impurities had been reported in the literature before. We spent more than a year to separate these impurities through preparative liquid phase, and confirmed the structure, and found that there is a structural formula of a degraded impurity as shown in formula (I):

Wherein, R is hydrogen, methyl, ethyl, isopropyl.

Subsequently, we conducted a synthetic study on the structure of the impurity.

Therefore, the object of the present invention is to provide a degraded impurity of the compound represented by formula (II), which has a structure represented by formula (I). Another object of the present invention is to provide a synthesis method of the structure shown in formula (I), which is characterized in that the compound shown in formula (II) is hydrolyzed with a base to obtain it;

Further, wherein the hydrolysis reaction temperature is -40-0°C.

Another object of the present invention is to provide a synthesis method of the structure shown in formula (I), which is obtained by hydrolyzing the compound shown in formula (II) with a base first, and then carrying out bromination reaction;

Further, the temperature of the hydrolysis reaction between the compound represented by formula (II) and the base is ~100°C;

Further, the bromination reagent of the bromination reaction is hydrogen bromide, acetyl bromide, bromine, NBS;

Further, the temperature of the bromination reaction is -10 to 110°C;

Further, the bromination reaction occurs in an aprotic solvent;

Further, the aprotic solvent of the bromination reaction is selected from any one of tetrahydrofuran, acetonitrile, DMF, acetone, methylene chloride, chloroform, ethylene dichloride, toluene, xylene or a combination thereof.

Further, in the above two methods, the hydrolysis reaction occurs in a protic solvent;

Further, the protic solvent of the hydrolysis reaction is any one of methanol, ethanol, isopropanol, butanol, water or a combination thereof;

Further, the alkali is any one of sodium hydroxide, potassium hydroxide, lithium hydroxide, calcium hydroxide, barium hydroxide, sodium carbonate, potassium carbonate or a combination thereof.

Further, the separation and purification method of the compound represented by formula (I) in the present invention is selected from any one or combination of column chromatography, recrystallization or preparative liquid phase separation and purification.

Further, the recrystallization method of the present invention is carried out in any one of methanol, ethanol, acetone, 2-butanone or a combination thereof with the organic acid salt of the compound represented by formula (I).

Further, the organic acid salt of the compound represented by formula (I) in the present invention refers to hydrochloride, hydrobromide, sulfate, nitrate, oxalate, maleate, p-toluenesulfonate or Tosylate.

Further, the conditions for separation and purification by column chromatography of the present invention are that the eluent is any one of ethyl acetate-petroleum ether, ethyl acetate-n-hexane or a combination thereof, and isocratic or gradient elution is carried out, and the collected The product fractions are concentrated or freeze-dried to obtain the obtained product.

Further, the conditions for the preparative liquid phase separation and purification of the present invention are that the mobile phase is any one of methanol-acetonitrile, methanol-acetonitrile-water, methanol-water or a combination thereof, and isocratic or gradient elution is performed, and the collected The product fractions are concentrated or freeze-dried to obtain the obtained product.

The compound represented by the formula (I) of the present invention is used as a standard or reference substance in the application of pharmaceutical analysis.

The inventors found through research on the compound represented by formula (I) that the compound represented by formula (I) of the present invention has better antithrombotic activity, and conducted preliminary research on its toxicity. Therefore, another object of the present invention is to provide the application of the compound described in formula (I) or its composition in antithrombotic drugs. The thrombotic disease is myocardial infarction, ischemic cardiomyopathy, cerebral thrombosis, cerebral embolism, cerebral infarction, transient ischemic attack or lacunar infarction. Another object of the present invention is to provide a novel antithrombotic composition, which contains an effective dose of the compound represented by formula (I), or its pharmaceutically acceptable salt, stereoisomer, tautomer Construct.

The pharmaceutically acceptable salt of formula (I) compound in the present invention refers to hydrochloride, hydrobromide, sulfate, nitrate, oxalate, maleate, p-toluenesulfonate or tosylate .

