Tryptamine indole alkaloids, processes for their preparation and uses thereof

Abstract

This invention discloses a tryptamine indole alkaloid, its preparation method, and its applications. The tryptamine indole alkaloid, its derivatives, organic and inorganic acid salts, pharmaceutical compositions containing this type of compound, and plant extracts can effectively treat or prevent diseases with pathological features of lysosomal pathway disorders, including but not limited to anxiety, depression, neurodegenerative diseases, and autoimmune diseases, by regulating TFEB. The tryptamine indole alkaloid of this invention can also act as an IDO and MAOA inhibitor, improving diseases closely related to IDO and MAOA, including but not limited to depression, neurodegenerative diseases, and autoimmune diseases.

Description

Technical Field

[0001] This invention belongs to the field of medicine and relates to tryptamine indole alkaloids, their preparation methods, and applications. Background Technology

[0002] Anxiety is a common mental illness characterized by unpleasant and distressing feelings, often accompanied by physical discomfort. 3.6-28.8% of people will experience it at some point in their lives, and over 90% of patients develop the condition before the age of 35. Women are more likely to suffer from anxiety disorders than men. Depression is a common mental illness characterized by prolonged low mood. Clinically, it often manifests as melancholy, and in severe cases, can lead to profound grief or even stupor. Depression and anxiety often coexist clinically, and many symptoms are similar, including poor sleep, decreased appetite, decreased energy, and irritability. Studies show that 60-90% of patients with depression also have anxiety symptoms, and over 50% of patients with depression have anxiety disorders that meet the diagnostic criteria. Due to increasing pressure in modern life, the probability of developing mental illnesses such as depression and anxiety is rising, requiring sufficient attention from society as a whole and the implementation of effective prevention and treatment measures.

[0003] During the immune response, both tolerance to self-components and rejection of "non-self" antigens occur under the control of the body's immune regulatory mechanisms. If immune regulation malfunctions: when the immune response is too strong, the immune system produces a high immune response to "foreign" antigens, leading to hypersensitivity reactions; or even the immune system's tolerance balance is disrupted, resulting in a strong attack on self-components and causing autoimmune diseases. Conversely, when the immune response is weak, the body cannot properly react to and clear invading substances, thus reducing the body's ability to fight infection and tumors, leading to immunosuppression. Therefore, the immune regulatory mechanism is crucial for maintaining the stability of the internal environment and has significant physiological importance for the normal functioning of various bodily functions.

[0004] Autophagy is a highly conserved lysosomal-mediated process of intracellular protein and organelle degradation. It maintains cellular survival and tissue homeostasis by clearing damaged intracellular material. Autophagy directly or indirectly participates in many important intracellular physiological activities, such as cell cycle regulation, cell proliferation, apoptosis, maintenance of stem cell stemness, establishment of induced pluripotent stem cells (iPSCs), and clearance of foreign pathogens. Recent studies have shown that autophagy is involved in lymphocyte development, regulation of innate and adaptive immune responses, aging, and various physiological and pathological processes related to aging, playing a crucial regulatory role in neurodegenerative diseases and the body's immune response. Autophagy is regulated by numerous intracellular and external factors at different stages, with multiple complexes involved in the formation of autophagosomes. Increasing evidence suggests that transcription factor EB (TFEB) is a major regulator of autophagy and lysosomal biosynthesis.

[0005] Indoleamine-2,3-dioxygenase (IDO) is an intracellular heme-containing enzyme and the rate-limiting enzyme for extrahepatic tryptophan metabolism in humans. It catalyzes the oxidative cleavage of the indole epoxide in the tryptophan molecule, leading to its catabolism via the kynurenic acid pathway. As an immunomodulatory enzyme, IDO is associated with the development of various diseases closely related to tryptophan deficiency, including cancer, depression, Alzheimer's disease, Parkinson's disease, and autoimmune diseases.

[0006] Monoamine oxidase (MAO) is a naturally occurring enzyme in the human body that catalyzes the oxidation and deamination of monoamines. It comprises two subtypes: monoamine oxidase A (MAOA) and monoamine oxidase B (MAOB), both of which can inactivate monoamine neurotransmitters. MAOA is mainly distributed in the gastrointestinal tract and liver, and is an important enzyme in the metabolism of monoamines ingested through the digestive tract, primarily involved in the deamination of norepinephrine, epinephrine, and serotonin. MAOB is mainly distributed in the central nervous system, responsible for metabolizing dopamine in the brain. Impaired MAO activity can lead to abnormal metabolism of monoamine neurotransmitters in the central nervous system, resulting in various neurological disorders. MAOA inhibitors are mainly used to treat depression and neurasthenia, while MAOB inhibitors are used to treat Parkinson's disease and Alzheimer's disease.

