Application of thienopyrimidine compound as dopamine D4 receptor allosteric modulator

Abstract

The invention discloses application of a thienopyrimidine compound to a dopamine D4 receptor allosteric antagonist, and belongs to the field of medicinal chemistry. The thienopyrimidine compound disclosed by the invention is shown as a formula 1, and the structure of the compound comprises a thiophene [3, 2-d] pyrimidine ring skeleton. Specifically, the thienopyrimidine compound can be used as an allosteric ligand of D4R to be combined with D4R so as to influence a downstream Gi signal channel and be used for treating diseases related to D4R. The functional activity of the compound on D4R is tested through a GloSensor cAMP content accumulation experiment, and whether the new compound has an allosteric regulation function on D4R or not is clarified. Research results show that most of the thienopyrimidine compounds have an allosteric regulation function on D4R, namely, the functional activity of D4R endogenous ligand dopamine can be subjected to negative allosteric regulation. Discovery of the thienopyrimidine D4R allosteric antagonist provides a new research and development thought and a chemical entity for research and development of a new drug targeting D4R.

Description

Technical Field

[0001] The present invention belongs to the technical field of medicinal chemistry, and specifically relates to the application of thienopyrimidine compounds as dopamine D4 receptor (D4R) allosteric modulators. Background Art

[0002] G protein-coupled receptors (GPCRs) are the largest transmembrane receptor protein family in the human genome. More than 800 GPCRs are expressed in the human body, distributed in different tissues and organs, mediating various physiological processes of the organism, and are also closely related to the formation and development of various diseases. GPCR has seven transmembrane helices (TM1-TM7), so GPCR is also called seven transmembrane receptor. GPCR can be activated by a variety of external signal molecules or endogenous ligands, and interact with downstream molecules to mediate a variety of physiological functions.

[0003] In the nervous system, dopamine receptors play an important physiological function. Dopamine (DA) is an important catecholamine neurotransmitter in the central nervous system. It is widely present in central and peripheral tissues. It mainly regulates the motor function, mental activity, secretion of pituitary hormones and cardiovascular function of the extrapyramidal system through dopamine receptors (DR). It also participates in the regulation of central emesis, gastrointestinal function, intraocular pressure and retinal information transmission. It is involved in the occurrence of Parkinson's disease, schizophrenia, Toumte syndrome, attention deficit hyperactivity syndrome and pituitary tumors (Physiol Rev. 1998, 78, 189-225). Dopaminergic innervation is mainly located in the brain and is divided into four main dopaminergic pathways, namely the mesolimbic pathway, the mesocortical pathway, the nigrostriatal pathway and the infundibular tubercle pathway. Dopaminergic neurons play a vital role in the movement, reward, sleep, emotion, feeding, memory and learning functions of the central nervous system (CNS). In the periphery, dopamine regulates physiological functions such as vomiting, hormone regulation, retinal projections, olfaction, cardiovascular and renal function. It mainly signals by binding to G protein-coupled dopamine receptors, mediating the regulation of a variety of neurophysiological functions, including movement, emotion regulation, motivation and cognition.

[0004] Dopamine is the most critical neurotransmitter in the dopaminergic system and drug addiction process, and its receptor is also an important target for studying the mechanism of drug addiction and finding anti-addiction drugs. Dopamine receptors (DR) can be divided into dopamine D1-like receptors and dopamine D2-like receptors. Dopamine D1-like receptors include D1R and D5R. After activation, they mainly act on the excitatory G protein-coupled receptors of the postsynaptic membrane, activate adenylate cyclase (AC), and lead to an increase in the concentration of cyclic adenosine monophosphate (cAMP). Dopamine D2-like receptors include D2R, D3R and D4R. After activation, they mainly act on the inhibitory G protein-coupled receptors, inhibit adenylate cyclase, and lead to a decrease in cAMP concentration. Among them, D4R is enriched in the prefrontal cortex area that is crucial for attention, cognition, decision-making and executive function, as well as the limbic area related to cognitive and emotional functions. It can not only regulate excitatory glutamatergic pyramidal neurons (corticostriatal pathway), but also regulate the inhibitory γ-aminobutyric acid (γ-aminobutyric acid, GABA) interneurons (Behav Brain Res, 2010, 208, 12-22). Abnormal dopamine and its mediated pathways can lead to a variety of diseases. Among them, D4R in the brain is mostly used to treat neuropsychiatric diseases related to impulse control. D4R levels are increased in schizophrenia.

