Pyrimido [2, 1-a: 4, 3-a '] diisoquinoline compound as well as preparation method and application thereof

Abstract

The invention provides pyrimido [2, 1-a: 4, 3-a '] diisoquinoline compounds as well as a preparation method and application thereof, and belongs to the technical field of biochemistry. The pyrimido [2, 1-a: 4, 3-a '] diisoquinoline compound has a structure as shown in a formula I: # imgabs0 #, a large number of substituent groups can be substituted through aryl formyl formaldehyde, so that the diverse substituted pyrimido [2, 1-a: 4, 3-a'] diisoquinoline compound is synthesized, and the compound has an inhibition effect on secretion of inflammatory factors.

Description

Technical Field

[0001] The present invention relates to the field of biochemistry, and particularly relates to a pyrimido[2,1-a:4,3-a']diisoquinoline compound, a preparation method thereof, and an application thereof. Background Art

[0002] Hydroisoquinoline compounds are an important class of organic molecules with a wide range of pharmacological activities. Their structural diversity and biological activities have attracted much attention in the field of medicinal chemistry. Research shows that such compounds exhibit significant effects in aspects such as anti-tumor, anti-viral, leukemia treatment, lipid-lowering, antibacterial, and anti-malaria. For example, tetrahydroprotoberberine alkaloids (THPBs), as derivatives of hydroisoquinoline, show effects such as analgesia, lipid regulation, and anti-neuropsychiatric diseases in the treatment of central nervous system diseases. In particular, their regulatory effect on dopamine receptors has attracted extensive research interest. In addition, compounds obtained through structural modification (such as DC371739) show good in vitro and in vivo activities in lipid-lowering and have completed phase I clinical trials, showing excellent tolerance. The alkaloid stepholidine isolated from Stephania japonica and its analogs (such as compounds 12a and 23b) have also been confirmed as highly efficient D1R and D3R ligands, further expanding the application potential of hydroisoquinoline compounds.

[0003] Although the pharmacological activities of hydroisoquinoline compounds have been widely studied, their structural complexity and limitations of synthetic methods have restricted their further development. Traditional synthetic methods are usually cumbersome, with low yields, and it is difficult to introduce diverse substituents, thus restricting structural modification and activity optimization.

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

[0005] In view of this, the purpose of the present invention is to provide a pyrimido[2,1-a:4,3-a']diisoquinoline compound, a preparation method thereof, and an application thereof.

[0006] To achieve the above purpose, the present invention provides the following technical solutions:

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

[0008]

[0009] Wherein, R 1= H, Cl or Br, and Ar is a substituted or unsubstituted benzene ring, naphthalene ring, biphenyl, fluorene ring, thiophene, pyridine, furan, benzothiophene or benzofuran; the substitution means being substituted by an alkyl group, an alkoxy group, a thiomethyl group or a halogen; the alkyl group is a C1-C4 alkyl group, and the alkoxy group is a C1-C4 alkoxy group.

[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 as follows:

[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 compound. After uniformly dispersing Compound 1, Compound 2 and acetic acid in a solvent, the reaction is carried out, 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 Compound 1 and Compound 2, R 1 , Ar are the same as R 1 and Ar in the above-mentioned pyrimido[2,1-a:4,3-a']diisoquinoline compound.

[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 mode is a sealed tube reaction, the reaction temperature is 60 °C, and the time is 6 h.

[0019] In the present invention, when R 1 = H, the general formula of the above reaction is:

[0020]

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

[0022]

[0023] The third technical solution of the present invention is an application of the above 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 pyrimido[2,1-a:4,3-a']diisoquinoline compound and pharmaceutically acceptable excipients.

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

[0026] The sixth technical solution of the present invention is a preparation method of a delayed fluorescence molecule, which comprises condensing and cyclizing the above pyrimido[2,1-a:4,3-a']diisoquinoline compound with hydrazine hydrate to obtain a pyrimidopyridazine compound; then carrying out a C-N coupling reaction between the pyrimidopyridazine compound and a donor unit under the catalysis of palladium acetate to obtain the delayed fluorescence molecule; the donor unit is phenoxazine or phenothiazine.