[0007] Passionflower ( Passiflora Linn ) is a member of the Passifloraceae family ( Passifloraceae Passion fruit (also known as egg fruit) is the largest genus in the family Cypripedium, containing approximately 500 species, of which about 50 are edible. Plants in this genus are a traditional Western remedy for anxiety disorders, and their medicinal effects are widely recognized in Europe and North America, with several countries approving them for clinical use. Passiflora edulia Passion fruit (Passiflora arvense) is a herbaceous vine belonging to the genus Passiflora in the family Passifloraceae. Also known as granadilla, it enjoys the reputation of being a "beverage MSG" and the "king of juices." It is usually divided into: yellow passion fruit (…P. edulis var. flavicarpaDegenerer) and purple passion fruit ( P. edulis Two varieties, *Sims*, were introduced to my country in 1913 and are mainly cultivated in the southern regions. They possess various functions and effects, including stimulating, strengthening, invigorating, stress-relieving, beautifying, preventing and treating gastroenteritis, lowering lipids, sedating and relieving pain, and enhancing immunity. They are widely used as a folk remedy in tropical countries and are particularly renowned in Europe, suitable for treating various ailments such as sleep disorders and anxiety. As early as the 20th century, this plant was a popular analgesic and sedative, a use that continues to this day.

[0008] *Peganum* is a genus of perennial herbaceous plants belonging to the family Zygophyllaceae. There are six species worldwide, mainly distributed in the former Soviet Union, Mongolia, the Mediterranean coast, and North America. In northwestern China, *Peganum* is primarily found in deserts, sandy areas, and arid grasslands of Xinjiang, Gansu, Qinghai, Ningxia, Inner Mongolia, and Shaanxi provinces. P. harmala L.), multi-lobed camel husk [ P. multisecrum (Maxim.) Bobr.] and camel wormwood ( P. nigellastrum There are 3 species of *Phyllostachys edulis* (Bge.). *Phyllostachys edulis* is a traditional medicinal herb used by the Uyghur and Mongolian peoples for a long time and has been included in the Uyghur medicine section of the Ministry of Health's drug standards. *Phyllostachys edulis* is neutral in nature, bitter and pungent in taste, and poisonous. It has functions such as strengthening tendons and veins, warming yin and yang, eliminating viscous body fluids, and dispelling cold and dampness. It is mainly used to treat symptoms such as weak tendons and veins, bone and joint pain, cough with excessive phlegm, hemiplegia, forgetfulness, dizziness, headache, and irregular menstruation. The main chemical components of *Phyllostachys edulis* are alkaloids, which have a wide range of pharmacological effects. Summary of the Invention

[0009] The purpose of this invention is to overcome the shortcomings and deficiencies of the prior art and provide a tryptamine indole alkaloid.

[0010] Another object of the present invention is to provide a method for preparing the above-mentioned tryptamine indole alkaloid.

[0011] Another object of the present invention is to provide the application of the above-mentioned tryptamine indole alkaloids. The objective of this invention is achieved through the following technical solution: A tryptophan indole alkaloid, the structural formula of which is shown in at least one of general formulas I, II, and III: .

[0012] The tryptophan indole alkaloid is at least one of the following compounds: Harmine (A-1), pegaharmine J (A-2), harman (A-3), banistecaines J (A-4), harmol (A-5), tetrahydroharmine (A-6), N2-methyltetrahydroharmine (A-7), 7-methoxy-1-methyl-2,3,4,9-tetrahydropyrido[3,4-b] indole-1-methanol (A-8)ketotetrahydronorharmine (A-9), 7-methoxy-1-methyl-1,3,4,9-tetrahydropyrido[3,4-b]indole -2-carboxylate (A-10), harmaline (A-11), harmalane (A-12), harmalan (A-13), harmalol (A-14), harmidol (A-15), pegaharmine I (A-16), banistenoside B (A-17), banistecaine A (A-18), banistecaine B (A-19), (±)-banistecaine I (A-20), banistecaine C (A-21), (±)-banistecaine E (A-22), banistecaine F (A-23), banistecaine G (A-24), banistecaine H (A-25).

[0013] The tryptamine indole alkaloids, wherein compounds A-1 to A-25 have the following structural formulas: .

[0014] A tryptamine indole alkaloid composition comprising at least one of the above-mentioned tryptamine indole alkaloids.