[0005] D2R antagonists have been used to treat schizophrenia, but they have serious side effects, namely extrapyramidal side effects and delayed dyskinesia. In contrast, D4R antagonists have no extrapyramidal side effects and delayed dyskinesia. Therefore, compounds with selective D4R antagonists will be safer and more effective in treating psychosis and schizophrenia. However, due to the high amino acid sequence homology in the transmembrane region, D1-like receptors have 78% homology with D2-like receptors, while D3R and D4R have 75% and 53% homology, respectively (J Med Chem. 2012, 55, 6689-6699), resulting in poor selectivity of therapeutic drugs for receptor subtypes and easy off-target effects. Therefore, the development of drugs targeting specific subtypes of dopamine receptors with both high affinity and high selectivity faces great challenges. Allosteric modulators have many advantages as potential drug candidates. First, it has potential targeting specificity, that is, targeting a specific subtype, mainly due to the low sequence conservation of allosteric sites. In addition, the effect of allosteric modulators may also reach a certain saturation limit, which depends on the synergistic effect of allosteric drugs and endogenous ligands or agonists, which makes allosteric drugs have different pharmacodynamic characteristics from orthosteric ligand drugs (Cell Biol. 2014, 27, 94-101.). For biological systems that need to target specific receptor subtypes, control the level of activation, or last for a long time, allosteric modulators are an ideal choice.

[0006] Therefore, candidate drugs that allosterically regulate D4R activity need to be further studied for the treatment of D4R-related central nervous system diseases. Summary of the invention

[0007] In order to solve the above technical problems, the present invention provides a use of a thienopyrimidine compound as a negative allosteric modulator of dopamine D4 receptor.

[0008] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0009] The first aspect of the present invention provides a thienopyrimidine compound, the compound structure of which is shown in Formula 1:

[0010]

[0011] R1 is one of the following structures: H or CH3-; R2 is one of the following structures:

[0012]

[0013] R3 is one of the following structures:

[0014]

[0015] Furthermore, the compound structure is specifically one of the following structural formulas:

[0016]

[0017] The second aspect of the present invention provides the use of the above-mentioned thienopyrimidine compounds as dopamine D4 receptor allosteric modulators.

[0018] Furthermore, the thienopyrimidine compound specifically acts as a D4R negative allosteric modulator or allosteric antagonist.

[0019] Furthermore, the final concentration of the thienopyrimidine compound is 100 μM.

[0020] Furthermore, the concentration gradient of the thienopyrimidine compound used to verify the allosteric mechanism is 150 μM, 100 μM, 50 μM, 25 μM, 12.5 μM, and 6.25 μM.

[0021] The third aspect of the present invention provides a GloSensor-based TMA method for detecting the biological activity of a D4R allosteric modulator by a cAMP cumulative content experiment, the method is applied to the above-mentioned application; the method comprises: constructing a D4R plasmid, using cells to overexpress D4R and 22F cAMP, stimulating the overexpressing cells with a thienopyrimidine compound to be tested, then stimulating the cells with the D4R endogenous ligand dopamine for 5-10 minutes, then adding 10 μM Forskolin, detecting the bioluminescent signal of the stimulated cells, and determining the content of cAMP produced by the strength of the bioluminescent signal.

[0022] Furthermore, the overexpression cells are HEK-293T cells.

[0023] The fourth aspect of the present invention provides a pharmaceutical composition, characterized in that the pharmaceutical composition comprises the above-mentioned compound, its pharmaceutically acceptable salt or prodrug, hydrate, solvate, crystal form, tautomer or their metabolite form; optionally, the pharmaceutical composition also comprises a pharmaceutically acceptable carrier or excipient.

[0024] The fifth aspect of the present invention provides use of the above-mentioned pharmaceutical composition as a dopamine D4 receptor allosteric modulator.

[0025] Compared with the prior art, the present invention has the following beneficial effects:

[0026] The present invention reveals the connection between thienopyrimidine compounds and dopamine D4R receptors, clarifies the relationship at the mechanism level, and provides a more reliable basis for the treatment of mental illnesses.

