A pyrimido[2,1-a:4,3-a']diisoquinoline compound and a preparation method and application thereof

Abstract

This invention provides a pyrimido[2,1-a:4,3-a']diisoquinoline compound, its preparation method, and its application, belonging to the field of biochemistry. The pyrimido[2,1-a:4,3-a']diisoquinoline compound has the structure shown in Formula I: This invention utilizes arylformylformaldehyde to incorporate a large number of substituents, thereby synthesizing diverse substituted pyrimido[2,1-a:4,3-a']diisoquinoline compounds, which have an inhibitory effect on the secretion of inflammatory factors.

Description

A pyrimido[2,1-a:4,3-a′]diisoquinoline compound, its preparation method and application Technical Field

[0001] This invention relates to the field of biochemistry, and in particular to a pyrimido[2,1-a:4,3-a′] diisoquinoline compound, its preparation method, and its application. Background Technology

[0002] Hydrogenated isoquinolines are an important class of organic molecules with broad pharmacological activities, and their structural diversity and biological activity have attracted much attention in the field of medicinal chemistry. Studies have shown that these compounds exhibit significant effects in antitumor, antiviral, leukemia treatment, lipid-lowering, antibacterial, and antimalarial applications. For example, tetrahydroproberberine alkaloids (THPBs), as derivatives of hydrogenated isoquinolines, have shown analgesic, lipid-regulating, and anti-neuropsychiatric effects in the treatment of central nervous system diseases, with their regulatory effects on dopamine receptors attracting particular research interest. Furthermore, compounds obtained through structural modification (such as DC371739) have shown good in vitro and in vivo lipid-lowering activity and have completed phase I clinical trials, demonstrating excellent tolerability. The alkaloid stepholidine isolated from *Euphorbia lathyris* and its analogues (such as compounds 12a and 23b) have also been proven to be highly efficient D1R and D3R ligands, further expanding the application potential of hydrogenated isoquinolines.

[0003] Although the pharmacological activities of hydrogenated isoquinoline compounds have been extensively studied, their structural complexity and limitations in synthetic methods restrict their further development. Traditional synthetic methods are typically cumbersome, yield-poor, and difficult to introduce diverse substituents, thus limiting structural modification and activity optimization.

[0004] Therefore, developing an efficient and simple synthetic method to construct novel and functionally diverse hydrogenated isoquinoline derivatives has become a current research focus. Summary of the Invention

[0005] In view of this, the present invention aims to provide a pyrimido[2,1-a:4,3-a′] diisoquinoline compound, its preparation method and application.

[0006] To achieve the above objectives, the present invention provides the following technical solution:

[0007] One of the technical solutions of this invention is a pyrimido[2,1-a:4,3-a′]diisoquinoline compound having the structure shown in Formula I:

[0008]

[0009] Among them, R 1 =H, Cl or Br, Ar is a substituted or unsubstituted benzene ring, naphthyl ring, biphenyl, fluorene ring, thiophene, pyridine, furan, benzothiophene or benzofuran; the substitution refers to being substituted by alkyl, alkoxy, thiomethyl or halogen; the alkyl is C1-C4 alkyl, the alkoxy is C1-C4 alkoxy.

[0010] In a preferred embodiment of the present invention, the halogen is F, Cl, Br or I.

[0011] In a preferred embodiment of the present invention, the structural formula of the pyrimido[2,1-a:4,3-a′]diisoquinoline compound is shown below:

[0012]

[0013]

[0014]

[0015] The second technical solution of the present invention is a method for preparing the above-mentioned pyrimido[2,1-a:4,3-a′]diisoquinoline compounds, wherein compound 1, compound 2 and acetic acid are uniformly dispersed in a solvent and then reacted, and the reaction process is monitored by thin-layer chromatography-mass spectrometry.

[0016] The structural formula of compound 1 is: The structural formula of compound 2 is

[0017] In compounds 1 and 2, R 1 Ar and R in the above-mentioned pyrimidine[2,1-a:4,3-a′]diisoquinoline compounds 1 Same as Ar.

[0018] In a preferred embodiment of the present invention, the molar ratio of compound 1 to compound 2 is 1:1.5; the solvent is acetonitrile; the reaction is a sealed tube reaction, the reaction temperature is 60°C, and the reaction time is 6 hours.