[0015] A method for preparing a tryptamine indole alkaloid composition includes the following steps: The raw material was crushed and percolated for extraction. The pH was adjusted to 2-3 and then extracted. After separation, an acidic aqueous layer was obtained. The pH was adjusted to 9-10 and extracted to obtain tryptamine indole alkaloids. or; The raw material was crushed and percolated for extraction. The pH was adjusted to 7, filtered, and then adsorbed using a resin column. After elution, tryptophan indole alkaloids were obtained.

[0016] The raw materials are plants belonging to the genus *Passiflora* of the family Passifloraceae or the genus *Peganum* of the family Zygophyllaceae; preferably, passion fruit (*Passiflora edulis*). Passiflora edulia ), Camel bonsai ( Passiflora harmala L.), multi-lobed camel husk [ Passiflora multisecrum (Maxim.) Bobr.], Camel Artemisia ( Passiflora nigellastrum At least one of Bge.).

[0017] The percolation extraction is performed using 5-10% hydrochloric acid.

[0018] The extraction method described is chloroform extraction.

[0019] The resin column is a D101 macroporous adsorption resin column.

[0020] The use of the above-mentioned tryptamine indole alkaloids or tryptamine indole alkaloid compositions in the preparation of medicaments for the treatment or prevention of diseases with pathological features of lysosomal pathway disorders; preferably at least one of anxiety, depression, neurodegenerative diseases, and autoimmune diseases.

[0021] The application of the above-mentioned tryptamine indole alkaloids or tryptamine indole alkaloid compositions in the preparation of TFEB activators.

[0022] The application of the above-mentioned tryptamine indole alkaloids or tryptamine indole alkaloid compositions in the preparation of IDO inhibitors.

[0023] The application of the above-mentioned tryptamine indole alkaloids or tryptamine indole alkaloid compositions in the preparation of MAOA inhibitors.

[0024] The application of the above-mentioned tryptamine indole alkaloids or tryptamine indole alkaloid compositions in the preparation of MAOB inhibitors.

[0025] The above-mentioned derivatives of tryptamine indole alkaloids, as well as their organic and inorganic acid salts, are all within the scope of protection of this invention.

[0026] Using the above-mentioned plants as raw materials, percolation extraction was performed using water, acidic water, ethanol, or an aqueous ethanol solution of any concentration, followed by separation and enrichment.

[0027] The separation and enrichment involve concentrating the extract, filtering or centrifuging it, and then loading the supernatant of the filtrate onto a macroporous adsorption resin or cation exchange resin chromatography column for separation and enrichment; or separating and enriching the concentrate using an extraction method. The present invention has the following advantages and effects compared with the prior art: This invention discloses a tryptamine indole alkaloid, its preparation method, and its applications. The tryptamine indole alkaloid, its derivatives, organic and inorganic acid salts, pharmaceutical compositions containing this type of compound, and plant extracts can effectively treat or prevent diseases with pathological features of lysosomal pathway disorders, including but not limited to anxiety, depression, neurodegenerative diseases, and autoimmune diseases, by regulating TFEB. The tryptamine indole alkaloid of this invention can also act as an IDO and MAOA inhibitor, improving diseases closely related to IDO and MAOA, including but not limited to depression, neurodegenerative diseases, and autoimmune diseases. Attached Figure Description

[0028] Figure 1 This is a schematic diagram of the structural formula of tryptamine indole alkaloid.

[0029] Figure 2 This is a high-throughput image analysis of the results of a TFEB nuclear translocation experiment in HeLa cells, which used total bases and monomeric compounds to promote transfection.

[0030] Figure 3 This is a high-throughput image analysis of the results of experiments on the promotion of endogenous TFEB nuclear translocation in Neuro-2a cells by total bases and monomeric compounds.

[0031] Figure 4 This is a graph showing the quantitative results of TFEB nuclear translocation. HeLa 3xFlag-TFEB cells were treated with the compound for 24 h in three independent replicates. Data are presented as mean ± SEM. Compared with the control group (0.1% DMSO), * P <0.05,*** P <0.001.

[0032] Figure 5 This is a diagram showing the escape latency results of mice in the Morris water maze test. Detailed Implementation

[0033] The present invention will be further described in detail below with reference to the embodiments and accompanying drawings, but the embodiments of the present invention are not limited thereto.

