Use of n-acyl dopamine family lipids in the prevention and treatment of drug-induced parkinsonian motor complications

Abstract

The application provides application of N-acyl dopamine family lipids in prevention and treatment of drug-induced Parkinson's disease motor complications, and belongs to the field of biological medicine. The application first proposes that the NADs compound can be used as a levodopa companion drug for preventing or treating levodopa-induced dyskinesia. The application takes 6-hydroxydopamine intracerebral injection and intraperitoneal injection of levodopa of an LID animal model as a research object, and studies the effect of NADs on improving LID. The results show that the NADs of the application can significantly improve the drug-induced PD motor complications LID, and it is proved that the NADs compound can be used as a companion drug to assist levodopa in treating PD.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of biological medicine, and particularly relates to application of N-acyl dopamine family lipids in prevention or treatment of drug-induced Parkinson's disease motor complications. BACKGROUND

[0002] Parkinson's disease (PD) is also known as paralysis agitans, which is a chronic neurodegenerative disease characterized by loss of dopaminergic neurons in the substantia nigra compacta of the midbrain and appearance of Lewy bodies in neurons. Epidemiological data show that the prevalence rate of the population over 60 years old is 2%, which is a heavy burden that the society with an increasingly aging population urgently needs to solve. For half a century, dopamine replacement therapy mainly with levodopa has been regarded as the gold standard for the treatment of Parkinson's disease. Levodopa can cross the blood-brain barrier and be metabolized to dopamine in the body, thereby supplementing this neurotransmitter. However, this treatment scheme is not a cure, and can only temporarily alleviate the behavioral symptoms of patients, and cannot stop the continuous loss and death of neurons. After 3-5 years of treatment, most patients will develop a series of motor complications induced by levodopa, such as motor fluctuations. These complications seriously affect the quality of life of patients and are one of the main causes of disability in patients with Parkinson's disease. Motor complications caused by levodopa mainly include motor fluctuations and dyskinesia (LID). According to the clinical manifestations, motor fluctuations can be divided into four types: end-of-dose phenomenon, on-period delay, on-off phenomenon and freezing gait, while dyskinesia includes three types: peak-dose dyskinesia, off-period dystonia and biphasic dyskinesia. Because levodopa has a short half-life and is rapidly absorbed and metabolized, fluctuations in its blood concentration can affect signal pathways such as dopamine D1 / D2 receptors, glutamate receptors and serotonin receptors, triggering a series of complex cascades involving neurons and non-neuronal cells, so its pathological mechanism is still not completely clear.

[0003] In terms of treatment, in addition to surgical methods such as deep brain stimulation and unilateral pallidotomy, the current available drug options are very limited, mainly including amantadine, monoamine oxidase B inhibitors, and catechol-O-methyltransferase inhibitors. However, the efficacy of these drugs is often difficult to maintain, and can cause new side effects. To date, there is still a lack of effective treatment for drug-induced motor complications, leading to a dilemma for patients in the use of anti-Parkinson's disease drugs.

[0004] N-acyldopamine lipids (NADs) are a family of endogenous cannabinoid compounds. The known family members include N-arachidonyl dopamine (NADA), N-oleoyl dopamine (OLDA), N-docosahexaenoyl dopamine (NDHA), N-palmitoyl dopamine (PALDA), N-stearoyl dopamine (STERDA) and N-octanoyl dopamine (NOD). As members of the cannabinoid family, they can activate transient receptor potential cation channel (TRPV1) and play an important role in nociception, or activate cannabinoid receptors CB1 and CB2 to support nerve activity. At present, the structural characteristics and physiological activity of NADs have not been well characterized. It is of great significance to develop a complete family of NADs and characterize their physiological activity. SUMMARY

[0005] To solve the problem of drug deficiency in the prior art in preventing or treating drug-induced Parkinson's disease motor complications, the present application proposes an application of NADs compounds in the field of preventing or treating drug-induced Parkinson's disease motor complications. The present application particularly points out that NADs compounds can be used as an auxiliary drug of levodopa for preventing or treating drug-induced Parkinson's disease motor complications.