In the present invention, the compound of formula (I) is embodied in the form of pharmaceutical preparations, medical devices, and daily chemical products; the forms of the pharmaceutical preparations, medical devices, and daily chemical products are hydrogels, foam gels, coating agents, and cataplasms , freeze-dried powder, liquid, aerosol, suppository, liniment for external use, ointment.

The pharmaceutical excipients of the pharmaceutical preparation of the compound of formula (I) in the present invention include traditional anti-adherents, binders, disintegrants, fillers, diluents, glidants, lubricants and preservatives.

Binders include, but are not limited to, gelatin, cellulose, modified cellulose (such as microcrystalline cellulose), methylcellulose, polyvinylpyrrolidone, starch, sucrose, and polyethylene glycol; polyvinylpyrrole is particularly preferred pyridone and/or microcrystalline cellulose. In a preferred embodiment, the content of anti-sticking agent is in the range from 0.001 to 30% by weight, more preferably 0.1 to 25% by weight.

Fillers and/or diluents are preferably selected from but not limited to the group consisting of cellulose, calcium diphosphate, lactose, sucrose, glucose, mannitol, sorbitol and calcium carbonate; microcrystalline cellulose and lactose are particularly preferred. In a preferred embodiment, the filler and/or diluent content ranges from 0.001 to 90% by weight, more preferably 0.1 to 80% by weight, most preferably 10 to 75% by weight.

Lubricants such as magnesium stearate, stearic acid, and stearic esters. In a preferred embodiment, the content of lubricant is in the range from 0.001 to 5.0% by weight.

Disintegrants such as croscarmellose sodium (croscarmellose sodium), crospovidone, and sodium starch glycolate. In a preferred embodiment, the content of the disintegrant is in the range from 0.001 to 5.0% by weight.

Antioxidants such as vitamin A, vitamin E, vitamin C, retinyl palmitate and selenium; cysteine, methionine, citric acid, sodium citrate, methyl and propyl benzoate.

Description of drawings

The HPLC figure of compound (R is methyl) shown in Fig. 1 formula (I),

The 1H-NMR figure of compound (R is methyl) shown in Fig. 2 formula (I)

The mass spectrum ([M-H]+) of compound (R is methyl) shown in Fig. 3 formula (I)

The mass spectrogram ([M+H]+) of the compound (R is methyl) shown in Fig. 4 formula (I)

The mass spectrum ([M+H]+) of the compound (R is hydrogen) shown in Fig. 5 formula (I)

The mass spectrum ([M-H]+) of the compound (R is ethyl) shown in Fig. 6 formula (I)

The mass spectrum ([M+H]+) of the compound (R is isopropyl) shown in Fig. 7 formula (I)

Detailed ways

The present invention will be further described in detail below in conjunction with specific embodiments, and the given examples are only for clarifying the present invention, not for limiting the scope of the present invention. For the specific techniques or conditions not indicated in the examples, the techniques or conditions described in the literature in this field or the product instructions shall be followed.

In an embodiment of the present invention,

HPLC: Analytical column: Diamonsil ^TM C ₁₈ (250mm×4.6mm); mobile phase: 0.01mol L ^-1 potassium dihydrogen phosphate (0.3mmol L ^-1 sodium dodecylsulfonate) - acetonitrile (65: 35); flow rate: 1ml·min-1; ultraviolet detection wavelength: 240nm; injection volume: 10μl.

1H-NMR: AVANCE III 500M fully digital superconducting nuclear magnetic resonance spectrometer

Mass Spectrometry: Bruker APEX IV Fourier Transform Cyclotron Resonance Mass Spectrometer.