[0034] Unless otherwise specified in the following implementation plan, the test conditions are generally as per standard test conditions or the test conditions recommended by the reagent company. Unless otherwise specified, all materials and reagents used are commercially available. Using the above-mentioned preferred plants as raw materials, percolation extraction was performed with water, acid water, ethanol, or an aqueous ethanol solution of any concentration, followed by separation and enrichment. Example 1: Preparation of egg fruit tryptamine indole alkaloid Take fresh passion fruit ( Passiflora edulia5 kg of the sample was crushed and extracted by percolation with 10% hydrochloric acid. The extracts were combined, and the pH was adjusted to 2-3. The extracts were then repeatedly extracted with chloroform to obtain chloroform and acidic aqueous layers. After separating the acidic aqueous layer, ammonia was added to the acidic aqueous layer to adjust the pH to 9-10. The extracts were then extracted with chloroform to obtain the total alkaloids. Example 2: Preparation of Camel's Feather Tritamine Indole Alkaloid Take camel husk ( Passiflora harmala 2 kg of L. medicinal material was dried, pulverized, and extracted by percolation with 5% hydrochloric acid water. The extracts were combined, the pH was adjusted to 7, filtered, and the supernatant was loaded onto a D101 macroporous adsorption resin column for adsorption. The column was then eluted sequentially with water, 50% ethanol, and 80% ethanol. The 80% ethanol eluent was collected, evaporated under reduced pressure, and finally the total alkaloids were obtained. Example 3: Isolation and structural identification of tryptamine indole alkaloid monomers The total alkaloids obtained in Example 1 were subjected to column chromatography using 200-300 mesh silica gel, with gradient elution using chloroform-methanol as the mobile phase to obtain 10 fractions, Fr.1 to Fr.10. Compounds A-1 to A-23 were separated by TLC and repeated Sephadex LH-20 column chromatography, low-pressure chromatography, and high-performance liquid chromatography. A-20 (methanol:water:diethylamine = 35:65:0.02) was isolated from Fr. 2. v / v / v , t R = 9.0 min, 6 mL / min), A-20 was separated into A-21 (acetonitrile: water = 50:50) by Phenomenex cellulose-2 column chromatography. v / v / v , t R = 4.9 min, 6 mL / min). A-17 (methanol:water:diethylamine = 63:37:0.02) was isolated from Fr. 3. v / v / v , t R = 11.5 min, 1 mL / min). A-1 (methanol:water:diethylamine = 40:60:0.02) was isolated from Fr. 4. v / v / v , t R = 10.5 min, 6 mL / min), A-2 (acetonitrile: water: diethylamine = 35 : 65 : 0.02, v / v / v , t R= 13.0 min, 6 mL / min), A-5 (acetonitrile: water: diethylamine = 70: 30: 0.02, v / v / v , t R = 21.6 min, 6 mL / min), A-7 (acetonitrile: water: diethylamine = 35:65:0.02, v / v / v , t R = 10.9 min, 6 mL / min), A-14 (acetonitrile: water: diethylamine = 20 : 80 : 0.02, v / v / v , t R = 25.2 min, 6 mL / min), A-16 (acetonitrile: water: diethylamine = 23 : 77 : 0.02, v / v / v , t R = 9.5 min, 6 mL / min). A-9 (acetonitrile: water: diethylamine = 40: 60: 0.02) was isolated from Fr. 5. v / v / v , t R = 10.5 min, 6 mL / min), A-12 (acetonitrile: water: diethylamine = 30: 70: 0.02, v / v / v , t R = 18.5 min, 6 mL / min), A-13 (acetonitrile: water: diethylamine = 30 : 70 : 0.02, v / v / v , t R = 15.3 min, 6 mL / min). A-10 (acetonitrile: water: diethylamine = 35 : 65 : 0.02, ) was isolated from Fr. 6. v / v / v , t R = 10.9 min, 6 mL / min), A-22 (acetonitrile: water: diethylamine = 25 : 75 : 0.02, v / v / v , t R = 14.3 min, 6 mL / min), A-23 (methanol: water: diethylamine = 55:45: 0.02, v / v / v , t R= 9.7 min, 6 mL / min). A-3 (methanol:water:diethylamine = 35:65:0.02) was isolated from Fr. 7. v / v / v , t R = 12.5 min, 6 mL / min), A-6 (acetonitrile: water: diethylamine = 35 : 65 : 0.02, v / v / v , t R = 14.0 min, 6 mL / min). A-15 (acetonitrile:water:diethylamine = 40:60:0.02) was isolated from Fr. 8. v / v / v , t R = 11.0 min, 6 mL / min), A-19 (acetonitrile: water: diethylamine = 30 : 70 : 0.02, v / v / v , t R = 12.5 min, 6 mL / min). A-4 (methanol:water:diethylamine = 60:40:0.02) was isolated from Fr. 9. v / v / v , t R = 16.0 min, 6 mL / min), A-8 (methanol: water: diethylamine = 45 : 55 : 0.02, v / v / v , t R = 25.0 min, 6 mL / min), A-11 (acetonitrile: water: diethylamine = 27 : 73 : 0.02, v / v / v , t R = 24.3 min, 6 mL / min), A-18 (acetonitrile: water: diethylamine = 30 : 70 : 0.02, v / v / v , t R = 11.5 min, 6 mL / min). Combined with infrared, ultraviolet, mass spectrometry, and nuclear magnetic resonance spectroscopy analysis, its structural formula was identified as follows:

