Nasal administration preparation of multi-target medicine and application of nasal administration preparation

Abstract

The invention relates to a nasal administration preparation of a multi-target drug and application of the nasal administration preparation. The nasal administration preparation of the multi-target medicine is prepared by dissolving or suspending the compound shown in the formula I or the chiral enantiomer or the pharmaceutically acceptable salt thereof in a solvent. The nasal administration preparation can smoothly reach the brain after nasal administration, and high exposure in the brain is achieved, so that the in-vivo effect of the nasal administration preparation for treating neurodegenerative diseases is improved. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the field of pharmaceutical technology and relates to pharmaceutical preparations, in particular to a nasal administration preparation of a multi-target drug and an application thereof. Background Art

[0002] The compound or its salt of the structure shown in Formula I is a multi-target drug developed by the research team of the present invention in the previous work, which can inhibit acetylcholinesterase and butyrylcholinesterase at the same time, and its biological activity is stronger than that of the currently commercially available cholinesterase inhibitor rivastigmine that simultaneously targets acetylcholinesterase and butyrylcholinesterase for the treatment of neurodegenerative diseases (Wu J et al., Bioorg Med Chem. 2020 Mar 1; 28 (5): 115324.).

[0003]

[0004] Based on the mechanism of action of this multi-target drug, its metabolites in the body can also stimulate β2 adrenergic receptors. β2 adrenergic receptors belong to G protein-coupled receptors and are widely involved in the regulation of physiological functions, including regulating vascular tension, regulating cardiac metabolism, and participating in the formation of memory. Studies have shown that stimulating β2 adrenergic receptors is beneficial for the treatment of neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease. Injection of β2 adrenergic receptor agonist salbutamol has a therapeutic effect on Parkinson's disease. The multi-target drug with the structure shown in Formula I also has the effect of treating neurodegenerative diseases.

[0005] Drugs for treating central nervous system diseases need to reach a certain exposure level in the brain to produce a therapeutic effect. During the research process, the research team of the present invention found that although the multi-target drug with the structure shown in Formula I has activity in treating neurodegenerative diseases in vitro, conventional administration methods, such as intravenous injection, cannot achieve sufficient exposure of the drug in the brain, thereby affecting its in vivo efficacy. Summary of the invention

[0006] Based on this, the purpose of the present invention is to develop an effective administration method (i.e., drug preparation) for the multi-target drug of the structure shown in Formula I, which can enable the drug to reach the brain and achieve a higher exposure amount in the brain, thereby improving its in vivo effect in treating neurodegenerative diseases.

[0007] In order to achieve the above-mentioned object of the invention, the present invention includes the following technical solutions.

[0008] In a first aspect, the present invention provides a nasal administration preparation of a multi-target drug, which is prepared by dissolving or suspending a compound of formula I or a chiral enantiomer thereof or a pharmaceutically acceptable salt thereof in a solvent;

[0009]

[0010] Among them, its pharmaceutically acceptable salts include sulfate, bisulfate, hydrochloride, hydrobromide, dihydrogen phosphate, methanesulfonate, bromide, acetate, oxalate, maleate, fumarate, succinate, 2-naphthyl sulfate, gluconate, citrate, tartrate, lactate, pyruvate, isethionate, benzenesulfonate, and p-toluenesulfonate.

[0011] Preferably, the solvent is a mixed solvent of 1,2-propylene glycol and water.

[0012] Preferably, the solvent is a mixed solvent of 1,2-propylene glycol and water in a volume ratio of 1:0.5-2.

[0013] Preferably, the solvent is a mixed solvent of 1,2-propylene glycol and water in a volume ratio of 1:0.8-1.2.

[0014] Preferably, the solvent is a mixed solvent of 1,2-propylene glycol and water in a volume ratio of 1:1.

[0015] Preferably, the concentration of the compound of formula I or its chiral enantiomer or its pharmaceutically acceptable salt in the solvent is 2 mg / mL-40 mg / mL.