[0006] Specifically, the present application is realized by the following technical solutions:

[0007] In the first aspect of the present application, a NADs compound (as shown in formula I) or a pharmaceutically acceptable acid addition salt thereof is provided:

[0008]

[0009] wherein R1 is selected from C 6～26 linear saturated hydrocarbon, C 6～26 branched saturated hydrocarbon, C 6～26 linear unsaturated hydrocarbon, C 6～26 branched unsaturated hydrocarbon, C 3～24 cyclic hydrocarbon, C 6～24 aromatic hydrocarbon; R2 is mono-substituted or poly-substituted, and the substituent is selected from hydrogen, halogen, nitro, hydroxyl, C 1～4 alkyl, C 1～4 halogenated alkyl, C 1～4 alkoxy, C 1～4 halogenated alkoxy; n is 1, 2 or 3, and the basic unit contains at least one carbon-carbon single bond or one carbon-carbon double bond; X is O or S.

[0010] Preferably, R1 is selected from C 12～26 linear saturated hydrocarbon, C 12～26 branched saturated hydrocarbon, C 12～26linear unsaturated hydrocarbons, C 12～26 branched unsaturated hydrocarbons, C 3～24 cyclic hydrocarbons, C 6～24 aromatic hydrocarbons. The R2is selected from one or more of hydrogen, hydroxyl, fluorine, chlorine, bromine, iodine, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, methoxy, ethoxy, n-propoxy, i-propoxy, difluoromethyl, trifluoromethyl, trifluoromethoxy, thiotrifluoromethyl, or trifluoroethyl. n is 1 or 2, whose basic unit contains at least one carbon-carbon single bond or one carbon-carbon double bond; X is O.

[0011] More preferably, the R1is selected from C 12～26 linear unsaturated hydrocarbons, C 12～26 branched unsaturated hydrocarbons; the R2group is hydrogen, 2-hydroxyl, 3-hydroxyl, 4-hydroxyl, 3,4-dihydroxyl, 2,6-dihydroxyl, 3-methoxyl, 4-methoxyl, 2-chloro, 3-chloro, 4-chloro, 2-fluoro, 3-fluoro, 4-fluoro, 4-bromo, 2,6-dichloro; n is 1, whose basic unit is a carbon-carbon single bond; X is O.

[0012] Specifically, the R1is selected from C 12～26 linear unsaturated hydrocarbons, whose corresponding carboxylic acid compound is

[0013] 2,5,8,11,14,17,20,23-octaoxahexacosane-26-oic acid, 2,5,8,11-tetraoxatetradecane-14-oic acid, cis-5,8,11,14,17-eicosapentaenoic acid, 5,8,11-trioxa-2-azatridecanedioic acid-1-tert-butyl ester, 15-bromopentadecanoic acid, (6Z,9Z,12Z)-octadeca-6,9,12-trienoic acid, heptadecanoic acid, (9Z,12Z)-octadeca-9,12-dienoic acid, (9Z)-9-octadecenoic acid, (5Z,8Z,11Z,14Z)-5,8,11,14-eicosatetraenoic acid, cis-9-hexadecenoic acid, (10E,12Z)-octadeca-10,12-dienoic acid, (9Z,12Z,15Z)-9,12,15-octadecatrienoic acid; the R2group is hydrogen, 3,4-dihydroxyl, 3-methoxyl, 4-methoxyl, 2-chloro, 3-chloro, 4-chloro, 4-fluoro, 4-bromo, 4-hydroxyl, 4-methyl, 3,4-dimethoxyl, 3-hydroxyl-4-methoxyl, 2,6-dichloro; n is 1, whose basic unit is a carbon-carbon single bond; X is O.

[0014] Further, the NADs compound can also be a pharmaceutically acceptable salt or solvate thereof.

[0015] The NADs compound of the present application can be prepared in various administration forms according to the conventional production method in the pharmaceutical field, for example, by mixing the active ingredient with one or more carriers, and then forming the desired dosage form. For example, the NADs compound itself or a mixture thereof with pharmaceutically acceptable excipients, diluents, etc. can be prepared into a pharmaceutical composition in the form of tablets, pills, oral solutions, capsules, syrups, dripping pills, granules, injection solutions, powder injections, sprays, aerosols, powder sprays, or suppositories, etc.

[0016] In addition, taking X as O and n = 1 as an example, the preparation process of the NADs compound is as follows:

[0017]

[0018] Further, the compound is mixed with a condensing agent, and then a solution of the compound is slowly added, and finally a base is added, and the reaction is carried out at room temperature for 1-4 hours to obtain the target product.

[0019] Further, the condensing agent is one of N,N'-carbonyldiimidazole, N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)urea hexafluorophosphate, benzotriazole-N,N,N',N'-tetramethyl urea hexafluorophosphate, N,N'-dicyclohexyl carbodiimide, 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide hydrochloride, and preferably N,N'-carbonyldiimidazole.

