Evodiamine compounds and pharmaceutical uses thereof

Abstract

The application belongs to the technical field of medicine, and discloses evodiamine compounds and pharmaceutical uses thereof, and provides evodiamine compounds shown in formula (I), and their physiologically acceptable salts and pharmaceutical compositions. The compounds have good inhibitory activity on HDAC6, and can be used for preventing or treating HDAC6 related diseases.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology and relates to a class of evodiamine compounds and their physiologically acceptable salts. It also relates to the use of these compounds in the preparation of treatments for HDAC6 inhibitor-related diseases, and pharmaceutical compositions containing said compounds. Background Technology

[0002] Histone deacetylases (HDACs) maintain homeostasis in the body by deacetylifying histones, regulating gene expression, cell survival, and differentiation. To date, 18 HDAC isoenzymes have been identified, classified into four classes: Class I (HDAC1, 2, 3, 8), Class IIa (HDAC4, 5, 7, 9), Class IIb (HDAC6, 10), and Class IV (HDAC11). The first three classes are zinc-dependent enzymes formed by the folding of arginine deacetylases, while Class III HDACs (sirtuins 1-7) are NAD+-dependent enzymes. HDAC6 is the largest member of the HDAC family, composed of 1215 amino acid residues, possessing two independent catalytic domains (CD1 and CD2) located at the N-terminus and central region, respectively, and a zinc finger domain at its C-terminus. HDAC6 is involved in a wide range of diseases, including tumors, neurodegenerative diseases, inflammation, bacterial infections, and autoimmune diseases. Given the unique structure and physiological function of HDAC6, the development of selective inhibitors of HDAC6 has attracted much attention.

[0003] To date, several HDAC6 inhibitors with different structural types have been reported in the literature, but they suffer from poor selectivity and unsatisfactory pharmacokinetic properties. Evodia rutaecarpa is an alkaloid and one of the main active components of the traditional important herb *Evodia rutaecarpa*, possessing various pharmacological activities. The inventors of this application, through structural analysis of evodia rutaecarpa and Tubastatin-A, have linked the hydroxamic acid fragment of Tubastatin-A to the structure of evodia rutaecarpa, aiming to provide a new and effective selective HDAC6 inhibitor, with the potential application in the treatment of tumors, neurodegenerative diseases, inflammation, bacterial infections, autoimmune diseases, etc. Summary of the Invention

[0004] The technical problem solved by the present invention is to provide an erythropoietin compound represented by general formula (I), a pharmaceutical composition thereof, and its use in the preparation of HDAC6 inhibitors and in the preparation of drugs for treating HDAC6-related diseases.

[0005] To solve the technical problem of this invention, the present invention provides the following technical solution:

[0006] The first aspect of the present invention is to provide a compound of general formula (I) or a physiologically acceptable salt thereof:

[0007]

[0008] Where X is a phenyl group or a chemical single bond, and n is 1, 3, 4, 5, 6, 7, or 8;

[0009] The most preferred compounds are those described below:

[0010]

[0011]

[0012] The second aspect of the present invention is to provide a method for preparing the compound described in the first aspect: Formula (II) reacts with a halogenated ester under the action of a base to obtain Formula (III); Formula (III) reacts with hydroxylamine under the action of a base to obtain Formula (I).

[0013]

[0014] A third aspect of the present invention is to provide a pharmaceutical composition containing the compound described in the first aspect and its physiologically acceptable salts.

[0015] To prepare a pharmaceutical preparation, a compound of general formula (I) may be mixed with a suitable pharmaceutical carrier substance, flavoring agent, flavoring agent and pigment by a known method and made into a tablet or coated tablet, or suspended or dissolved in water or oil with other additives.

[0016] The present invention also relates to a pharmaceutical composition comprising an effective dose of a compound as described in general formula I and a pharmaceutically acceptable carrier.

[0017] The compounds of this invention can be administered orally or via non-gastrointestinal routes. Oral administration may include tablets, capsules, or coated formulations; non-gastrointestinal dosage forms include injections and suppositories. These formulations are prepared according to methods well known to those skilled in the art. The excipients used in the manufacture of tablets, capsules, and coated formulations are conventionally used adjuvants, such as starch, gelatin, gum arabic, silica, and polyethylene glycol; solvents used in liquid dosage forms include, for example, water, ethanol, propylene glycol, and vegetable oils such as corn oil, peanut oil, and olive oil. Formulations containing the compounds of this invention may also contain other adjuvants, such as surfactants, lubricants, disintegrants, preservatives, flavoring agents, and colorants.

