Azulenes, methods for their preparation and use

Abstract

The application discloses an azulene compound, a preparation method and application thereof. The azulene compound is a compound of formula I. The compound has one or more advantages of good stability, antioxidant activity, anti-inflammatory activity and high safety, and has a good application prospect.

Description

Technical Field

[0001] This invention belongs to the field of synthetic medicinal chemistry, and specifically relates to an azurite compound, its preparation method and application. Background Technology

[0002] Oxidation is a crucial process in living systems, but it can also be harmful to health. For example, hydrogen peroxide generated during oxidation can damage cell membranes and cause mutagenic effects. Oxidation also produces reactive free radicals, such as reactive oxygen species (ROS) and reactive nitrogen species (RNS), which play a role in regulating bodily functions. However, when the human body is exposed to external stimuli such as X-rays, air pollutants, or chemicals, ROS and RNS may increase uncontrollably, leading to a redox state (known as oxidative stress). Oxidative stress can damage cellular structures such as DNA, proteins, and lipids. Prolonged exposure to this state can lead to chronic inflammation, cancer, endothelial dysfunction, atherosclerosis, and neurodegenerative diseases (Querio, G. et al. Front. Physiol. 2018, 9, 246; Rekka, E. et al. Chem. Pharm. Bull. 2002, 50, 904). Therefore, there is a close relationship between free radical-induced oxidative stress and acute or chronic inflammation.

[0003] Azulene is a blue, non-benzene aromatic compound with a unique molecular structure. It consists of a fused seven-membered ring in a positive ionic state and a five-membered ring in a negative ionic state. Its ionic state is more stable than the azulene ring itself, and it is an isomer of naphthalene. The unique structure of azulene endows it with high chemical activity and provides more possibilities for the chemical modification of azulene molecules. More importantly, azulene compounds are widely found in various plant essential oils in nature, such as ylang-ylang oil, cypress oil, patchouli oil, vetiver oil, and guaiac oil, possessing advantages such as low toxicity, wide availability, and high activity. Chamazulene and guaiazulene (GA) are the two most common azurite natural products. They have good therapeutic effects on various inflammations, ulcers, and burns occurring in the human digestive, respiratory, urinary, skin, and conjunctiva systems (Pratsinis, H.; Haroutounian, S. Nat. Prod. Lett. 2002, 16, 201.; Cao, T. et al. Chem. Biol. Drug Des. 2016, 88, 264-271.; Wada, T. et al. Vivo. 2018, 32, 479.; Ayaz, F.; Yuzer, A.; Innce, T.; Innce, M. Inflammation. 2020, 43, 1009.). It is worth noting that by introducing a sodium sulfonate group at the 3 position of the five-membered ring of the GA azulene parent compound through a sulfonation reaction, a water-soluble, dark blue, flaky powder, namely sodium guaiacol sulfonate (5-isopropyl-3,8-dimethylazine-1-sulfonate, abbreviated as GAS-Na), can be obtained. Compared with the lipid-soluble GA, the hydrophilicity of GAS-Na is more conducive to its application, and it is currently widely used in clinical practice as an anti-inflammatory and anti-ulcer agent (Yanagisawa, T. et al. Chem. Pharm. Bull. (Tokyo), 1990, 38, 3355.; Guarrera, M. et al. J. Eur. Acad. Dermatol. Venereol. 2001, 15, 481; Andersen, FAInt. J. Toxicol. 1999, 18, 27.). The molecular structures of azurite, chamomile azurite, guaiac azurite, and guaiac azurite sodium sulfonate are shown in the following formulas:

[0004]

[0005] However, currently, azurites, including azurite, guaiac azurite, and guaiac azurite sodium sulfonate, suffer from poor light and heat stability, hindering their widespread application. Therefore, there is an urgent need to develop azurite parent molecules with high chemical activity, low biotoxicity, and diverse biological activities. Summary of the Invention

[0006] The technical problem to be solved by this invention is to overcome the defects of existing azurite biofunctional molecules, such as instability, difficulty in preservation, or moderate activity, and to provide an azurite derivative with antioxidant and anti-inflammatory activity, its preparation method, and its application. The compounds of this invention have at least one of the following advantages: good stability, antioxidant and anti-inflammatory activity, and high safety.

[0007] The present invention solves the above-mentioned technical problems through the following technical solutions.

[0008] This invention provides a compound as shown in Formula I:

[0009] YLZ style I;

[0010] In Formula I, each substituent is defined as defined in group (a) or group (b).

[0011] Group (a): Y is

[0012] Z is

[0013] R 1a R 1g R 2a R 2g R 3a R 3g R 4a and R 4g Independently for H and C 1-12 Alkyl or -SO3Q;

[0014] Q is an alkali metal;

[0015] R 1b R 1c R 1d R 1e R 1f R 2b R 2c R 2d R 2e R 2f R 3b R 3c R 3d R 3e R 3f R 4b R 4c R 4d R 4e and R 4f Independently for H and C 1-12 Alkyl, R 1b-1 Replacement C 1-12 Alkyl (R) 1b-1The number can be multiple, for example, 1, 2, 3, or 4; when R 1b-1 When there are multiple, R 1b-1 (can be the same or different), -COOH, -OC(O)-C 1-12 Alkyl, C 1-12 oligoglycol or C 1-12 oligomeric glycol ethers;

[0016] R 1b-1 It can be independently hydroxyl, halogen, nitro or amino;

[0017] L represents a non-existent (Y and Z are directly connected, i.e., equation I is YZ) or a connector;

[0018] The connector has the following structure

[0019]

[0020] Among them, X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 and X 10 Independently O, S, or N;

[0021] m1, m2, m3, m4, m5 and m6 are independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0022] n1, n2, n3, n4, n5, n6, n7, n8, n9 and n 10 Independently, they can be 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;

[0023] The ring B is a 4-6 membered heterocyclic alkane with two nitrogen atoms (N).

[0024] Group (b): Y is

[0025] Z is

[0026] R 11a R 11g Independently for H and C 1-12 Alkyl group, -SO3Q 1 Or SO2Q';

[0027] Q 1 Alkali metals, NH4,

[0028] Q' is C 1-12 Alkoxy, -NH-C 1-12 Alkyl, -NH-C6-C 10 Aryl, -NH- (5-7 quinone heteroaryl), C 1-12 oligoglycol groups or C 1-12 The oligomeric glycol ether group; in the 5-7 membered heteroaryl group, the number of heteroatoms is 1 or 2, and the heteroatoms are selected from one or two of O, N, and S;

[0029] R 11b R 11c R 11d R 11e R 11f Independently for H and C 1-12 Alkyl, R 11b-1 Replacement C 1-12 Alkyl (R) 11b-1 The number can be multiple, for example, 1, 2, 3, or 4; when R 11b-1 When there are multiple, R 11b-1 (can be the same or different), -COOH, -OC(O)-C 1-12 Alkyl, C 1-12 oligoglycol or C 1-12 oligomeric glycol ethers;

[0030] R 11b-1 It can be independently hydroxyl, halogen, nitro or amino;

[0031] R 10a Independently for H and C 1-12 Alkyl, C6-C 10 Aryl, R 10a-1 Replacement C6-C 10 Aryl (R) 10a-1 The number can be multiple, for example, 1, 2, 3, or 4; when R 10a-1 When there are multiple, R 10a-1 (Can be the same or different);

[0032] R 10a-1 C 1-12 Alkyl, C 1-12 Halogenated or C 1-12 Alkoxy;

[0033] R 10b R 10c R 10d R 10e Independently -H, -CN, halogen, or C substituted with one or more halogens. 1-12 Alkyl, C 1-12 Alkyl, hydroxyl, nitro, amino, -NH-C1-12 Alkyl, -OC(O)-C 1-12 Alkyl or C 1-12 Alkoxy;

[0034] L represents non-existence (Y and Z are directly connected, i.e., equation I represents YZ). -NH-、 S atom, O atoms or linkers;

[0035] The connector has the following structure

[0036] Among them, X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 and X 10 Independently O, S, or N;

[0037] m1, m2, m3, m4, m5 and m6 are independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0038] n1, n2, n3, n4, n5, n6, n7, n8, n9 and n 10 Independently, they can be 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;

[0039] The ring B is a 4-6 membered heterocyclic alkane with two N heteroatoms.

