Deuterated acetylclidine medication and use thereof

Abstract

Disclosed in the present invention is deuterated acetylclidine, as shown in the following formula (I). The present invention relates to deuterated acetylclidine, a pharmaceutical composition thereof, and the use thereof.

Description

Deuterated acetylcholine drug and use thereof TECHNICAL FIELD

[0001] The present application belongs to the field of biological medicine, and particularly relates to deuterated acetylcholine and its medical use. BACKGROUND

[0002] With the growth of age, the human eyeball lens hardens and thickens, and after the eye muscle regulation ability decreases, in people with good naked-eye far vision, the minimum focal length of the eyeball gradually increases, and objects at close range become blurred, which is a physiological phenomenon closely related to aging and is called presbyopia, also known as hyperopia. Presbyopia usually occurs after the age of 45, and the main symptoms include blurred vision at close range, eye fatigue, soreness, tearing, photophobia, dry eye, etc. The current treatment methods for presbyopia are mainly two types, one is to solve the problem of hyperopia by wearing appropriate glasses, and the other is to correct the problem of hyperopia through surgery, mainly laser surgery and corneal implantation. Cholinergic receptor agonists can cause pupil constriction, thereby producing pinhole effect to improve vision. Pilocarpine, as an M choline receptor agonist, was approved for the treatment of glaucoma, dry eye, etc. in 1965, and for the treatment of presbyopia in 2021. Acetylcholine developed by LENZ Therapeutics is a small molecule muscarinic acetylcholine receptor agonist, which has been approved for the treatment of glaucoma, and the presbyopia indication is currently in clinical phase III, which has been introduced into China by Shanghai Jixing Pharmaceutical Co., Ltd. and is carrying out phase I clinical trials.

[0003] Deuterated drugs, which are new drug molecules in which one or more carbon-hydrogen bonds of a drug molecule are replaced with carbon-deuterium bonds, can improve the pharmacokinetic properties of the original drug, thereby overcoming the defects of the original drug such as easy metabolism and large side effects.

[0004] The present application is a deuterated acetylcholine drug, which can further improve the pharmacokinetic properties of acetylcholine, reduce the dosage and possible side effects of drug administration. SUMMARY

[0005] The deuterated acetylcholine or pharmaceutically acceptable salt thereof provided by the present application can further improve the pharmacokinetic properties of deuterated acetylcholine, reduce the dosage and possible side effects of drug administration.

[0006] In order to achieve the above-mentioned purpose, the present application provides a deuterated acetylcholine or a pharmaceutically acceptable salt thereof as shown in the following formula I:

[0007] wherein R1, R2, R3, R4, R5, R6, R7 and R8 are independently selected from H or deuterium, and are not H at the same time.

[0008] Further, the deuterated acetylcholine provided by the present application has the following structure:

[0009] The deuterated acetylclivine or the pharmaceutically acceptable salt thereof according to the present application is selected from hydrochloride, methanesulfonate, maleate or phosphate.

[0010] The deuterated acetylclivine or the pharmaceutically acceptable salt thereof according to the present application includes the application thereof in the treatment of ophthalmic diseases such as presbyopia, glaucoma or dry eye.

[0011] The deuterated acetylclivine or the pharmaceutically acceptable salt thereof according to the present application includes the deuterated acetylclivine or the pharmaceutically acceptable salt thereof as an active ingredient and a pharmaceutically acceptable carrier.

[0012] The pharmaceutical composition of the deuterated acetylclivine or the pharmaceutically acceptable salt thereof according to the present application is selected from a capsule, a powder, a tablet, a granule, a pill, an injection, a syrup, an oral liquid, an inhalant, an ointment, a suppository or a patch.

[0013] Advantages: Compared with the prior art, the present application has the following advantages:

[0014] The present application provides a class of deuterated acetylclivine drugs, which further improve the pharmacokinetic properties of acetylclivine, reduce the dosage and possible side effects of drug administration. DETAILED DESCRIPTION

[0015] The present application will be further described below in conjunction with examples, but the present application is not limited in the scope of the described examples. The experimental methods in the following examples are not specified, which are selected according to the conventional methods and conditions, or according to the instructions of the commodity.

