CN107721925B - Novel acetylcholinesterase inhibitor and preparation method and application thereof - Google Patents

Abstract

The invention provides a novel acetylcholinesterase inhibitor, a preparation method and application thereof, and particularly discloses substituted acridine compounds with double AChE binding sites, which are shown as a formula A, a preparation method and application thereof as an acetylcholinesterase inhibitor. The novel compound prepared by the invention shows good effect of inhibiting acetylcholinesterase, and has application value of treating and preventing Alzheimer's Disease (AD).

Description

Novel acetylcholinesterase inhibitor and preparation method and application thereof

Technical Field

The invention belongs to the technical field of pharmaceutical chemical industry, and particularly relates to a substituted acridine compound with double AChE binding sites, and a preparation method and application thereof. The compound can effectively inhibit the activity of acetylcholinesterase (AChE), and can be used for preparing medicines for treating and preventing Alzheimer's Disease (AD).

Background

Alzheimer's disease is called AD for short, and is discovered and described by Alois Alzheimer for the first time, and the neurodegenerative disease is mainly characterized by cognitive decline. At present, the number of people suffering from AD in the whole world tends to rise year by year, and the number of people suffering from AD in the whole world is estimated to exceed 1 hundred million and 2 million by 2050. With the aging of the global population, the number of patients with AD and the economic losses caused by AD are increasing. Therefore, the design and research of novel compounds for treating AD is urgently needed.

AD has been shown to be a multifactorial disease. According to researches, a plurality of factors such as heredity, environment, life style and the like can influence the pathogenesis of the AD. There are therefore a wide variety of hypotheses about the pathogenesis of AD, such as: cholinergic hypotheses, amyloid hypotheses, tau hypotheses, glutamatergic hypotheses, immune dysregulation hypotheses, oxidative stress hypotheses, and the like. Among the numerous hypotheses, the cholinergic and amyloid hypotheses are currently the predominant hypothesis for the pathogenesis of AD, supported by numerous evidences.

Rossor, the first clear link the cholinergic hypothesis to AD, suggests that impairment of the ascending cholinergic system of the brain leads to dysfunctional manifestations in AD patients, specifically Alzheimer's disease, whereas the amyloid protein hypothesis states that amyloid A β, which aggregates into fibrillar structures, can produce neurotoxicity by affecting intracellular calcium homeostasis, leading to AD-related pathological changes.

Acetylcholinesterase (AChE) is present at the neuromuscular junction and between cholinergic synapses and its primary function is to hydrolyze acetylcholine, thereby terminating signal transmission. Inhibition of AChE can increase synaptic cleft choline concentration, promoting signaling. AChE is complex in structure and mainly includes: a catalytically active site CAS region, an active pocket entrance PAS region, a fragrance canyon, and a backdoor region, among others. Since AChE is the theoretical basis of the cholinergic and amyloid hypothesis of AD, it is, of course, one of the popular targets for anti-AD drug research.

At present, AChE inhibitors taking AChE as a target point are classified according to action sites and mainly classified into three types: inhibitors that act only on the CAS site, inhibitors that act only on the PAS site, and inhibitors that act simultaneously on the dual binding site of CAS and PAS. However, in the currently marketed AChE inhibitors, all inhibitors act only on the CAS site of AChE except for the weak interaction of donepezil with PAS. Theoretically, a dual site inhibitor can increase the binding capacity of a compound to AChE compared to an AChE single site inhibitor.

Therefore, research and development of novel AChE inhibitors acting on the dual binding sites of CAS and PAS are currently a new direction in the art, and it is expected that the efficacy of existing anti-AD drugs can be greatly enhanced.

Disclosure of Invention

The invention aims to overcome the defects in the prior art and provides a substituted acridine compound with double AChE binding sites, and a preparation method and application thereof.

The first aspect of the invention provides a substituted acridine compound of a double AChE binding site, or a pharmaceutically acceptable salt or prodrug thereof, wherein the compound has a structure shown as a compound in a formula A:

wherein n is an integer of 2 to 12.

In another preferred embodiment, the compounds of the present invention have the formula shown in formula (I):

in another preferred embodiment, the compounds of the present invention have the formula shown in formula (II):

in another preferred embodiment, the compounds of the present invention have a formula as shown in formula (III):

in another preferred embodiment, the compound of formula a inhibits the IC of AChE enzyme activity₅₀Less than or equal to 5 μ M, preferably IC₅₀Less than or equal to 1 mu M, preferably IC₅₀Less than or equal to 0.2. mu.M, optimally IC₅₀≤0.05μM。

In another preferred embodiment, the maximum inhibition of AChE enzyme activity by said compound of formula a is > 95%.

In a second aspect, the present invention provides a pharmaceutical composition, which comprises (a) the substituted acridine compound of the double AChE binding site according to the first aspect of the present invention, or a pharmaceutically acceptable salt or prodrug thereof as an active ingredient and (b) a pharmaceutically acceptable carrier.

A third aspect of the present invention provides a process for preparing a pharmaceutical composition according to the second aspect of the present invention, said process comprising the steps of:

mixing a pharmaceutically effective amount of a compound of formula a, or a pharmaceutically acceptable salt or prodrug thereof, with a pharmaceutically acceptable carrier to form a pharmaceutical composition.