[0040] In a preferred embodiment of the present invention, the compound as shown in Formula I is:

[0041] YLZ style I;

[0042] Where Y is

[0043] Z is

[0044] R 1a R 1g R 2a R 2g R 3a R 3g R 4a and R 4g Independently for H and C 1-12 Alkyl or -SO3Q;

[0045] Q is an alkali metal;

[0046] R 1b R 1c R 1d R 1e R 1f R 2b R 2c R 2d R 2e R 2f R 3b R 3c R 3d R 3e R 3f R 4b R 4c R 4d R 4e and R 4f Independently for H and C 1-12 Alkyl, R 1b-1 Replacement C 1-12 Alkyl (R) 1b-1 The number can be multiple, for example, 1, 2, 3, or 4; when R 1b-1 When there are multiple, R 1b-1 (can be the same or different), -COOH, -OC(O)-C 1-12 Alkyl, C 1-12 oligoglycol or C 1-12 oligomeric glycol ethers;

[0047] R 1b-1 It can be independently hydroxyl, halogen, nitro or amino;

[0048] L represents a non-existent (Y and Z are directly connected, i.e., equation I is YZ) or a connector;

[0049] The connector has the following structure

[0050] Among them, X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 and X 10 Independently O, S, or N;

[0051] m1, m2, m3, m4, m5 and m6 are independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0052] n1, n2, n3, n4, n5, n6, n7, n8, n9 and n 10 Independently, they can be 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;

[0053] The ring B is a 4-6 membered heterocyclic alkane with two N heteroatoms.

[0054] In a preferred embodiment of the present invention, the alkali metal may be Li, Na, or K, for example, Na.

[0055] In a preferred embodiment of the present invention, Q 1 In the middle, the said C in 1-12 Alkyl groups can independently be C10-3 ... 1-6 alkyl.

[0056] In a preferred embodiment of the present invention, Q 1 In the middle, the said C in 6-10 The aryl group can independently be a phenyl group.

[0057] In a preferred embodiment of the present invention, in Q', the C 1-12 C in alkoxy 1-12 Alkyl, -NH-C 1-12 C in alkyl 1-12 Alkyl group, the aforementioned C 1-12 C in oligoglycol 1-12 Alkyl groups and the C 1-12 C in oligoglycol ethers 1-12 Alkyl groups can independently be C10-3 ... 1-6 alkyl.

[0058] In a preferred embodiment of the present invention, in Q', the -NH-C6-C 10 C in aryl 6-10 The aryl group can be phenyl or naphthyl independently.

[0059] In a preferred embodiment of the present invention, R 1a R 1g R 2a R 2g R 3a R 3g R 4a and R 4g In the context, C 1-12 The alkyl group can be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl, such as methyl, ethyl, or isopropyl.

[0060] In a preferred embodiment of the present invention, R 1b R 1c R1d R 1e R 1f R 2b R 2c R 2d R 2e R 2f R 3b R 3c R 3d R 3e R 3f R 4b R 4c R 4d R 4e and R 4f In the context, -OC(O)-C 1-12 C in alkyl 1-12 Alkyl group, the aforementioned C 1-12 C in oligoglycol 1-12 Alkyl groups and the C 1-12 C in oligoglycol ethers 1-12 Alkyl groups can independently be C10-3 ... 1-6 alkyl.

[0061] In a preferred embodiment of the present invention, R 1b R 1c R 1d R 1e R 1f R 2b R 2c R 2d R 2e R 2f R 3b R 3c R 3d R 3e R 3f R 4b R 4c R 4d R 4e and R 4f In the context, C 1-12 Alkyl groups and the R- 1b-1 Replacement C 1-12 C in alkyl 1-12 The alkyl group can independently be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl, such as methyl, ethyl, or isopropyl.

[0062] In a preferred embodiment of the present invention, R 10a In the context, C 1-12 The alkyl group can be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl, such as methyl, ethyl, or isopropyl.

[0063] In a preferred embodiment of the present invention, R 10a In the context of C6-C 10 The aryl group described by R 10a-1 Replacement C6-C 10 C6-C in aryl 10 The aryl group can be phenyl or naphthyl.

[0064] In a preferred embodiment of the present invention, R 10b R 10c R 10d R 10e In the context, C 1-12 The alkyl group can be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl, such as methyl, ethyl, or isopropyl.

[0065] In a preferred embodiment of the present invention, R 10b R 10c R 10d R 10e In, the -NH-C 1-12 C in alkyl 1-12 Alkyl group, the -OC(O)-C 1-12 C in alkyl 1-12 Alkyl and C 1-12 C in alkoxy 1-12 Alkyl groups can independently be C10-3 ... 1-6 alkyl.

[0066] In a preferred embodiment of the present invention, R 11a and R 11g In the context, C 1-12 The alkyl group can be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl, such as methyl, ethyl, or isopropyl.

[0067] In a preferred embodiment of the present invention, R 11b R 11c R 11d R 11e and R 11f In the context, C 1-12 Alkyl groups and the R- 1b-1 Replacement C 1-12 C in alkyl 1-12 The alkyl group can independently be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl, such as methyl, ethyl, or isopropyl.

[0068] In a preferred embodiment of the present invention, R 11b R 11c R 11d R 11e and R11f In the context, -OC(O)-C 1-12 C1- in alkyl groups 12 Alkyl group, the aforementioned C 1-12 C in oligoglycol 1-12 Alkyl groups and the C 1-12 C in oligoglycol ethers 1-12 Alkyl groups can independently be C10-3 ... 1-6 alkyl.

[0069] In a preferred embodiment of the present invention, X in the connector 7 X 8 X 9 and X 10 S or N are preferred independently.

[0070] In a preferred embodiment of the present invention, m4 and m5 are preferably 0 in the connector.

[0071] In a preferred embodiment of the present invention, in the connector, n7 and n 10 The preferred value is 2.

[0072] In a preferred embodiment of the present invention, ring B in the linker is preferably a piperazine ring.

[0073] In a preferred embodiment of the present invention, the connector preferably has any of the following general structural forms:

[0074]

[0075] Preferably, the connector is any of the following general structural forms:

[0076] More preferably, the connector is

[0077] In a preferred embodiment of the present invention, R 1a and R 1g Among them, one of the preferred options is C. 1-12 Alkyl group, another preferably H or -SO3Q.

[0078] In a preferred embodiment of the present invention, R 2a and R 2g Among them, one of the preferred options is C. 1-12 Alkyl group, another preferably H or -SO3Q.

[0079] In a preferred embodiment of the present invention, R 3a and R 3g Among them, one of the preferred options is C.1-12 Alkyl group, another preferably H or -SO3Q.

[0080] In a preferred embodiment of the present invention, R 4a and R 4g Among them, one of the preferred options is C. 1-12 Alkyl group, another preferably H or -SO3Q.

[0081] In a preferred embodiment of the present invention, R 1d R 2d R 3d and R 4d H is preferred.