Embodiment 1: the preparation of compound (R is methyl) shown in formula (I)

Dissolve 5 g of the compound represented by formula (II) (R is methyl) in 50 ml of methanol, cool down to -40°C, add dropwise 10% aqueous sodium hydroxide solution to adjust the pH to 9-10, and keep the reaction solution at this level all the time. pH value, keep the reaction temperature at -40～-30°C and stir until the reaction is complete, wash with dilute hydrochloric acid aqueous solution to pH 6～7, extract with 200ml dichloromethane, dry the dichloromethane layer with anhydrous magnesium sulfate overnight, concentrate, and Column (eluent is ethyl acetate:petroleum ether=1:15), the eluent is concentrated under reduced pressure, the residue is heated to 60°C with a small amount of ethanol, 1g of oxalic acid is added, stirred for 30min, cooled to 0-5°C and analyzed crystallized, filtered, and the filter cake was neutralized with sodium bicarbonate, and dried to obtain 0.96 g of the compound shown in formula (I). According to the HPLC area normalization method, the content was 99.7%.

Mp: 210.2℃～211.9℃

MS: 425.0[M+H] ⁺ , 423.1[MH] ⁺

¹ H-NMR (500MHz, DMSO-d ₆ )

δ: 0.91～1.08(m, 1H), 2.30(s, 3H), 2.50～2.60(m, 3H), 3.56(s, 1H), 4.04～4.06(d, 1H), 6.81(d, 1H), 7.47～7.49(m, 1H), 7.61～7.65(m, 2H), 7.85～7.87(m, 1H), 7.91～7.95(m, 1H), 8.10～8.11(m, 1H), 8.53～8.54(m , 1H) 12.07(s, 1H).

Embodiment 2: the preparation of compound (R is hydrogen) shown in formula (I)

Dissolve 5g of the compound represented by formula (II) (R is hydrogen) in 35ml of ethanol, cool down to -30～-20°C, add dropwise 5% potassium hydroxide aqueous solution to adjust the pH to 10～12, and keep the pH during the reaction. pH value, keep warm at -20～-10°C for reaction, add 200ml of dichloromethane after the reaction is completed, wash with acetic acid aqueous solution to pH 6～7, dry over anhydrous magnesium sulfate, concentrate, pass through the column (eluent is ethyl acetate: n-Hexane=1:20), the eluent was concentrated under reduced pressure, the residue was heated to 45°C with a small amount of acetone, 1.6g maleic acid was added, stirred for 30min, cooled to 0-5°C, stirred and crystallized, filtered, and used for filter cake Sodium bicarbonate neutralized and dried to obtain 1.57 g of the compound represented by formula (I).

Mp: 203.4℃～205.1℃

MS: 411.2[M+H] ⁺

Embodiment 3: the preparation of compound (R is ethyl) shown in formula (I)

Dissolve 5g of the compound represented by formula (II) (R is ethyl) in 60ml of isopropanol, cool down to -10～-5°C, add 0.5g of lithium hydroxide monohydrate in batches, stir at room temperature until the reaction is complete, add 200ml of dichloromethane, washed with dilute hydrochloric acid solution to pH 6-7, dried over anhydrous magnesium sulfate, concentrated, passed through the column (eluent: ethyl acetate: petroleum ether = 1:10), the eluent was concentrated under reduced pressure , the residue was separated with a preparative liquid phase, the mobile phase was methanol: acetonitrile: water: triethylamine = 45:25:30:0.01, the fractions were received and concentrated to obtain 1.15 g of the compound represented by formula (I).

Mp: 213.4℃～215.1℃

MS: 437.1[MH] ⁺

Embodiment 4: the preparation of compound (R is isopropyl) shown in formula (I)

Dissolve 5g of the compound represented by formula (II) (R is isopropyl) in 30ml of n-butanol and 10ml of water down to 0°C, add 1.58g of potassium carbonate in batches, stir at room temperature until the reaction is complete, add 200ml of dichloromethane, and use Wash with hydrochloric acid aqueous solution to pH 6-7, dry over night with anhydrous magnesium sulfate, concentrate, and pass through the column (eluent is ethyl acetate:cyclohexane=1:15), the eluent is concentrated under reduced pressure, and the residue is washed with a small amount of methanol Heat to 50°C, add 1.6g of fumaric acid, stir for 30min, cool down to 0-5°C, stir and crystallize, filter, and neutralize the filter cake with sodium bicarbonate to obtain 1.48g of the compound represented by formula (I).

