CN101311171B - Compound with dual restraining activities to acetyl cholinesterase and butyryl cholinesterase and use thereof - Google Patents

Abstract

The present invention uses virtual drug screening technology and high-throughput screening technology to evaluate the AChE inhibitory activity of more than 10,000 samples in the compound sample library by establishing a virtual acetylcholinesterase inhibitor screening method and an AChE activity detection method, and found that A batch of compounds with high inhibitory activity. According to the pathological characteristics of senile dementia, the inhibitory activity of butyrylcholinesterase was detected for the compounds with higher AChE inhibitory activity. The research results show that the bimolecular 3-piperidinyl-propiophenone not only has strong AChE inhibitory activity, but also has a strong BuChE inhibitory activity; animal experiments prove that the bimolecular 3-piperidinyl-propiophenone can significantly improve Dementia caused by scopolamine and sodium nitrite can enhance learning and memory functions.

Description

Compounds with dual inhibitory activity on B- and butyrylcholinesterase and uses thereof

technical field

The invention relates to a class of bimolecular 3-piperidinyl-propiophenone compounds, which not only have strong AChE inhibitory activity, but also have strong BuChE inhibitory activity; The application in dementia belongs to the technical field of medicine.

Background technique

Alzheimer's is closely related to aging. As the world population ages, so does the incidence of dementia. According to statistics, the annual incidence rate of the elderly aged 65-69 is 1.4‰, that of 70-74 years old is 3.9‰, that of 75-79 years old is 16.7‰, and that it increases to 45.4‰ at the age of 85. Alzheimer's is a disease that seriously affects the quality of life and health of the elderly after heart disease, tumors, and diabetes. This disease can damage memory and decrease the ability of daily life. Its gradually declining functions include: (1) memory and cognitive function-showing memory decline, often forgetting the names of relatives and friends including children; unable to distinguish time, Seasons, getting lost in familiar places and so on. (2) Behavioral psychiatric symptoms - including hallucinations, delusions of persecution, irritability and aggressive tendencies, etc. (3) Ability of daily living——As the disease progresses, the patient gradually loses the ability of daily living, unable to participate in public activities, eating and dressing abnormally, and even unable to take care of himself. Therefore, the research on anti-senile dementia drugs not only has huge social benefits, but also can obviously reduce the economic burden of the country and the family, so it is very necessary.

Alzheimer's disease can be divided into Alzheimer's disease (AD), vascular dementia (vascular dementia, VD) and mixed dementia. AD is more common and better studied. Alzheimer's disease is a complex disease whose typical pathological changes include extensive neuronal loss in the cortex and hippocampus, cholinergic dysfunction in the basal forebrain, numerous neurofibrillary tangles (NFTs) and β-amyloid in the brain (Aβ) deposition forms senile plaques (SP). According to the pathological features and pathogenesis of senile dementia, the treatment approaches for senile dementia patients include: (1) Drugs that improve the function of the choline system. (2) Drugs that correct calcium homeostasis disorders and antioxidation. (3) Neurotrophic factors. (4) Drugs that interfere with the formation and deposition of Aβ. (5) Anti-neurocyte apoptosis agent.

So far, the cholinergic system has been considered the most important neurotransmitter system in normal cognitive function, whether through the action of acetylcholinesterase (AChE) inhibitors or through the action of M, N receptor agonists Both can improve cognitive function. AD is a disease mainly involving memory impairment, and its cholinergic impairment conclusively demonstrates the key role of the cholinergic system in memory function.

In the past ten years, acetylcholinesterase inhibitors have become the first-line treatment drugs for AD, confirming their effective role in alleviating cognitive dysfunction. The product sales account for the largest share of anti-dementia drugs. The main drugs are Tacrine ), donepezil hydrochloride, rivastigmine, galantamine and huperzine A have become representative drugs in this field, thus promoting the development of AD treatment drugs development process. Tacrine can effectively improve the symptoms of AD patients clinically, but due to its greater damage to the liver, its use has been reduced. Several other AChE inhibitors have low toxicity and can effectively improve the cognitive dysfunction of patients, but their clinical efficacy is relatively short. Therefore, finding new and effective anti-AD drugs is still an arduous task for drug researchers.

Contents of the invention

The object of the present invention is to provide a kind of bimolecular 3-piperidinyl-propiophenone compound.

Another object of the present invention is to provide a pharmaceutical composition comprising a bimolecular 3-piperidinyl-propiophenone compound and a pharmaceutically acceptable carrier.

Another object of the present invention is to provide the application of the bimolecular 3-piperidinyl-propiophenone compound in the preparation of drugs for treating learning and memory disorders and anti-senile dementia drugs.

