TCP derivative as well as preparation method and application thereof
By modifying phenylcyclopropylamine, TCP derivatives with strong LSD1 inhibitory activity were developed, which solved the optimization space for existing LSD1 inhibitors in the treatment of Parkinson's disease and achieved significant therapeutic effects and improvements.
Patent Information
- Application Number
- CN202510290653.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-03-12
- Publication Date
- 2025-06-13
AI Technical Summary
There is room for optimization and improvement in the treatment of Parkinson's disease, especially in improving the efficiency and safety of the drug.
By structural modification of phenylcyclopropylamine, a variety of structurally novel TCP derivatives have been developed, which have strong LSD1 inhibitory activity and are used to treat Parkinson's disease.
The developed TCP derivatives are significantly better than their precursor compounds, showing obvious therapeutic effects in animal models, and have a significant improvement in motor dysfunction in Parkinson's disease.
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Abstract
Description
Technical Field
[0001] The present invention belongs to the field of drug synthesis, and relates to the synthesis and preparation method of new compounds and their use in the preparation of drugs for treating or preventing PD. Specifically, it relates to a TCP derivative, its preparation method and use. Background Art
[0002] Neurodegenerative diseases are a spectrum of diseases mainly characterized by the progressive degeneration and eventual death of neurons, including Parkinson's disease (PD) and Alzheimer's disease (AD), etc. These diseases seriously affect the quality of life of patients and the incidence rate is increasing with the aging of the global population, bringing a heavy burden to society and families.
[0003] PD is a common neurodegenerative disease that mainly affects the motor function of the central nervous system. Its characteristic symptoms include tremors, muscle stiffness, bradykinesia and postural instability. The pathological feature of PD is the progressive loss of dopaminergic neurons in the substantia nigra of the midbrain and the formation of Lewy bodies. Although the exact cause of PD is not fully understood, genetic factors and environmental factors are considered to play important roles in its pathogenesis. Currently, the treatment of PD mainly focuses on symptom management, which can only relieve symptoms to a certain extent and cannot prevent the progression of the disease. Therefore, the development of new, safe and effective therapeutic drugs remains an urgent medical challenge.
[0004] Histone demethylase LSD1 (Lysine Specific Demethylase 1) is an important epigenetic regulatory enzyme. By removing the methylation of histone H3K4 and H3K9, it affects the chromatin structure and thus regulates the transcriptional activity of genes. LSD1 plays a role in a variety of biological processes, including cell differentiation, proliferation and development. Its abnormal expression or dysfunction is closely related to the occurrence and development of various diseases. In recent years, studies have shown that LSD1 also plays an important role in the occurrence and development of neurodegenerative diseases. LSD1 affects the health and function of neurons by regulating the gene expression of neurons. In addition, LSD1 plays an important role in regulating the expression of inflammation-related genes, and its abnormality may lead to the out-of-control of inflammatory response and exacerbate the pathological process of related diseases. LSD1 has become a potential target for treating related diseases.
[0005] Tranylcypromine (TCP) is a known monoamine oxidase inhibitor, which is mainly used in the clinical treatment of depression and other diseases. Its cyclopropylamine structure provides an important structural template for its use as an LSD1 inhibitor and plays an important role in the design and development of LSD1 inhibitors. A variety of TCP-based LSD1 inhibitors have been reported. Currently, several LSD1 inhibitors of this type have entered the clinical research stage and are mainly used in the clinical treatment of tumors through single or combined drug use. Only ORY-2001 is used to treat neurodegenerative diseases. Although certain progress has been made in the research of ORY-2001, there is still room for further optimization and improvement. Conducting new structural optimization of TCP to develop new and highly efficient LSD1 inhibitors is of great significance for the drug development of PD.
[0006] The purpose of the present invention is to provide a novel TCP-based compound and its preparation method, which are used as LSD1 inhibitors and for the treatment of PD, providing more and better treatment options for patients. Summary of the Invention
[0007] The first object of the present invention is to provide a compound with a novel structure, the second object is to provide a preparation method of the compound, and the third object is to provide the pharmaceutical use of the compound.