[0027] The present invention has experimentally found that thienopyrimidine compounds can act on D4R, causing changes in downstream cAMP. In-depth mechanism studies have shown that the target compounds in the present invention are all D4R allosteric antagonists. At present, drugs on the market for treating mental illness act on dopamine receptors, but due to the homology between various subtypes of dopamine receptors, the specificity is not high and it is easy to be off-target. Long-term use of such drugs can cause obvious side effects. Allosteric modulators have specific targeting to receptors, and the regulatory function of allosteric modulators has a certain saturation limit, and will not over-activate or completely block the function of the receptor itself, so it is expected to develop safer and more effective D4R allosteric antagonist candidate drugs.

[0028] The present invention relates to one or more thienopyrimidine compounds that have the potential to be used as active pharmaceutical ingredients. In this case, these compounds can be combined with a pharmaceutically acceptable carrier and then converted into a pharmaceutical preparation. More specifically, these thienopyrimidine compounds are widely used to prepare D4R allosteric antagonists, which is a new targeted therapy strategy. Through this technology, specific receptors, such as D4R, can be effectively targeted to develop drugs with potential clinical value and specificity, thereby potentially solving unmet medical needs in the current market. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 is the structural formula of lead compound 2;

[0030] Figure 2 is the structural formula of thienopyrimidine compound 6q;

[0031] Figure 3 This is the concentration dependence curve of the allosteric regulation of DA by the thienopyrimidine compound 6q;

[0032] Figure 4 is the structural formula of thienopyrimidine compound 10d;

[0033] Figure 5 This is the concentration dependence curve of the allosteric regulation of DA by the thienopyrimidine compound 10d;

[0034] Figure 6 is the structural formula of thienopyrimidine compound 6o;

[0035] Figure 7 This is the concentration-dependency curve of the allosteric regulation of DA by thienopyrimidine compound 6o. DETAILED DESCRIPTION

[0036] According to existing technologies, G protein-coupled receptors are the largest family of membrane proteins in the human genome, playing a key role in regulating various physiological and pathological processes. Dopamine D4 receptor is one of the GPCRs. Activating this receptor can activate the D4R downstream pathway, release Gi signals, and cause a decrease in cellular cAMP content. Due to its important physiological function, D4R has become a potential target for the development of new drugs for schizophrenia.

[0037] Since new reagents and drugs that can act on dopamine receptors are still under research and development, the purpose of the present invention is to provide thienopyrimidine compounds for use as dopamine D4 receptor allosteric antagonists.

[0038] The present invention is described in detail below in conjunction with specific implementation modes.

[0039] It should be noted that the following detailed description of the contents of the present disclosure is illustrative, but the present disclosure is not limited to these embodiments and specific examples.

[0040] According to the present disclosure, the term "G protein coupled receptor" or "GPCR" refers to a transmembrane receptor capable of transmitting signals from the outside of a cell to the inside of a cell through a G protein pathway and / or an arrestin pathway.

[0041] According to the present disclosure, the term "ligand" or "receptor ligand" means a molecule that specifically binds to a GPCR intracellularly or extracellularly. Without limiting the purpose, the ligand can be a protein, (poly)peptide, lipid, small molecule, protein scaffold, antibody, antibody fragment, nucleic acid, carbohydrate. The ligand can be synthetic or naturally occurring. The term "ligand" includes "natural ligands", which are endogenous, natural ligands of natural GPCRs. In most cases, the ligand is a "modulator" that increases or decreases the intracellular response when it contacts (e.g., binds) to the GPCR expressed by the cell. Examples of ligands as modulators include agonists, partial agonists, inverse agonists, and antagonists. Among them, "agonist" refers to a ligand that increases the signaling activity of a receptor by binding to the receptor. A full agonist can stimulate the receptor to the maximum extent; a partial agonist cannot induce full activity even at saturating concentrations. Partial agonists can also function as "blockers" by preventing the binding of more powerful agonists. "Antagonists" refer to ligands that bind to receptors without stimulating any activity. "Antagonists" are also known as "blockers" because of their ability to prevent other ligands from binding and thus block agonist-induced activity. In addition, "inverse agonists" refer to antagonists that, in addition to blocking the effects of agonists, also reduce the basal or constitutive activity of a receptor below that of a receptor that does not bind a ligand.