[0020] Further, the diluent is at least one of inert organic solvents, in particular, aliphatic, alicyclic or aromatic, optionally halogenated hydrocarbons, which can be selected from benzene, toluene, xylene, chlorobenzene, dichlorobenzene, petroleum ether, hexane, cyclohexane, dichloromethane, chloroform, carbon tetrachloride, diethyl ether, diisopropyl ether, dioxane, tetrahydrofuran, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, acetone, butanone, methyl isobutyl ketone, acetonitrile, propionitrile, butyronitrile, N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-formanilide, N-methylpyrrolidone, hexamethylphosphoric triamide, methyl acetate, ethyl acetate, dimethyl sulfoxide, methanol, ethanol, n-propanol, isopropanol, ethylene glycol monomethyl ether, ethylene glycol monoethyl ether, diethylene glycol monomethyl ether, and diethylene glycol monoethyl ether; and preferably at least one of N,N-dimethylformamide, dimethyl sulfoxide, and tetrahydrofuran.

[0021] Further, the presence of a base facilitates the reaction, which base is preferably sodium hydroxide, potassium carbonate, sodium ethoxide, triethylamine, trimethylamine, tributylamine, pyridine, N,N-dimethylaniline, N,N-dimethylbenzylamine, N-methylpiperidine, N-methylmorpholine, N,N-dimethylaminopyridine, diazabicyclooctane, diazabicyclononene or diazabicycloundecene. Preferably, the base is triethylamine.

[0022] The present application provides the use of NADs compounds in preventing or treating drug-induced motor complications of Parkinson's disease.

[0023] As an alternative, in the above use, the motor complications of Parkinson's disease are levodopa-induced, that is, the drug is levodopa.

[0024] As an alternative, in the above use, the motor complications of Parkinson's disease include motor fluctuations and dyskinesia.

[0025] As an alternative, in the above use, the motor complications include axial, limb and orofacial abnormal involuntary movements, wherein the axial abnormal involuntary movement is a twisting movement of the neck and upper body to the contralateral side of the injury, the limb abnormal involuntary movement is a repetitive jumping of the contralateral forelimb or dystonic posture, and the orofacial abnormal involuntary movement is a jaw movement and protrusion of the contralateral tongue.

[0026] As an alternative, in the above use, the NADs compound is used as a companion drug of levodopa to prevent or treat levodopa-induced dyskinesia (LID).

[0027] The present application uses LID behavioral data as the main indicator for evaluating the improvement effect of LID, detects and calculates the score of abnormal involuntary movement (AIM), and proves that NADs has a significant improvement effect on LID.

[0028] The AIM score includes the evaluation of three parts of axial, limb and orofacial. The observation method is to observe for 1 minute every 30 minutes after levodopa administration, and the whole process lasts for 120 minutes.

[0029] The pathogenesis of LID is diverse, the present application uses 6-hydroxydopamine (6-OHDA) to simulate neurotoxicity to cause nerve cell damage to cause PD, and uses levodopa to induce animal models to appear LID symptoms.

[0030] As an alternative, in the above use, the dose of the NADs compound for preventing or treating drug-induced motor complications of Parkinson's disease is 1-10 mg / kg.

[0031] Preferably, the NADs compound for preventing or treating drug-induced Parkinson's disease motor complications is in a dosage of 5 mg / kg.

[0032] The present application has the following beneficial effects over the prior art:

[0033] The present application first proposes that NADs compounds can be used as levodopa companion drugs for preventing or treating levodopa-induced dyskinesia. The present application takes 6-hydroxydopamine intracerebral injection and intraperitoneal injection of levodopa to model LID animal models as the research object, and studies the effect of NADs on improving LID. The results show that the NADs of the present application can significantly improve drug-induced PD motor complications LID, indicating that NADs compounds can be used as companion drugs to assist levodopa in treating PD. BRIEF DESCRIPTION OF DRAWINGS

[0034] The accompanying drawings are included to provide a further understanding of the present application, and constitute a part of the specification, illustrate the present application and explain the principles of the present application, and do not constitute a limitation of the present application. In the drawings:

[0035] Figure 1 LID modeling and grouping drug administration process is shown;

[0036] Figure 2 The modeling and administration method has no obvious effect on the body weight of mice;

[0037] Figure 3 The effect of drugs in each group on the axial score of abnormal involuntary movement of mice is shown;