[0016] Preferably, the concentration of the compound of formula I or its chiral enantiomer or its pharmaceutically acceptable salt in the solvent is 10 mg / mL-30 mg / mL.

[0017] Preferably, the concentration of the compound of formula I or its chiral enantiomer or its pharmaceutically acceptable salt in the solvent is 15 mg / mL-25 mg / mL.

[0018] Preferably, the concentration of the compound of formula I or its chiral enantiomer or its pharmaceutically acceptable salt in the solvent is 18 mg / mL-22 mg / mL.

[0019] Preferably, the concentration of the compound of formula I or its chiral enantiomer or its pharmaceutically acceptable salt in the solvent is 20 mg / mL.

[0020] The nasal administration preparation of the multi-target drug may also include other excipients acceptable in nasal administration preparations.

[0021] Preferably, the nasal administration preparation is nasal drops, nasal emulsion, nasal spray and the like.

[0022] In a second aspect, the present invention provides the use of the nasal administration preparation of the multi-target drug in the preparation of drugs for treating central nervous system diseases; the central nervous system diseases can be neurodegenerative diseases (such as Alzheimer's disease and Parkinson's disease, etc.) and mental diseases (such as epilepsy, migraine, hysteria, etc.).

[0023] Specifically, the drug is administered through the nasal cavity during use, such as nasal administration through nasal drops, or nasal administration through nasal sprays.

[0024] The present invention has the following beneficial effects:

[0025] The research team of the present invention found in the process of developing the multi-target drug shown in Formula I that although the drug has good in vitro activity, it is not possible to achieve sufficient exposure of the drug in the brain through conventional administration methods (such as intravenous administration). In the further study process, it was unexpectedly found that dissolving it in a simple solvent (especially a mixed solvent of propylene glycol and water) and administering it by nasal administration can make the drug have brain targeting, can smoothly reach the brain, can effectively increase the exposure of the drug in the brain, and thus can improve the in vivo therapeutic effect of the multi-target drug on neurodegenerative diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 Schematic diagram of brain tissue partitioning (A) and sample placement (B) in Example 3.

[0027] Figure 2 This is the detection result of multi-target drugs in rat brain tissue using desorption electrospray ionization mass spectrometry in Example 3. DETAILED DESCRIPTION

[0028] In order to facilitate the understanding of the present invention, the present invention will be described more fully below. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present invention more thorough and comprehensive.

[0029] The experimental methods in the following examples without specifying specific conditions are usually carried out under conventional conditions or conditions recommended by the manufacturers. The various commonly used chemical reagents used in the examples are all commercially available products.

[0030] Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as those commonly understood by those skilled in the art to which the present invention belongs. The terms used in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used in the present invention includes any and all combinations of one or more of the related listed items.

[0031] In addition, as used in the present invention, the term "or" is an inclusive "or" symbol and is equivalent to the term "and / or", unless the context clearly dictates otherwise. The term "based on" is not exclusive and allows for being based on other factors not described, unless the context clearly dictates otherwise. In addition, throughout the specification, the meanings of "a", "an", and "the" include plural referents. The meaning of "in..." includes "in..." and "on...".

[0032] Drugs for the treatment of central nervous system diseases need to reach a certain exposure in the brain to produce a therapeutic effect. The inventor team of the present invention found in the research process that although the multi-target drug of the structure shown in Formula I has the activity of treating neurodegenerative diseases in vitro, conventional administration methods, such as intravenous administration, cannot make the drug reach sufficient exposure in the brain, which will affect its good therapeutic effect in vivo. It is speculated that it may be due to the presence of the blood-brain barrier (BBB), which prevents the drug from entering the brain tissue. The presence of the blood-brain barrier (BBB) ​​prevents some small molecules and macromolecules from entering the brain tissue, including drugs entering the brain through the blood circulation. Therefore, some drugs are difficult to reach the exposure amount that produces a therapeutic effect in the brain through conventional administration methods, which will affect its in vivo therapeutic effect; or when the therapeutic drug reaches the brain tissue through the BBB, a higher systemic dose is usually required to reach the therapeutic level, which is easy to cause adverse reactions in the body.

