Oxacocarpon compounds and their pharmaceutical uses

Abstract

The present invention discloses novel oxaflavine compounds represented by formula I, and solvates and crystal forms thereof, pharmaceutical preparations containing the compounds, and applications of the compounds in preventing and treating diseases associated with neurodegeneration.

Description

Technical Field

[0001] The present invention relates to novel oxaflavine compounds of general formula I, and solvates and crystalline forms thereof, the use of these compounds in the treatment of neurodegenerative diseases, methods for their use in treatment, and pharmaceutical compositions containing the compounds. Background Art

[0002] Neurodegenerative diseases are multifactorial diseases, and their pathogenesis is still unclear. The pathophysiological basis of neurodegenerative diseases is secondary damage to neurons and nerve cells, which leads to cellular and molecular changes. Compounds with good neuroprotective effects show good application prospects in neurodegenerative diseases.

[0003] Fructus luteolin is an isoquinoline alkaloid, which exists in plants such as Menispermaceae, Rutaceae and Ranunculaceae. It is the main active ingredient of many commonly used Chinese medicines in my country and has been reported to have multiple pharmacological activities. Fructus luteolin has a certain neuroprotective effect in cells and shows certain effects in dementia mice, involving multiple mechanisms such as cholinesterase inhibition, amyloid formation, TAU phosphorylation, anti-inflammatory, and antioxidant. However, it also has disadvantages such as low activity, poor solubility, and poor oral bioavailability, resulting in poor drugability. At present, its structural modification is mainly concentrated on individual positions on the ring, and its skeleton is rarely changed. The present invention adopts the strategy of skeleton transition, inserts oxygen atoms at position 13, and focuses on the transformation of the skeleton of Fructus luteolin.

[0004] The present invention aims to provide a new oxaflavine compound, which has high neuroprotective activity and can be used to prevent or treat neurodegenerative diseases. Summary of the invention

[0005] The object of the present invention is to provide a new oxaflavine compound represented by formula I, and its solvate and crystalline form.

[0006] Another object of the present invention is to provide the use of the compound represented by formula I in neuroprotection, and in preventing or treating neurodegenerative diseases.

[0007] In order to achieve the purpose of the present invention, the present invention adopts the following technical solution:

[0008] Wherein, n is 1, 3, 4, 5, 6, 7, 8, A is phenyl or a chemical single bond, and R is selected from substituted or unsubstituted C1-C3 alkyl.

[0009] Another object of the present invention is to provide the compound, and its solvate and crystalline form, characterized in that the compound is selected from:

[0010]

[0011] For the preparation of pharmaceutical preparations, the compounds of general formula I can be mixed with suitable pharmaceutical carrier materials, aromatics, flavoring agents and pigments in a known manner and made into tablets or coated tablets, or suspended or dissolved in water or oil with other additives.

[0012] The present invention also relates to a pharmaceutical composition comprising a pharmaceutically effective dose of the compound of formula I and a pharmaceutically acceptable carrier.

[0013] Pharmacological studies have shown that the compounds of the general formula I of the present invention have strong neuroprotective activity, thereby achieving the purpose of treatment.

[0014] The compounds of the present invention can be administered orally or parenterally. Oral administration can be tablets, capsules, coatings, and parenteral administration forms include injections and suppositories. These preparations are prepared according to methods well known to those skilled in the art. The adjuvants used to make tablets, capsules, and coatings are conventional adjuvants, such as starch, gelatin, gum arabic, silica, polyethylene glycol, and the solvents used in liquid dosage forms include water, ethanol, propylene glycol, and vegetable oils such as corn oil, peanut oil, olive oil, etc. The preparations containing the compounds of the present invention may also contain other adjuvants, such as surfactants, lubricants, disintegrants, preservatives, flavoring agents, pigments, etc. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 Cytotoxicity of compounds L1-L14;

[0016] Figure 2 Neuroprotective effects of compounds L1-L14 on SH-SY5Y cells. DETAILED DESCRIPTION

[0017] The invention will be further described below with reference to the embodiments, but these embodiments do not limit the scope of the invention.