[0019] In this invention, when R 1 When H = H, the general formula for the above reaction is:

[0020]

[0021] When R 1 When Cl or Br is present, the general formula for the above reaction is:

[0022]

[0023] The third technical solution of the present invention is the application of the above-mentioned pyrimido[2,1-a:4,3-a′]diisoquinoline compound in the preparation of anti-inflammatory drugs.

[0024] The fourth technical solution of the present invention is an anti-inflammatory drug, the raw materials of which include the above-mentioned pyrimidine[2,1-a:4,3-a′]diisoquinoline compounds and pharmaceutically acceptable excipients.

[0025] The fifth technical solution of the present invention is the application of the above-mentioned pyrimido[2,1-a:4,3-a′]diisoquinoline compound in the preparation of delayed fluorescence molecules.

[0026] The sixth technical solution of this invention is a method for preparing a delayed fluorescence molecule, which involves condensing and cyclizing the above-mentioned pyrimido[2,1-a:4,3-a′]diisoquinoline compound with hydrazine hydrate to obtain a pyrimidopyridazine compound; then, the pyrimidopyridazine compound undergoes a CN coupling reaction with a donor unit under palladium acetate catalysis to obtain the delayed fluorescence molecule; the donor unit is a phenothiazine or a phenoxazine.

[0027] The seventh technical solution of the present invention is a delayed fluorescence molecule prepared by the above preparation method.

[0028] Delayed fluorescence materials generally possess long luminescence lifetimes and high luminescence quantum yields, and have wide applications in fields such as bioimaging (including lifetime imaging), temperature / oxygen sensing, and photodynamic therapy. Based on this synthetic strategy, this invention derivatizes the target molecule to prepare a series of organic light-emitting molecules, demonstrating that the reaction strategy of derivatizing the pyrimido[2,1-a:4,3-a′]diisoquinoline compound to synthesize organic light-emitting molecules is feasible and can be further used to construct organic light-emitting molecules.

[0029] The present invention discloses the following technical effects:

[0030] This invention utilizes arylformylformaldehyde to incorporate a large number of substituents, thereby synthesizing diverse substituted pyrimido[2,1-a:4,3-a′]diisoquinoline compounds, which have an inhibitory effect on the secretion of inflammatory factors.

[0031] The preparation method of pyrimido[2,1-a:4,3-a′]diisoquinoline compounds in this invention is simple and can be synthesized in one pot in one step.

[0032] The pyrimido[2,1-a:4,3-a′]diisoquinoline compounds provided by this invention can be used to synthesize a series of organic light-emitting molecules through derivatization. Attached Figure Description

[0033] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0034] Figure 1 shows the effect of different drug concentrations on the cell viability of Raw264.7.

[0035] Figure 2 shows the effect of different drug concentrations on the cell viability of Raw 264.7 cells after LPS induction;

[0036] Figure 3 shows the effect of different drug concentrations on the NO secretion content of Raw 264.7 after LPS induction;

[0037] Figure 4 shows the effect of different drug concentrations on the TNF-α secretion content of Raw264.7 after LPS induction;

[0038] Figure 5 shows the effect of different drug concentrations on the IL-6 secretion level of Raw264.7 after LPS induction;

[0039] Figure 6 shows the effect of different drug concentrations on the IL-1β secretion level of Raw264.7 after LPS induction;

[0040] Figure 7 shows the UV-Vis absorption and fluorescence spectra of compounds 6(a) and 7(b) in o-xylene;

[0041] Figure 8 shows the fluorescence emission spectra of compounds 6(a) and 7(b) in o-xylene under argon and air atmospheres at room temperature;

[0042] Figure 9 shows the fluorescence emission of compounds 6(a) and 7(b) with the reaction of THF / water mixed solution (f w The curve of change. Detailed Implementation

[0043] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0044] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0045] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0046] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0047] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0048] Unless otherwise specified, the technical solutions described in this invention are all conventional solutions in the field, and the reagents or raw materials used are all purchased from commercial channels or are publicly available unless otherwise specified.

[0049] To better understand the present invention, the following embodiments further illustrate the content of the present invention, but the content of the present invention is not limited to the following embodiments.