[0016] DETAILED DESCRIPTION

[0017] Example 1, preparation of compound 1

[0018] Synthetic route of example 1 compound (compound 1):

[0019] Synthesis of intermediate 1-B:

[0020] Dissolve intermediate 1-A (1.29 g, 10 mmol) with EtOH (25 mL), and add dichlorosulfoxide (12 mmol) dropwise at 0℃. Stir the reaction at room temperature overnight, then warm to reflux for 4 hours. After TLC detection of the complete reaction, concentrate the solvent and directly proceed to the next step.

[0021] Synthesis of intermediate 1-C:

[0022] Intermediate 1-B (5 mmol) and ethyl chloroacetate (7.5 mmol) were dissolved in 25 mL of acetonitrile, and potassium carbonate (10 mmol) was added portionwise to the reaction solution at 0 °C. The reaction was continued to stir overnight, and after TLC monitoring of the reaction completion, it was filtered, the filtrate was concentrated, and column chromatography (petroleum ether: ethyl acetate = 10: 1) was performed to obtain intermediate 1-C.

[0023] Synthesis of intermediate 1-D:

[0024] Potassium tert-butoxide (10 mmol) was dissolved in 25 mL of toluene, and intermediate 1-C (4 mmol) was dissolved in 10 mL of toluene and added dropwise to the above solution. Under nitrogen atmosphere, it was warmed to 80 °C, and the reaction was stirred for 3 hours, and after TLC detection of the reaction completion, the toluene solvent was concentrated, 10 N hydrochloric acid solution (25 mL) was added, and the reaction was refluxed for 10 hours. After cooling to 0 °C, potassium carbonate was added to adjust the pH to 10, dichloromethane was added to extract, the organic phase was collected and dried over Na2SO4, and concentration was performed to obtain 1-D.

[0025] Synthesis of intermediate 1-E:

[0026] Intermediate 1-D (5 mmol) was dissolved in oxygen-free water (10 mL), and NaBH4 was added portionwise to the above solution at 0 °C. The reaction was continued to stir for half an hour at room temperature, sodium bicarbonate solution was added, dichloromethane was further added to extract, and column chromatography was performed to obtain intermediate 1-E.

[0027] Synthesis of example 1:

[0028] Intermediate 1-E (2 mmol) was added to deuterated acetic anhydride (10 mL), and 0.5 mL of concentrated sulfuric acid was slowly added. It was warmed to 75 °C, and the reaction was stirred for half an hour. After cooling to room temperature, 50 mL of water was added, and after stirring for 5 min, it was suction filtered, the filter cake was dissolved in dichloromethane, and column chromatography was performed to obtain the compound of example 1. 1 H NMR (400 MHz, Cholroform-d) δ 4.85 (d, J = 7.8 Hz, 1H), 3.21-3.18 (m, 1H), 2.91-2.88 (m, 3H), 2.63 (d, J = 8.3 Hz, 2H), 2.03-1.97 (m, 1H), 1.92-1.89 (m, 1H), 1.68-1.65 (m, 1H), 1.60-1.52 (m, 2H).

[0029] Preparation of example 2, compound 2

[0030] Synthetic route of example 2 compound (compound 2):

[0031] Synthesis of intermediate 2-B:

[0032] D2-A (5 mmol) and potassium carbonate (10 mmol) were dissolved in deuterium water (15 mL), palladium carbon (1 mmol) was added, and the mixture was heated at 80 °C under deuterium gas (200 psi) for 24 h. TLC indicated that the reaction was complete. The mixture was filtered and the filtrate was concentrated to give intermediate 2-B.

[0033] Intermediate 2-C, 2-D can be prepared according to the synthetic method of 1-B, 1-C.