In another preferred embodiment, the pharmaceutical composition further comprises: donepezil, memantine, rivastigmine and galantamine.

In another preferred embodiment, the pharmaceutical composition comprises 0.01-99 wt%, preferably 10-80 wt%, more preferably 30-75 wt% of the compound of formula a, or its pharmaceutically acceptable salt, based on the total weight of the composition.

In another preferred embodiment, the pharmaceutically acceptable carrier is selected from the group consisting of: saline, buffer, glucose, water, glycerol, ethanol, dimethyl sulfoxide, or a combination thereof.

In another preferred embodiment, the pharmaceutical composition is an oral preparation or an injection.

In a fourth aspect of the invention, there is provided a method of non-therapeutically inhibiting AChE enzymatic activity in vitro, said method comprising: administering to a subject an inhibitory effective amount of a substituted acridine compound of the double AChE binding site according to the first aspect of the invention or a pharmaceutically acceptable salt or prodrug thereof, and/or a pharmaceutical composition according to the second aspect of the invention.

The fifth aspect of the invention provides a preparation method of substituted acridine compounds with double AChE binding sites having a structure shown as compound XIII, which comprises the following steps:

reacting compound VIII and compound XII in the presence of an organic solvent to form compound XIII,

in the formula, n is an integer of 2-12; the organic solvent is selected from the following group: methanol, acetonitrile, acetone, or combinations thereof; the reaction is carried out at a temperature of 50 ℃ to 90 ℃.

A sixth aspect of the present invention provides a use of a substituted acridine compound of a double AChE binding site according to the first aspect of the present invention, or a pharmaceutically acceptable salt or prodrug thereof, for a use selected from the group consisting of:

(i) for preparing AChE enzyme activity inhibitor;

(ii) for the preparation of a pharmaceutical composition for the treatment or prevention of a neurodegenerative disease; or

(iii) For non-therapeutic inhibition of AChE enzymatic activity in vitro.

In another preferred embodiment, the neurodegenerative disease includes alzheimer's disease and parkinson's disease.

It is to be understood that within the scope of the present invention, the above-described features of the present invention and those specifically described below (e.g., in the examples) may be combined with each other to form new or preferred embodiments. Not to be reiterated herein, but to the extent of space.

Detailed Description

The inventor of the invention has long and intensive research and unexpectedly found that the substituted acridine compound with double AChE binding sites is a novel, safe and efficient AChE inhibitor, wherein acridine is used as an aromatic ring head, fatty chains with different lengths are introduced to be used as a linker, and benzyl is used as an aromatic ring tail. The AChE enzyme activity inhibition test confirms that the novel compound prepared by the invention has good AChE enzyme activity inhibition effect, and can be used for preparing medicines for treating and preventing neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease and the like. On the basis of this, the present invention has been completed.

Active ingredient

As used herein, the term "compound of the invention" refers to a compound of formula a. The term also includes various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compounds of formula a, as well as tautomers, racemates, enantiomers and diastereomers of the compounds of formula a.

The pharmaceutically acceptable solvates include (but are not limited to): and (3) solvates of the compound of the formula A and solvents such as water, ethanol, isopropanol, diethyl ether, acetone and the like.

As used herein, the term "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention with an acid or base that is suitable for use as a pharmaceutical. Pharmaceutically acceptable salts include inorganic or organic acid salts; the inorganic acid is selected from one or more of the following groups: hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid; the organic acid is selected from one or more of the following groups: formic acid, acetic acid, propionic acid, trifluoroacetic acid, benzoic acid, maleic acid, fumaric acid, succinic acid, tartaric acid, citric acid, alkylsulfonic acids (e.g., methanesulfonic acid, ethanesulfonic acid, etc.), arylsulfonic acids (e.g., benzenesulfonic acid, p-toluenesulfonic acid, etc.).

Pharmaceutical compositions and methods of administration

The compound has excellent inhibitory activity on AChE, so the compound and various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates thereof, and a pharmaceutical composition containing the compound as a main active ingredient can be used for treating, preventing and relieving diseases mediated by AChE.

The compound can be used for preparing medicaments for treating and preventing Alzheimer's Disease (AD).

The pharmaceutical composition of the present invention comprises the compound of the present invention or a pharmacologically acceptable salt thereof in a safe and effective amount range and a pharmacologically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects.

"pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances which are suitable for human use and must be of sufficient purity and sufficiently low toxicity. By "compatible" is meant herein that the components of the composition are capable of intermixing with and with the compounds of the present invention without significantly diminishing the efficacy of the compounds. Examples of pharmaceutically acceptable carrier moieties are cellulose and its derivatives (e.g., sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (e.g., stearic acid, magnesium stearate), calcium sulfate, vegetable oils (e.g., soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (e.g., propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (e.g., propylene glycol, glycerin, mannitol, sorbitol, etc.), and the like) Wetting agents (e.g., sodium lauryl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, and the like.

The mode of administration of the compounds or pharmaceutical compositions of the present invention is not particularly limited, and representative modes of administration include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia; (c) humectants, for example, glycerol; (d) disintegrating agents, for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) slow solvents, such as paraffin; (f) absorption accelerators, e.g., quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glycerol monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage forms may also comprise buffering agents.

Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shells such as enteric coatings and other materials well known in the art. They may contain opacifying agents and in such compositions, the active compound or compounds may be released in a delayed manner in a portion of the digestive tract. Examples of embedding components which can be used are polymeric substances and wax-like substances. If desired, the active compound may also be in microencapsulated form with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly employed in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1, 3-butylene glycol, dimethylformamide and oils, in particular, cottonseed, groundnut, corn germ, olive, castor and sesame oils or mixtures of such materials and the like.

In addition to these inert diluents, the compositions can also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

Suspensions, in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances, and the like.

Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols and suitable mixtures thereof.

Dosage forms for topical administration of the compounds of the present invention include ointments, powders, patches, sprays, and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants which may be required if necessary.

The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds.

The use of pharmaceutical compositions in which a safe and effective amount of a compound of the invention is administered to a mammal (e.g., a human) in need of treatment is within the skill of the skilled practitioner in a dosage amount that is pharmaceutically effective for administration, the particular dosage amount being determined by the route of administration, the health of the patient, and the like.

AChE inhibitors

As used herein, the term "AChE inhibitor" refers to a compound or composition that can be used to inhibit AChE enzymatic activity.

AChE is present at the neuromuscular junction as well as between cholinergic synapses and has the primary function of hydrolyzing acetylcholine, thereby terminating the transmission of signals. Inhibition of AChE can increase synaptic cleft choline concentration, promoting signaling. AChE is a complex mechanism of catalysis, is the theoretical basis of the cholinergic and amyloid hypothesis of alzheimer's disease, and administration of AChE inhibitors to subjects suffering from the above diseases can effectively treat or ameliorate the diseases.

Wherein "treating" or "treatment" refers to reducing, preventing, or reversing the disease or condition or at least one discernible symptom thereof, ameliorating, preventing, or reversing at least one measurable physical parameter associated with the disease or condition being treated, inhibiting or slowing the progression of the disease or condition, or delaying the onset of the disease or condition.

The term "ameliorating" a symptom of a particular disorder as used herein refers to any reduction, prevention, or reversal of the reduction, whether permanent, temporary, long-term, transient, or at least one discernible symptom of the disorder or condition.

A compound of formula A

The invention provides a substituted acridine compound of a double AChE binding site, and particularly provides a compound shown as a formula A:

the purpose of the invention is realized by the following technical scheme:

the invention relates to substituted acridine compounds of double AChE binding sites, which have the structural formula as follows:

in the formula (A), n is any one of integers of 2-12.

Preferably, the structural formula is shown as formula (I), formula (II) and formula (III):

process for the preparation of the compounds of the invention

The synthetic route of the substituted acridine compound with the double AChE binding sites is shown as follows:

the invention also relates to a method for preparing the bis 2-substituted ethylene sulfonate compound with the structure shown in the formula (III), which comprises the following steps:

(1) to a 500ml eggplant-shaped flask was added 2-chlorobenzoic acid (23.9g,153mmol), aniline (9.8g,105mmol), copper powder (3.2g,50mmol) and potassium carbonate (34.0g,246mmol), and finally DMF (250ml) was added, stirred in an oil bath at 130 ℃ overnight, TLC was used to detect the complete disappearance of aniline, the oil bath was removed, allowed to cool naturally to room temperature, water (750ml) was added to the reaction mixture, and the pH was adjusted to weak acidity with dilute hydrochloric acid. The precipitated precipitate is filtered, washed with water and drained to obtain the N-phenyl anthranilic acid.

(2) In a 100ml eggplant-shaped flask, N-phenylanthranilic acid (5.0g, 23.5mmol), phosphorus oxychloride (50ml, 537mmol) were added, stirred in an oil bath at 110 ℃ for 3 hours, TLC (developing solvent: dichloromethane/methanol ═ 20/1) detected the complete disappearance of compound III-14, the oil bath was removed, allowed to cool to room temperature naturally, and the reaction mixture was slowly added to 150g of ice and stirred vigorously. After the exotherm was complete, the pH was adjusted with saturated sodium bicarbonate solution until no more bubbles were formed. And filtering, leaching the filter cake with water for 2-3 times, and pumping to obtain a solid, namely 9-chloroacridine.

(3) 9-chloroacridine (0.201g,0.94mmol) and phenol (1.30g,13.8mmol) were added to the test tube, heated in an oil bath at 80 ℃ for 1 hour, the reaction mixture was removed from the oil bath, and after cooling to room temperature, the reaction mixture was dissolved in dichloromethane (50ml) and washed with 1mol/L sodium hydroxide solution (30ml) in portions until the phenol disappeared. The organic phase was washed with saturated sodium chloride solution and dried over sodium sulfate. Evaporating and pumping to dry to obtain the target product 9-phenoxyacridine.

(4) 1, 9-nonanediol (5.0g, 31.2mmol), dichloromethane (100ml) and DMF (3 drops) were added to a 500ml eggplant-type bottle. Thionyl chloride (11.7g, 98.3mmol) was dissolved in dichloromethane (155 ml). Under ice bath, the thionyl chloride solution was added dropwise to the eggplant-shaped bottle and reacted overnight. Washing with saturated sodium bicarbonate solution, washing with saturated sodium chloride solution, drying the organic phase with anhydrous sodium sulfate, and evaporating to obtain 1, 9-dichlorononane.