[0033] Nasal administration preparations refer to preparations that deliver drugs to the nasal cavity and then absorb them through the nasal mucosa to exert local or systemic therapeutic effects. Some drugs can reach the brain through the blood-brain barrier through multiple pathways, ultimately achieving the goal of delivering drugs to the brain while reducing the exposure of the drug to the whole body. However, affected by the differences in the physical and chemical properties of specific drugs, many studies have found that many drugs cannot be absorbed by the nasal mucosa through nasal administration or other various reasons cause them to be unable to enter the brain tissue better through nasal administration. For example, studies have shown that after nasal administration, estradiol and progesterone are rapidly absorbed by the systemic circulation, non-protein-bound hormones may enter the cerebrospinal fluid through the blood-brain barrier, and no direct transport phenomenon of the nose to the cerebrospinal fluid has been found (Mascha P van den Berg et al. Eur J Pharm Biopharm. 2004 Jul; 58 (1): 131-5.). Other studies have shown that in rats, there is no additional melatonin uptake in the cerebrospinal fluid after nasal administration compared with intravenous administration, which is consistent with the results found in humans (Mascha P van den Berg et al. Pharm Res. 2004 May; 21(5): 799-802.).

[0034] However, during the research and development process, the inventor team of the present invention unexpectedly discovered that by dissolving the compound of formula I of the present invention in a simple solvent and administering it nasally, the drug can be made brain-targeted, so that it can reach the brain smoothly, and the exposure of the multi-target drug in the brain can be effectively increased, thereby improving its in vivo therapeutic effect on neurodegenerative diseases, thereby successfully solving the problem of difficulty in administering the multi-target drug shown in formula I.

[0035] Based on the above unexpected discovery, some embodiments of the present invention relate to a nasal administration preparation of a multi-target drug, which is prepared by dissolving or suspending a compound of formula I or a chiral enantiomer thereof or a pharmaceutically acceptable salt thereof in a solvent;

[0036]

[0037] Among them, its pharmaceutically acceptable salts include sulfate, bisulfate, hydrochloride, hydrobromide, dihydrogen phosphate, methanesulfonate, bromide, acetate, oxalate, maleate, fumarate, succinate, 2-naphthyl sulfate, gluconate, citrate, tartrate, lactate, pyruvate, isethionate, benzenesulfonate, and p-toluenesulfonate.

[0038] The nasal preparation provided by the present invention can enable the multi-target drug represented by Formula I or its chiral enantiomer or its pharmaceutically acceptable salt to reach the brain smoothly after nasal administration, and achieve a higher exposure amount in the brain, thereby improving its in vivo effect of treating neurodegenerative diseases.

[0039] In some preferred embodiments, the solvent is a mixed solvent of 1,2-propylene glycol and water.

[0040] In some of the more preferred embodiments, the solvent is a mixed solvent of 1,2-propylene glycol and water in a volume ratio of 1:0.5-2.

[0041] In some of the more preferred embodiments, the solvent is a mixed solvent of 1,2-propylene glycol and water in a volume ratio of 1:0.8-1.2.

[0042] In some more preferred embodiments, the solvent is a mixed solvent of 1,2-propylene glycol and water in a volume ratio of 1:1.

[0043] In some preferred embodiments, the concentration of the compound of formula I or its chiral enantiomer or a pharmaceutically acceptable salt thereof in the solvent is 2 mg / mL-40 mg / mL.

[0044] In some of the more preferred embodiments, the concentration of the compound of formula I or its chiral enantiomer or its pharmaceutically acceptable salt in the solvent is 10 mg / mL-30 mg / mL.

[0045] In some more preferred embodiments, the concentration of the compound of formula I or its chiral enantiomer or a pharmaceutically acceptable salt thereof in the solvent is 15 mg / mL-25 mg / mL.