[0042] According to the present disclosure, there are also allosteric regulatory sites on GPCRs, which do not bind to endogenous ligands, but can react specifically with allosteric modulators (AMs) to change the receptor conformation. Allosteric modulators can exhibit a certain range of activity on the target protein. Among them, positive allosteric modulators (PAMs) or allosteric agonists may not have any intrinsic efficacy, but when they bind to receptors, they enhance the binding affinity or efficacy (or both) of agonists. Negative allosteric modulators (NAMs) or allosteric antagonists do not have any intrinsic efficacy, but when they bind to receptors, they inhibit the binding affinity or efficacy (or both) of agonists.

[0043] In this specification, the term "treatment" refers to reversing, alleviating, delaying the onset or inhibiting the progression of a disease. In some embodiments, treatment can be administered after one or more signs or symptoms of the disease have developed or have been observed. In other embodiments, treatment can be administered in the absence of signs or symptoms of the disease.

[0044] The term "pharmaceutically acceptable" refers to molecular entities and compositions that do not produce adverse reactions, allergic reactions or other untoward reactions when administered to animals or humans, as appropriate. As used herein, the term "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial agents, isotonic and absorption delaying agents, buffers, excipients, binders, lubricants, gels, surfactants, etc. that can be used as media for pharmaceutically acceptable substances.

[0045] The term "pharmaceutically acceptable salt" refers to a salt that possesses the potency of the parent agent and is not biologically or otherwise undesirable (e.g., is neither toxic nor otherwise harmful to the recipient thereof). Suitable salts include acid addition salts, which can be formed, for example, by mixing a solution of the parent compound with a solution of a pharmaceutically acceptable acid such as hydrochloric acid, sulfuric acid, acetic acid, trifluoroacetic acid or benzoic acid. If the drug carries an acidic moiety (e.g., COOH or a phenolic group), pharmaceutically acceptable salts thereof may include alkali metal salts (e.g., sodium or potassium salts), alkaline earth metal salts (e.g., calcium or magnesium salts) and salts formed with suitable organic ligands (e.g., quaternary ammonium salts).

[0046] In a specific embodiment of the present invention, the thienopyrimidine or a pharmaceutically acceptable salt thereof is a negative allosteric modulator of the dopamine D4 receptor.

[0047] In a specific embodiment of the present invention, the thienopyrimidine or a pharmaceutically acceptable salt thereof activates dopamine D4R via the Gi-AC-cAMP pathway.

[0048] In one embodiment of the present invention, a thienopyrimidine compound or a pharmaceutically acceptable salt thereof is provided for use in the preparation of a D4R allosteric antagonist.

[0049] Wherein, the thienopyrimidine compound is an allosteric antagonist of D4R.

[0050] In a specific embodiment of the present invention, the allosteric antagonist of D4R includes ligands and small molecule compounds that inhibit D4R.

[0051] In a specific embodiment of the present invention, the ligand is the endogenous ligand DA of the dopamine receptor.

[0052] In another specific embodiment of the present invention, there is provided the use of thienopyrimidine compounds with different concentration gradients as D4R antagonist ligands.

[0053] In another specific embodiment of the present invention, a pharmaceutical composition is provided, which comprises the above-mentioned thienopyrimidine or its solvate or pharmaceutically acceptable salt.

[0054] The technical solution of the present disclosure is described in detail below.

[0055] Example 1

[0056] HEK 293T cells expressing human D4R that were in good growth condition and had a cell confluence of more than 80% were plated at 6.0×10 5 The cells were seeded into a 3.5 cm cell culture dish, shaken crosswise to evenly cover the surface, and then cultured overnight in a constant temperature and humidity cell culture incubator containing 5% CO2 at 37°C.

[0057] After 24 hours, the GloSensor-22F cAMP plasmid was transiently transfected into adherent HEK 293T cells using FuGENE HD transfection reagent (Promega) and opti-MEM medium, and the cells were cultured in an incubator for another 24 hours.

[0058] HEK 293T cells transfected with the expression plasmid for 24 hours were harvested, diluted with culture medium according to the calculated amount, and then evenly seeded into a transparent 96-well plate coated with poly-lysine (PDL) using a spray gun, and cultured for another 24 hours.