Mp: 214.6℃～216.4℃

MS: 453.4[M+H] ⁺

Embodiment 5: Preparation of compound (R is isopropyl) shown in formula (I)

Dissolve 5g of the compound represented by formula (II) (R is isopropyl) in 15ml of methanol and 10ml of water, add 0.45g of sodium hydroxide at room temperature, stir at 45°C until the reaction is complete, add 300ml of dichloromethane, and wash with aqueous hydrochloric acid to pH To 6~7, dry over night with anhydrous magnesium sulfate, concentrate, dissolve the residue with 50ml of toluene, heat to 110°C, add 8ml of 40% hydrogen bromide acetic acid solution dropwise, react until complete at 110~120°C, reduce to 60°C The solvent was concentrated under pressure, the residue was dissolved in 100ml of dichloromethane, neutralized to neutral with saturated sodium bicarbonate solution, the dichloromethane layer was dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was passed through the column (eluent was acetic acid Ethyl ester: petroleum ether = 1:30), the eluent was concentrated under reduced pressure, the residue was heated to 45°C with a small amount of 2-butanone, 2g of tartaric acid was added, stirred for 30min, cooled to 0-5°C, stirred and crystallized, filtered, The filter cake was neutralized with sodium bicarbonate and dried to obtain 0.36 g of the compound represented by formula (I).

Embodiment 6: the preparation of compound (R is ethyl) shown in formula (II)

Dissolve 5g of the compound represented by formula (II) (R is ethyl) in 25ml of ethanol and 5ml of water, add 1.5g of sodium carbonate at room temperature, stir at 65°C until the reaction is complete, add 300ml of dichloromethane, wash with aqueous hydrochloric acid to pH 6 ～7, dry over night with anhydrous magnesium sulfate, concentrate, dissolve the residue with 50ml of xylene, heat to 110°C, add 10ml of acetyl bromide dropwise, react at 110～120°C until complete, concentrate under reduced pressure at 80°C to remove the solvent, and the residue Dissolve in 100ml of dichloromethane, neutralize to neutral with saturated sodium bicarbonate solution, dry the dichloromethane layer with anhydrous sodium sulfate, filter, concentrate, and separate the residue with a preparative liquid phase. The mobile phase is methanol: acetonitrile: Water: triethylamine=45:25:30:0.01, the fractions were received and concentrated to obtain 0.47 g of the compound represented by formula (I).

Embodiment 7: Pharmacological experiment methods and results of the compound (R is methyl) shown in formula (I) on platelet aggregation activity

Experimental method: Take 2 rabbits, use lidocaine to local anesthesia, surgically separate the common carotid artery, take blood, take 3.8% sodium citrate 1:9 anticoagulation, and centrifuge at 500r/min for 10min to prepare platelet-rich plasma (PRP ), and the remaining part was centrifuged at 3000r/min to prepare platelet-poor plasma (PPP), and the platelet aggregation test was carried out according to the turbidimetric method. Add 240 μL of PRP and 30 μL of different concentrations of the test drug into the measurement tube, incubate at 37°C for 5 minutes, use 30 μL of adenosine diphosphate sodium salt (ADP) (final concentration: 10 μmol/L) as the inducer, observe and record the maximum aggregation within 5 minutes Rate. DMSO was used as a blank control and aspirin was used as a positive control to calculate the platelet aggregation inhibition rate (AIR) of the target compound.