[0008] The above objects of the present invention are achieved through the following technical solutions:
[0009] A phenylcyclopropylamine derivative represented by the general structural formula (I), or a pharmaceutically acceptable salt or solvate thereof:
[0010]
[0011] Specifically selected from the following structures:
[0012]
[0013] The use of the above phenylcyclopropylamine derivative or a pharmaceutically acceptable salt or solvate thereof for the preparation of a drug for treating a disease that can be treated or alleviated by inhibiting histone demethylase LSD1.
[0014] Furthermore, the disease that can be treated or alleviated by inhibiting histone demethylase LSD1 is a neurodegenerative disease.
[0015] Even further, the neurodegenerative disease is Parkinson's disease.
[0016] A pharmaceutical preparation for treating a disease that can be treated or alleviated by inhibiting histone demethylase LSD1, using the above phenylcyclopropylamine derivative or a pharmaceutically acceptable salt or solvate thereof as the active ingredient.
[0017] Furthermore, it also includes a pharmaceutically acceptable carrier or excipient, and is made into a pharmaceutically acceptable dosage form.
[0018] Furthermore, the carrier or excipient is solid, semi-solid or liquid.
[0019] Furthermore, the dosage form includes tablets, capsules, injections, granules, pills and powders.
[0020] Advantageous effects:
[0021] In the present invention, structural modification is carried out on phenylcyclopropylamine to obtain a variety of TCP derivatives with novel structures. Activity studies have shown that the TCP derivatives provided by the present invention have potent LSD1 inhibitory activity, and are significantly superior to their precursor compound phenylcyclopropylamine. Those skilled in the art know that LSD1 plays an important role in the occurrence and development of various diseases such as neurodegenerative diseases, and has become a potential target for the treatment of related diseases. Animal experiments have shown that the compounds provided by the present invention have obvious therapeutic effects on Parkinson's disease and have the prospect of being developed into drugs for the treatment of Parkinson's disease. Specific embodiments
[0022] The following specifically introduces the substantial content of the present invention in combination with examples, but does not limit the protection scope of the present invention thereby.
[0023] Example 1: Preparation of Compound I-1
[0024]
[0025] Dissolve (E)-4-(2-(2-methyl-[1,1'-biphenyl]-3-yl)vinyl)benzaldehyde (50 mmol) in 5 mL of dichloromethane, add phenylcyclopropylamine hydrochloride (50 mmol) and triethylamine (60 mmol), and then add a drop of acetic acid. Stir at room temperature for 1 h. Then add sodium triacetoxyborohydride (80 mmol) and stir at room temperature overnight. After the reaction is completed, add the reaction solution to 5 mL of ethyl acetate, wash three times with water, dry the organic phase with anhydrous sodium sulfate, and then perform column chromatography. After passing through the column, a white solid is obtained with a yield of 58%. The spectral data of the compound shown in Formula I-1 are as follows: 1 HNMR(500MHz,Chloroform-d)δ7.61(m,1H),7.51(m,2H),7.46-7.38(m,4H),7.33(m,4H),7.28-7.24(m,3H),7.18(m,2H),7.03(m,3H),3.93(m,2H),2.43(m,1H),2.33(s,3H),2.08(m,1H),1.98(m,1H),1.16(m,1H),1.01(m,1H).
[0026] Experimental Example 2: Preparation of Compound I-2
[0027]
[0028] Dissolve 3-((4-formyl-3,5-dimethoxyphenoxy)methyl)-[1,1'-biphenyl]-2-carbonitrile (50 mmol) in 5 mL of dichloromethane, add phenylcyclopropylamine hydrochloride (50 mmol) and triethylamine (60 mmol), and then add one drop of acetic acid. Stir at room temperature for 1 h. Then add sodium triacetoxyborohydride (80 mmol) and stir at room temperature overnight. After the reaction is completed, add the reaction solution to 5 mL of ethyl acetate, wash with water three times, dry the organic phase with anhydrous sodium sulfate, and then perform column chromatography. The white solid is obtained by passing through the column with a yield of 71%. The spectral data of the compound shown in Formula I-2 are as follows: 1 HNMR(500MHz,Chloroform-d)δ7.73 - 7.68(m,2H),7.62 - 7.49(m,6H),7.24(m,2H),7.13(m,1H),7.01(m,2H),6.20(s,2H),5.31(s,2H),3.92(m,2H),3.68(s,6H),2.32(m,1H),1.92(m,1H),1.27(m,1H),1.15(m,1H),0.98 - 0.89(m,1H).