[0059] Wash the cells once with CO2 independent medium, and then add 10 mL of equilibration medium (2% GloSensor TMcAMP Reagent, 10% FBS and 88% CO2independent medium) in a 96-well plate, incubate in a cell culture incubator for 1 hour, place on a clean bench at room temperature for 1 hour, and immediately collect bioluminescent signals with a multifunctional microplate reader until a stable background signal is obtained. Then, the thienopyrimidine compound to be tested is added to the 96-well plate cells containing the substrate at the same time (the final concentration is 100 μM. The present invention tests the maximum allosteric effect of the target compound under the condition of 100 μM. If the concentration continues to increase, the cells may be intolerant or toxic, so this concentration is selected). After incubation at 37°C for 30 minutes, a concentration gradient negative control DA (the final concentration gradient is 1nM, 10nM, 100nM, 1μM, 10μM, 100μM) is added, incubated at room temperature for 5 minutes, and then 10μM Forskolin is added to the cells, and the changes in bioluminescence are read with a microplate reader. Compare the differences in the allosteric antagonistic activity of thienopyrimidine compounds and the control.

[0060] The thienopyrimidine compound of the present invention is shown in Formula 1, and the compound structure contains a thieno[3,2-d]pyrimidine ring skeleton. Specifically, the thienopyrimidine compound can be used as an allosteric ligand of D4R, bind to D4R, thereby affecting the downstream Gi signaling pathway, and is used to treat diseases related to D4R. The test results in Table 1 show that most thienopyrimidine compounds have different degrees of allosteric antagonism to D4R, and some of the compounds have significantly better allosteric antagonism activity to D4R than the lead compound 2, and its structural formula is as follows Figure 1 As shown, 6e, 6o, 6s, 6g, 6q, and 10d are 1.27 times, 1.51 times, 1.12 times, 1.46 times, 2.32 times, and 1.81 times of the lead compound, respectively.

[0061] Table 1 Statistical comparison of the allosteric antagonistic activity of thienopyrimidine compounds and compound 2

[0062]

[0063]

[0064]

[0065] Example 2

[0066] The GloSensor cAMP accumulation experiment was used to further study the allosteric antagonistic mechanism of this class of compounds. The target compounds were tested to see whether they could allosterically regulate the functional activity of the endogenous ligand DA of D4R to study its mechanism of negative allosteric regulation of D4R, and further confirm that this class of compounds is a negative allosteric modulator (NAM) of D4R. After waiting for the background signal of bioluminescence to stabilize, thienopyrimidine compounds with good antagonistic activity were added to the cells expressing the receptors. The concentrations were in multiples. After incubation at 37°C for 30 minutes, the temperature was restored to room temperature, and different concentration gradients of endogenous ligand DA were added to the cells (the final concentration gradient was 1nM, 10nM, 100nM, 1μM, 10μM, 100μM), incubated at room temperature for 5 minutes, and then 10μM Forskolin was added to the cells to test whether the change in the bioluminescence signal at this time showed a concentration-dependent limited downward trend. From the concentration-dependence curve, it can be found that the EC of DA red 50 After adding different concentrations of the same compound, the curve clearly showed a limited downward shift in a concentration-dependent manner.

[0067] Compound 6q( Figure 2 ) as an example, Figure 3 As shown, the red DA curve shows a slight downward shift as the concentration gradient of compound 6q increases by two times from 6.26 μM (brown curve), 12.5 μM (green curve), to 25 μM (blue curve), until the concentration of 6q increases to 50 μM and then shows a sharp downward trend. When the concentration of 6q increases to 100 μM (rose red curve), the DA curve shows a sharp downward shift. This is because high concentrations of 6q enhance the allosteric regulatory ability of endogenous ligand DA on D4R, which is manifested as a sudden drop in the concentration curve. When the concentration of 6q increased from 100.0 μM (rose-red curve) to 150.0 μM (lavender curve), the concentration curve of DA shifted downward very slightly. This phenomenon explains that with the increase in the concentration of compound 6q, the concentration-dependent curve of DA showed a limited downward shift, indicating that the thienopyrimidine compound 6q can negatively allosterically regulate the functional activity of the D4R endogenous ligand DA, that is, the newly synthesized 6q is a negative allosteric modulator or allosteric antagonist of D4R.