[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 have long luminescence lifetimes and high luminescence quantum yields, and have very wide applications in the fields of bioimaging (including lifetime imaging), temperature / oxygen sensing, photodynamic therapy, etc. On the basis of this synthesis strategy, the present invention has carried out derivatization synthesis on the target molecule to prepare a series of organic light-emitting molecules, indicating that the reaction strategy of derivatizing the pyrimido[2,1-a:4,3-a']diisoquinoline compound into organic light-emitting molecules in the present invention is feasible and can continue to construct organic light-emitting molecules.

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

[0030] In the present invention, a large number of substituents can be introduced through arylformyl formaldehyde, so as to synthesize diversely substituted pyrimido[2,1-a:4,3-a']diisoquinoline compounds, and these compounds have an inhibitory effect on the secretion of inflammatory factors.

[0031] The preparation method of the pyrimido[2,1-a:4,3-a']diisoquinoline compound in the present invention is simple and can be synthesized by a one-pot one-step method.

[0032] The pyrimido[2,1-a:4,3-a']diisoquinoline compound provided by the present invention can be derivatized to synthesize a series of organic light-emitting molecules. Description of the Drawings

[0033] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required in the embodiments. Obviously, the drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0034] Figure 1 Effect of drugs with different concentrations on the cell viability of Raw264.7;

[0035] Figure 2 Effect of drugs with different concentrations on the cell viability of Raw 264.7 induced by LPS;

[0036] Figure 3 Effect of drugs with different concentrations on the NO secretion content of Raw 264.7 induced by LPS;

[0037] Figure 4 Effect of drugs with different concentrations on the TNF-α secretion content of Raw264.7 induced by LPS;

[0038] Figure 5 Effect of drugs with different concentrations on the IL-6 secretion content of Raw264.7 induced by LPS;

[0039] Figure 6 Effect of drugs with different concentrations on the IL-1β secretion content of Raw264.7 induced by LPS;

[0040] Figure 7 Ultraviolet-visible absorption spectra and fluorescence spectra of compound 6(a) and compound 7(b) in o-xylene;

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

[0042] Figure 9 Fluorescence emission of compound 6(a) and compound 7(b) as a function of the THF / water mixed solution (f w ) change curve. Detailed implementation manners

[0043] Now, various exemplary implementation manners of the present invention will be described in detail. This detailed description should not be considered as a limitation of the present invention, but should be understood as a more detailed description of certain aspects, characteristics, and implementation schemes of the present invention.

[0044] It should be understood that the terms used in the present invention are only for describing specific embodiments and are not intended to limit the present invention. Additionally, for the numerical ranges in the present invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any intermediate value within any stated value or stated range, as well as each smaller range between any other stated value or intermediate value within the stated range, is also included in the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0045] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Although the present invention only describes preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials related to the documents. In case of conflict with any incorporated document, the content of this specification shall prevail.

[0046] Without departing from the scope or spirit of the present invention, various modifications and variations can be made to the specific embodiments of the present invention specification, which are obvious to those skilled in the art. Other embodiments obtained from the specification of the present invention are obvious to those skilled in the art. The specification and examples of the present invention are merely exemplary.

[0047] Regarding the use of "comprising", "including", "having", "containing", etc. herein, they are all open-ended terms, meaning including but not limited to.

[0048] The technical solutions described in the present invention are all conventional solutions in the art unless otherwise specified. 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 content of the present invention will be further clarified below in conjunction with examples, but the content of the present invention is not limited to the following examples only.