[0046] In some more preferred embodiments, the concentration of the compound of formula I or its chiral enantiomer or a pharmaceutically acceptable salt thereof in the solvent is 18 mg / mL-22 mg / mL.

[0047] In some more preferred embodiments, the concentration of the compound of formula I or its chiral enantiomer or a pharmaceutically acceptable salt thereof in the solvent is 20 mg / mL.

[0048] The nasal administration preparation of the multi-target drug of the present invention may also include other excipients acceptable to nasal administration preparations, such as vitamin E, benzyl alcohol, dodecyl maltoside, tetradecyl maltoside, etc.

[0049] In some preferred embodiments, the nasal administration preparation is nasal drops, nasal emulsion, nasal spray, etc.

[0050] Some of the embodiments also involve the use of the nasal administration preparation of the multi-target drug in the preparation of drugs for treating central nervous system diseases; the central nervous system diseases can be neurodegenerative diseases (such as Alzheimer's disease and Parkinson's disease, etc.) and mental diseases (such as epilepsy, migraine, hysteria, etc.).

[0051] In some embodiments, the drug is administered via the nasal cavity, such as by nasal drops or nasal sprays.

[0052] The present invention is further described in detail below with reference to specific embodiments.

[0053] Example 1: Preparation of the compound of formula I (preparation of racemic hydrochloride)

[0054]

[0055] (1) Preparation of intermediate B

[0056] Take 6g of 3-hydroxyacetophenone, 5.7g of potassium carbonate with crystallization water, 1.31g of potassium carbonate with anhydrous content, 0.25g of pyridine, 7.1g of N,N-dimethylaminoformyl chloride and 100mL of ethyl acetate and place them in a reaction bottle. Stir the reaction at 70°C and monitor with TLC. After the reaction is completed, add 60mL of water and continue stirring for 1.5h. Then wash the organic phase with water twice and with dilute sulfuric acid twice, dry the organic phase with anhydrous magnesium sulfate, filter, and concentrate the filtrate in vacuo. The obtained crude product is directly subjected to the next step of reaction.

[0057] (2) Preparation of intermediate C

[0058] 6 g of intermediate B, 12.94 g of copper bromide and 100 mL of a mixed solvent of ethyl acetate and chloroform in a volume ratio of 1:1 were placed in a reaction bottle, reacted at 60°C, and monitored by TLC. After the reaction was completed, the filtrate was filtered, washed with water 5 times, dried with anhydrous magnesium sulfate, filtered, and concentrated. Recrystallized with petroleum ether and ethyl acetate to obtain solid intermediate C.

[0059] (3) Preparation of intermediate D

[0060] Take 4g of intermediate C, 1.59g of sodium borohydride and 80mL of dichloromethane and place them in a reaction bottle. Add 20mL of methanol under an ice bath, then raise the temperature to 40°C and monitor the reaction by TLC. After the reaction is completed, add 60mL of saturated ammonium chloride solution and stir at room temperature for 2h. Extract the aqueous phase twice with dichloromethane. Combine the organic phases, dry with anhydrous magnesium sulfate, filter, and concentrate to obtain an oil. The crude product is directly used in the next step reaction.

[0061] (4) Preparation of intermediate E

[0062] Take 0.5g of intermediate D, 0.32g of tert-butylamine and 20mL of isopropanol and place them in a reaction bottle. React in the dark under nitrogen protection at 80°C and monitor by TLC. After the reaction is completed, concentrate to obtain an oil. Add water and dichloromethane to the oil, and extract the aqueous phase with dichloromethane. Combine the organic phases, dry with anhydrous magnesium sulfate, filter, and concentrate to obtain an oil. The crude product is used directly in the next step.

[0063] (5) Preparation of target compound F

[0064] The intermediate E was dissolved in 10 mL of anhydrous ethanol, and a 10.0 mol / L hydrogen chloride ethanol solution was added dropwise under ice bath conditions to adjust the pH value of the system to 1. After reacting for 4 hours, the reaction solution was concentrated and purified by column chromatography using dichloromethane and methanol as eluents to obtain compound F with a yield of 49%.