[0082] In a preferred embodiment of the present invention, R 1b R 1c R 1e R 1f R 2b R 2c R 2e R 2f R 3b R 3c R 3e R 3f R 4b R 4c R 4e and R 4f Independently preferably H or C 1-12 alkyl.

[0083] In a preferred embodiment of the present invention, R 1b R 1c R 1e R 1f R 2b R 2c R 2e R 2f R 3b R 3c R 3e R 3f R 4b R 4c R 4e and R 4f H, methyl, ethyl or isopropyl are preferred independently.

[0084] In a preferred embodiment of the present invention, R 11a and R 11g In this process, one of the components is preferably H, and the other is preferably C. 1-12 Alkyl or H. In a preferred embodiment of the invention, R 11b R 11c R 11e and R11f Independently preferably H or C 1-12 alkyl.

[0085] In a preferred embodiment of the present invention, R 10a R 10b R 10e H is preferred independently.

[0086] In a preferred embodiment of the present invention, R 10c and R 10d H and C are preferred independently. 1-12 Alkoxy or halogen.

[0087] In a preferred embodiment of the present invention Preferred

[0088] In a preferred embodiment of the present invention Preferred

[0089] In a preferred embodiment of the present invention Preferred In a preferred embodiment of the present invention Preferred

[0090] In a preferred embodiment of the present invention Preferred

[0091] In a preferred embodiment of the present invention for

[0092] In a preferred embodiment of the present invention, the compound represented by Formula I,

[0093] R 1a and R 1g In the middle, one of them is C 1-12 Alkyl group, the other being H or -SO3Q;

[0094] R 2a and R 2g In the middle, one of them is C 1-12 Alkyl group, the other being H or -SO3Q;

[0095] R 3a and R 3g In the middle, one of them is C 1-12 Alkyl group, the other being H or -SO3Q;

[0096] R 4a and R 4g In the middle, one of them is C 1-12 Alkyl group, the other being H or -SO3Q; R 1d R 2d R 3d and R 4d For H;

[0097] R 1b R 1c R 1e R 1f R 2b R 2c R 2e R 2f R 3b R 3c R 3e R 3f R 4b R 4c R 4e and R 4f Independently H or C 1-12 alkyl;

[0098] L represents non-existence.

[0099] In a preferred embodiment of the present invention, the compound represented by Formula I is IA:

[0100]

[0101] Among them, R 1g and R 3g Independently H or -SO3Q;

[0102] R 1a R 1b R 1c R 1d R 1e R 1f R 3a R 3b R 3c R 3d R 3e and R 3f Independently H or C 1-12 alkyl.

[0103] In a preferred embodiment of the present invention, the compound represented by Formula I is IB:

[0104]

[0105] Among them, R 11g and R 10a For H;

[0106] R 11a R 11b R 11c R 11d R 11e R 11f Independently H or C 1-12 alkyl;

[0107] R 10b R 10c R 10d and R 10e Independent of H, halogen or C 1-12 Alkoxy;

[0108] Preferably, in the general formula IB,

[0109] R 10a For H;

[0110] R 11a R 11b R 11c R 11d R 11e R 11f and R 11g Not both H;

[0111] R 10b R 10c R 10d and R 10e Independent of H, halogen or C 1-12 Alkyl group.

[0112] In a preferred embodiment of the present invention, the compound represented by Formula I is any of the following general formulas:

[0113]

[0114]

[0115] In a preferred embodiment of the present invention, the compound represented by Formula I is any of the following general formulas:

[0116]

[0117]

[0118]

[0119] In a preferred embodiment of the present invention, the compound represented by Formula I is any of the following compounds:

[0120]

[0121]

[0122] The present invention also provides a method for preparing the compound shown in Formula I, comprising method 1, method 2, method 3, method 4, method 5, method 6 or method 7.

[0123] When L does not exist, R in Y and Z 1a R 1g R 2a R 2g R 3a R 3g R 4a and R 4g Independently H or C 1-12 When alkyl, the preparation method of the compound as shown in Formula I is Method 1 or Method 2:

[0124] Method 1: It includes the following steps: in the presence of an alkaline reagent, a palladium catalyst and a phosphine ligand, a coupling reaction is carried out in a solvent between a compound of formula II and a compound of formula III to obtain the compound shown in formula I.

[0125]

[0126] Wherein, -Bpin is pinacol borate ester;

[0127] Method 2: It includes the following steps: in the presence of an alkaline reagent and a palladium catalyst, a compound of formula II and a compound of formula IV are subjected to a cross-coupling reaction in a solvent to obtain the compound shown in formula I.

[0128]

[0129] In equation IV, R 9 Halogens (e.g., Cl, Br, or I);

[0130] When "R" is in Y and Z 1a R 1g R 3a and R 3g At least one substituent is -SO3Q, and the remaining substituents are independently C. 1-12 Alkyl", "R" 1a R 1g R 4a and R 4g At least one substituent is -SO3Q, and the remaining substituents are independently C. 1-12 Alkyl", "R" 2a R 2g R3a and R 3g At least one substituent is -SO3Q, and the remaining substituents are independently C. 1-12 Alkyl", "R" 2a R 2g R 4a and R 4g At least one substituent is -SO3Q, and the remaining substituents are independently C. 1-12 Alkyl or R 11a R 11g R 10a R 10c and R 10d At least one substituent is -SO3Q, and the remaining substituents are independently C. 1-12 When "alkyl" is used, the preparation method of the compound shown in Formula I is Method 3:

[0131] Method 3: It includes the following steps: reacting an alkali metal base and compound V in a solvent to obtain the compound shown in Formula I:

[0132]

[0133] The compound V is one of the following compounds: V-1, V-2, V-3, V-4, or V-5:

[0134]

[0135] In compound V-1, R 5a R 5g R 7a and R 7g At least one substituent is -SO3H, and the remaining substituents are independently C. 1-12 Alkyl; R 1b R 1c R 1d R 1e R 1f R 3b R 3c R 3d R 3e and R 3f The definitions are the same as those described above;

[0136] In compound V-2, R 5a R 5g R 8a and R 8g At least one substituent is -SO3H, and the remaining substituents are independently C. 1-12 Alkyl; R 1b R 1c R 1d R1e R 1f R 4b R 4c R 4d R 4e and R 4f The definitions are the same as those described above;

[0137] In compound V-3, R 6a R 6g R 7a and R 7g At least one substituent is -SO3H, and the remaining substituents are independently C. 1-12 Alkyl; R 2b R 2c R 2d R 2e R 2f R 3b R 3c R 3d R 3e and R 3f The definitions are the same as those described above;

[0138] In compound V-4, R 6a R 6g R 8a and R 8g At least one substituent is -SO3H, and the remaining substituents are independently C. 1-12 Alkyl; R 2b R 2c R 2d R 2e R 2f R 4b R 4c R 4d R 4e and R 4f The definitions are the same as those described above;

[0139] In compound V-5, R 12a and R 12g R 10a R 10c and R 10d At least one substituent is -SO3H, and the remaining substituents are independently C. 1-12 Alkyl; R 11b R 11c R 11d R 11e R 11f R 10b and R 10e The definitions are the same as those described above;

[0140] When L is a linker, the preparation method of the compound shown in Formula I is method 4 or method 5:

[0141] Method 4: It includes the following steps: in the presence of cuprous halide, alkali metal base and N-methylglycine, the compound shown in Formula IV, the compound shown in Formula VI and the compound shown in Formula VII are reacted in a solvent to obtain the compound shown in Formula I.