[0050] Example 1

[0051] 1,2,3,4-Tetrahydroisoquinoline 1a (1.0 mmol), benzoylformaldehyde 2a (1.5 mmol) and 5 equiv of acetic acid were dispersed in MeCN (6 mL) solvent, and the reaction was carried out in a sealed tube at 60 °C for 6 hours. The reaction of the raw materials was monitored by TLC-MS. After the reaction was completed, the reaction solution was concentrated by rotary evaporation under reduced pressure, and then saturated sodium carbonate solution was added to quench the remaining acetic acid. Equal volume of ethyl acetate was added and extracted with a 150 mL separatory funnel. The aqueous phase was continuously extracted with an equal volume of ethyl acetate twice. The combined organic phases were extracted with an equal volume of saturated sodium chloride solution, and the aqueous phase was discarded. After extraction three times, the organic phase was dried over anhydrous sodium sulfate, allowed to stand for one hour and then concentrated by rotary evaporation under reduced pressure. Purification by silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 10:1 (v / v)) gave yellow solid 3aa (186 mg, 5%).

[0052]

[0053] For the synthesis of the remaining compounds, the method of Example 1 was referred to, and only the corresponding raw materials were replaced to prepare the compounds in Table 1.

[0054] The synthesized representative pyrimido[2,1-a:4,3-a']diisoquinoline compounds and their 1H and 13C NMR data are shown in Table 1.

[0055] Table 1 1H and 13C NMR data of representative compounds

[0056]

[0057]

[0058]

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

[0060] The anti-inflammatory inhibitory ability of the 7 representative compounds 3aa - 3ba in Table 1 in Example 1 at the cellular level was studied, and the regulatory effect of the drugs on the RAW 264.7 macrophage inflammation model was systematically analyzed through a three-dimensional evaluation system:

[0061] (1) Basic cytotoxicity assessment: The CCK-8 method was used to detect the effects of different concentrations of drugs on the cell viability of normal and lipopolysaccharide (LPS)-stimulated cells, and to reveal the drug safety window;

[0062] Under normal conditions, the cell survival rate is as Figure 1 shown. The drugs at each concentration have a high cell viability, indicating that the drugs are almost non-toxic to the cells. Based on the cell experiment results under normal conditions, we further evaluated the survival rate after lipopolysaccharide induction.

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

[0064] (2) Evaluation of inflammatory stress response: Combine the Griess method to detect the inhibitory effect of drugs on NO production induced by LPS;

[0065] Compared with the blank control group, the NO release amount in the LPS stimulation group increased significantly, and a macrophage inflammation activation model ( Figure 3 ) was successfully established. After intervention with the positive control drug dexamethasone, the NO level decreased significantly. In the 7-molecule drug administration groups, as the drug concentration gradient increased, the NO secretion amount decreased in a dose-dependent manner, indicating that the drugs had a certain inhibitory effect on the secretion of inflammatory factors and showed concentration dependence, and further research on the regulation of key inflammatory mediators could be carried out.

[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 pro-inflammatory factor index of cells decreased with the increase of concentration after drug treatment.

[0067] Compared with the blank control group, in the RAW 264.7 macrophage inflammation model stimulated by LPS, the secretion levels of pro-inflammatory factors TNF-α, IL-6 and IL-1β were significantly higher than those in the blank group, indicating that the inflammation model was successfully established ( Figures 4 - 6 ). After intervention with the positive control dexamethasone, the levels of pro-inflammatory factors decreased significantly. All 7 drug administration groups showed a dose-dependent inhibitory effect and were expected to become potential anti-inflammatory molecules.

[0068] Example 3

[0069] Taking compound 3ap as an example, a two-step method was used to construct a delayed fluorescence molecule 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) as the solvent. The reaction was carried out in a sealed tube at 60 °C for 6 hours. After separation and purification, the bromopyrimidine-fused bis-tetrahydroisoquinoline product 3ap was obtained. 3ap (0.5 mmol) was condensed and cyclized with hydrazine hydrate (25.0 mmol). After separation and purification, the pyrimidine-fused pyridazine product 5 was obtained. Compound 5 (0.2 mmol), phenoxazine PXZ (0.5 mmol), Pd(OAc)2 (0.008 mmol), Cs2CO3 (2 mmol), [(t-Bu)3PH]BF4 (0.024 mmol) were dispersed in o-Tol (15 mL), and the fluorescent molecule 6 was obtained after separation and purification by using the C-N coupling reaction. Compound 5 (0.2 mmol), phenoxazine PTZ (0.5 mmol), Pd(OAc)2 (0.008 mmol), Cs2CO3 (2 mmol), [(t-Bu)3PH]BF4 (0.024 mmol) were dispersed in o-Tol (15 mL), and the fluorescent molecule 7 was obtained after separation and purification by using the C-N coupling reaction.