(5) To a 200ml stopcock were added 1, 9-dichlorononane (4.0g,20.3mmol), phthalimide potassium salt (12.5g,68.5mmol) and DMF (100 ml). The reaction was carried out in an oil bath at 130 ℃ overnight. The reaction mixture was poured into water (800ml) and the precipitate was collected as 2, 2' - (nonane-1, 9-diyl) bis (isoindole-1, 3-dione).

(6) 2, 2' - (nonane-1, 9-diyl) bis (isoindole-1, 3-dione) (4.2g, 10mmol), 80% (10ml) hydrazine hydrate and absolute ethanol (100-200 ml) were added to an eggplant-shaped bottle, and the mixture was refluxed overnight. And (3) treatment: filtering, and washing the filter residue with anhydrous ethanol for 3 times; evaporating the filtrate, filtering again, washing the filter residue with absolute ethyl alcohol again, and evaporating the filtrate to obtain the 1, 9-nonanediamine crude product.

(7) 1, 9-nonanediamine (5.0ml,51mmol) and anhydrous methanol (38ml) were added to a 100ml eggplant-shaped flask, benzaldehyde (1000. mu.L, 9.8mmol) was dissolved in anhydrous methanol (5ml), the solution was added dropwise to the eggplant-shaped flask, after the addition was completed, stirring was continued for 30min, then sodium borohydride (0.371g,11mmol) was gradually added under ice bath, the reaction was allowed to stand overnight, and after the reaction mixture was evaporated, dichloromethane (50ml) was added, and the mixture was washed twice with saturated saline (5 ml). The organic phase was dried over sodium sulfate and evaporated to give N-benzyl-1, 9-nonanediamine.

(8) To an eggplant type flask were added 9-phenoxyacridine (54mg, 0.2mmol) and 1, 9-nonanediamine monobenzylamine (55mg, 0.22 mmol). Add the appropriate amount of methanol and reflux for 24-48 hours until disappearance of starting material a or 48 hours of reaction time as monitored by TLC. The eggplant-shaped bottle was removed from the oil bath. The reaction mixture was evaporated to dryness and purified by column chromatography or preparative TLC to give N- (9-acridine) -N' -benzyl-1, 9-nonanediamine.

The main advantages of the invention include

(1) The invention provides substituted acridine compounds with double AChE binding sites with novel structures, and the substituted acridine compounds have reasonable design of synthetic routes, easily obtained raw materials and suitability for application.

(2) The compound of formula A provided by the invention can be used for preparing a series of medicines for treating AChE enzyme activity related diseases. Has application value in preventing or treating the Alzheimer disease with the AChE as the target point, and the like, and has wide application.

(3) The compound of formula A provided by the invention has high activity and minimum IC as AChE enzyme inhibitor₅₀The value can be less than or equal to 0.05 mu m, and the inhibitory activity can be shown under extremely low concentration.

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out under conventional conditions or conditions recommended by the manufacturers. Unless otherwise indicated, percentages and parts are by weight.

Example 1

Synthesis of N-Phenylanthranilic acid (VI): to a 500ml eggplant type bottle were added 2-chlorobenzoic acid (23.9g,153mmol), aniline (9.8g,105mmol), copper powder (3.2g,50mmol) and potassium carbonate (34.0g,246mmol), and finally DMF (250ml) was added and stirred in an oil bath at 130 ℃ overnight. The disappearance of aniline was detected by TLC. The oil bath was removed, the temperature was naturally lowered to room temperature, water (750ml) was added to the reaction mixture, and the pH was adjusted to weak acidity with dilute hydrochloric acid. The precipitated precipitate was filtered, washed with water, and then drained to obtain 14.2g of N-phenylanthranilic acid; (grey solid, yield 63.0%).¹H NMR(CDCl₃,400MHz):δ11.55(br s, 1H),9.34(br s,1H),8.06(s,1H),7.36(q,3h),7.26-7.28(m,3H),7.13(t,J＝6.9 Hz,1H),6.78(s,1H)；

Example 2

Synthesis of 9-chloroacridine (VII): in a 100ml eggplant type bottle, N-phenylanthranilic acid (5.0g, 23.5mmol), phosphorus oxychloride (50ml, 537mmol) were added. After stirring in an oil bath at 110 ℃ for 3 hours, compound III-14 was detected by TLC (developer: dichloromethane/methanol: 20/1) as having disappeared completely. The oil bath was removed, allowed to cool to room temperature naturally, and the reaction mixture was slowly added to 150g of ice and stirred vigorously. After the exotherm was complete, the pH was adjusted with saturated sodium bicarbonate solution until no more bubbles were formed. Filtering, leaching a filter cake with water for 2-3 times, and pumping to dry to obtain a solid, namely 2.8g of 9-chloroacridine; (brown solid, yield 56%).¹H NMR(CDCl₃,400MHz)δ8.44 (d,J＝8.8Hz,2H),8.25(d,J＝8.8Hz,2H),δ8.44(d,J＝8.8Hz),8.25(d,J＝8.8 Hz),7.86-7.79(m,2H),7.68-7.61(m,2H)；13C NMR(CDCl₃,100MHz)δ148.8, 133.4,130.6,129.7,126.9,124.6,124.3。