[0018] The fourth aspect of this invention provides the use of the compounds described in the first aspect and their physiologically acceptable salts in the preparation of HDAC6 inhibitors, and their use in the preparation of medicaments for the prevention or treatment of HDAC6-related diseases. The diseases are selected from tumors, neurodegenerative diseases, inflammation, bacterial infections, and autoimmune diseases.

[0019] Beneficial technical effects:

[0020] The compounds of this invention exhibit good inhibitory activity against HDAC6, reaching the nM level, and have a higher selectivity index compared to HDAC1, which is expected to provide a safe and effective selective inhibitor of HDAC6 for the treatment of related diseases. Detailed Implementation

[0021] The invention will be further described below with reference to the embodiments, but these embodiments do not limit the scope of the invention.

[0022] The structure of the compound was determined by nuclear magnetic resonance (NMR), mass spectrometry (MS), or high-resolution mass spectrometry (HRMS). NMR shifts (δ) are given in parts per million (ppm). mp is the melting point given in °C, uncorrected for temperature. Column chromatography typically uses 200–300 mesh silica gel as the support. NMR measurements were performed using an INOVA-400 analyzer with CDCl3 and DMSO-D6 as solvents and TMS as the internal standard. Chemical shifts are given in ppm. MS measurements were performed using an Agilent LC / MSD TOF liquid chromatography-mass spectrometry system.

[0023] Example 1: Preparation of TM-1

[0024]

[0025] 1) Tryptophan (4.8 g, 30 mmol) was dissolved in ethyl formate (11.1 g, 15 mmol), and the mixture was refluxed at 60 °C for 20 h. After the reaction was complete, the solvent was evaporated, and the product was obtained by column chromatography, yielding 3.8 g of the product (79% yield).

[0026] 2) N-(2-(1H-indol-3-yl)ethyl)formamide (3.8 g, 20 mmol) was dissolved in 50 mL of anhydrous dichloromethane. The solution was cooled to approximately 5 °C using a cold hydrazine filter. POCl3 (15.5 g, 101 mmol) was slowly added dropwise using a constant pressure funnel. The reaction was carried out in an ice bath for 2 h, followed by a further increase in temperature to room temperature for 2 h. After the reaction was complete, most of the solvent was evaporated, and 10% glacial acetic acid was added dropwise until no more bubbles were observed. The filtrate was collected by suction filtration, and ammonia was added. The solid was collected by suction filtration again. The product was washed several times with water and dried to obtain 2.9 g of product, with a yield of 76%.

[0027] 3) 4,9-dihydro-3H-pyrido[3,4-b]indole (510 mg, 3 mmol), salicylic acid (414 mg, 3 mmol), and EDCI (864 mg, 4.5 mmol) were dissolved in 30 mL of anhydrous dichloromethane and reacted at room temperature for 12 h. After the reaction was complete, salicylic acid was removed with NaHCO3, and EDCI was removed with water. After extraction with dichloromethane and water, the product was obtained by column chromatography, yielding 444 mg of the product (87% yield).

[0028] 4) 7,8,13,13b-tetrahydro-5H-benzo[5',6'][1,3]oxazino[3',2':1,2]pyrido[3,4-b]indol-5-one (290 mg, 1 mmol) was dissolved in 10 mL of anhydrous DMF, followed by the addition of NaH (100 mg, 4.2 mmol). The mixture was kept under argon protection at room temperature for 10 min, then methyl 4-bromomethylbenzoate (458 mg, 2 mmol) was added, and the reaction was carried out at 100 °C for 6 h. After the reaction was complete, most of the solvent was evaporated, and the mixture was extracted with ethyl acetate and water. The product was obtained by column chromatography, yielding 246 mg of product (85% yield).