[0018] The structure of the compound was determined by nuclear magnetic resonance (NMR). NMR was measured using INOVA-300 and the solvent was DMSO- d 6. The internal standard is TMS, and the chemical shift is given in ppm. Column chromatography generally uses 200-300 mesh silica gel as the carrier.

[0019] Example 1: L-1

[0020]

[0021] a) Add 6,7-dimethoxy-3,4-dihydroisoquinoline (1 mmol), 5-hydroxysalicylic acid (1 mmol), EDCI (2 mmol) and DMF (15 mL) into a 100 mL round-bottom flask and react at room temperature for 12 h. After the reaction is completed, most of the DMF is evaporated, and the residue is added with dilute hydrochloric acid solution, filtered, and dried for later use.

[0022] c) The above product, ethyl bromoacetate (1.5 mmol), K2CO3 (2 mmol), and CH3CN (10 mL) were added to a 100 mL round-bottom flask in sequence and reacted at 60°C for 12 h. After the reaction was completed, the solvent was evaporated and the product was separated by column chromatography to obtain the target compound with a yield of 67.8%. 1 H NMR (400 MHz, DMSO- d 6) δ 7.29 (d, J = 3.1 Hz, 1H), 7.19 (dd, J =8.9, 3.1 Hz, 1H), 7.14 – 7.06 (m, 2H), 6.89 (s, 1H), 6.29 (s, 1H), 4.82 (s,2H), 4.35 (dt, J = 12.8, 4.1 Hz, 1H), 4.18 (q, J = 7.1 Hz, 2H), 3.79 (d, J =2.1 Hz, 6H), 3.27 – 3.16 (m, 1H), 2.94 – 2.75 (m, 2H), 1.22 (t, J = 7.1 Hz, 3H).

[0023] Example 2: L-2

[0024]

[0025] The preparation method is similar to that of Example 1, except that ethyl bromoacetate in Example 1 is replaced by methyl 6-bromohexanoate. 1 H NMR (400 MHz, DMSO- d6) δ 7.30 (d, J = 3.0 Hz, 1H), 7.14 (dd, J =8.9, 3.0 Hz, 1H), 7.09 (d, J = 8.6 Hz, 2H), 6.89 (s, 1H), 6.28 (s, 1H), 4.35(dt, J = 2.34 (t, J = 7.3Hz, 2H), 1.72 (p, J = 6.6 Hz, 2H), 1.60 (p, J = 7.4 Hz, 2H), 1.43 (h, J =7.0, 6.1 Hz, 2H).

[0026] Example 3: L-3

[0027]

[0028] The preparation method is similar to that of Example 1, except that ethyl bromoacetate is used instead of ethyl 4-bromobutyrate. 1 H NMR (400 MHz, DMSO- d 6) δ 7.30 (d, J = 2.9 Hz, 1H), 7.15 (dd, J =8.8, 3.0 Hz, 1H), 7.12 – 7.06 (m, 2H), 6.89 (s, 1H), 6.28 (s, 1H), 4.35 (dt,J = 12.9, 4.2 Hz, 1H), 4.09 (d, J = 7.1 Hz, 1H), 4.02 (q, J = 7.2, 6.4 Hz, 3H), 3.79 (d, J = 1.9 Hz, 6H), 3.27 – 3.11 (m, 1H), 2.93 – 2.75 (m, 2H), 2.46(d, J = 7.3 Hz, 2H), 1.98 (p, J = 6.9 Hz, 2H), 1.19 (t, J = 7.1 Hz, 3H).

[0029] Example 4: L-4

[0030]

[0031] The preparation method is similar to that of Example 1, except that ethyl bromoacetate in Example 1 is replaced by methyl 4-bromomethylbenzoate. 1 H NMR (400 MHz, DMSO- d 6) δ 7.99 (d, J = 7.9 Hz, 2H), 7.61 (d, J= 7.9 Hz, 2H), 7.47 – 7.21 (m, 2H), 7.21 – 7.05 (m, 2H), 6.89 (s, 1H), 6.28(s, 1H), 5.26 (s, 2H), 4.35 (dt, J = 13.0, 3.9 Hz, 1H), 3.86 (s, 3H), 3.79 (d, J = 2.4 Hz, 6H), 3.27 – 3.12 (m, 1H), 2.98 – 2.73 (m, 2H).