[0142]

[0143] R 9 and R 10 It can be a halogen on its own (e.g., Cl, Br, or I);

[0144] In Formula VII, H in "HLH" represents a hydrogen atom;

[0145] Method 5: It includes the following steps: in the presence of a base, cuprous halide, palladium catalyst and phosphine ligand, the compound shown in Formula IV and the compound shown in Formula VIII are coupled in a solvent to obtain the compound shown in Formula I.

[0146]

[0147] In equation IV, R 9 Halogens (e.g., Cl, Br, or I);

[0148] In Formula VIII, H in “ZLH” is a hydrogen atom; the definitions of Y, Z and L are the same as those described above;

[0149] When L is Then, the method for preparing the compound as shown in Formula I is Method 6:

[0150] Method 6: It includes the following steps: reacting the compound shown in Formula IX, the compound shown in Formula X, and the compound XI in a solvent to obtain the compound shown in Formula I;

[0151]

[0152] R 11 and R 12 Independently -COOH, -S(O)2OH, -COOR 13 or -S(O)2R 14 ;

[0153] R 13 and R 14 It can be a halogen on its own (e.g., Cl, Br, or I);

[0154] The compound XI is Ring B, X 9 X 10 The definitions of n and n10 are the same as those described above;

[0155] When L does not exist, in group (b), R in Y 11a and R 11g Independently H or C 1-12 When alkyl, the method for preparing the compound as shown in Formula I is Method 7:

[0156] Method 7: It comprises the following steps: in the presence of a base, a palladium catalyst and a phosphine ligand, a compound as shown in Formula XII and a compound as shown in Formula VI are coupled in a solvent to obtain the compound as shown in Formula I.

[0157]

[0158] In equation VI, R 10 Halogens (e.g., Cl, Br, or I);

[0159] In formula XII, H in “ZH” is a hydrogen atom; the definitions of Y and Z are the same as those described above.

[0160] In Method 1, the conditions and operations of the coupling reaction can be conventional conditions and operations for such reactions in the art. The present invention particularly prefers the following conditions and operations:

[0161] The basic reagent can be an alkali metal carbonate (e.g., potassium carbonate). The molar ratio of the basic reagent to the compound of formula II can be (2-7):1 (e.g., 5:1). The palladium catalyst can be Pd(OAc)₂. The molar ratio of the palladium catalyst to the compound of formula II can be (0.05-0.15):1 (e.g., 0.1:1). The phosphine ligand can be PPh₃. The molar ratio of the phosphine ligand to the compound of formula II can be (0.1-0.3):1 (e.g., 0.2:1). The solvent can be an alcohol solvent (e.g., ethanol). The reaction temperature can be 50-70°C, for example, 60°C.

[0162] In Method 2, the conditions and operations for the cross-coupling reaction can be conventional conditions and operations for such reactions in the art. The present invention particularly prefers the following conditions and operations:

[0163] The basic reagent can be an alkali metal carbonate (e.g., Cs₂CO₃). The molar ratio of the basic reagent to the compound of formula II can be (1.5-3.5):1 (e.g., 3:1). The palladium catalyst can be Pd(dppf)Cl₂. The molar ratio of the palladium catalyst to the compound of formula II can be (0.02-0.07):1 (e.g., 0.05:1). The solvent can be an ether solvent (e.g., tetrahydrofuran). The reaction temperature can be room temperature.

[0164] In method 3, the reaction conditions and operations can be conventional conditions and operations for such reactions in the art. The present invention particularly prefers the following conditions and operations:

[0165] The alkali metal base may be potassium hydroxide. The reaction temperature may be -5 to 5°C, for example, 0°C.

[0166] In method 4, the reaction conditions and operations can be conventional conditions and operations for such reactions in the art. The present invention particularly prefers the following conditions and operations:

[0167] The cuprous halide may be cuprous iodide. The molar ratio of the cuprous halide to the compound of formula IV may be (0.1-0.3):1 (e.g., 0.2:1). The alkali metal base may be potassium hydroxide. The molar ratio of the alkali metal base to the compound of formula IV may be (3-7):1 (e.g., 5:1). The molar ratio of N-methylglycine to the compound of formula IV may be (0.03-0.05):1 (e.g., 0.04:1). The solvent may be an ether solvent (e.g., 1,4-dioxane). The reaction temperature may be 80-120°C, for example, 100°C.

[0168] In method 5, the conditions and operations of the coupling reaction can be conventional conditions and operations for such reactions in the art. The present invention particularly prefers the following conditions and operations:

[0169] The base is a tertiary amine base (e.g., triethylamine). The molar ratio of the base to the compound of formula IV can be (2-7):1 (e.g., 5:1). The cuprous halide can be cuprous iodide. The molar ratio of the cuprous halide to the compound of formula IV can be (0.05-0.15):1 (e.g., 0.1:1). The palladium catalyst can be Pd(PPh3)2Cl2. The molar ratio of the palladium catalyst to the compound of formula II can be (0.01-0.1):1 (e.g., 0.05:1). The phosphine ligand can be PPh3. The molar ratio of the phosphine ligand to the compound of formula II can be (0.05-0.15):1 (e.g., 0.1:1). The solvent can be an aromatic solvent (e.g., toluene). The reaction temperature can be 50-70°C, for example, 60°C.

[0170] In method 6, the reaction conditions and operations can be conventional conditions and operations for such reactions in the art. The present invention particularly prefers the following conditions and operations:

[0171] When the R 11 and R 12 When the reaction is independently -COOH or -S(O)2OH, the reaction can be carried out in the presence of a condensing agent and a base conventional in the art. The condensing agent can be a combination of EDCI and HOBT. The base can be a tertiary amine base, such as triethylamine.

[0172] When the R 11 and R 12 Independently for -COOR 13 or -S(O)2R 14 The reaction can be carried out in the presence of a base conventional in the art. The base can be a tertiary amine base, such as triethylamine.

[0173] In method 7, the reaction conditions and operations can be conventional conditions and operations for such reactions in the art. The present invention particularly prefers the following conditions and operations:

[0174] The alkaline reagent can be an alkali metal alkoxide (e.g., NaO). t Bu). The molar ratio of the basic reagent to the compound of formula VI can be (1.5-3.5):1 (e.g., 3:1). The palladium catalyst can be Pd2(dba)3. The molar ratio of the palladium catalyst to the compound of formula VI can be (0.02-0.07):1 (e.g., 0.05:1). The solvent can be an aromatic solvent (e.g., toluene). The reaction temperature can be 90-110°C, for example, 110°C.

[0175] The present invention also provides a compound V-1, V-2, V-3, V-4 or V-5:

[0176]

[0177] In compound V-1, R 5a R 5g R 7a and R 7g At least one substituent is -SO3H, and the remaining substituents are independently C. 1-12 Alkyl; R 1b R 1c R 1d R 1e R 1f R 3b R 3c R 3d R 3e and R 3f The definitions are the same as those described above;

[0178] In compound V-2, R 5a R 5g R 8a and R 8g At least one substituent is -SO3H, and the remaining substituents are independently C. 1-12 Alkyl; R 1b R 1c R 1d R 1e R 1f R 4b R 4c R 4d R 4e and R 4f The definitions are the same as those described above;

[0179] In compound V-3, R 6a R 6g R 7a and R 7g At least one substituent is -SO3H, and the remaining substituents are independently C. 1-12 Alkyl; R 2b R 2c R 2d R 2e R 2f R 3b R 3c R 3d R 3e and R 3f The definitions are the same as those described above;

[0180] In compound V-4, R 6a R6g R 8a and R 8g At least one substituent is -SO3H, and the remaining substituents are independently C. 1-12 Alkyl; R 2b R 2c R 2d R 2e R 2f R 4b R 4c R 4d R 4e and R 4f The definitions are the same as those described above;

[0181] In compound V-5, R 12a and R 12g R 10a R 10c and R 10d At least one substituent is -SO3H, and the remaining substituents are independently C. 1-12 Alkyl; R 11b R 11c R 11d R 11e R 11f R 10b and R 10e The definitions are the same as those described above.