Result of the test: the compound of formula (I) of the present invention has been listed in table 1 to ADP-induced rabbit platelet aggregation activity data, positive control drug is aspirin

Table 1 The inhibitory activity of some compounds of the present invention on ADP-induced platelet aggregation in rabbits

The above pharmacological data show that the compound of formula (I) of the present invention can inhibit platelet aggregation, and has stronger platelet aggregation inhibitory effect than aspirin.

Similar results were also obtained when R was hydrogen, ethyl, or isopropyl.

Claims (16)

1. A 3-benzodiazepine Compounds, or pharmaceutically acceptable salts, stereoisomers, and tautomers thereof, the compound has a structure shown in formula (I): Wherein, R is hydrogen, methyl, ethyl, isopropyl. 2. The compound according to claim 1, characterized in that the pharmaceutically acceptable salt of the compound of formula (I) refers to hydrochloride, hydrobromide, sulfate, nitrate, oxalate, horse tosylate, p-toluene sulfonate or tosylate. 3. a method for preparing the compound shown in formula (I) described in claim 1, is characterized in that, the compound shown in formula (II) is hydrolyzed with alkali to obtain Wherein, R is hydrogen, methyl, ethyl, isopropyl. 4. The method according to claim 3, characterized in that the temperature of the hydrolysis reaction between the compound represented by formula (II) and the base is -40-0°C. 5. A method for preparing the compound represented by formula (I) according to claim 1, characterized in that the compound represented by formula (II) is hydrolyzed with a base, and then brominated. 6. according to the synthetic method described in claim 5, it is characterized in that, the bromination reagent of this bromination reaction is hydrogen bromide, acetyl bromide, bromine, NBS. 7. according to the described synthetic method of claim 3 or 5, it is characterized in that, described alkali is any of sodium hydroxide, potassium hydroxide, lithium hydroxide, calcium hydroxide, barium hydroxide, sodium carbonate, salt of wormwood one or a combination thereof. 8. The synthesis method according to claim 5, characterized in that the temperature of the hydrolysis reaction between the compound represented by the formula (II) and the base is room temperature -40-100°C. 9. The synthesis method according to any one of claims 5 and 6, characterized in that the temperature of the bromination reaction is -10 to 110°C. 10. A 3-benzodiazepine Compounds, or pharmaceutically acceptable salts, stereoisomers, tautomers thereof, in the preparation of antithrombotic drugs, the compound has a structure shown in formula (I): Wherein, R is hydrogen, methyl, ethyl, isopropyl. 11. The application according to claim 10, characterized in that the pharmaceutically acceptable salt of the compound of the formula (I) refers to hydrochloride, hydrobromide, sulfate, nitrate, oxalate, malate salt, p-toluenesulfonate or toluenesulfonate. 12. The application of an antithrombotic composition in the preparation of drugs for the prevention and treatment of cardiovascular and cerebrovascular diseases, the composition contains an effective amount of the compound described in formula (I) according to claim 1, or its Pharmaceutically acceptable salts, stereoisomers, tautomers. 13. The application according to claim 12, characterized in that said cardiovascular and cerebrovascular diseases refer to thrombotic diseases. 14. The use according to claim 13, wherein the thrombotic disease is myocardial infarction, ischemic cardiomyopathy, cerebral thrombosis, cerebral embolism, cerebral infarction, transient ischemic attack or lacunar infarction. 15. The application according to claim 12, characterized in that: the application is embodied in the form of pharmaceutical preparations, medical equipment, and daily chemical products; the form of the pharmaceutical preparation, medical equipment, and daily chemical products is hydrogel, foam Gels, coatings, cataplasms, freeze-dried powders, liquids, aerosols, suppositories, liniments for external use, ointments. 16. A pharmaceutical composition for preventing and treating cardiovascular and cerebrovascular diseases, characterized in that: the pharmaceutical composition contains a therapeutically effective amount of the compound of formula (I) according to claim 1, or a pharmaceutically acceptable salt thereof , stereoisomers, tautomers, pharmaceutical excipients, pharmaceutical carriers or excipients.