[0032] Example 5: L-5

[0033]

[0034] The preparation method is similar to that of Example 1, except that ethyl bromoacetate is used instead of ethyl bromoacetate in Example 1. 1 H NMR (400 MHz, DMSO- d 6) δ 7.31 (d, J = 3.0 Hz, 1H), 7.15 (dd, J =8.9, 3.0 Hz, 1H), 7.09 (d, J = 9.3 Hz, 2H), 6.89 (s, 1H), 6.28 (s, 1H), 4.35(dt, J = 12.7, 4.2 Hz, 1H), 4.05 (q, J = 7.1 Hz, 2H), 4.00 (t, J = 6.0 Hz,2H), 3.79 (d, J = 1.8 Hz, 6H), 3.24 – 3.17 (m, 1H), 2.93 – 2.77 (m, 2H), 2.37(t, J = 7.0 Hz, 2H), 1.77 – 1.65 (m, 4H), 1.18 (t, J = 7.1 Hz, 3H).

[0035] Example 6: L-6

[0036]

[0037] The preparation method is similar to that of Example 1, except that 4-hydroxysalicylic acid is used instead of 5-hydroxysalicylic acid in Example 1. 1 H NMR (400 MHz, DMSO- d 6) δ 7.77 (d, J = 8.7 Hz, 1H), 7.08 (s, 1H), 6.89 (s, 1H), 6.77 (dd, J = 8.7, 2.5 Hz, 1H), 6.70 (d, J = 2.4 Hz, 1H), 6.34(s, 1H), 4.89 (s, 2H), 4.37 – 4.30 (m, 1H), 4.18 (q, J = 7.1 Hz, 2H), 3.79 (d, J = 1.7 Hz, 6H), 3.18 (ddd, J = 12.7, 10.9, 4.0 Hz, 1H), 2.92 – 2.72 (m,2H), 1.23 (d, J = 7.1 Hz, 3H).

[0038] Example 7: L-7

[0039]

[0040] The preparation method is similar to that of Example 1, except that 4-hydroxysalicylic acid is used instead of 5-hydroxysalicylic acid in Example 1, and ethyl 4-bromobutyrate is used instead of ethyl bromoacetate in Example 1. 1 H NMR (400 MHz, DMSO- d 6) δ7.75 (d, J = 8.6 Hz, 1H), 7.09 (s, 1H), 6.89 (s, 1H), 6.76 – 6.67 (m, 2H), 6.32 (s, 1H), 4.33 (ddd, J = 12.7, 4.8, 3.3 Hz, 1H), 4.10 – 4.04 (m, 4H), 3.79 (s, 6H), 3.18 (ddd, J = 12.8, 10.9, 3.9 Hz, 1H), 2.88 (ddd, J = 15.8,10.9, 4.9 Hz, 1H), 2.82 – 2.73 (m, 1H), 2.46 (t, J = 7.3 Hz, 2H), 1.98 (p, J = 6.9 Hz, 2H), 1.18 (t, J = 7.1 Hz, 3H).

[0041] Example 8: L-8

[0042]

[0043] The preparation method is similar to that of Example 1, except that 4-hydroxysalicylic acid is used instead of 5-hydroxysalicylic acid in Example 1, and ethyl 5-bromovalerate is used instead of ethyl bromoacetate in Example 1. 1 H NMR (400 MHz, DMSO- d 6) δ7.76 (s, 1H), 7.09 (s, 1H), 6.89 (s, 1H), 6.77 – 6.66 (m, 2H), 6.32 (s, 1H), 4.33 (dt, J = 12.8, 4.1 Hz, 1H), 4.05 (td, J = 7.1, 6.5, 4.0 Hz, 4H), 3.80(s, 6H), 3.18 (ddd, J = 12.7, 10.9, 4.0 Hz, 1H), 2.91 – 2.71 (m, 2H), 2.37(t, J = 7.2 Hz, 2H), 1.76 – 1.65 (m, 4H), 1.17 (d, J = 7.1 Hz, 3H).