Example 3

Synthesis of 9-phenoxyacridine (VIII): 9-chloroacridine (0.201g,0.94mmol) and phenol (1.30g,13.8mmol) were added to the tube and heated in an oil bath at 80 ℃ for 1 hour. The reaction mixture is removed from the oil bath, cooled to room temperature, dissolved in dichloromethane (50ml) and treated with 1 mol-L sodium hydroxide solution (30ml) was washed in portions until the phenol disappeared. The organic phase was washed with saturated sodium chloride solution and dried over sodium sulfate. After evaporation and suction drying, the target product, namely the 9-phenoxyacridine, is 230mg (light brown solid, yield is 90%). 1H-NMR (400MHz, CDCl)₃)δ8.27(d,J＝8.9Hz,2H),8.10(d,J＝8.7Hz,2H), 7.81-7.74(m,2H),7.46(t,J＝7.6Hz,2H),7.27(dd,J＝8.3,4.4Hz,2H),7.05(t,J ＝7.3Hz,1H),6.85(d,J＝8.1Hz,2H).13C-NMR(CDCl₃,100MHz)δ159.5, 155.2,150.5,130.6,129.9,129.6,125.8,122.7,122.6,120.3,115.5。

Example 4

Synthesis of 1, 9-dichlorononane (IX): 1, 9-nonanediol (5.0g, 31.2mmol), dichloromethane (100ml) and DMF (3 drops) were added to a 500ml eggplant-type bottle. Thionyl chloride (11.7g, 98.3mmol) was dissolved in dichloromethane (155 ml). Under ice bath, the thionyl chloride solution was added dropwise to the eggplant-shaped bottle and reacted overnight. The reaction mixture was washed with saturated sodium bicarbonate solution and then with saturated sodium chloride solution, and the organic phase was dried over anhydrous sodium sulfate and evaporated to dryness to give 4.0g of 1, 9-dichlorononane (pale yellow oily substance, yield 65%). 1H NMR (CDCl)₃,400MHz)δ3.53(t,J＝6.8Hz,4H),1.77(quint,J＝6.8Hz,4H),1.43(quint, J＝8Hz,4H),1.314(m,6H)；13C NMR(CDCl₃,100MHz)δ46.1,32.6,29.2,28.7, 26.8。

Example 5

2, 2' - (nonane-1, 9-diyl) bis (isoindole-1, 3-dione) (X₁) The synthesis of (2): 1, 9-Dichlorononane (4.0g,20.3mmol), phthalimide potassium salt (12.7g,68.6mmol) and DMF (100ml) were added to a 250ml eggplant-shaped flask, heated in an oil bath at 130 ℃ overnight, the reaction was poured into water, and the precipitate was collected and dried to give 12.0g (white solid, yield 91%) of 2,2 "- (nonane-1, 9-diyl) bis (isoindole-1, 3-dione).¹H NMR(CDCl₃,400MHz)δ7.85-7.82(m,4H),7.74-7.68(m,4H),3.67(t,J＝7.3 Hz),1.69-1.63(m,4H),1.31-1.30(m,10H)；13CNMR(CDCl₃,100MHz)δ168.4, 133.8,132.1,123.1,38.0,29.3,29.0,28.5,26.8。

Example 6

1, 9-nonanediamine (XI)₁) The synthesis of (2): in eggplant shapeAdding 10g of 2, 2' - (nonane-1, 9-diyl) bis (isoindole-1, 3-dione), 80% (10ml) of hydrazine hydrate and 100-200 ml of absolute ethyl alcohol into a bottle, refluxing overnight, filtering the reaction mixed solution, washing filter residues for 3 times by using the absolute ethyl alcohol, evaporating the filtrate, filtering again, washing the filter residues by using the absolute ethyl alcohol again, and evaporating the filtrate to obtain 2.4g of 1, 9-nonanediamine (pale yellow semisolid, yield 44%).¹H NMR(CDCl₃,400MHz)δ2.68(t,J＝7.0Hz,4H),1.49-1.37(m,4H),1.29(m, 10H)。

Example 7

1, 5-Pentanediamine (XI)₂) The synthesis of (2): referring to example 7, the yield was 38%.¹H NMR(CDCl₃,400 MHz)δ2.80-2.59(m,4H),1.47-1.35(m,4H),1.25-1.23(m,2H)。

Example 8

N-benzylethylenediamine (XII)₁) The synthesis of (2): in a 100ml eggplant-shaped flask were added ethylenediamine (3.4ml,51 mmol) and anhydrous methanol (38 ml). Benzaldehyde (1000. mu.L, 9.8mmol) was dissolved in anhydrous methanol (5ml), added dropwise to an eggplant-shaped flask, after the addition was completed, stirring was continued for 30min, and then sodium borohydride (0.371g,11mmol) was gradually added under ice bath, and the reaction was allowed to proceed overnight. After the reaction mixture was evaporated, methylene chloride (50ml) was added, and the mixture was washed twice with saturated brine (5 ml). The organic phase was dried over sodium sulfate and evaporated to yield N-benzylethylenediamine 938mg (colorless liquid, yield 64%). 1H NMR (CDCl)₃,400MHz)δ7.33-7.31(m, 4H),7.26-7.22(m,1H),3.81(s,2H),2.83(t,J＝5.7Hz,2H),2.71(t,J＝5.8Hz, 2H)。