[0035]

[0036]

[0037]

[0038]

[0039] Among the above compounds, compounds A-4 and A-16 to A-23 are new compounds, and their NMR data are as follows: Compound A-4: 1 H NMR (600 MHz, CD3OD) δ H 8.05 (d, J = 8.7 Hz, 1H), 7.74 (s,1H), 7.00 (d, J = 2.1 Hz, 1H), 6.82 (dd, J = 8.7, 2.2 Hz, 1H), 4.06 (s, 3H), 3.87(s, 3H), 2.66 (s, 3H); 13 C NMR (150 MHz, CD3OD) δ C 161.8, 151.7, 143.2, 137.0,135.8, 125.7, 120.0, 119.4, 115.9, 110.8, 95.2, 56.4, 55.9, 18.9; HR-ESI-MS m / z : 243.1130 [M+H] + (calcd for C 14 H 15 N2O2, 243.1128).

[0040] Compound A-16: 1 H NMR (600 MHz, CD3OD) δ H 7.33 (d, J = 8.6 Hz, 1H), 6.93 (d, J = 2.2 Hz, 1H), 6.68 (dd, J= 8.6, 2.3 Hz, 1H), 4.89 (overlapped, 1H), 4.84(overlapped, 1H), 4.64 (d, J = 8.3 Hz, 1H), 3.82 (s, 3H), 3.80 (overlapped,1H), 3.60 (dd, J = 9.3, 6.7 Hz, 1H), 3.50 (t, J = 7.9 Hz, 1H), 3.13 (td, J = 12.4,4.2 Hz, 1H), 2.83 (m, 1H), 2.76 (m, 1H); 13 C NMR (150 MHz, CD3OD) δ C 173.5,157.8, 139.2, 129.5, 121.7, 119.4, 110.1, 109.7, 95.7, 79.5, 77.0, 74.5,72.3, 56.0, 54.4, 40.3, 21.7; HR-ESI-MS m / z : 371.1211 [M+Na] + (calcd forC 17 H 20 N2NaO2, 371.1274)。

[0041] Compound A-17: 1 H NMR (600 MHz, CD3OD) δ H 7.31 (d, J = 8.6 Hz, 1H), 6.89 (d, J =2.2 Hz, 1H), 6.69 (dd, J = 8.6, 2.2 Hz, 1H), 5.22 (s, 1H), 5.05 (s, 1H), 4.90(overlapped, 1H), 4.22 (t, J = 4.0 Hz 1H), 4.18 (d, J = 3.1 Hz, 1H), 3.81 (s,3H), 3.69 (dt, J = 4.4, 1.4 Hz, 1H), 3.55 (s, 3H), 3.46 (td, J = 12.0, 4.2,1H), 2.83 (m, 1H), 2.70 (m, 1H);13 C NMR (150 MHz, CD3OD) δ C 174.0, 157.7,139.1, 130.3, 122.4, 119.2, 111.4, 109.8, 95.8, 87.4, 76.2, 70.1, 69.4, 61.0,56.0, 52.1, 42.6, 21.4; HR-ESI-MS m / z 385.1382 [M+Na] + (calcd for C 18 H 22 N2NaO2, 385.1370).

[0042] Compound A-18: 1 H NMR (400 MHz, CD3OD) δ H 7.20 (d, J = 8.3 Hz, 1H), 6.67 (dd, J = 8.3, 2.3 Hz, 1H), 6.58 (d, J = 2.3 Hz, 1H), 4.32 (m, 2H), 3.83 (s, 3H), 2.66(ddd, J = 13.8, 8.8, 5.2 Hz, 1H), 2.45 (ddd, J = 13.2, 9.4, 7.3 Hz, 1H), 1.72(s, 3H); 13 C NMR (100 MHz, CD3OD) δ C 179.6, 163.0, 149.0, 144.5, 126.0, 122.1,109.2, 98.8, 62.0, 61.8, 56.0, 29.9, 9.9; HR-ESI-MS m / z : 247.1084 [M+H] + (calcdfor C 13 H 15 N2O3, 247.1077).