[0065] 1 H NMR (400MHz, DMSO) δ = 7.39 (t, J = 7.9Hz, 1H, H arom),7.28(d,J=7.7Hz,1H,H arom ),7.18(d,J=1.7Hz,1H,H arom ),7.03-7.09(m,1H,H arom ),5.00(d,J=8.5Hz,1H,CH),2.86-3.08(m,8H,CH2 and CON(CH3)2),1.31(s,9H,C(CH3)3)ppm. 13 C NMR (101MHz, DMSO)δ=154.43,151.82,143.92,129.62,123.24,121.74,119.87,68.92,56.63,49.04,36.76,36.57,25.57ppm.TOF MS ES(+)(m / z):calculated for C 15 H 25 N2O3([M+H] + )281.1860; found 281.1856.

[0066] Example 2: Preparation of multi-target drug nasal drops and injections

[0067] Preparation of multi-target drug nasal drops: Compound F prepared in Example 1 was dissolved in a mixed solvent of 1,2-propylene glycol: water = 1:1 (volume ratio) to prepare a 20 mg / mL nasal drops solution for nasal administration and stored at 4°C for use.

[0068] Preparation of intravenous injection solution of multi-target drugs: Compound F prepared in Example 1 was dissolved in a mixed solvent of 1,2-propylene glycol: water = 1:1 (volume ratio) to prepare a 2 mg / mL intravenous injection solution for injection and stored at 4°C for use.

[0069] Example 3: Dosing, sample preparation and testing

[0070] 1) Preparation of standard products: After cardiac perfusion of anesthetized rats, the brain was taken out and weighed. Two volumes of physiological saline were added for grinding, and the drug solution prepared in Example 2 was prepared with a mixed solvent (volume ratio) of 1,2-propylene glycol: water = 1:1 to prepare drug solutions with concentrations of 200 μg / mL, 20 μg / mL, 2 μg / mL and 200 ng / mL, respectively. 40 μL of brain homogenate was taken and placed in 5 centrifuge tubes, and then 40 μL of drug solutions of different concentrations and a mixed solvent (volume ratio) of 1,2-propylene glycol: water = 1:1 were added to prepare standard products for use.

[0071] 2) Sample preparation for nasal administration: After anesthetizing rats, nasal administration was performed. The nasal drops prepared in Example 2 were dripped into the nasal cavity of the rats in small amounts and multiple times. The dosage was 4 mg / kg. Brain tissue was obtained 30 minutes after administration and frozen sections were made. The obtained rat brain was divided into four regions (such as Figure 1 Each area was cut into a slice with a thickness of 20 μm.

[0072] 3) Sample preparation for intravenous administration: After anesthesia, rats were administered via the tail vein at a dose of 4 mg / kg. Brain tissue was obtained 30 minutes after administration and frozen sections were made. The obtained rat brain was divided into four regions (eg Figure 1 Each area was cut into a slice with a thickness of 20 μm.

[0073] 4) Sample placement: Place the brain slices of untreated mice and the tissue slices of the same area under different drug administration methods on a glass slide; take 2.5 μL of different concentrations of standard solution and apply it on the slide containing brain tissue slices (apply on Figure 1 B's 100ug / mL, 10ug / mL, 1ug / mL, etc.), dry, and wait for detection. Brain frozen sections and standards are placed as follows Figure 1 As shown in Figure B.

[0074] 5) Detection of multi-target drugs in rat brain tissue using desorption electrospray ionization mass spectrometry

[0075] Liquid-mass spectrometry (LC-MS) is often used for drug detection in animal tissues. Traditional LC-MS methods require complex pretreatment of tissue samples for drug analysis, and cannot obtain spatial distribution information of drug molecules in tissues. Desorption electrospray ionization technology is a technology for ionization at atmospheric pressure. It has low sample requirements, does not require complex pretreatment and pre-separation, and can also obtain spatial distribution information of target ions in tissues. Therefore, in this embodiment, desorption electrospray ionization mass spectrometry is used to detect slice samples, which can directly analyze the spatial distribution of drug molecules in tissues. The instrument is set to positive ion mode, the spray capillary voltage is set to 4.5kV, the sample cone voltage is 60V, the ion source temperature is 100°C, and a mixed solvent of 95% methanol, 5% water, and 1‰ formic acid is selected as the desorption solvent, containing the internal standard leucine enkephalin, with a concentration of 6μg / mL and a flow rate of 2μL / min. The ion scan range is set to 50-1000Da.