[0044] Example 9: L-9

[0045]

[0046] The preparation method is similar to that of Example 1, except that 4-hydroxysalicylic acid is used to replace 5-hydroxysalicylic acid in Example 1, and 6-bromohexanoic acid methyl ester is used to replace ethyl bromoacetate in Example 1. 1 H NMR (400 MHz, DMSO- d 6) δ 7.74 (d, J =8.6 Hz, 1H), 7.09 (s, 1H), 6.89 (s, 1H), 6.77 – 6.65 (m, 2H), 6.31 (s, 1H), 4.40 – 4.24 (m, 1H), 4.03 (t, J = 6.5 Hz, 2H), 3.80 (s, 6H), 3.58 (s, 3H), 3.18 (ddd, J = 12.8, 10.9, 3.9 Hz, 1H), 2.94 – 2.73 (m, 2H), 2.33 (t, J = 7.3Hz, 2H), 1.72 (p, J = 6.7 Hz, 2H), 1.62 – 1.55 (m, 2H), 1.42 (td, J = 8.5, 4.3 Hz, 2H).

[0047] Example 10: L-10

[0048]

[0049] The preparation method is similar to that of Example 1, except that 4-nitrosalicylic acid is used instead of 5-nitrosalicylic acid in Example 1, and methyl 4-bromomethylbenzoate is used instead of ethyl bromoacetate in Example 1. 1 H NMR (400 MHz, DMSO- d 6) δ 8.04 – 7.96 (m, 2H), 7.78 (d, J = 8.6 Hz, 1H), 7.62 – 7.56 (m, 2H), 7.08 (s, 1H), 6.89 (s, 1H), 6.87 – 6.80 (m, 2H), 6.34 (s, 1H), 5.30 (s, 2H), 4.34 (dt, J = 12.9, 4.4 Hz, 1H), 3.86 (s, 3H), 3.79 (d, J = 1.5 Hz, 6H), 3.23– 3.13 (m, 1H), 2.92 – 2.69 (m, 2H).

[0050] Example 11: L-11

[0051]

[0052] The preparation method is similar to that of Example 1, except that ethyl bromoacetate is used instead of ethyl 7-bromoheptanoate. 1 H NMR (400 MHz, DMSO- d6) δ 7.30 (d, J = 3.0 Hz, 1H), 7.14 (dd, J =8.9, 3.0 Hz, 1H), 7.09 (d, J = 8.2 Hz, 2H), 6.89 (s, 1H), 6.27 (s, 1H), 4.35(ddd, J = 12.8, 4.7, 3.3 Hz, 1H), 4.05 (q, J = 7.2 Hz, 2H), 3.97 (t, J = 6.5Hz, 2H), 3.79 (d, J = 1.8 Hz, 6H), 3.22 (ddd, J = 12.8, 10.9, 4.0 Hz, 1H),2.95 – 2.72 (m, 2H), 2.29 (t, J = 7.4 Hz, 2H), 1.71 (p, J = 6.6 Hz, 2H), 1.59– 1.52 (m, 2H), 1.42 (t, J = 8.1 Hz, 2H), 1.37 – 1.28 (m, 2H), 1.17 (t, J =7.1 Hz, 3H).

[0053] Example 12: L-12

[0054]

[0055] The preparation method is similar to that of Example 1, except that ethyl bromoacetate is used instead of ethyl bromoacetate in Example 1. 1 H NMR (400 MHz, DMSO- d6) δ 7.30 (d, J = 3.0 Hz, 1H), 7.14 (dd, J =8.9, 3.0 Hz, 1H), 7.12 – 7.05 (m, 2H), 6.89 (s, 1H), 6.27 (s, 1H), 4.35 (ddd,J = 12.8, 4.7, 3.3 Hz, 1H), 4.04 (q, J = 7.1 Hz, 2H), 3.98 (t, J = 6.5 Hz, 2H), 3.79 (d, J = 1.8 Hz, 6H), 3.22 (ddd, J = 12.7, 10.8, 4.0 Hz, 1H), 2.95 –2.75 (m, 2H), 2.28 (t, J = 7.4 Hz, 2H), 1.71 (p, J = 6.6 Hz, 2H), 1.54 (q, J= 7.2 Hz, 2H), 1.44 – 1.37 (m, 2H), 1.36 – 1.27 (m, 4H), 1.17 (t, J = 7.1 Hz,3H).