[0043] Compound A-19: 1 H NMR (600 MHz, CD3OD) δ H 7.15 (d, J = 8.2 Hz 1H), 6.60 (dd,J = 8.2, 2.3 Hz, 1H), 6.52 (d, J = 2.3 Hz, 1H), 3.79 (s, 3H), 3.28 (q, J = 6.6 Hz, 1H), 3.19 (m, 2H), 2.27 (m, 2H), 0.99 (d, J = 6.6 Hz, 3H); 13 13C NMR (150 MHz, CD3OD) δ C 183.7, 161.8, 144.5, 125.9, 124.2, 108.4, 98.2, 64.8, 59.0, 55.9, 46.3, 39.2, 13.4.; HR-ESI-MS m / z : 233.1293 [M+H]+ + (calcd for C 13 H 17 N2O2, 233.1285).

[0044] Compound A-20 (3 R ,1′ R -A-20 and 3 S ,1′ S- -A-20): 1 1H NMR (600 MHz, CD3OD) δ H 8.23 (s, 1H), 6.98 (d, J = 8.3 Hz, 1H), 6.62 (dd, J = 8.4, 2.3 Hz, 1H), 6.57 (d, J = 2.3 Hz, 1H), 4.06 (q, J = 6.4 Hz, 1H), 3.82 (m, 2H), 3.80 (s, 3H), 2.38 (dt, J = 12.8, 9.3 Hz, 1H), 2.12 (m, 1H), 1.10 (d, J = 6.5 Hz, 3H); 13 13C NMR (150 MHz, CD3OD) δ C180.8, 163.0, 162.1, 144.1, 126.0, 122.2, 108.2, 98.6, 61.1, 58.3,55.9, 44.1, 33.8, 16.0; HR-ESI-MS m / z 283.1050 [M+Na] + (calcd for C 14 H 16 N2NaO3, 283.1053).

[0045] Compound A-21: 1 H NMR (600 MHz, CD3OD) δ H 8.23 (s, 1H), 7.16 (d, J = 8.3 Hz, 1H), 6.63 (dd, J = 8.3, 2.3 Hz, 1H), 6.53 (d, J = 2.3 Hz, 1H), 4.08 (q, J = 6.5 Hz,1H), 3.90 (m, 1H), 3.80 (s, 3H), 3.78 (m, 1H), 2.28 (m, 2H), 1.22 (d, J = 6.5Hz, 3H); 13 C NMR (150 MHz, CD3OD) δ C 181.2, 162.8, 162.2, 144.5, 124.7, 122.1,108.4, 98.2, 62.0, 57.9, 55.9, 44.3, 33.4, 14.8; HR-ESI-MS m / z 283.1054 [M+Na] + (calcd for C 14 H 16 N2NaO3, 283.1053).

[0046] Compound A-22 (3 R ,1′ S- A-22 and 3 S ,1′ R -A-22): 1 H NMR (600 MHz, DMSO- d 6) δ H 7.05 (d,J = 8.3 Hz, 1H), 6.52 (dd, J = 8.3, 2.3 Hz, 1H), 6.40 (d, J = 2.3 Hz,1H), 3.85 (q, J = 6.4 Hz, 1H), 3.73 (overlapped, 1H), 3.72 (s, 3H), 3.61 (s,3H), 3.66 (m, 1H), 2.10 (m, 2H), 1.09 (d, J = 6.4 Hz, 3H); 13 C NMR (125 MHz, DMSO) δ C 178.5, 159.8, 154.7, 143.0, 123.6, 122.8, 106.8, 96.5, 61.0, 55.4, 55.2, 51.9, 45.2, 33.6, 15.0; HR-ESI-MS m / z : 313.1180 [M+H] + (calcd for C 15 H 18 N2NaO4, 313.1159)。