[0182] The preferred compounds for compound V-1 are as follows:

[0183]

[0184] The present invention also provides a pharmaceutical composition comprising the compound shown in Formula I above and a pharmaceutically acceptable carrier.

[0185] The present invention also provides the use of the above-described compound as shown in Formula I or the above-described pharmaceutical composition in the preparation of antiviral drugs, antibacterial drugs, anti-inflammatory drugs, anti-allergic drugs, anti-ulcer drugs or drugs for treating burns or scalds.

[0186] The present invention also provides a cosmetic composition comprising an active ingredient and a cosmetically acceptable medium; said active ingredient comprising the compound shown in Formula I above.

[0187] The present invention also provides the use of the compound shown in Formula I above in the preparation of cosmetics for preventing or isolating radiation or for preventing the oxidation of protein materials.

[0188] In structural fragments This means that the structural segment is connected to the rest of the molecule through this site.

[0189] The "-" at the end of a group indicates that the group is connected to the rest of the molecule through that site.

[0190] In this invention, room temperature refers to 10-35℃.

[0191] The term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0192] The term "alkyl" refers to a saturated monovalent hydrocarbon group that has a specified number of carbon atoms (e.g., C1 to C6), is straight-chain or branched. Alkyl groups include, but are not limited to: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, etc.

[0193] The term "heteroalkyl" refers to a cyclic group having a specified number of ring atoms (e.g., 5 to 6), a specified number of heteroatoms (e.g., 1 or 2), and a specified type of heteroatoms, which is monocyclic and saturated.

[0194] The term "pharmaceuticalally acceptable carrier" refers to all substances contained in a pharmaceutical preparation other than the active pharmaceutical ingredient, generally divided into two main categories: excipients and additives. For details, please refer to the *Pharmacopoeia of the People's Republic of China (2020 Edition)* and the *Handbook of Pharmaceutical Excipients* (Paul J Sheskey, Bruno C Hancock, Gary PMoss, David J Goldfarb, 2020, 9th Edition).

[0195] The term "cosmetic acceptable medium" refers to a medium that is compatible with any keratinous material such as skin, hair, nails, eyelashes, eyebrows, and lips, as well as any other area of ​​the skin on the body or face.

[0196] The positive and progressive effects of this invention are that the compounds of this invention have at least one of the following advantages: good stability, antioxidant activity, anti-inflammatory activity and high safety. Attached Figure Description

[0197] Figure 1 The bar chart shows the relative antioxidant capacity of the compounds determined by the FRAP method in Example 2; where "**" indicates P<0.01, which is a significant difference.

[0198] Figure 2 The bar chart shows the scavenging rates of some compounds on DPPH free radicals at different reaction times in Example 3; where "**" indicates P<0.01, "***" indicates P<0.001, and "****" indicates P<0.0001, representing significant differences.

[0199] Figure 3 The bar chart shows the inhibitory effects of some compounds in Example 4 on NO production in LPS-activated RAW 264.7 cells; where "*" indicates P<0.05, "**" indicates P<0.01, "***" indicates P<0.001, and "****" indicates P<0.0001, representing significant differences.

[0200] Figure 4 The bar chart shows the inhibitory effects of some compounds in Example 4 on NO production in LPS-activated RAW 264.7 cells; where "*" indicates P<0.05, "**" indicates P<0.01, "***" indicates P<0.001, and "****" indicates P<0.0001, representing significant differences. Detailed Implementation

[0201] The present invention will be further illustrated by way of embodiments below, but the present invention is not limited to the scope of the embodiments described herein.

[0202] Synthesis of sodium 5-isopropyl-3,8-dimethylazine-1-sulfonate (GAS-Na).

[0203] References (Peet, J.; Selyutina, A.; Bredihhin, A. Biorg. Med. Chem. 2016, 24, 1653.). Guaiacomo (793.2 mg, 4 mmol) was added to a 50 mL three-necked flask, along with 2 mL of acetic anhydride. The mixture was stirred at room temperature until completely dissolved. The flask was then placed in an ice bath, and a mixture of 1 mL concentrated H₂SO₄ and 2 mL acetic anhydride was slowly added dropwise. The mixture was allowed to heat naturally for 4 h, and the reaction was monitored by TLC until complete. The reaction flask was then placed in an ice bath, and NaOH aqueous solution was slowly added dropwise to adjust the pH to 8–9. A deep blue solid precipitated. The blue solid was filtered, and the lipophilic impurities and excess inorganic base were washed away with petroleum ether and a small amount of water. Finally, recrystallization with anhydrous ethanol yielded 1.0565 g of the blue solid product, with a yield of 88%.

[0204] 1H NMR spectrum: 1 H NMR(400MHz,CD3OD)δ:8.26(d,J=1.9Hz,1H),8.12(s,1H),7.54(dd,J=11.0,1.8Hz,1H),7.28 (d,J=11.1Hz,1H),3.43(s,3H),3.12(hept,J=6.8Hz,1H),2.58(s,3H),1.38(d,J=6.9Hz,6H).

[0205] Example 1: Synthesis of 2-(7-isopropyl-1,4-dimethylazin-2-yl)-4,4,5,5-tetramethyl-1,3,2-dioxoborane (1).

[0206]

[0207] Reference (Narita, M. et al. J. Org. Chem. 2018, 83, 1298.). Guaiac (993 mg, 5.0 mmol), pinacol diboronate (1.32 g, 5.2 mmol), 4,7-dimethyl-1,10-phenanthroline (10 mg, 0.05 mmol), and a stir bar were weighed into a 50 mL Schlenk flask. The flask was purged with N2 three times. In a glove box, catalyst [Ir(OMe)(cod)]2 (17 mg, 0.025 mmol) was weighed into a reaction tube. 15 mL of ultra-dry n-heptane was added under N2 atmosphere, and the mixture was refluxed at 110 °C for 3 h. The reaction was monitored by TLC until complete. After returning to room temperature, the solution was quenched with water, extracted three times with ethyl acetate, dried over anhydrous Na2SO4, concentrated, and then subjected to silica gel column chromatography [V(PE):V(EA)=20:1] to give 1.56 g of blue solid 1, yield 96%.

[0208] 1H NMR spectrum: 1 H NMR (400MHz, CDCl3) δ: 8.46 (s, 1H), 7.87 (s, 1H), 7.49 (dd, J = 10.5, 1.7Hz, 1H), 7.06 (d, J = 10.6H) z,1H),3.20(hept,J=6.8Hz,1H),3.09(s,3H),3.01(s,3H),1.57(s,12H),1.53(d,J=6.9Hz,6H).

[0209] Example 2: Synthesis of 7,7′-diisopropyl-1,1′,4,4′-tetramethyl-2,2′-diazine (BGA).

[0210]

[0211] Step A: Synthesis of 2-bromo-7-isopropyl-1,4-dimethyl azulene (2a) and 2-iodo-7-isopropyl-1,4-dimethyl azulene (2b).