[0029] Example 3: Preparation of Compound I-3
[0030]
[0031] Dissolve 2,6-dimethoxy-4-((2-methyl-[1,1'-biphenyl]-3-yl)methoxy)benzaldehyde (50 mmol) in 5 mL of dichloromethane, add phenylcyclopropylamine hydrochloride (50 mmol) and triethylamine (60 mmol), and then add one drop of acetic acid. Stir at room temperature for 1 h. Then add sodium triacetoxyborohydride (80 mmol) and stir at room temperature overnight. After the reaction is completed, add the reaction solution to 5 mL of ethyl acetate, wash with water three times, dry the organic phase with anhydrous sodium sulfate, and then perform column chromatography. The white solid is obtained by passing through the column with a yield of 61%. The spectral data of the compound shown in Formula I-3 are as follows: 1HNMR(500MHz,Chloroform-d)δ7.45(m,3H),7.38(m,5H),7.24(m,2H),7.13(m,1H),7.01(m,2H),6.22(s,2H),5.09(s,2H),3.94(s,2H),3.68(s,6H),3.53(s,1H),2.34(m,1H),2.29(s,3H),1.95(m,1H),1.17(m,1H),0.95(m,1H).
[0032] Example 4: Preparation of Compound I-4
[0033]
[0034] Dissolve methyl 3-((4-formyl-3,5-dimethoxyphenoxy)methyl)-[1,1'-biphenyl]-2-carboxylate (50 mmol) in 5 mL of dichloromethane, add phenylcyclopropylamine hydrochloride (50 mmol) and triethylamine (60 mmol), then add a drop of acetic acid, and stir at room temperature for 1 h. Then add sodium triacetoxyborohydride (80 mmol) and stir at room temperature overnight. After the reaction is completed, add the reaction solution to 5 mL of ethyl acetate, wash it three times with water, dry the organic phase with anhydrous sodium sulfate, and then perform column chromatography. After passing through the column, a white solid is obtained with a yield of 78%. Spectral data of the compound shown in Formula I-4: 1 HNMR(500MHz,Chloroform-d)δ7.55-7.49(m,3H),7.43-7.36(m,5H),7.23(m,2H),7.15-7.11(m,1H),7.00(m,2H),6.15(s,2H),5.19(s,2H),3.90(m,1H),3.80(s,1H),3.64(s,6H),3.54(s,3H),2.31(m,1H),1.91(m,1H),1.14(m,1H),0.93(m,1H).
[0035] Example 5: LSD1 Inhibitory Activity Test
[0036] Compounds I-1 to I-4 were prepared according to Examples 1 to 4, with a purity of not less than 98%, and used as test samples. The positive control was phenylcyclopropylamine. Weigh an appropriate amount of the sample and prepare a solution with a concentration of 20 mM in DMSO, store it at 4 °C, and dilute it with DMSO according to the required concentration during the experiment.
[0037] Incubate the test samples at different concentrations (1.25 μL) with LSD1 recombinant protein (0.25 μM) in HEPES buffer at room temperature for 10 min. Then add the LSD1 reaction substrate H3K4me2 (2.5 μL, final concentration 25 μM) and incubate at 37 °C for 30 min. Finally, add the fluorescent dye AmplexRed solution (0.1 μL) and horseradish peroxidase HRP (1 μL) and incubate at room temperature for 5 min. Then measure the fluorescence intensity on a fluorescence microplate reader at an excitation wavelength of 530 nm and an emission wavelength of 590 nm. Set up blank control wells and 100% control wells on each plate simultaneously. In the blank wells, the samples and H3K4me2 are replaced with 1.25 μL of DMSO and 2.5 μL of buffer respectively, and in the 100% control wells, the samples are replaced with 1.25 μL of DMSO.
[0038] Calculate the inhibition rate of the test samples at different concentrations on LSD1 protein according to the following formula:
[0039]
[0040] Based on the inhibition rate of the test samples at different concentrations on LSD1 protein, use SPSS software to calculate the IC 50 value. The experimental results are shown in Table 1.