[0047] Compound A-23 (3 R ,1′ R -A-23 and 3 S ,1′ S- A-23): 1 H NMR (500 MHz, CD3OD) δ H 7.20 (d, J [[ID=4O]]= 8.3 Hz, 1H), 6.60 (dd, J = 8.4, 2.4 Hz, 1H), 6.53 (d, J = 2.3 Hz, 1H), 4.00 (q, J = 6.6 Hz, 1H), 3.87 (dt, J = 10.5, 7.7 Hz, 1H), 3.80 (s, 3H), 3.73 (s, 3H), 3.66 (m, 1H), 2.26 (m, 1H), 2.18 (m, 1H), 1.18 (d, J = 6.6 Hz, 3H); 13C NMR (125MHz, CD3OD) δ C 183.1, 162.0, 157.6, 144.3, 127.1, 122.0, 107.9, 98.2, 60.2,57.5, 55.9, 53.0, 46.1, 34.4, 17.9; HR-ESI-MS m / z 313.1158 [M+Na] + (calcd forC 15 H 18 N2NaO4, 313.1159). Example 4: The role of transcription factor EB (TFEB) Hela TFEB-3xFlag cells (a gift from Professor Song Juxian of Guangzhou University of Chinese Medicine, see the literature Xu J, Ao YL, Huang C, et al. Harmol promotes α-synuclein degradation and improves motor impairment in Parkinson's models via regulating autophagy-lysosome pathway[J]. npj Parkinson's Disease[2023-06-19].DOI:10.1038 / s41531-022-00361-4) and Neuro-2a cell suspensions were seeded into 96-well plates and cultured in a 37 ℃, 5% CO2 incubator. After 24 h of adhesion, different test samples were added, and after 24 h of culture, the cells were fixed with 4% paraformaldehyde for 15 min, washed 3 times with HBSS, permeated with 0.2% Triton X-100 for 10 min, washed 3 times with HBSS, blocked with 10% FBS at room temperature for 1 h, washed once with HBSS, and Flag cells were then cultured. Incubate M2 (dilution ratio 1:500) primary antibody solution overnight at 4 ℃, wash 3 times with HBSS, incubate Alexa Fluor 594 (dilution ratio 1:800) secondary antibody and DAPI (dilution ratio 1:2000) at room temperature in the dark for 2 h, and use a high-content cell imaging analysis system to photograph, detect and calculate the percentage of nuclear translocation data.

[0048] The above samples are as follows: Control group: DMSO; Group A-1: ​​Compound A-1 at concentrations of 10, 50, and 100 μM; Group A-6: Compound A-6 at concentrations of 10, 50, and 100 μM; Group A-11: Compound A-11 at concentrations of 10, 50, and 100 μM; Group A-14: Compound A-14 at concentrations of 10, 50, and 100 μM; Group A-16: Compound A-16 at concentrations of 10, 50, and 100 μM; Group A-20: Compound A-20 at concentrations of 10, 50, and 100 μM; Total alkaloids group: The total alkaloids prepared in Example 1 had a concentration of 50 μg / mL; Rapamycin group: 0.25 μM rapamycin. Each group had 3 replicates.

[0049] Under normal conditions, phosphorylated TFEB binds to the 14-3-3 protein and remains in the cytoplasm. However, under conditions such as starvation or lysosomal stress, TFEB is no longer phosphorylated, and unphosphorylated TFEB is transported to the nucleus, promoting the transcription of its target genes, leading to the upregulation of autophagy and lysosomal genes. High-throughput imaging analysis was used to determine the subcellular localization of TFEB and to screen for TFEB activators. The results are as follows: Figure 2 As shown, total bases, A-1, A-16, and A-20 can induce TFEB nuclear translocation.