[0212]

[0213] Reference (Narita, M. et al. J. Org. Chem. 2018, 83, 1298.). CuBr (473.4 mg, 3.3 mmol) was weighed into a 250 mL single-necked flask, and 20 mL of DMF was added. The mixture was heated and stirred to 90 °C. The solution changed from light green to brownish-red. After maintaining the temperature at 90 °C for 5 min, 20 mL of DMF solution (972.8 mg, 3.0 mmol) was slowly added. The reaction was carried out at 90 °C for 3 h. After cooling to room temperature, the reaction solution was poured into water and filtered through diatomaceous earth to obtain the metal salt. Simultaneously, DMF was washed away with a large amount of water. The filtrate was extracted with ethyl acetate. The organic phase was blue-purple. The blue-purple product remaining on the diatomaceous earth was washed with ethyl acetate. The organic phases were combined, dried over anhydrous Na₂SO₄, concentrated, and then subjected to silica gel column chromatography with PE as the eluent. 704.8 mg of a blue-purple oil, 2a, was finally obtained, with a yield of 85%. 1H NMR spectrum: 1 H NMR (400MHz, CDCl3) δ: 8.16 (d, J = 1.5Hz, 1H), 7.46 (dd, J = 10.6, 1.6Hz, 1H), 7.26 (s, 1H), 7.09 (d,J=10.6Hz,1H),3.10(hept,J=6.9Hz,1H),2.78(s,3H),2.60(s,3H),1.36(d,J=6.9Hz,6H).

[0214] The synthesis of compound 2b follows the same steps as 2a, yielding a blue-purple oily substance in 53% yield. (H1N NMR spectrum follows.) 1 H NMR (400MHz, CDCl3) δ: 8.44 (d, J = 1.8 Hz, 1H), 7.67 (s, 1H), 7.59 (dd, J = 10.6, 1.9 Hz, 1H), 7.17 (d,J=10.6Hz,1H),3.28(hept,J=6.9Hz,1H),2.91(s,3H),2.89(s,3H),1.60(d,J=6.9Hz,4H).

[0215] Step B: Synthesis of 7,7′-diisopropyl-1,1′,4,4′-tetramethyl-2,2′-diazine (BGA)

[0216] Weigh 1 (340.5 mg, 1.05 mmol), 2a (277.2 mg, 1 mmol), Pd(dppf)Cl2 (36.6 mg, 0.05 mmol), and Cs2CO3 (977.5 mg, 3 mmol) into a 50 mL Schlenk tube. Replace the tube with N2 three times. Add 12 mL of freshly distilled THF and 3 mL of H2O bubbled with N2 under an N2 atmosphere. React at room temperature for 24 h. After the reaction is complete as monitored by TLC, pour the reaction solution into water, extract with petroleum ether, dry over anhydrous Na2SO4, concentrate, and perform silica gel column chromatography [V(PE):V(EA) = 50:1] to obtain 387.9 mg of a green solid, with a yield of 98%.

[0217] 1H NMR spectrum: 1 H NMR (400MHz, CDCl3) δ: 8.30 (d, J = 1.7Hz, 1H), 7.44 (s, 1H), 7.42 (dd, J = 10.6, 1.8Hz, 1H), 7.07 (d, J = 10.6 Hz, 1H), 3.13 (hept, J = 6.9 Hz, 1H), 2.89 (s, 3H), 2.72 (s, 3H), 1.40 (d, J = 6.9 Hz, 6H).

[0218] The results showed that not only in step A, the yield of bromide 2a (85%) was higher than that of iodide 2b (53%), but also in the Suzuki coupling reaction in step B, the conversion of bromide 2b to BGA was higher than that of iodide 2a. Specific yields are shown in the table below:

[0219]

[0220] a The yields of generating 2a and 2b from 1, respectively; b The yields of BGA produced from 2a and 2b via Suzuki coupling reactions are given. c The overall yield is obtained by multiplying the yields from steps A and B.

[0221] Example 3: Synthesis of 7,7′-diisopropyl-1,1′,4,4′-tetramethyl-2,2′-diazine (BGA).

[0222]

[0223] Compound 1 (1.4961 g, 4.6 mmol), Pd(OAc)2 (51.6 mg, 0.23 mmol), PPh3 (120.7 mg, 0.46 mmol), and K2CO3 (1.5894 g, 11.5 mmol) were weighed into a three-necked flask, and 46 mL of ultradry ethanol was added. Two of the flask necks were sealed with flap stoppers, and the remaining neck was evacuated three times using an oxygen bulb connected to a three-way connector. The reaction was then carried out at 60 °C for 30 min. After the reaction was completed, the mixture was cooled to room temperature, and the ethanol was removed by rotary evaporation. The mixture was then extracted with ethyl acetate, dried over anhydrous Na2SO4, concentrated, and subjected to silica gel column chromatography [V(PE):V(EA) = 50:1]. The final product was a blackish-green solid of 355.8 mg, with a yield of 37%. The characterization results were the same as in Example 3.

[0224] Example 4: Sodium 5,5′-diisopropyl-3,3′,8,8′-tetramethyl-[2,2′-diazine]-1,1′-disulfonate (A) 2-2 Synthesis of ).

[0225] A 2-2 The product was obtained by sulfonation of BGA. 394.6 mg (1 mmol) of BGA was added to a 50 mL three-necked flask, along with 4 mL of acetic anhydride. The mixture was stirred at room temperature until completely dissolved. The flask was placed in an ice bath, and a mixture of 2 mL concentrated H₂SO₄ and 2.5 mL acetic anhydride was slowly added dropwise. The mixture was allowed to react naturally for 8 hours, and the reaction was monitored by TLC until complete. NaOH aqueous solution was then slowly added dropwise to adjust the pH to 8–9, resulting in the precipitation of a deep blue solid. The blue solid was filtered, and the product was washed with petroleum ether and a small amount of water to remove fat-soluble impurities and excess inorganic base. Finally, recrystallization with anhydrous ethanol yielded 335.2 mg of a blue-purple solid product, with a yield of 56%.

[0226] 1H NMR spectrum: 1 ¹H NMR (400MHz, CD₃OD) δ: 8.25 (d, J = 2.0Hz, 1H), 7.44 (dd, J = 11.2, 2.0Hz, 1H), 7.26 (d, J = 11.2Hz, 1H), 3.48 (s, 3H), 3.09 (hept, J = 6.8Hz, 1H), 2.14 (s, 3H), 1.37 (d, J = 6.9Hz, 6H); ¹H NMR spectrum: 13 C NMR (101MHz, CD3OD) δ: 152.0, 148.0, 143.1, 138.4, 135.7, 135.3, 133.8, 131.8, 127.1, 124.0, 38.8, 29.0, 24.9, 11.8; High-resolution mass spectrometry: HRMS (ESI) m / z: [M+H] + calcd for C30 H 33 O6Na2S2 599.1508, found 599.1506.

[0227] Example 5: Synthesis of compound B-1

[0228] Synthesis of compound 1′:

[0229]

[0230] Specific steps: The standard Sonogashira coupling reaction was performed. Compound 2a (654.3 mg, 2.3 mmol), Pd(PPh3)2Cl2 (163.5 mg, 0.23 mmol), PPh3 (122.2 mg, 0.46 mmol), and CuI (88.7 mg, 0.46 mmol) were weighed into a 100 mL Schlenk reaction flask. The flask was purged with N2 three times. Under N2 atmosphere, redistilled toluene (9.7 mL), ultra-dry triethylamine (9.7 mL), and trimethylethynylsilane (1 mL, 7 mmol) were added. The reaction was carried out at 60 °C for 2 h. After the reaction was complete, the mixture was extracted with dichloromethane, concentrated, and then subjected to silica gel column chromatography with n-hexane as the eluent, yielding 664.7 mg of a blue oily product, with a yield of 97%. Next, 328.5 mg (1.1 mmol) of the product was weighed and dissolved in 5.5 mL of DMF, followed by the addition of 1.1 mL of an aqueous solution of KF (127.8 mg, 2.2 mmol). The reaction was carried out at room temperature for 5 h. After the reaction was completed, the product was extracted with ethyl acetate, concentrated, and then subjected to neutral alumina column chromatography. Hexane elution was used to give product 1′187.8 mg, with a yield of 77%.