Example 9

N-benzyl-1, 3-propanediamine (XII)₂) The synthesis of (2): reference example 8, yield 9%. 1H NMR (CDCl)₃,400MHz)δ7.31(m,4H),7.25(m,1H),3.79(s,2H),2.78(t,J＝6.7Hz),2.70(t,J ＝6.8Hz),1.70-1.65(m,2H)；13C NMR(CDCl₃,100MHz)δ140.2,128.4,128.1, 126.9,54.1,47.3,40.5,33.5。

Example 10

N-benzyl-1, 4-butanediamine (XII)₃) The synthesis of (2): referring to example 8, the yield was 46%. 1H NMR (CDCl)₃,400MHz)δ7.27-7.28(m,4H),7.20-7.21(m,1H),3.75(s,2H),2.59-2.67(m, 4H),1.31-1.53(m,4H)；13C NMR(CDCl₃,100MHz)δ140.4,128.3,128.0,126.8, 54.0,49.2,42.1,31.5,27.4。

Example 11

N-benzyl-1, 5-pentanediamine (XII)₄) The synthesis of (2): referring to example 8, the yield was 22%.¹H NMR(CDCl₃,400MHz)δ7.28-7.30(m,5H),3.76(s,2H),2.59-2.67(m,4H),1.44-1.51(m,4H), 1.23-1.27(m,2H)。

Example 12

N-benzyl-1, 6-hexanediamine (XII)₅) The synthesis of (2): referring to example 8, the yield was 31%.¹H NMR(CDCl₃,400MHz)δ7.29-7.30(m,5H),3.77(s,2H),2.59-2.67(m,4H),1.47-1.50(m,4H), 1.23-1.27(m,4H)。

Example 13

N-benzyl-1, 7-heptanediamine (XII)₆) The synthesis of (2): referring to example 8, the yield was 25%. 1H NMR (CDCl)₃,400MHz)δ7.29-7.30(m,4H),7.22-7.24(m,1H),3.76(s,2H),2.58-2.66(m,4H), 1.47-1.50(m,4H),1.23-1.28(m,6H)。

Example 14

N-benzyl-1, 8-octanediamine (XII)₇) The synthesis of (2): referring to example 8, the yield was 22%. 1H-NMR (400MHz, CDCl)₃)δ7.32-7.31(m,4H),7.30-7.24(m,1H),3.78(s,2H),2.69-2.60(m, 4H),1.43-1.40(m,2H),1.33-1.26(m,10H)；13C-NMR(101MHz,CDCl₃)δ140.5, 128.4,128.1,126.8,54.1,49.5,42.2,33.8,30.1,29.5,29.4,27.3,26.8。

Example 15

N-benzyl-1, 9-nonanediamine (XII)₈) The synthesis of (2): referring to example 8, yield 24%. 1H NMR (CDCl)₃,400MHz)δ7.35-7.32(m 4H),7.24-7.22(m,1H),3.19(s,2H),2.68(t,J＝6.4Hz, 2H),2.62(t,J＝7.2Hz,2H),1.53-1.49(m,6H),；13C NMR(CDCl₃,100MHz)δ 140.4,128.4,128.1,126.8,54.1,49.5,42.2,33.8,30.0,29.7,29.5,29.5,29.4,27.3, 26.8。

Example 16

N-benzyl-1, 10-sunflower diamine (XII)₉) The synthesis of (2): referring to example 8, the yield was 30%. 1H-NMR (400MHz, CDCl)₃)δ7.32-7.31(m,4H),7.25-7.21(m,1H),3.78(s,2H),2.69-2.60(m, 4H),1.61(m,4H),1.52-1.47(m,2H),1.44-1.41(m,2H),1.33-1.28(m,8H)； 13C-NMR(100MHz,CDCl₃)δ140.5,128.3,129.1,126.8,54.1,49.5,42.2,33.8, 30.1,29.5,19.4,27.3,26.8。

Example 17

N- (9-acridine) -N' -benzyl-1, 2-ethylenediamine (XIII)₁) The synthesis of (2): 9-Phenoxyacridine (1eq) and N-benzylethylenediamine (1-1.2 eq) were added to an eggplant-shaped flask. Add the appropriate amount of methanol and reflux for 24-48 hours until disappearance of starting material a or 48 hours of reaction time as monitored by TLC. The eggplant-shaped flask was taken out of the oil bath and the reaction solution was evaporated to dryness and purified by column chromatography or preparative TLC to give 15mg of N- (9-acridine) -N' -benzyl-1, 2-ethylenediamine (yellow solid, yield 69%).¹H NMR(CDCl₃,400MHz)δ8.18-8.14(m,2H),8.03(d,J＝4.4Hz, 2H),7.52-7.46(m,4H),7.39-7.36(m,2H),7.32-7.29(m,1H),7.16-7.12(m,2H), 4.256(t,J＝5Hz,2H),4.03(s,2H),3.28(t,J＝5.6Hz,2H)；13C NMR(CDCl₃,100 MHz)δ155.2,140.1,138.4,133.8,128.7,128.7,127.7,123.8,123.5,120.4,112.3, 53.0,46.9,46.64,29.7。