[0050] High-throughput image analysis showed consistent results in determining the subcellular localization of transfected and endogenous TFEB. Figure 3 The TFEB nuclear translocation results showed that both total bases and monomers could activate the TFEB target, with monomer compounds A-1, A-16, and A-20 promoting EC50 nuclear translocation of TFEB. 50 The values ​​were 28.32, 46.26, and 31.65 μM, respectively. Figure 4 ). Example 5 Effects on depressed mice A mouse model of depression was established using chronic unpredictable mild stress (CUMS) combined with isolation. Sixty 10-month-old adult male C57BL / 6J mice were randomly divided into five groups: control group (n=10), model group (n=10), total alkaloid group (n=10), A-1 group (n=10), A-16 group (n=10), and positive control group (n=10) using fluoxetine hydrochloride. The choice of CUMS method could be adjusted according to the actual situation. Mice were housed individually and randomly exposed to two different stressors daily, with the same stimuli not appearing consecutively, thus preventing the mice from predicting which stimulus would be applied and avoiding adaptation. The total modeling period was 28 days. The model design was as follows: Monday, restraint for 2 hours combined with 12 hours of moist bedding; Tuesday, ice water platform for 2 hours combined with 24 hours of day-night reversal; Wednesday, noise stimulation (110 dB, 30 min) combined with foot shocks (4 mA, 50 times, 5 s intervals); Thursday, light stimulation (100,000 lx, 30 min) combined with tail clamping (approximately 1 cm from the rat's tail tip, 2 min); Friday, restraint for 2 hours combined with noise stimulation (110 dB, 30 min); Saturday, moist bedding for 12 hours combined with 24 hours of day-night reversal; Sunday, tail clamping (approximately 1 cm from the rat's tail tip, 2 min) combined with foot shocks (4 mA, 50 times, 5 s intervals). This cycle was repeated for a total of 4 weeks. During the modeling period, mice were administered medication daily via gavage. The control and model groups received only 0.5% CMC-Na solution, while the total alkali group, A-1 group, and A-16 group received 20 mg / kg. The positive control group (fluoxetine hydrochloride) received 10 mg / kg. Successful modeling was assessed behaviorally; we used the sucrose preference test (SPT) to evaluate depressive behaviors in these mice. Successful model groups, compared to the control group, exhibited characteristics such as weight loss (Table 1) and reduced sucrose consumption (Table 2), mimicking the symptoms of human patients with depression. Table 1. Changes in body weight of mice in each group (m ± SD, g) Table 2. Sugar water preference index (%) of mice in each group CUMS combined with solitary rearing significantly reduced spatial exploration ability, loss of interest, anhedonia, and increased disappointment in mice, mimicking the clinical manifestations of depression. In this experiment, total alkaloids, A-1, and A-16 shortened the escape latency in mice. Figure 5 These drugs counteract despair behavior, reducing the time spent immobile in experimental animals and demonstrating an antidepressant effect. Example 6: Inhibitory effect on the immunomodulatory enzyme indoleamine-2,3-dioxygenase (IDO) Mix 100 mM potassium phosphate buffer (pH 6.5), 20 μM methylene blue, 200 μg / mL catalase, 300 mM substrate L-tryptophan, and 100 mM of each test sample. 1-Methyltryptophan is used as a positive control. Incubate the mixture at 37°C for 5 minutes, then add 0.75 U / mL IDO enzyme and react at 37°C for 30 minutes. Add 200 μL of 30% (w / v) trichloroacetic acid to terminate the reaction and heat at 65°C for 15 minutes to complete the conversion from formylkynurenine to kynurenine. Centrifuge at high speed for 10 minutes, and mix the supernatant with an equal volume of 2% (w / v) p-dimethylaminobenzaldehyde acetic acid solution. Detect the result using a microplate reader at 492 nm and calculate the inhibition rate of IDO enzyme based on the absorbance.

[0051] Table 3. Inhibitory effects of compounds A1-A23 on IDO enzymes The results are shown in Table 3: Total alkaloids and 23 monomeric alkaloids inhibited IDO activity to varying degrees. Compared with the positive control 1-methyltryptophan, total alkaloids and 15 monomeric alkaloids showed higher IDO inhibitory activity, with 8 of them exhibiting inhibition rates exceeding 50%. Example 7 Inhibitory effect on monoamine oxidase A (MAOA) and monoamine oxidase B (MAOB) Moclobemide and the monomeric compound to be tested were diluted with PBS containing 2% DMSO to prepare sample solutions with a specific concentration gradient. 50 μL of MAOA / MAOB (2.5 U / mL) and 100 μL of sample solution were added sequentially to each well of a 96-well plate. After incubation at 37°C for 10 min, 50 μL of kynurenilamide (0.2 mM) was added, and incubation continued for 30 min. Finally, 40 μL of NaOH (2 M) was added to terminate the reaction. The fluorescence intensity of each well was measured using a microplate reader at excitation wavelengths of 310 nm and emission wavelengths of 400 nm, and the inhibition rate was calculated. Based on the experimental results, the IC50 of the monomeric compound was calculated using Graphpad Prism 5. 50 value.

[0052] Table 4. Inhibitory activity of compounds against MAOA and MAOB

[0053] Note: "-" indicates no test; SI = MAOB / MAOA, indicating selectivity for MAOA. The results are shown in Table 4: 21 monomeric alkaloids inhibited MAOA activity to varying degrees. Compared with the positive control drug moclobemide, 13 monomeric alkaloids showed higher MAOA inhibitory activity, of which 7 had a selectivity for MAOA exceeding 10. The above experimental results reveal that the tryptamine indole alkaloid compounds described in this invention can be used for anti-anxiety, anti-depression, prevention and treatment of neurodegenerative diseases, and immune regulation. The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. A tryptamine indole alkaloid, characterized in that... The following compounds: banistecaines J (A-4); Its structural formula is shown below: 。 2. The use of the tryptamine indole alkaloid according to claim 1 in the preparation of IDO / MAOA inhibitors.

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