[0041] Table 1 Inhibitory effects of compounds I-1 to I-4 on LSD1 protein
[0042]
[0043] The above results indicate that compounds I-1 to I-4 have potent LSD1 inhibitory activity, and compound I-2 is the strongest, with an IC 50 value of 0.021 μM, significantly superior to the positive control pCPA.
[0044] Experimental Example 6: Testing of motor coordination and balance ability in a chronic PD mouse model induced by MPTP
[0045] I. Experimental materials
[0046] Two-month-old SPF-grade C57BL / 6 male mice weighing approximately 25 ± 5 g were selected for the experiment. Five mice were housed in each cage, with free access to water and food. The light-dark cycle was 12 / 12 h, and the room temperature was maintained at 22 ± 2 °C. One week of adaptive feeding was carried out before the experiment, and those that did not meet the experimental requirements such as diseases and accidental deaths were excluded. All animal breeding and tissue collection processes involved in this experiment complied with the approval of the Medical Ethics Committee of Henan University of Chinese Medicine.
[0047] II. Grouping and administration
[0048] Thirty mice were randomly divided into 3 groups, with 10 mice in each group.
[0049] Normal group: Normal saline;
[0050] Model group: MPTP administration dose was 20 mg / kg + probenecid administration dose was 250 mg / kg;
[0051] Compound group: After administration of MPTP (administration dose was 20 mg / kg) combined with probenecid (administration dose was 250 mg / kg), compound I-2 was given, with an administration dose of 10 mg / kg;
[0052] MPTP combined with probenecid was administered continuously for 35 days, once every 3.5 days, for a total of 10 times. Compound I-2 was administered once a day.
[0053] III. Behavioral experiment methods
[0054] 1. Open field test
[0055] The open field test can evaluate the spontaneous activity ability and exploratory ability of PD experimental animals in a novel environment. This experiment referred to the method of Rial. First, the experimental animals were placed in the middle of the test box to adapt to the activity for 10 min, and then the open field analysis system was used to record the movement trajectory (crossing line) and the number of rearings of the mice within 5 minutes. After each animal was tested, the feces of the animal on the bottom surface were cleaned and disinfected with 75% alcohol and dried, and then the next animal was tested. Each animal was tested 3 times, and the average value was taken for statistical analysis.
[0056] 2. Pole climbing test
[0057] The pole climbing test is often used to detect the degree of bradykinesia and limb coordination and balance ability of PD experimental animals. This experiment referred to the method of Park et al. A self-made simple climbing pole was 55 cm high, and the diameter of the spherical top was 1 cm. The experimental animals were placed on the spherical top, and the time for the mice to turn from head up to head down was recorded as T-turn (turn time), and the time to climb to the bottom of the pole was recorded as T-LA (locomotor activity time). If the mice did not move on the ball for more than 30 seconds, they were guided to climb down the pole head down, and T-turn was recorded as 30 seconds. Each animal was tested 3 times, and the average value was taken for statistical analysis.
[0058] 3. Rotarod test
[0059] The rod rotation test is used to evaluate the limb balance and coordination ability of mice and is an important method for drug screening in terms of motor coordination. Three days before the test, the mice were placed on a roller with a set rotation speed of 20 rmp and trained for 3 minutes every day. During the test, the time the mice stayed on the roller, that is, the falling latency, was recorded. After each animal was tested, the feces of the animal on the roller of the rod rotation instrument were cleaned, disinfected with 75% alcohol to remove the odor, and dried before testing the next animal. Each animal was tested 3 times repeatedly, and the average value was taken for statistical analysis.
[0060] 4. Statistical data analysis
[0061] All experimental data were expressed as mean ± standard error (mean ± SEM). All experiments were independently repeated 3 times. All data were analyzed and processed using GraphPad Prism 8.0 software. One-way analysis of variance (One-Way ANOVA) or two-way analysis of variance (Two-Way ANOVA) was used for comparison of means among multiple groups, and a significant difference was indicated by P < 0.05.