[0231] 1H NMR spectrum: 1 H NMR(400MHz, CDCl3)δ:8.30(d,J=1.6Hz,1H),7.49(s,1H),7.46(dd,J=10.6,1.7Hz,1H),7.09(d, J=10.6Hz,1H),3.75(s,1H),3.17(p,J=6.9Hz,1H),2.88(d,J=4.7Hz,6H),1.49(d,J=7.0Hz,6H).

[0232] Synthesis of compound B-1

[0233]

[0234] Compound 1′ was reacted with compound 2a to give compound B-1, using the same synthetic procedure as the Sonogashira coupling reaction described above. The yield was 64%. The 1H NMR spectrum of compound 1 is shown below. 1H NMR (400MHz, CDCl3) δ8.16(d,J=1.5Hz,1H),7.43(s,1H),7.37(dd,J=10.6,1.6Hz,1H),7.0 4(d,J=10.6Hz,1H), 3.09(hept,J=6.9Hz,1H), 2.82(d,J=5.4Hz,6H), 1.38(d,J=6.9Hz,6H).

[0235] Example 6: Synthesis of compound B-2

[0236]

[0237] Specific steps: Compound 2b (214 mg, 0.66 mmol), N-methylglycine (11.8 mg, 0.132 mmol), CuI (6.3 mg, 0.033 mmol), and KOH (92.6 mg, 1.65 mmol) were weighed into a 10 mL sealed tube. Nitrogen gas was purged three times. Then, under a nitrogen atmosphere, 2 mL of ultra-dry solvent 1,4-dioxane and 48.8 μL of 3,6-dioxo-1,8-octanedithiol (0.3 mmol) were added. The reaction was carried out at 100 °C for 24 hours. After the reaction was completed, the mixture was cooled to room temperature, extracted with ethyl acetate, dried over anhydrous Na₂SO₄, and the solvent was removed by rotary evaporation. The mixture was then subjected to silica gel column chromatography [V(PE):V(EA) = 10:1], finally yielding 48.4 mg of a blue oil, with a yield of 28%.

[0238] 1H NMR spectrum: 1 ¹H NMR (400MHz, CDCl₃) δ: 8.02 (d, J = 1.5Hz, 1H), 7.32 (dd, J = 10.7, 1.5Hz, 1H), 7.11–7.01 (m, 2H), 3.85 (t, J = 7.1Hz, 2H), 3.71 (s, 2H), 3.40 (t, J = 7.1Hz, 2H), 3.07 (hept, J = 6.8Hz, 1H), 2.79 (s, 3H), 2.53 (s, 3H), 1.36 (d, J = 6.9Hz, 6H); ¹H NMR spectrum: 13 C NMR(101MHz, CDCl3)δ:144.65,141.55,140.30,137.18,136.23,133.01,130. 37,126.76,121.77,108.71,70.59,70.29,38.51,31.90,24.91,24.29,10.80.

[0239] Example 7: Synthesis of compound B-3

[0240]

[0241] Specific steps: Compound 2b (365.9 mg, 1.13 mmol), Pd(OAc)2 (7.6 mg, 0.034 mmol), Xantphos (19.6 mg, 0.034 mmol), and DCC (466.3 mg, 2.26 mmol) were weighed into a 50 mL Schlenk reaction flask. The flask was purged with N2 three times. Ultra-dry triethylamine (0.34 mL, 2.26 mmol) and DMF (7 mL) were added under N2 atmosphere, and the reaction was carried out at 80 °C for 12 h. After the reaction was complete, the DMF solvent was washed off with water, and the mixture was extracted with ethyl acetate. The concentrate was then subjected to silica gel column chromatography [V(PE):V(EA) = 5:1]. After drying, 231.4 mg of green solid 3′ was obtained, with a yield of 85%.

[0242] 1H NMR spectrum: 1 H NMR(400MHz, CDCl3)δ:8.38(d,J=2.0Hz,1H),7.77(s,1H),7.49(dd,J=10.5,2.0Hz,1H),7.0 4(d,J=10.5Hz,1H),3.08(p,J=6.9Hz,1H),2.91(s,3H),2.85(s,3H),1.37(d,J=6.9Hz,6H).

[0243]

[0244] Specific steps: Compound 3′ (167.2 mg, 0.69 mmol), EDCl (172.5 mg, 0.9 mmol), and HOBt (121.6 mg, 0.9 mmol) were weighed into a 50 mL Schlenk reaction flask. The flask was purged with N2 three times. Under N2 atmosphere, ultra-dry triethylamine (125 μL, 0.9 mmol) and DMF (3 mL) were added. After reacting at room temperature for 40 min, 44 μL (0.3 mmol) of 1,8-diamino-3,6-dioxaoctane was added, and the reaction was continued at room temperature for 24 h. After the reaction was complete, the mixture was extracted with dichloromethane, concentrated, and then subjected to silica gel column chromatography [V(DCM):V(MeOH) = 15:1]. After drying, 172.9 mg of blue solid product B-3 was obtained, with a yield of 97%.

[0245] 1H NMR spectrum: 1¹H NMR (400MHz, CDCl₃) δ: 8.28 (d, J = 1.8Hz, 2H), 7.44 (dd, J = 10.6, 1.7Hz, 2H), 7.34 (s, 2H), 7.02 (d, J = 10.6Hz, 2H), 6.63 (s, 2H), 3.71 (d, J = 4.8Hz, 12H), 3.07 (p, J = 6.8Hz, 2H), 2.78 (d, J = 4.1Hz, 12H), 1.64 (s, 4H), 1.35 (d, J = 6.9Hz, 12H); High-resolution mass spectrometry: HRMS (ESI) m / z: [M+Na] + calcd for C 38 H 48 N2O4Na 619.35063,found619.35122.

[0246] Example 8: Synthesis of compound B-4

[0247]

[0248] Specific steps: In a 250 mL round-bottom flask, add GAS-Na (300.4 mg, 1 mmol) and 10 mL dichloromethane, stir thoroughly, and slowly add 0.4 mL DMF, 0.4 mL pyridine, and a mixed solution of oxaloyl chloride (320 mg, 2.5 mmol) and dichloromethane dropwise under an ice-water bath. React for 30 min under ice bath conditions. Next, add piperazine (42.2 mg, 0.49 mmol) to the reaction solution, and slowly add a mixed solution of pyridine (1 mL) and triethylamine (2 mL). Allow the mixture to react naturally at elevated temperature for 8 h. Finally, quench with water, extract with dichloromethane, concentrate, and then perform silica gel column chromatography [V(PE):V(EA) = 5:1]. After drying, 88.7 mg of green solid B-4 is obtained, with a yield of 30%.

[0249] 1H NMR spectrum: 1 H NMR (400MHz, CDCl3) δ: 8.28 (d, J = 2.0Hz, 2H), 8.11 (s, 2H), 7.62 (dd, J = 11.1, 2.0Hz, 2H), 7.42 (d, J = 1 1.2Hz,2H),3.32(s,6H),3.27(s,8H),3.14(hept,J=6.9Hz,2H),2.56(s,6H),1.38(d,J=6.9Hz,12H); 13C NMR (101MHz, CDCl3) δ: 149.40, 145.50, 143.33, 141.56, 137.64, 135.71, 135.52, 133.07, 123.60, 116.78, 44.69, 38.00, 28.21, 24.63, 13.05; High-resolution mass spectrometry: HRMS (ESI) m / z: [M+H] + calcd for C 34 H 43 N2O4S2 607.2659, found 607.2664.