Shown in compound general formula (I) of the present invention:

in,

R1, R2, R3 and R4 can be the same or different substituents, independently selected from C _1-10 alkyl groups;

R1 and R2, R3 and R4 may also form a multi-component heterocyclic ring.

Preferably R1, R2, R3 and R4 are independently selected from CH ₃ —, C ₂ H ₅ —, C ₃ H ₇ —, C ₄ H ₉ —.

Or the substituents R1, R2 and the nitrogen atom may form a five-membered ring or a six-membered ring.

More preferred substituents R1, R2 and the five-membered ring or six-membered ring formed by the nitrogen atom are selected from

Or the substituents R3, R4 and the nitrogen atom may form a five-membered ring or a six-membered ring.

More preferred substituents R3, R4 and the five-membered ring or six-membered ring formed by the nitrogen atom are selected from

Most preferred compounds include but are not limited to

         

         

Bimolecular 3-piperidinyl propiophenone (bimolecular Bimolecular 3-morpholino propiophenone (bimolecular

3-piperidino propiophenone, compound 1) 3-morpholin propiophenone, compound 2)

              

              

Bimolecular 3-Diethylaminopropiophenone (bimolecular Bimolecular 3-Dimethylaminopropiophenone (bimolecular

3-diethylamino propiophenone, compound 3) 3-dimethylamino propiophenone, compound 4)

The inventors found that the compound of the present invention has higher AchE inhibitory activity, further experimental results show that the compound of the present invention has dual inhibitory activity of acetylcholinesterase and butyrylcholinesterase. The compounds of the present invention improve the learning and memory of dementia animal models in vivo.

The present invention therefore also relates to pharmaceutical compositions comprising a compound according to the invention as active ingredient. The pharmaceutical composition can be prepared according to methods known in the art. Any dosage form suitable for human or animal use can be prepared by combining the compounds of the present invention with one or more pharmaceutically acceptable solid or liquid excipients and/or adjuvants. The content of the compound of the present invention in its pharmaceutical composition is usually 0.1-95% by weight.

The compound of the present invention or the pharmaceutical composition containing it can be administered in the form of a unit dosage, and the route of administration can be enteral or parenteral, such as oral, intravenous injection, intramuscular injection, subcutaneous injection, nasal cavity, oral mucosa, eye, lung and Respiratory tract, skin, vagina, rectum, etc.

The dosage form for administration may be a liquid dosage form, a solid dosage form or a semi-solid dosage form. Liquid dosage forms can be solutions (including true solutions and colloid solutions), emulsions (including o/w type, w/o type and double emulsion), suspensions, injections (including aqueous injections, powder injections and infusion solutions), eye drops Agents, nasal drops, lotions and liniments, etc.; solid dosage forms can be tablets (including ordinary tablets, enteric-coated tablets, buccal tablets, dispersible tablets, chewable tablets, effervescent tablets, orally disintegrating tablets), capsules ( Including hard capsules, soft capsules, enteric-coated capsules), granules, powders, pellets, dripping pills, suppositories, films, patches, gas (powder) aerosols, sprays, etc.; semi-solid dosage forms can be ointments, Gels, pastes, etc.

The compound of the present invention can be made into common preparations, sustained-release preparations, controlled-release preparations, targeted preparations and various microparticle drug delivery systems.

To form the compound of the present invention into tablets, various excipients known in the art can be widely used, including diluents, binders, wetting agents, disintegrants, lubricants, glidants. Diluents can be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.; wetting agents can be water, ethanol, iso Propanol, etc.; binders can be starch slurry, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, arabic mucilage, gelatin slurry, sodium carboxymethylcellulose, methylcellulose, hypromellose Base cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol, etc.; disintegrants can be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, cross-linked poly Vinylpyrrolidone, croscarmellose sodium, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitan fatty acid ester, sodium dodecylsulfonate, etc.; lubricant and flow aid The agent can be talc, silicon dioxide, stearate, tartaric acid, liquid paraffin, polyethylene glycol and the like.

Tablets can also be further made into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double-layer tablets and multi-layer tablets.

In order to make the administration unit into a capsule, the active ingredient compound of the present invention can be mixed with a diluent and a glidant, and the mixture can be directly placed in a hard capsule or a soft capsule. The active ingredient compound of the present invention can also be made into granules or pellets with diluents, binders, and disintegrants, and then placed in hard capsules or soft capsules. Various diluents, binders, wetting agents, disintegrants, and glidants used in the preparation of tablets of the compound of the present invention can also be used in the preparation of capsules of the compound of the present invention.

In order to make the compound of the present invention into injection, water, ethanol, isopropanol, propylene glycol or their mixtures can be used as solvent and an appropriate amount of commonly used solubilizers, cosolvents, pH regulators and osmotic pressure regulators in this field can be added. The solubilizer or co-solvent can be poloxamer, lecithin, hydroxypropyl-β-cyclodextrin, etc.; the pH regulator can be phosphate, acetate, hydrochloric acid, sodium hydroxide, etc.; the osmotic pressure regulator can be Sodium chloride, mannitol, glucose, phosphate, acetate, etc. For preparation of freeze-dried powder injection, mannitol, glucose, etc. can also be added as proppants.