[0076] Test results such as Figure 2As shown. For zone 1 of nasal and intravenous administration, the results showed that after nasal administration, the multi-target drug can effectively enter the brain tissue and reach a higher drug concentration. The drug is mainly distributed in the black circle of the brain tissue; while no drug distribution was detected in the brain tissue sections of the intravenous administration group. This result shows that the nasal drops of the present invention can achieve brain targeting of the multi-target drug of the present invention through nasal administration, and can better deliver it to the brain, thereby better exerting the in vivo therapeutic effect.

[0077] Based on the above research results, it can be found that the multi-target drug of formula I can enter the rat's nasal cavity and reach a higher drug concentration (10 μg / mL), and has the potential to become a new method for treating neurodegenerative diseases.

[0078] The above-mentioned embodiments only express several implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. A multi-target drug preparation for nasal administration, characterized in that: Prepared by dissolving or suspending a compound of formula I or its chiral enantiomer or a pharmaceutically acceptable salt thereof in a solvent; 2. The multi-target drug nasal administration preparation according to claim 1, characterized in that: The solvent is a mixed solvent of 1,2-propylene glycol and water.

3. The multi-target drug nasal administration preparation according to claim 2, characterized in that: The solvent is a mixed solvent of 1,2-propylene glycol and water in a volume ratio of 1:0.5-2; Preferably, the solvent is a mixed solvent of 1,2-propylene glycol and water in a volume ratio of 1:0.8-1.2; Preferably, the solvent is a mixed solvent of 1,2-propylene glycol and water in a volume ratio of 1:

1.

4. The multi-target drug preparation for nasal administration according to any one of claims 1 to 3, characterized in that: The concentration of the compound of formula I or its chiral enantiomer or its pharmaceutically acceptable salt in the solvent is 2 mg / mL-40 mg / mL.

5. The multi-target drug nasal administration preparation according to claim 4, characterized in that: The concentration of the compound of formula I or its chiral enantiomer or its pharmaceutically acceptable salt in the solvent is 10 mg / mL-30 mg / mL, preferably 15 mg / mL-25 mg / mL, and more preferably 18 mg / mL-22 mg / mL.

6. The multi-target drug preparation for nasal administration according to any one of claims 1 to 3, characterized in that: The pharmaceutically acceptable salts thereof include sulfate, bisulfate, hydrochloride, hydrobromide, dihydrogen phosphate, methanesulfonate, bromide, acetate, oxalate, maleate, fumarate, succinate, 2-naphthyl sulfate, gluconate, citrate, tartrate, lactate, pyruvate, isethionate, benzenesulfonate, and p-toluenesulfonate.

7. The multi-target drug preparation for nasal administration according to any one of claims 1 to 3, characterized in that: The nasal administration preparation also includes other adjuvants acceptable to nasal administration preparations.

8. The multi-target drug preparation for nasal administration according to any one of claims 1 to 3, characterized in that: The nasal administration preparation is nasal drops, nasal emulsion, and nasal spray.

9. Use of the nasal administration preparation of the multi-target drug according to any one of claims 1 to 8 in the preparation of drugs for treating central nervous system diseases; Preferably, the central nervous system diseases include neurodegenerative diseases and psychiatric diseases; Preferably, the neurodegenerative diseases include Alzheimer's disease and Parkinson's disease; Preferably, the mental illness includes epilepsy, migraine, and hysteria.

10. The use according to claim 9, characterized in that: The drug is administered via the nasal cavity.

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