[0034] Synthesis of intermediate 2-E:

[0035] Intermediate 2-D (1 mmol) was dissolved in 0.1 M sodium methoxide (1 mmol) in methanol, and the mixture was heated at reflux for 30 h. TLC indicated that the reaction was complete. The solvent was concentrated, and water and dichloromethane were added. The organic phase was collected and purified by column chromatography to give intermediate 2-E.

[0036] Example 2 can be prepared according to the synthetic method of Example 1, by replacing 1-D with 2-E, replacing sodium borohydride with sodium borodeuteride, and replacing deuterated acetic anhydride with acetic anhydride. 1 H NMR (400 MHz, Cholroform-d) δ 2.07 (s, 3H).

[0037] Preparation of Example 3, compound 3

[0038] Synthetic route of Example 3 compound (compound 3):

[0039] Example 3 compound can be prepared according to the synthetic route of Reference Example 2, by replacing 2-B with 1-A. 1 H NMR (400 MHz, Cholroform-d) δ 3.19 (dd, J = 14.9, 7.9 Hz, 1H), 2.93-2.86 (m, 3H), 2.08 (s, 3H), 2.01 (q, J = 2.9 Hz, 1H), 1.94-1.91 (m, 1H), 1.65-1.63 (m, 1H), 1.61-1.55 (m, 2H).

[0040] Preparation of Example 4, compound 4

[0041] Synthetic route of Example 4 compound (compound 4):

[0042] Example 4 compound can be prepared according to the synthetic method of Example 1, by replacing 1-A with 2-B, and replacing deuterated acetic anhydride with acetic anhydride. 1H NMR (400 MHz, Cholroform-d) δ 4.86 (d, J = 7.7 Hz, 1H) 2.62 (dt, J = 8.1, 3.4 Hz, 2H), 2.07 (s, 3H).

[0043] Example 5, Preparation of Compound 5

[0044] Synthetic route of Example 5 compound (Compound 5):

[0045] The Example 5 compound can be prepared by replacing acetic anhydride with deuterated acetic anhydride according to the synthetic method of Example 2. (ESI) m / z: [M+H] + : 185.2.

[0046] Example 6, Preparation of Compound 6

[0047] Synthetic route of Example 6 compound (Compound 6):

[0048] The Example 6 compound can be prepared by replacing acetic anhydride with deuterated acetic anhydride according to the synthetic method of Example 4. 1 H NMR (400 MHz, Cholroform-d) δ 4.83 (dt, J = 7.7, 3.4 Hz, 1H) 2.62 (d, J = 7.7 Hz, 2H).

[0049] Example 7, Preparation of Compound 7

[0050] Synthetic route of Example 7 compound (Compound 7):

[0051] The Example 7 compound can be prepared by replacing acetic anhydride with deuterated acetic anhydride according to the synthetic method of Example 3. 1 H NMR (400 MHz, Cholroform-d) δ 3.10 - 3.06 (m, 1H), 2.93 - 2.85 (m, 3H), 2.05 (q, J = 3.3 Hz, 1H), 1.95 - 1.92 (m, 1H), 1.66 - 1.62 (m, 1H), 1.59 - 1.53 (m, 2H).

[0052] Test Example 1: Phospholipid Turnover Assay Test

[0053] A9L cells expressing M1 and M3 type cholinergic receptors were incubated with [3H]-inositol (0.5 μCi / well) in 24-well plates at a cell density of 10 5Cells were incubated for 48 hours at 37°C. After incubation, cells were washed twice with 10 mM LiCl in EMEM. Cells were incubated for another 20 minutes at 37°C. The agonist was added at the determined concentration and incubation was continued for half an hour. After the incubation period, the reaction was stopped by adding 0.5 mL of 5% (v / v) trichloroacetic acid (TCA). The wells were rinsed with 0.5 mL of distilled water and then the TCA extract was added. The TCA extract was applied to a Dowex-formate column (Biorad AG1-X8 resin, formate form, 100-200 mesh). The column was washed three times with 3 mL of 5 mM myo-inositol and then the total [3H]myo-inositol phosphate was eluted with 1 mL of 1.0 M ammonium formate / 0.1 M formic acid. Then, 0.5 mL of the eluate was counted in 10 mL of CytoScint on a TM Analytic beta counter.