[0062] IV. Results of behavioral data
[0063] 1. Results of the open field test
[0064] In the open field test, the number of times the mice stood on their hind feet (Rearing Number) and the time spent in the central area (Time In Center Zone) represented the exploratory behavior of the mice, and the total moving distance (Total Distance) reflected the motor function of the mice. One-way analysis of variance was used for comparison between groups. The results are shown in Table 2: Compared with the normal group, the number of times the mice in the model group stood on their hind feet, the total moving distance, and the time spent in the central area decreased (P < 0.001, P < 0.01); compared with the model group, the number of times the mice in the compound group stood on their hind feet, the total moving distance (P < 0.001), and the time spent in the central area increased.
[0065] Table 2 Results of the open field test for mice in each group (Mean ± SEM, n = 10)
[0066]
[0067] Note: Compared with the normal group, ### P < 0.001, ## P < 0.01; compared with the model group, *** P < 0.001
[0068] 2. Results of the pole climbing test
[0069] In the pole climbing experiment, the time required for the mouse to turn from head-up to head-down is the turning time (T-turn), which is used to reflect the bradykinesia of the mouse. The time when the hind feet touch the ground when the mouse climbs to the bottom of the pole after turning (T-LA) is used to judge the motor coordination ability of the mouse. The results are shown in Table 3: Compared with the normal group, the turning time and the time when the hind feet touch the ground of the mice in the model group were prolonged (P < 0.0001); compared with the model group, the turning time and the time when the hind feet touch the ground of the mice in the compound group were shortened (P < 0.0001).
[0070] Table 3 Results of the pole climbing experiment of mice in each group (Mean±SEM, n = 10)
[0071]
[0072] Note: Compared with the normal group, #### P < 0.0001; compared with the model group, **** P < 0.0001,
[0073] 3. Results of the rotarod test
[0074] In the rotarod test, by recording the latency to fall of the mouse on the rotating rod, the muscle endurance and body balance ability of the mouse were judged. The results of one-way ANOVA are shown in Table 4: Compared with the normal group, the latency to fall of the mice in the model group was shortened (P < 0.0001); compared with the model group, the latency to fall of the mice in the compound group was prolonged (P < 0.001).
[0075] Table 4 Results of the rotarod test of mice in each group (Mean±SEM, n = 10)
[0076]
[0077] Note: Compared with the normal group, #### P < 0.0001; compared with the model group, *** P < 0.001
[0078] In summary, it can be seen that: Compounds I-1 to I-4 have potent LSD1 inhibitory activity, significantly superior to the positive control tranylcypromine; Compound I-2 can effectively improve the exploratory behavior, motor coordination ability, muscle endurance and body balance ability of Parkinson's disease PD model mice, and improve the motor dysfunction of Parkinson's disease PD model mice, and has the prospect of being developed into an anti-PD drug.
[0079] The function of the above embodiments is to specifically introduce the substantial content of the present invention. However, those skilled in the art should know that the protection scope of the present invention should not be limited to this specific embodiment.
Claims
1. A phenylcyclopropylamine derivative represented by the general structural formula (I), or a pharmaceutically acceptable salt or solvate thereof: Specifically selected from the following structures:
2. Use of the phenylcyclopropylamine derivative or a pharmaceutically acceptable salt or solvate thereof according to claim 1 for preparing a drug for treating a disease that can be treated or alleviated by inhibiting histone demethylase LSD1.
3. The use according to claim 2, characterized in that: The disease to be treated or alleviated by inhibiting histone demethylase LSD1 is a neurodegenerative disease.
4. The use according to claim 3, characterized in that: The neurodegenerative disease is Parkinson's disease.
5. A pharmaceutical preparation for treating a disease that can be treated or alleviated by inhibiting histone demethylase LSD1, comprising the phenylcyclopropylamine derivative according to claim 1 or a pharmaceutically acceptable salt or solvate thereof as an active ingredient.
6. The pharmaceutical preparation according to claim 5, characterized in that: It also includes pharmaceutically acceptable carriers or excipients to prepare pharmaceutically acceptable dosage forms.
7. The pharmaceutical preparation according to claim 6, wherein the carrier or excipient is solid, semisolid or liquid.
8. The pharmaceutical preparation according to claim 6, characterized in that: The dosage forms include tablets, capsules, injections, granules, pills and powders.