In addition, coloring agents, preservatives, fragrances, flavoring agents or other additives can also be added to the pharmaceutical preparations, if necessary.

In order to achieve the purpose of medication and enhance the therapeutic effect, the medicine or pharmaceutical composition of the present invention can be administered by any known administration method.

The dosage of the pharmaceutical composition of the compound of the present invention can vary widely depending on the nature and severity of the disease to be prevented or treated, individual conditions of the patient or animal, administration route and dosage form, etc. Generally speaking, the suitable daily dosage range of the compound of the present invention is 0.001-150 mg/Kg body weight, preferably 0.1-100 mg/Kg body weight, more preferably 1-60 mg/Kg body weight, most preferably 2-30 mg/Kg body weight. The above-mentioned dosage can be administered in one dosage unit or divided into several dosage units, depending on the clinical experience of the doctor and the dosage regimen including the use of other therapeutic means.

The compound or composition of the present invention can be taken alone, or used in combination with other therapeutic drugs or symptomatic drugs. When the compound of the present invention has a synergistic effect with other therapeutic drugs, its dose should be adjusted according to the actual situation.

Description of drawings

The structure of Fig. 1 compound of the present invention

The dose-effect curve that Fig. 2 compound of the present invention inhibits AChE

The dose-effect curve of Fig. 3 compound inhibiting BChE

Fig. 4 Compound (1) influences on platform jumping test of scopolamine model mice

Fig. 5 Compound (1) influences on platform jumping test of scopolamine model mice

Fig. 6 Compound (1) influences on water maze test of scopolamine model mice

Figure 7 Effect of compound (1) on sodium nitrite model mice avoiding the dark test

Figure 8 compound (1) on the platform jumping test of sodium nitrite model mice

Figure 9 Effect of compound (1) on sodium nitrite model mice avoiding the dark test

Detailed ways

Example 1: Virtual screening of AchE inhibitors. Based on the crystal structures of the two complexes of donepezil and huperzine A with AChE, the virtual screening program FlexX and the consistent scoring method were used to add the results of the two virtual screenings, and a total of 2855 compounds were selected with higher scores. as the test compound. The prediction results were analyzed comprehensively, and some compounds were selected for activity detection.

Example 2: Inhibitory activity on AchE.

According to the literature, the AchE inhibitor screening model was established, and the activity of the selected 2885 compounds was tested. It was found that compound 1, compound 2, compound 3, and compound 4 had high AChE inhibitory activity. The activity values are shown in Table 1. Its IC ₅₀ ranges from 1 μM to 10 μM.

AChE Inhibitory Activity of Compounds in Table 1 in Vitro

Note: The above data is the average value of 3 experimental data.

Example 3: Inhibitory activity against butyrylcholinesterase

Establish a butyrylcholinesterase (Butyrocholinesterase, BChE) inhibitor screening model according to the literature, and use this model to detect the activity of the compound,

The experimental results show that compounds (1), (2), (3) and (4) have strong BChE inhibitory activity, and the activity values are shown in Table 2. Its IC ₅₀ ranges from 0.1 μM to 1 μM. The positive control drug huperzine A has no inhibitory activity on BChE, and the positive drug donepezil (donepezil) has lower inhibitory activity on BChE than the active compounds (1), (2), (3) and (4).

BChE Inhibitory Activity of Compounds in Table 2

Note: The above data are the average value of 3 experimental data.

Embodiment 4: The influence of compound on learning and memory impairment of mice caused by scopolamine

Scopolamine was used to construct mouse learning and memory impairment models, and huperzine A and donepezil were used as positive control drugs to observe the effect of compound (1). The learning and memory function of the mice was compared with the normal control group, the model group and the drug group with behavioral indicators such as autonomous activities, hole boards, diving platforms, and water mazes. The dosage for the pharmacodynamic evaluation of the compound is determined according to the conventional dosage calculation method of the drug combined with the results of the acute toxicity test. In the pharmacodynamic evaluation of compound (1), medium dose and low dose were selected: the medium dose group was 5 mg/kg, and the low dose group was 1 mg/kg.

90 mice were used in the experiment for pharmacodynamic evaluation of animal models of dementia. The mice were randomly divided into 6 groups, normal group, model group, huperzine A group, donepezil group, compound (1) medium dose group, compound (1) Low-dose group, 15 mice in each group.

Effects of compound (1) on locomotor activity in mice. After intragastric administration of the experimental mice, compared with the normal group, there was no significant change in the number of autonomous activities of the mice in each dose group.