[0054] Table 1. A9L cell assay of compounds for agonist activity at M1 and M3 cholinergic receptors

[0055] From the data in Table 1, it can be seen that the deuterated example compounds are significantly superior to the non-deuterated positive control compound, Aceclidine, in agonist activity at cholinergic receptors.

[0056] Test Example 2: Preparation of an ophthalmic preparation

[0057] An appropriate amount of sterilized water for injection was taken, hydroxypropyl methylcellulose (HPMC) was added, dispersed, a small amount of hydrochloric acid was added, boiled and dissolved, the dissolved material was cooled, the example compound was added, and menthol, sodium dihydrogen phosphate monohydrate, sodium hydrogen phosphate, sodium chloride, disodium edetate, benzalkonium chloride were added, stirred and dissolved, the pH was adjusted to 6.0 with hydrochloric acid, and the volume was made up to 100 mL with sterilized water for injection. The relevant ophthalmic preparations were prepared according to the following table.

[0058] Table 2. Preparation of ophthalmic preparation

[0059] Test Example 3: Comparison of irritation of ophthalmic preparation administration

[0060] Test animals: healthy New Zealand rabbits (purchased from the Experimental Animal Center of Yangzhou University), both male and female, 10 in total.

[0061] Test method: One day before the test, the test animals were examined for eye irritation, corneal defects and conjunctival damage symptoms, and test animals with relevant symptoms were excluded. Two drops of the preparation were instilled into the left eye of the test animal, and then the eyelids were gently closed for 10 seconds. Two drops of sterile water for injection were instilled into the right eye, and then the eyelids were gently closed for 10 seconds. Three times a day, for 28 days.

[0062] The example was scored for eye irritation according to the following scoring table

[0063] Table 3 Irritation score of formulation examples on rabbit eyes

[0064] The test results show that the formulation examples in Table 2 have low irritation to rabbit eyes after administration, and the formulation examples have low irritation and can increase patient compliance, which is better than the positive control compound acetycholine (Aceclidine) which is not deuterated.

[0065] Finally, it should be noted that the above detailed description of the specific embodiments of the present application is only exemplary, and the present application is not limited to the above described specific embodiments. Any equivalent modifications and alternatives to the present application made by those skilled in the art are also within the scope of the present application. Therefore, any equivalent transformations and modifications made without departing from the spirit and scope of the present application should be included in the scope of the present application.

Claims

1. A deuterated acetylchlidine represented by Formula I, or a pharmaceutically acceptable salt thereof, wherein R1, R2, R3, R4, R5, R6, R7, R8are independently selected from H or deuterium, and are not H at the same time.

2. The deuterated acetylcridin of claim 1, wherein selected from the group consisting of:

3. The deuterated acetylcridin or pharmaceutically acceptable salt thereof according to claim 1 or 2, characterized in that The pharmaceutically acceptable salt is selected from a hydrochloride salt, a methanesulfonate salt, a maleate salt, or a phosphate salt.

4. Use of the deuterated acetylclivine or a pharmaceutically acceptable salt thereof according to claim 1 or 2 for the manufacture of a medicament for treating an ophthalmic disease.

5. Use according to claim 4, characterized in that, The ophthalmic disease is selected from presbyopia, glaucoma, or dry eye.

6. The pharmaceutical composition of deuterated acetylclivperine or a pharmaceutically acceptable salt thereof according to claim 1 or 2, characterized in that, The pharmaceutical composition consists of the deuterated acetylclivine or a pharmaceutically acceptable salt thereof as an active ingredient and a pharmaceutically acceptable carrier.

7. The pharmaceutical composition of deuterated acetylclivperine according to claim 6, characterized in that, The pharmaceutical composition is selected from a capsule, a powder, a tablet, a granule, a pill, an injection, a syrup, an oral solution, an inhalant, an ointment, a suppository, or a patch.

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