[0029] 5) Weigh 20 mg of NaOH into a two-necked flask and add 0.1 ml of NH₂OH. Under argon protection at 0°C, add 44 mg (0.2 mmol) of methyl 4-((5-oxo-8,13b-dihydro-5H-benzo[5',6'][1,3]oxazine[3',2':1,2]pyridin[3,4-b]indol-13(7H)-yl)methyl)benzoate in 5 ml of a mixed solution (THF:MeOH = 1:1). React at room temperature. After the reaction is complete, evaporate the solvent, adjust the pH to 7-8 at 0°C, and allow to stand. A white solid precipitates. Filter to obtain a white solid with a yield of approximately 59%. mp: 165-166°C. 1 H NMR(400MHz,DMSO-d6)δ10.96(s,1H),8.97(s,1H),7.86–7.79(m,1H),7.69– 7.57(m,3H),7.52–7.47(m,1H),7.45–7.39(m,1H),7.21–7.06(m,5H),6.79– 6.71(m,1H),6.69(s,1H),5.66–5.52(m,2H),4.77–4.65(m,1H),3.23–3.12( m,1H),3.07–2.82(m,1H),2.93–2.82(m,1H).HR-ESI-MS: m / z=440.1606[M+H] + calcd for C 26 H 22 O4N3: 440.1605.

[0030] Example 2: Preparation of TM-2

[0031]

[0032] Its synthesis method and operation are the same as TM-1, except that methyl 4-bromomethylbenzoate is replaced with methyl 7-bromoheptanoate, with a yield of approximately 98%. mp: 173-175℃. 1 H NMR(400MHz,DMSO-d6)δ10.27(s,1H),8.60(s,1H),7.94–7.81(m,1H),7.62–7.47 (m,3H),7.25–7.18(m,2H),7.11–7.03(m,2H),6.78(s,1H),4.73–4.66(m,1H),4. 23–4.15(m,2H),3.23–3.09(m,1H),2.98–2.79(m,2H),1.95–1.84(m,2H),1.82–1 .72(m,2H),1.52–1.40(m,2H),1.39–1.20(m,4H).HR-ESI-MS:m / z=432.1908[M+H] + calcd for C 25 H 26 O4N3:432.1918.

[0033] Example 3: Preparation of TM-3

[0034]

[0035] The synthesis method and operation are the same as TM-1, except that methyl 4-bromomethylbenzoate is replaced with methyl 5-bromopentanoate, with a yield of approximately 22%. mp: 125-126℃. 1 H NMR(400MHz,DMSO-d6)δ10.31(s,1H),8.62(s,1H),7.89–7.83(m,1H),7.59–7.53 (m,2H),7.52–7.48(m,1H),7.24–7.16(m,3H),7.09–7.03(m,1H),6.77(s,1H),4. 75–4.65(m,1H),4.29–4.09(m,2H),3.16–3.07(m,1H),2.95–2.76(m,2H),2.04–1 .92(m,2H),1.82–1.71(m,2H),1.62–1.52(m,2H).HR-ESI-MS:m / z=404.1598[M+H] + calcd for C 23 H22 O4N3: 404.1605.

[0036] Example 4: Preparation of TM-4

[0037]

[0038] The synthesis method and operation are the same as TM-1, except that methyl 4-bromomethylbenzoate is replaced with methyl 4-bromobutyrate, with a yield of approximately 46%. mp: 190-191℃. 1 H NMR (400MHz, DMSO-d6) δ10.38(s,1H),8.70(s,1H),7.89–7.86(m,1H),7.59–7.51(m,3H),7.26–7.16(m,3H),7.10–7.05(m,1H),6.77( s,1H),4.77–4.64(m,1H),4.25–4.14(m,2H),3.22–3.08(m,1H),2.94–2.82(m,2H),2.13–1.96(m,4H).HR-ESI-MS: m / z=390.1447[M+H] + calcd for C 22 H 20 O4N3: 390.1448.

[0039] Example 5: Preparation of TM-5

[0040]

[0041] The synthesis method and operation are the same as TM-1, except that methyl 4-bromomethylbenzoate is replaced with methyl 6-bromohexanoate, with a yield of approximately 72%. mp: 112-113℃. 1 H NMR(400MHz,DMSO-d6)δ10.29(s,1H),8.62(s,1H),7.91–7.85(m,1H),7.60–7.53(m,2 H),7.54–7.48(m,1H),7.25–7.18(m,2H),7.12–7.03(m,2H),6.79(s,1H),4.77–4.64( m,1H),4.27–4.16(m,2H),3.16–3.08(m,1H),2.96–2.80(m,2H),1.94–1.89(m,2H),1. 82–1.73(m,2H),1.56–1.47(m,2H),1.34–1.28(m,2H).HR-ESI-MS: m / z=390.1447[M+H] + calcd for C 24H 24 O4N3: 418.1761.