The effect of compound (1) on hole plate test in mice. After intragastric administration of the experimental mice, compared with the normal group, the number of times the mice in each dose group did not change significantly.

Effect of compound (1) on platform jumping test of scopolamine model mice. After the mice were injected with scopolamine, compared with the blank group, the number of platform avoidance reaction errors in the model group increased significantly (P<0.05). After the mice in the scopolamine model group were shocked by electric shock, the time to find the platform was significantly prolonged. After 24 hours, the time on the platform (latency) was very short, and the latency was significantly different from that in the blank group (P<0.05). The medium and small doses of compound (1) can significantly prolong the incubation period, compared with the model group, the difference is significant (P<0.05). (see Figure 4 for the number of errors, and Figure 5 for the latency period)

The effect of compound (1) on the water maze test of scopolamine model mice. After the mice were injected with scopolamine, compared with the blank group, the time for the mice to reach the end point in the model group was significantly increased (P<0.05), and a small dose of compound (1) could resist the learning and memory impairment caused by scopolamine. Compared with the model group, the difference Significantly (P<0.05) (see Figure 6).

Embodiment 5: Compound is to the effect of sodium nitrate-induced mouse learning, memory impairment model

Sodium nitrite was used to build mouse learning and memory impairment models, and huperzine A and donepezil were used as positive control drugs to observe the effect of compound (1). 90 mice were used in the experiment for pharmacodynamic evaluation of animal models of dementia. The mice were randomly divided into 6 groups, normal group, model group, huperzine A group, donepezil group, compound (1) medium dose group, compound (1) Low-dose group, 15 mice in each group. The learning and memory function of the mice was compared with the normal control group, the model group and the drug group with behavioral indicators such as autonomous activities, hole boards, platform jumping, and dark avoidance.

Effects of compound (1) on locomotor activity in mice. After intragastric administration to the experimental mice, compared with the normal group, the number of autonomous activities of the mice in the middle dose group of compound (1) was significantly increased (P<0.05), and the number of autonomous activities of the mice in the other dose groups had no significant change.

The effect of compound (1) on hole plate test in mice. After intragastric administration of the experimental mice, compared with the normal group, the number of burrowing in the huperzine A group was significantly reduced (P<0.05), and the number of burrowing in the other dose groups had no significant change.

Effect of compound (1) on platform jumping test of sodium nitrite model mice. After the mice were injected with sodium nitrite, compared with the blank group, the number of platform-avoidance reaction errors in the model group increased significantly (P<0.05), and the medium dose of compound (1) could significantly reduce the number of errors (see Figure 8). It can resist learning and memory impairment caused by sodium nitrite, and compared with the model group, the difference is significant (P<0.05). After the mice in the sodium nitrite model group were shocked by electric shock, the time to find the platform was significantly prolonged. After 24 hours, the time on the platform was very short. Compared with the blank group, the latency period was significantly different (P<0.05). The medium dose of compound (1) can significantly prolong the incubation period, compared with the model group, the difference is significant (P<0.05).

The effect of compound (1) on the dark avoidance test of sodium nitrite model mice. After the mice were injected with sodium nitrite, compared with the blank group in the model group, the number of avoidance reaction errors in the mice increased, and both medium and small doses of compound (1) could resist the learning and memory impairment caused by sodium nitrite. Compared with the model group, Significant difference. Compared with the blank group, the incubation period was significantly different. Both medium and small doses of compound (1) can significantly prolong the incubation period (see Figure 7) and reduce the number of errors (see Figure 9), compared with the model group, the difference is significant.

Claims (8)

1. A compound of the formula (I):

wherein,

r1, R2, R3 and R4 may be the same or different and are independently selected from C_1-10Alkyl groups of (a);

the substituents R1 and R2 and the nitrogen atom, and the substituents R3 and R4 and the nitrogen atom form a six-membered ring;

but does not include the following compounds

2. The compound of claim 1, wherein R1, R2, R3 and R4 are independently selected from CH₃-，C₂H₅-、C₃H₇-、C₄H₉-; but does not include the following compounds

3. A compound according to claim 1, wherein the six-membered ring formed by the substituents R1, R2 and the nitrogen atom is selected from

4. A compound according to claim 1, wherein the six-membered ring formed by the substituents R3, R4 and the nitrogen atom is selected from

5. The compound according to claim 1, characterized in that said compound is selected from

6. A pharmaceutical composition comprising at least one compound of any one of claims 1-5 and a pharmaceutically acceptable carrier.

7. Use of a compound according to any one of claims 1 to 5 for the manufacture of a medicament for the treatment of learning and memory disorders.

8. Use of a compound according to any one of claims 1 to 5 for the manufacture of a medicament for the treatment of senile dementia.

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