[0056] Example 13: L-13

[0057]

[0058] The preparation method is similar to that of Example 1, except that 4-nitrosalicylic acid is used instead of 5-nitrosalicylic acid in Example 1, and ethyl 7-bromoheptanoate is used instead of ethyl bromoacetate in Example 1. 1 H NMR (400 MHz, DMSO- d6) δ7.74 (d, J = 8.6 Hz, 1H), 7.09 (s, 1H), 6.89 (s, 1H), 6.77 – 6.66 (m, 2H), 6.31 (s, 1H), 4.33 (dt, J = 12.7, 4.0 Hz, 1H), 4.04 (d, J = 7.3 Hz, 4H), 3.80(s, 6H), 3.18 (td, J = 12.8, 11.8, 3.9 Hz, 1H), 2.92 – 2.70 (m, 2H), 2.28(td, J = 7.4, 4.8 Hz, 3H), 1.73 (dt, J = 14.3, 7.0 Hz, 2H), 1.58 – 1.52 (m,2H), 1.40 (q, J = 11.9, 9.2 Hz, 3H), 1.18 – 1.15 (m, 3H).

[0059] Example 14: L-14

[0060]

[0061] The preparation method is similar to that of Example 1, except that 4-nitrosalicylic acid is used instead of 5-nitrosalicylic acid in Example 1, and ethyl 8-bromooctanoate is used instead of ethyl bromoacetate in Example 1. 1 H NMR (400 MHz, DMSO- d 6) δ7.74 (d, J = 8.7 Hz, 1H), 7.09 (s, 1H), 6.89 (s, 1H), 6.73 (dd, J = 8.7, 2.4Hz, 1H), 6.68 (d, J = 2.4 Hz, 1H), 6.31 (s, 1H), 4.33 (ddd, J = 12.8, 4.8,3.3 Hz, 1H), 4.10 – 3.95 (m, 5H), 3.80 (s, 6H), 3.18 (ddd, J = 12.7, 10.9,4.0 Hz, 1H), 2.90 – 2.74 (m, 2H), 2.27 (t, J = 7.4 Hz, 2H), 1.71 (t, J = 7.3Hz, 2H), 1.56 – 1.50 (m, 2H), 1.40 (s, 1H), 1.34 – 1.27 (m, 4H), 1.18 (d, J =7.1 Hz, 3H).

[0062] Pharmacological experiments

[0063] Experimental Example 1: The cytotoxicity of the compound of the present invention is low

[0064] Human neuroblastoma cells SH-SY5Y and human liver cancer cells HepaG2 were seeded in 96-well plates, with 10,000 cells per well, and the cells were cultured overnight. 14 compounds were added to the cells at final concentrations of 1, 10, and 50 μg / mL, with the addition of solvent DMSO as a negative control and the same concentration of evodiamine as a positive control (6 replicates for each drug). After the cells were cultured for 24 hours, the cell activity was analyzed using the CCK-8 kit ( Figure 1 AB). The results showed that many compounds showed low toxicity in SH-SY5Y and HepaG2.

[0065] Experimental Example 2: Neuroprotective effect of the compounds of the present invention on SH-SY5Y cells

[0066] Human neuroblastoma SH-SY5Y cells were seeded in 96-well plates, with 10,000 cells per well, and the cells were cultured overnight. 100 μM hydrogen peroxide and 14 compounds were added to the cells, with final concentrations of 0.001, 0.01, and 0.1 μg / mL, respectively. The addition of solvent DMSO was used as a negative control, and the same concentration of evodiamine was used as a positive control (6 replicates for each drug). After the cells were cultured for another 24 hours, the cell activity was analyzed using the CCK-8 kit ( Figure 2 ). The results showed that a variety of compounds exhibited antioxidant capacity and played a neuroprotective role.

Claims

1. An oxaflavine compound selected from: 。 2. A pharmaceutical composition comprising an effective dose of the compound as claimed in claim 1 and a pharmaceutically acceptable carrier.

3. The pharmaceutical composition according to claim 2, characterized in that The pharmaceutical composition is selected from tablets, capsules, pills, injections, sustained-release preparations, controlled-release preparations or various microparticle delivery systems.

4. Use of the compound according to claim 1 in the preparation of a medicament for preventing or treating a neurodegenerative disease.

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