[0072] Example 4

[0073] Optical Property Test of Delayed Fluorescent Molecules

[0074] The performance tests were carried out on the delayed fluorescent molecules 6 and 7 prepared in Example 3:

[0075] (1) Ultraviolet / Visible Absorption Spectra and Fluorescence Spectra

[0076] As Figure 7 shown, compound 6 has a strong absorption peak at 325 nm, and compound 7 has a strong absorption peak at 320 nm, belonging to π→π * transition. The fluorescence spectra of compound 6 and compound 7 at room temperature show yellow-green emission, and the maximum emission peaks are 530 nm and 540 nm respectively.

[0077] (2) Delayed Fluorescence Property Test

[0078] The fluorescence intensities of the two molecules, compound 6 and compound 7, in o-xylene solution under an argon atmosphere are significantly stronger than those under an air atmosphere ( Figure 8 ), which preliminarily proves that triplet excitons participate in luminescence and have the property of delayed fluorescence.

[0079] (3) Aggregation-Induced Emission (AIE) Property Test

[0080] These two molecules emit light due to their own molecular structures in pure tetrahydrofuran solution, but their luminescence is weak. In the mixed system, the water component (f w) When the percentage increases to 70%, the fluorescence intensity of the compound gradually increases. It may be that when the molecules form an aggregated state, the intramolecular motion is restricted, blocking the non-radiative decay channel, resulting in an increase in the intensity of the photoluminescence spectrum. Both of these two molecules have obvious AIE characteristics( Figure 9 ).

[0081] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope 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 Formula I: Among them, R 1 = H, Cl or Br, and Ar is a substituted or unsubstituted benzene ring, naphthalene ring, biphenyl, fluorene ring, thiophene, pyridine, furan, benzothiophene or benzofuran; the substitution means being substituted by an alkyl group, an alkoxy group, a thiomethyl group or a halogen; the alkyl group is a C1-C4 alkyl group, and the alkoxy group is a C1-C4 alkoxy group.

2. The pyrimido[2,1-a:4,3-a']diisoquinoline compound according to claim 1, wherein The structural formula of the pyrimido[2,1-a:4,3-a']diisoquinoline compound is as follows:

3. A method for preparing the pyrimido[2,1-a:4,3-a']diisoquinoline compound according to claim 1 or 2, characterized in that, 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; The structural formula of the said Compound 1 is: The structural formula of Compound 2 is In Compounds 1 and 2, R 1 , Ar are the same as R 1 and Ar in Claim 1.

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 mode is a sealed tube reaction, the reaction temperature is 60 °C, and the time is 6 h.

5. Use of the pyrimido[2,1-a:4,3-a']diisoquinoline compound according to 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 compound according to claim 1 and pharmaceutically acceptable excipients.

7. Use of the pyrimido[2,1-a:4,3-a']diisoquinoline compound according to claim 1 in the preparation of a delayed fluorescence molecule.

8. A method for preparing a delayed fluorescence molecule, characterized in that, The pyrimido[2,1-a:4,3-a']diisoquinoline compound according to claim 1 is condensed and cyclized with hydrazine hydrate to obtain a pyrimidopyrazine compound; then the pyrimidopyrazine compound undergoes a C-N coupling reaction with a donor unit under the catalysis of palladium acetate to obtain the delayed fluorescence molecule; the donor unit is phenoxazine or phenothiazine.

9. A delayed fluorescence molecule prepared by the preparation method according to claim 8.

Images