[0050] Example 1

[0051] 1,2,3,4-Tetrahydroisoquinoline 1a (1.0 mmol), benzoylcarbaldehyde 2a (1.5 mmol), and 5 equiv acetic acid were dispersed in MeCN (6 mL) solvent. The reaction mixture was sealed in a tube at 60 °C for 6 hours, and the reaction was monitored by thin-layer chromatography-mass spectrometry. After the reaction was completed, the reaction solution was concentrated by vacuum distillation in a rotary evaporator. The remaining acetic acid was quenched by adding saturated sodium carbonate solution, and an equal volume of ethyl acetate was added for extraction using a 150 mL separatory funnel. The aqueous phase was extracted twice more with an equal volume of ethyl acetate. The combined organic phases were extracted with an equal volume of saturated sodium chloride solution, and the aqueous phase was discarded. After three extractions, the organic phase was dried with anhydrous sodium sulfate, allowed to stand for one hour, concentrated by vacuum distillation, and purified by silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 10:1 (v / v)) to give a yellow solid 3aa (186 mg, 5%).

[0052]

[0053] The other compounds were synthesized using the same method as in Example 1, with only the corresponding raw materials replaced to prepare the compounds listed in Table 1.

[0054] Representative synthesized pyrimido[2,1-a:4,3-a′]diisoquinoline compounds and their 1H and 1C NMR spectra are shown in Table 1.

[0055] Table 1. 1H and 1C NMR spectra of representative compounds

[0056]

[0057]

[0058]

[0059] Example 2: Verification of anti-inflammatory activity

[0060] The inflammatory inhibitory capabilities of the seven representative compounds 3aa-3ba in Table 1 of Example 1 at the cellular level were investigated. A three-dimensional evaluation system was used to systematically analyze the regulatory effects of the drugs on the RAW 264.7 macrophage inflammation model.

[0061] (1) Basic cytotoxicity assessment: The CCK-8 assay was used to detect the effect of different drug concentrations on the survival rate of normal and lipopolysaccharide (LPS) stimulated cells, revealing the drug safety window;

[0062] Under normal conditions, cell survival rates are shown in Figure 1. The high survival rates at various drug concentrations indicate that the drugs are almost non-toxic to the cells. Based on the results of cell experiments under normal conditions, we further evaluated the survival rates after lipopolysaccharide induction.

[0063] Under LPS stimulation-induced conditions (Figure 2), the cell survival rate of the inflammation model group (LPS group) was slightly higher than that of the blank control group. After intervention with gradient concentrations of drugs, the cell survival rate of the 7-molecule inflammation model remained at a high level and was basically the same as that of normal cells, indicating that the drugs did not exhibit significant cytotoxicity within the experimental concentration range.

[0064] (2) Evaluation of inflammatory stress response: The inhibitory effect of drugs on LPS-induced NO production was detected by combining the Griess method;

[0065] Compared with the blank control group, the LPS stimulation group showed a significant increase in NO release, successfully establishing a macrophage inflammation activation model (Figure 3). After intervention with the positive control drug dexamethasone, NO levels significantly decreased. In the seven drug administration groups, NO secretion decreased in a dose-dependent manner with increasing drug concentration gradient, indicating that the drugs have a certain inhibitory effect on the secretion of inflammatory factors in a concentration-dependent manner, which can be further investigated in the regulation of key inflammatory mediators.

[0066] (3) Regulation of key inflammatory mediators: Quantitative analysis of the expression of pro-inflammatory factors such as TNF-α, IL-6 and IL-1β by ELISA showed that the cell pro-inflammatory factor index decreased with increasing drug concentration after treatment.

[0067] Compared with the blank control group, the secretion levels of pro-inflammatory factors TNF-α, IL-6, and IL-1β in the LPS-stimulated RAW 264.7 macrophage inflammation model were significantly increased, indicating that the inflammation model was successfully established (Figures 4-6). In the positive control group, dexamethasone intervention significantly reduced the levels of pro-inflammatory factors. All seven treatment groups showed dose-dependent inhibitory effects, suggesting they may be potential anti-inflammatory molecules.

[0068] Example 3

[0069] With compound 3ap For example, a two-step method can be used to construct delayed fluorescent molecules with aggregation-induced emission properties.