Example 18

N- (9-acridine) -N' -benzyl-1, 3-propanediamine (XIII)₂) The synthesis of (2): referring to example 17, the yield was 38%. 1HNMR (CD)₃OD,400MHz)δ8.41(d,J＝8.6Hz,2H),7.89(t,J＝7.7Hz,2H), 7.78(d,J＝8.6Hz,2H),7.43-7.31(m,7H),4.33(t,J＝6.3Hz,2H),3.98(s,2H), 3.31(s,2H),3.10(t,J＝6.2Hz,2H),2.22-2.16(m,2H)；13C NMR(CD₃OD,100 MHz)δ159.2,141.6,138.1,136.1,130.2,129.9,129.1,127.0,124.8,119.7,114.2。

Example 19

N- (9-acridine) -N' -benzyl-1, 4-butanediamine (XIII)₃) The synthesis of (2): referring to example 17, the yield was 59%.¹HNMR(d6-DMSO,400MHz):δ8.37(d,J＝8.0Hz,2H),7.75(d,J＝8.0Hz,2H), 7.64-7.68(m,2H),7.41-7.43(m,2H),7.27-7.28(m,5H),3.89(s,4H),2.72-2.75(m, 2H),1.79-1.80(m,2H),1.66-1.68(m,2H)；13C NMR(DMSO,100MHz)δ154.2, 143.9,143.9,135.3,132.2,129.8,128.8,128.4,126.0,122.4,122.3,115.7,51.2, 49.8,47.1,28.1,24.6；

Example 20

N- (9-acridine) -N' -benzyl-1, 5-pentanediamine (XIII)₄) The synthesis of (2): referring to example 17, the yield was 45%.¹HNMR(d6-DMSO,400MHz)δ8.53(d,J＝8.0Hz,2H),7.84-7.89(m,4H), 7.50-7.51(m,2H),7.43(t,J＝8.0Hz,2H),7.35-7.38(m,3H),4.02(s,2H),3.99(t, J＝7.2Hz,2H)2.79(t,J＝7.2Hz,2H),1.82-1.87(m,2H),1.65-1.68(m,2H),1.37 -1.41(m,2H)。

Example 21

N- (9-acridine) -N' -benzyl-1, 6-hexanediamine (XIII)₅) The synthesis of (2): reference example 17, yield 17%. 1HNMR (CD)₃OD,400MHz)δ8.54(br,2H),7.88-7.51(m,8H),7.45-7.50(m,5H), 4.03-4.05(m,4H),1.88-2.01(m,4H),1.85-1.87(m,4H),1.21-1.26(m,2H)；13C NMR(CD₃OD,100MHz)δ158.2,135.0,132.7,130.7,129.5,128.8,123.5,123.5, 118.2,48.9,31.6,30.4,29.3,29.1,26.0,22.3。

Example 22

N- (9-acridine) -N' -benzyl-1, 7-heptanediamine (XIII)₆) The synthesis of (2): referring to example 17, the yield was 29%. 1HNMR (CD)₃OD,400MHz)δ9.84(s,1H),9.10(s,2H),8.57(br,2H),7.86-7.94 (m,5H),7.49-7.53(m,5H),7.38-7.39(m,3H),4.06(s,4H),2.80-2.81(m,2H), 1.84-1.86(m,2H),1.59-1.60(m,2H),1.26-1.27(m,2H),1.19-1.20(m,4H)；13C NMR(CD₃OD,100MHz)δ157.6,135.1,132.6,130.5,129.1,129.0,123.7,123.6, 123.6,123.6,119.1,50.2,49.1,46.7,29.2,28.5,26.4,26.3,25.5。

Example 23

N- (9-acridine) -N' -benzyl-1, 8-octanediamine (XIII)₇) The synthesis of (2): referring to example 17, the yield was 26%. 1HNMR (CD)₃OD,400MHz)δ9.83(s,2H),8.54(br,2H),7.87-7.95(m,8H), 7.48-7.53(m,5H),4.02-4.04(m,4H),1.26-1.27(m,2H),1.22-1.23(m,2H),1.20 -1.21(m,4H)。

Example 24

N- (9-acridine) -N' -benzyl-1, 9-nonanediamine (XIII)₈) The synthesis of (2): referring to example 17, the yield was 60%. 1HNMR (CD)₃OD,400MHz)δ8.53(d,J＝8.2Hz,2H),7.96(t,J＝7.6Hz,2H), 7.85(d,J＝8.4Hz,2H),7.58-7.52(m,4H),7.45-7.44(m,3H),4.20(s,2H),3.04 -3.00(m,2H),2.01-1.98(m,2H),1.72(m,2H),1.48-1.47(m,2H),1.35-1.28(m, 10H)；13C NMR(CD₃OD,100MHz)δ159.5,136.4,132.7,132.6 131.1,130.9, 130.6,130.3,125.0,119.7,119.6,52.4,50.5,48.7,30.6,30.3,30.1,30.0,27.8,27.6, 27.1。