[0038] Figure 4 The effect of drugs in each group on the limb score of abnormal involuntary movement of mice is shown;

[0039] Figure 5 The effect of drugs in each group on the oral facial score of abnormal involuntary movement of mice is shown;

[0040] Figure 6 The effect of drugs in each group on the total score of abnormal involuntary movement of mice is shown. DETAILED DESCRIPTION

[0041] The present application provides the use of NADs to improve PD motor complications, and those skilled in the art can refer to the content herein to appropriately improve the process parameters. It is particularly pointed out that all similar substitutions and changes are obvious to those skilled in the art, and are considered to be included in the present application. The method and application of the present application have been described by the preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the method and application herein without departing from the content, spirit and scope of the present application, to realize and apply the present application technology.

[0042] Unless otherwise specified, the specific techniques or conditions in the examples are in accordance with the techniques or conditions described in the literature in the field, or in accordance with the product instructions. Unless otherwise specified, the reagents or instruments used are conventional products that can be purchased through regular channels.

[0043] Unless otherwise specified, the experimental methods in the following examples are conventional methods. Unless otherwise specified, the test materials used in the following examples are commercially available products.

[0044] The present application is further illustrated by the following examples:

[0045] Example 1

[0046] I. Experimental materials and instruments

[0047] 1. Experimental animals

[0048] C57BL / 6J mice, male, 6 weeks old, body weight 20-22 g, purchased from the Experimental Animal Management Center of South Medical University (License: SCXK (Yue) 2021-0041), the animals were raised in the Medical Experimental Animal Center of Jinan University, the temperature was 23±2℃, the humidity was 50±10%, and the standard mouse feed was used. The experimental animals in each group were free to eat and drink.

[0049] 2. Main experimental reagents

[0050] 6-OHDA (Sigma, H4381), levodopa (Sigma, D9628), benserazide (Sigma, B7283), apomorphine (Sigma, A4393), dopamine hydrochloride (Anjie Chemical, E0801830250), arachidonic acid (Anjie, A875622), docosahexaenoic acid (Anjie, BD121651), oleic acid (Anjie, O815203). N-arachidonoyl dopamine (Cayman, 90057), N-oleoyl dopamine (Cayman, 10115), N-docosahexaenoyl dopamine (Cayman, 9001394).

[0051] 3. Main experimental instruments

[0052] Mouse stereotaxic apparatus (Shenzhen Ruivode Life Science and Technology Co., Ltd.), mouse rotarod apparatus (Ugo Basile, Italy).

[0053] II. Experimental methods and results

[0054] 1. Preparation of LID model

[0055] First, use 6-OHDA to build a PD model: after the mice were adaptively fed for seven days, they were weighed and deeply anesthetized with isoflurane gas, the head hair was shaved with a shaving knife and the mice were fixed in a brain stereotaxic instrument, and the left and right ear bars were adjusted to make it more stable. The mouse head skin was cut along the midline to expose the mouse skull, and hydrogen peroxide was used to wipe the bregma and lambda. With the bregma as the coordinate origin, refer to the mouse brain stereotaxic atlas, the injection coordinate site is AP: +0.3, L: +2.3, DV: -2.9, carefully drill a small hole with a skull drill (avoid damaging brain tissue), slowly inject 1.5 μL of 5 μg / μL 6-OHDA at a speed of 0.1 μL / min, leave the needle for 5 min after injection, slowly pull out the needle, suture the scalp, and complete the brain positioning injection. After 21 days of injection, apomorphine (0.5 mg / kg) was injected intraperitoneally to induce the mouse to rotate to the healthy side, and the rotation was detected three times, each for 10 min, and the average value was taken. The number of rotations was more than 70, and the other mice were eliminated.

[0056] Second, after the PD model of the mice was successfully established, levodopa was used. When administered, levodopa and benserazide were mixed in a ratio of 4:1 to build an LID model. The above-mentioned PD model mice were injected intraperitoneally with levodopa (20 mg / kg) every day. The successful PD modeling was verified by 6-OHDA as Day 0, and lasted for 42 days in total. The experimental procedure is shown in Figure 1

[0057] 2. Experimental grouping and body weight monitoring

[0058] The experimental mice were divided into 6 groups: 6 untreated mice were taken as the blank group (Sham). The PD model mice were randomly divided into 5 groups, 6 mice in each group, respectively: 6-OHDA group, 6-OHDA+levodopa group, 6-OHDA+levodopa+NADA (5 mg / kg, tail vein injection) group, 6-OHDA+levodopa+NDHA (5 mg / kg, tail vein injection) group and 6-OHDA+levodopa+OLDA (5 mg / kg, tail vein injection) group.