[0070]

[0071] 1a (1.0 mmol), p-bromobenzoylformaldehyde 2p (1.5 mmol), and 5 equiv of acetic acid were dispersed in MeCN (6 mL) solvent and reacted in sealed tubes at 60 °C for 6 hours. After separation and purification, the bromopyrimidine ditetrahydroisoquinoline product 3ap was obtained. 3ap (0.5 mmol) was condensed and cyclized with hydrazine hydrate (25.0 mmol), and after separation and purification, the pyrimidine pyridazine product 5 was obtained. Compound 5 (0.2 mmol), phenoxazine PXZ (0.5 mmol), Pd(OAc)2 (0.008 mmol), Cs2CO3 (2 mmol), and [(t-Bu)3PH]BF4 (0.024 mmol) were dispersed in o-Tol (15 mL) and purified by CN coupling reaction to obtain fluorescent molecule 6. Compound 5 (0.2 mmol), phenoxazine PTZ (0.5 mmol), Pd(OAc)2 (0.008 mmol), Cs2CO3 (2 mmol), and [(t-Bu)3PH]BF4 (0.024 mmol) were dispersed in o-Tol (15 mL) and purified by CN coupling reaction to obtain fluorescent molecule 7.

[0072] Example 4

[0073] Delayed fluorescence molecular optical property testing

[0074] Performance tests were performed on delayed fluorescence molecules 6 and 7 prepared in Example 3:

[0075] (1) Ultraviolet / Visible absorption spectroscopy and fluorescence spectroscopy

[0076] As shown in Figure 7, compound 6 has strong absorption peaks at 325 nm and compound 7 has strong absorption peaks at 320 nm, which belong to the π→π group. * Transitions. The fluorescence spectra of compounds 6 and 7 at room temperature show yellow-green emission, with maximum emission peaks at 530 nm and 540 nm, respectively.

[0077] (2) Delayed fluorescence performance test

[0078] The fluorescence intensity of compounds 6 and 7 in o-xylene solution under an argon atmosphere was significantly stronger than that under an air atmosphere (Figure 8), which preliminarily proves that triplet excitons participate in luminescence and have delayed fluorescence properties.

[0079] (3) Aggregation-induced emission (AIE) performance test

[0080] These two molecules emit light in pure tetrahydrofuran solution due to their molecular structure, but the light emission is weak. The water component (f) in the mixed system... wWhen the percentage increases to 70%, the fluorescence intensity of the compound gradually increases. This may be because the intramolecular motion is restricted after the molecules form an aggregated state, which blocks the non-radiative decay channel and leads to an increase in the photoluminescence spectrum intensity. Both molecules have obvious AIE characteristics (Figure 9).

[0081] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A pyrimido[2,1-a:4,3-a']diisoquinoline compound, characterized in that, It has the structure shown in Equation I: Equation I; where R 1 =H, Cl or Br, Ar is a substituted or unsubstituted benzene ring, naphthyl ring, thiophene; the substitution refers to being substituted by alkyl, alkoxy, thiomethyl or halogen; the alkyl is C1-C4 alkyl, and the alkoxy is C1-C4 alkoxy.

2. The pyrimido[2,1-a:4,3-a']diisoquinoline compound according to claim 1, characterized in that, The structural formula of the pyrimido[2,1-a:4,3-a']diisoquinoline compound is shown below: 。 3. A method for preparing the pyrimido[2,1-a:4,3-a']diisoquinoline compound of claim 1, characterized in that, Compound 1, compound 2, and acetic acid were uniformly dispersed in a solvent and then reacted. The reaction process was monitored by thin-layer chromatography-mass spectrometry. The structural formula of compound 1 is: The structural formula of compound 2 is In compounds 1 and 2, R 1 Ar and R in claim 1 1 Same as Ar.

4. The preparation method according to claim 3, characterized in that, The molar ratio of compound 1 to compound 2 is 1:1.5; the solvent is acetonitrile; the reaction is a sealed tube reaction, the reaction temperature is 60°C, and the reaction time is 6 hours.

5. The use of the pyrimido[2,1-a:4,3-a']diisoquinoline compound of claim 1 in the preparation of an anti-inflammatory drug.

6. An anti-inflammatory drug, characterized in that, The raw materials include the pyrimido[2,1-a:4,3-a']diisoquinoline compounds as described in claim 1, and pharmaceutically acceptable excipients.

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