Example 25

N- (9-acridine) -N' -benzyl-1, 10-sunflower diamine (XIII)₉) The synthesis of (2): referring to example 17, the yield was 19%. 1HNMR (CD)₃OD,400MHz)δ9.26(s,1H),9.23(s,2H),8.59(br,2H),7.93-7.96(m, 4H),7.52-7.54(m,4H),7.38-7.41(m,3H),4.07(s,4H),2.77-2.81(m,2H), 1.84-1.88(m,2H),1.58-1.61(m,2H),1.32-1.35(m,2H),1.17-1.26(m,10H)；13C NMR(CD₃OD,100MHz)δ157.8,135.4,135.3,132.5,130.4,129.3,129.0,118.9, 50.2,49.2,46.8,29.1,29.1,29.0,28.9,28.9,26.5,26.4,25.6。

Example 26

AChE inhibitory activity assay: the method is characterized in that the mouse intracerebral cholinesterase is used as an enzyme source, an Ellman colorimetric method is used for determination, the cholinesterase is hydrolyzed by acetylcholine to generate choline and acetic acid, the choline reacts with a mercapto color developing agent to generate a yellow compound, the quantity of the choline is detected by the colorimetric method, and therefore the quantity of the hydrolysate choline reflects the experimental principle of the activity of the acetylcholinesterase, Kunming mouse forebrain homogenate is adopted, and the inhibitory activity of a medicament on the mouse intracerebral cholinesterase is determined according to the AChE kit (A026) provided by Nanjing Biotechnology Limited.

TABLE 1 results of AChE inhibitory Activity test of substituted acridine compounds with double AChE binding sites

The results of the tests in Table 1 show that most of the tested compounds showed better AChE inhibitory activity, among which compound XIII₈，XIII₈Good inhibitory activity on AChE enzyme, IC₅₀The values were 0.023. mu.M and 0.020. mu.M, respectively. In addition, the experiment also discovers a phenomenon that: when the chain length of the compound is 2 to 5, the compound is poor in activity. This is because the molecules bind to AChE with a strong steric hindrance effect, resulting in a decrease in binding force. And when the chain length is 6, the compound (i.e. compound XIII)₅) A leap in AChE inhibitory activity was 0.063. mu.M, since II-1e produced hydrogen bonding while maintaining the pi-pi interaction of the benzene ring with the W286 residue in AChE protein. Whereas when the chain length is 8, 9 and 10, the activity of the compound is higher, IC₅₀Reaching around 0.023 μ M, which may be related to cationic-pi interaction and pi-pi interaction of the benzylamine fragment in the molecule with PAS.

In conclusion, the substituted acridine compound with double AChE binding sites and the derivatives thereof have the function of inhibiting the activity of AChE enzyme, can be used for preparing medicines for inhibiting the activity of AChE enzyme, and can be particularly used for preparing medicines for treating or preventing the Alzheimer disease with the AChE as a target.

All documents referred to herein are incorporated by reference into this application as if each were individually incorporated by reference. Furthermore, it should be understood that various changes and modifications of the present invention can be made by those skilled in the art after reading the above teachings of the present invention, and these equivalents also fall within the scope of the present invention as defined by the appended claims.

Claims (10)

1. A substituted acridine compound of a double AChE binding site or a pharmaceutically acceptable salt thereof is characterized by having a structure shown as a formula A:

wherein n is an integer of 6 to 12.

2. The substituted acridine compound of the double AChE binding site of claim 1, or a pharmaceutically acceptable salt thereof, selected from the group consisting of:

3. the substituted acridine compound of the double AChE binding site of claim 1, or a pharmaceutically acceptable salt thereof, having a structural formula as shown in formula (II):

4. the substituted acridine compound of the double AChE binding site of claim 1, or a pharmaceutically acceptable salt thereof, having a structural formula as shown in formula (III):

5. a pharmaceutical composition comprising (a) a substituted acridine compound of the double AChE binding site as defined in claim 1, or a pharmaceutically acceptable salt thereof, as an active ingredient and (b) a pharmaceutically acceptable carrier.

6. A process for preparing the pharmaceutical composition of claim 5, wherein the process for preparing the pharmaceutical composition comprises the steps of:

mixing a pharmaceutically effective amount of a compound of formula a or a pharmaceutically acceptable salt thereof with a pharmaceutically acceptable carrier to form a pharmaceutical composition.

7. A method of non-therapeutically inhibiting AChE enzymatic activity in vitro comprising: administering to a subject an inhibitory effective amount of a substituted acridine compound of the double AChE binding site of claim 1 or a pharmaceutically acceptable salt thereof, and/or a pharmaceutical composition of claim 5.

8. A method of preparing substituted acridine compounds of double AChE binding sites as claimed in claim 1, comprising the steps of:

reacting compound VIII and compound XII in the presence of an organic solvent to form compound XIII,

in the formula, n is an integer of 6-12; the organic solvent is selected from the following group: methanol, acetonitrile, acetone, or combinations thereof.

9. Use of substituted acridines of the double AChE binding site of claims 1-4, or pharmaceutically acceptable salts thereof, for use selected from the group consisting of:

(i) for preparing AChE enzyme activity inhibitor;

(ii) for the preparation of a pharmaceutical composition for the treatment or prevention of a neurodegenerative disease; or

(iii) For non-therapeutic inhibition of AChE enzymatic activity in vitro.

10. The neurodegenerative disease of claim 9, comprising alzheimer's disease and parkinson's disease.