[0059] Body weight monitoring: as shown in Figure 2 The body weight data for 6 consecutive weeks showed that the modeling and administration methods used in this experiment had no significant effect on the body weight of the mice.

[0060] 3. Behavioral methods and results: abnormal involuntary movement score (AIM)

[0061] ​Method description: AIM score is divided into 3 parts, including axial, limb and orofacial. Each observation is 1 min, every 30 min after levodopa administration, lasting for 120 min. (1) Axial AIM: neck and upper body twisting movement to the contralateral side of injury; (2) Limb AIM: contralateral forelimb repetitive jumping or dystonic posture; (3) Orofacial AIM: jaw opening movement and contralateral tongue protrusion. According to the intensity of movement disorder during the observation period, each subtype is scored from 1-4, 0: no movement disorder; 1: occasional (less than 30 s); 2: often (more than 30 s); 3: symptoms persist, stop with external stimulus; 4: persist, stimulus does not stop. Total AIM score is the cumulative score of each part of each observation period.

[0062] Experimental results: According to the abnormal AIM principle, the effects of NADA, NDHA and OLDA on levodopa-induced LID were evaluated, as shown in Table 1, including axial AIM score (Table 1 (a)), orofacial AIM score (Table 1 (b)), limb AIM score (Table 1 (c)) and total AIM score (Table 1 (d)). Figures 3-6 Figure 3 Figure 4 Figure 5 Figure 6 Compared with the 6-OHDA group alone, the AIM score of mice in the levodopa administration group was significantly increased on the 7th day, 21st day and 42nd day, and the longer the administration time, the higher the AIM score, indicating that long-term administration of levodopa induced LID behavior disorder in the PD model. There was no significant difference in the improvement of each AIM score on the 7th day between NADA, NDHA and OLDA treatment, but both on the 21st day and the 42nd day, each AIM score of mice could be significantly alleviated (Table 1 (a) (b) (c) (d)). * p<0.05,

[0063] ** p<0.01, *** p<0.001 vs. 6-OHDA group, # p<0.05, ## p<0.005, ### p<0.001 vs. 6-OHDA+levodopa group. N=6 mice in each group). The above data prove that NADs treatment can improve levodopa-induced LID.

[0064] ​​​​​The foregoing description of the embodiments has been presented for the purpose of illustration and description. It is not intended to be exhaustive or to limit the application to the precise form disclosed. Modifications and variations are possible in light of the above teachings or can be acquired from practice of the application. As well, the description is presented in the context of the preferred embodiments as of their onset and might not represent the only or even the dominant form of the application. It is intended that the scope of the application embrace all techniques capable of attaining the same results caused by the embodiments described herein, including all equivalents thereof within the scope of the appended claims.

Claims

1. Use of N-acyl dopamine family lipids selected from N-arachidonoyl dopamine, N-oleoyl dopamine, N-docosahexaenoyl dopamine in the preparation of a medicament for preventing and treating drug-induced Parkinson's disease motor complications.

2. The use according to claim 1, wherein the drug inducing Parkinson's disease motor complications is levodopa.

3. The use according to claim 1, wherein the N-acyl dopamine family lipids are selected from N-docosahexaenoyl dopamine.

3. The use according to claim 1, wherein 4. The use according to claim 1, wherein the dosage of the N-acyl dopamine family lipids is 1-10 mg / kg.

4. The use according to claim 1, wherein 5. The use according to claim 1, wherein the dosage of the N-acyl dopamine family lipids is 5 mg / kg.

5. The use according to claim 4, wherein the compound is ###0002### 6. The medicament for preventing and treating drug-induced Parkinson's disease motor complications is N-docosahexaenoyl dopamine.

6. The use according to claim 1, wherein 7. The drug inducing Parkinson's disease motor complications is levodopa.

8. The dosage of N-docosahexaenoyl dopamine is 1-10 mg / kg.

7. Use according to claim 6, wherein 9. The dosage of N-docosahexaenoyl dopamine is 5 mg / kg.

8. Use according to claim 7, wherein the compound is ###0002### 10. The medicament further comprises a pharmaceutically acceptable excipient, and the dosage form of the medicament is one of a pill, an oral liquid, a capsule, a granule, an injection, a powder injection, a spray, or a suppository.

9. The use according to claim 1, wherein ​

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