[0068] Compound 10d ( Figure 4 ) as an example, Figure 5As shown in the figure, the red DA curve shows a significant downward shift as the concentration gradient of compound 10d increases from 6.25μM (yellow curve) to 12.5μM (green curve). When the concentration of 10d increases to 100μM (purple curve), the DA curve shows a significant downward shift. This is because the high concentration of 10d enhances the allosteric regulation of the endogenous ligand DA on D4R, which is manifested as a sudden drop in the concentration curve. When the concentration of 10d increases from 100.0μM (purple curve) to 150.0μM (rose red curve), the downward shift of the DA concentration curve is very weak. This phenomenon explains that with the increase in the concentration of compound 10d, the concentration-dependent curve of DA shows a limited downward shift, indicating that the thienopyrimidine compound 10d can negatively allosterically regulate the functional activity of the endogenous ligand DA of D4R, that is, the newly synthesized 10d is also a negative allosteric modulator or allosteric antagonist of D4R.

[0069] Compound 6o( Figure 6 ) as an example, Figure 7 As shown in the figure, the red DA curve increases 4 times from 6.26μM (brown curve) to 25μM (purple curve) with the concentration gradient of compound 10d. When the concentration of 6o increases to 100.0μM (blue curve), the DA curve shows a significant downward shift. This is because the high concentration of 6o enhances the allosteric regulation of the endogenous ligand DA on D4R, which is manifested as a sudden drop in the concentration curve. When the concentration of 6o increases from 100.0μM (blue curve) to 150.0μM (rose red curve), the concentration curve of DA moves downward very slightly. This phenomenon explains that with the increase of the concentration of compound 10d, the concentration-dependent curve of DA shows a limited downward shift, indicating that the thienopyrimidine compound 6o can negatively allosterically regulate the functional activity of the endogenous ligand DA of D4R, that is, the newly synthesized 6o is also a negative allosteric modulator or allosteric antagonist of D4R. This allosteric regulation phenomenon is consistent with the reported allosteric antagonist regulation mechanism (J.Med.Chem.2019,62,174-206.), indicating that the target compound can negatively allosterically regulate the functional activity of the D4R endogenous ligand DA, that is, the thienopyrimidine compounds in the present invention are all D4R allosteric antagonists or negative allosteric modulators (NAM).

[0070] Those skilled in the art can understand that the above-mentioned embodiments are specific examples for implementing the present application, and in practical applications, various changes can be made to them in form and detail without departing from the spirit and scope of the present application. Any person skilled in the art can make their own changes and modifications without departing from the spirit and scope of the present application, so the scope of protection of the present application shall be based on the scope defined in the claims.

Claims

1. A thienopyrimidine compound, characterized in that: The compound structure is shown in Formula 1: R1 is one of the following structures: H or CH3-; R2 is one of the following structures: R3 is one of the following structures:

2. The thienopyrimidine compound according to claim 1, characterized in that: The compound structure is specifically one of the following structural formulas:

3. Use of the thienopyrimidine compound according to claim 1 or 2 as a dopamine D4 receptor allosteric modulator.

4. The use according to claim 3, characterized in that The thienopyrimidine compounds are specifically used as D4R negative allosteric modulators or allosteric antagonists.

5. The use according to claim 4, characterized in that The final concentration of the thienopyrimidine compound is 100 μM.

6. The use according to claim 4, characterized in that The concentration gradient of the thienopyrimidine compound used to verify the allosteric mechanism is 150 μM, 100 μM, 50 μM, 25 μM, 12.5 μM, and 6.25 μM.

7. A GloSensor-based TM The method for detecting the biological activity of D4R allosteric modulators by cAMP cumulative content experiment is characterized in that: The method is applied to the application as claimed in claim 3; the method comprises: constructing a D4R plasmid, using cells to overexpress D4R and 22F cAMP, stimulating the overexpressing cells with the thienopyrimidine compound to be tested, then stimulating the cells with the D4R endogenous ligand dopamine for 5-10 minutes, then adding 10 μM Forskolin, detecting the bioluminescent signal of the stimulated cells, and determining the content of cAMP produced by the strength of the bioluminescent signal.

8. The method according to claim 7, characterized in that The overexpression cells are HEK-293T cells.

9. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the compound according to claim 1 or 2, its pharmaceutically acceptable salt or prodrug, hydrate, solvate, crystal form, tautomer or their metabolite form; optionally, the pharmaceutical composition also comprises a pharmaceutically acceptable carrier or excipient.

10. Use of the pharmaceutical composition according to claim 9 as a dopamine D4 receptor allosteric modulator.

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