[0042] Example 6: Preparation of TM-6

[0043]

[0044] The synthesis method and operation are the same as TM-1, except that methyl 4-bromomethylbenzoate is replaced with methyl 8-octylheptanoate, with a yield of approximately 97%. mp: 108-109℃. 1 H NMR(400MHz,DMSO-d6)δ10.25(s,1H),8.58(s,1H),7.90–7.83(m,1H),7.61–7.54(m,2H),7 .52–7.45(m,1H),7.24–7.15(m,2H),7.10–7.03(m,2H),6.77(s,1H),4.77–4.64(m,1H),4. 32–4.11(m,2H),3.17–3.06(m,1H),2.95–2.77(m,2H),1.89–1.82(m,2H),1.80–1.73(m,2H ),1.49–1.35(m,2H),1.34–1.24(m,4H),1.23–1.15(m,2H).HR-ESI-MS: m / z=448.2242[M+H] + calcd for C 26 H 30 O4 N3:448.2231.

[0045] Example 7: Preparation of TM-7

[0046]

[0047] The synthesis method and operation are the same as TM-1, except that methyl 4-bromomethylbenzoate is replaced with methyl 9-bromononanoate, with a yield of approximately 58%. mp: 153-154℃. 1H NMR(400MHz,DMSO-d6)δ10.25(s,1H),8.59(s,1H),7.90–7.85(m,1H),7.61–7.54 (m,2H),7.51–7.46(m,1H),7.25–7.17(m,2H),7.10–7.04(m,2H),6.78(s,1H),4.7 9–4.56(m,1H),4.26–4.10(m,2H),3.17–3.09(m,1H),2.95–2.77(m,2H),1.90–1. 83(m,2H),1.82–1.74(m,2H),1.45–1.14(m,10H).HR-ESI-MS:m / z=462.2378[M+H] + calcd for C 27 H 32 O4N3:462.2387418.1761.

[0048] Pharmacological experiments:

[0049] Experimental Example 1: In vitro enzyme inhibitory activity test

[0050] Use a 1X experimental buffer containing 50 mM Tris (pH 7.5), 0.01% Tween-20, and 50 mM NaCl. Gradient dissolution of the compound: Transfer 250 nL of the 100X concentration of the drug dissolved in 100% DMSO to the experimental plate. After sonication to dissolve the compound, dilute it in the experimental plate to a final DMSO concentration of 1%. Enzyme solution preparation: Prepare enzyme solutions using the 1X experimental buffer: HDAC1: 10 nM, HDAC6: 5 nM. Substrate solution preparation: Prepare an HDAC6 substrate solution containing 0.125 μM trypsin and 20 μM Ac-peptide using the 1X experimental buffer. Prepare an HDAC6 substrate solution containing 0.025 μM trypsin and 27.5 μM Ac-peptide using the 1X experimental buffer.

[0051] Add 15 μL of enzyme solution to each well of the experimental plate, and 15 μL of 1X experimental buffer to the control group. Incubate at room temperature for 15 min, then add 10 μL of substrate solution to each well of the experimental plate to start the reaction. Dynamically monitor fluorescence intensity using a microplate reader: excitation light: 355 nm, emission light: 460 nm.

[0052] Data calculation: Inhibition rate was calculated using Excel: Inh% = (Max - Signal) / (Max - Min) * 100. IC was calculated using XL-Fit. 50 :Y=Bottom+(Top-Bottom) / (1+(IC50 / X)*HillSlope). Where Y is the inhibition rate and X is the compound concentration. Table 1 shows the in vitro enzyme inhibitory activity results of the compounds of the present invention.

[0053] Table 1. In vitro enzyme inhibitory activities of the compounds

[0054]

[0055]

Claims

1. A rutinoid compound represented by the following general formula (I) and its physiologically acceptable salt, in, X is a phenyl group or a chemical single bond, and n is 1, 3, 4, 5, 6, 7, or 8.

2. The compound according to claim 1 and its physiologically acceptable salt, characterized in that, The compounds mentioned are selected from:

3. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises an effective dose of any compound as described in any one of claims 1 to 2, and a physiologically acceptable salt and a pharmaceutically acceptable carrier thereof.

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

5. The use of the compound of any one of claims 1 to 2 or a physiologically acceptable salt thereof in the preparation of an HDAC6 inhibitor.

6. The use of the compound of any one of claims 1 to 2 or a physiologically acceptable salt thereof in the preparation of a medicament for the prevention or treatment of HDAC6-related diseases.

7. The application according to claim 6, characterized in that, The diseases mentioned are selected from tumors, neurodegenerative diseases, inflammation, bacterial infections, and autoimmune diseases.

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