[0250] Example 9: Synthesis of CAS-Na

[0251]

[0252] Specific steps: Add 40 mg (0.22 mmol) of CA to a 50 mL single-necked flask, add 1 mL of acetic anhydride, and stir at room temperature until CA is completely dissolved. Place the single-necked flask in an ice bath and slowly add a mixed solution of 0.35 mL concentrated H₂SO₄ and 1 mL acetic anhydride. After the addition is complete, allow the mixture to heat naturally and react at room temperature for 4 hours. Monitor the reaction for completeness using TLC. Then, place the single-necked flask in an ice bath and slowly add a concentrated aqueous solution of NaOH to adjust the pH to 8-9. Add a small amount of methanol, and a yellow precipitate forms. Filter to remove the precipitate. Add a small amount of methanol and a very small amount of acetone again, and a yellow precipitate forms. Filter to remove the precipitate. Repeat this process several times until no yellow precipitate forms. After removing a large amount of solvent by rotary evaporation, continue to add a small amount of methanol and a very small amount of acetone until no yellow precipitate forms. Add a small amount of acetone and sonicate. A blue precipitate forms. Filter the precipitate and wash away impurities with a small amount of water and petroleum ether to obtain 13 mg of blue solid CAS-Na, with a yield of 20%.

[0253] 1H NMR spectrum: 1 ¹H NMR (400MHz, Methanol-d⁴) δ: 8.22(s, 1H), 8.10(s, 1H), 7.49(d, J = 10.8Hz, 1H), 7.22(d, J = 11.0Hz, 1H), 3.41(s, 3H), 2.85(q, J = 7.5Hz, 2H), 2.56(s, 3H), 1.34(t, J = 7.6Hz, 3H); ¹H NMR spectrum: 13C10 NMR (100MHz, Methanol-d4) δ: 149.21, 141.62, 140.08, 139.06, 138.64, 136.78, 134.06, 130.95, 129.41, 123.15, 33.93, 28.11, 17.39, 12.87; High-resolution mass spectrometry: HRMS (ESI) m / z: [M+H] + calcd for C 14 H 15 NaO3S 263.0747,found263.0740.

[0254] Example 10: Synthesis of 2-(7-ethyl-1,4-dimethylazin-2-yl)-4,4,5,5-tetramethyl-1,3,2-dioxoborane (4')

[0255]

[0256] Specific steps: Take CA (860 mg, 4.7 mmol), pinacol diboronic acid (1193.8 mg, 4.7 mmol), and 4,7-dimethyl-1,10-phenanthroline (9.4 mg, 0.047 mmol) in a 100 mL Schlenk flask, replace with N2 three times, and weigh the catalyst [Ir(OMe)(cod)]2 (15.98 mg, 0.0235 mmol) into a reaction tube in a glove box. Add 15 mL of ultra-dry n-heptane under N2 atmosphere, heat to reflux at 110 °C for 3 h, and monitor the reaction until complete by TLC. After returning to room temperature, quench with water, extract three times with ethyl acetate, dry with anhydrous Na2SO4, concentrate, and perform silica gel column chromatography [V(PE):V(EA) = 10:1] to obtain 750 mg of blue-black solid 4′, yield 51%.

[0257] 1H NMR spectrum: 1 ¹H NMR (400MHz, CD₂Cl₂) δ: 8.20 (s, 1H), 7.51 (s, 1H), 7.38 (d, J = 10.4 Hz, 1H), 6.93 (d, J = 10.5 Hz, 1H), 2.86–2.75 (m, 8H), 1.38 (s, 12H), 1.32 (t, J = 7.6 Hz, 3H); ¹H NMR spectrum: 13 C10 NMR (125MHz, CDCl3) δ: 146.23, 137.94, 137.67, 137.13, 136.46, 135.69, 133.33, 124.85, 120.47, 83.34, 33.89, 25.08, 24.36, 17.44, 12.75; High-resolution mass spectrometry: HRMS (ESI) m / z: [M+H]+ calcd for C 20 H 27 BO2 330.2213, found 310.2210.

[0258] Example 11: Synthesis of 7,7′-diethyl-1,1′,4,4′-tetramethyl-2,2′-diazine (BCA)

[0259]

[0260] Step A: Synthesis of 2-bromo-7-ethyl-1,4-dimethylazine (5a) and 2-iodo-7-ethyl-1,4-dimethylazine (5b).

[0261]

[0262] CuBr (104.7 mg, 0.55 mmol) was added to a 50 mL single-necked flask, along with 5 mL of DMF. The mixture was heated to 90 °C and stirred openly for 5 min. The solution changed from light green to brownish-red. 5 mL of DMF solution (4′, 155.12 mg, 0.5 mmol) was slowly added, and the mixture was reacted at 90 °C for 3 h. The reaction was monitored by TLC until complete. After the reaction solution cooled to room temperature, a small amount of water was added to quench the reaction. The mixture was filtered through diatomaceous earth, and the blue-purple product remaining on the diatomaceous earth was washed away with ethyl acetate, yielding a blue-purple filtrate. After extraction with ethyl acetate and a large amount of water, the upper organic phase was collected, dried over anhydrous Na₂SO₄, concentrated, and then subjected to silica gel column chromatography with PE as the eluent. 100 mg of a blue-purple oil, 5a, was finally obtained, with a yield of 76%. (1H NMR spectrum:) 1 ¹H NMR (400MHz, CD₂Cl₂) δ: 8.16 (s, 1H), 7.46 (d, J = 10.5 Hz, 1H), 7.25 (s, 1H), 7.09 (d, J = 10.5 Hz, 1H), 2.86 (q, J = 7.6 Hz, 2H), 2.77 (s, 3H), 2.57 (s, 3H), 1.34 (t, J = 7.2 Hz, 3H); ¹H NMR spectrum: 13 C NMR (125MHz, CDCl3) δ: 143.21, 137.71, 136.41, 135.35, 134.31, 128.23, 126.63, 122.97, 114.45, 33.96, 24.24, 17.46, 11.81; High-resolution mass spectrometry: HRMS (ESI) m / z: [M+H] + calcd for C 14 H 15 Br263.0430,found263.0432.

[0263] The synthesis of compound 5b is the same as that of 5a; it is a blue-purple oil with a yield of 55%. (H NMR spectrum:) 1 ¹H NMR (400MHz, CDCl₃) δ: 8.16 (s, 1H), 7.47–7.40 (m, 2H), 7.04 (d, J = 10.5 Hz, 1H), 2.85 (q, J = 7.6 Hz, 2H), 2.77 (s, 3H), 2.61 (s, 3H), 1.34 (t, J = 7.5 Hz, 2H); ¹H NMR spectrum: 13 C10 NMR (100MHz, CDCl3) δ: 143.00, 137.59, 137.50, 136.84, 134.79, 134.37, 127.45, 126.46, 120.24, 103.34, 33.96, 24.28, 17.48, 14.71; High-resolution mass spectrometry: HRMS (ESI) m / z: [M+H] + calcd for C 14 H 15 I 311.0291, found 311.0291.

[0264] Step B: Synthesis of 7,7′-diethyl-1,1′,4,4′-tetramethyl-2,2′-diazine (BCA)

[0265] 5a (92.1 mg, 0.35 mmol), 1 (108.6 mg, 0.35 mmol), Pd(dppf)Cl2 (14.3 mg, 0.0175 mmol), and Cs2CO3 (342.1 mg, 1.05 mmol) were placed in a 25 mL Schlenk tube. The tube was purged with N2 three times. Under an N2 atmosphere, 4 mL of redistilled THF and 1 mL of H2O bubbled with N2 were added, and the reaction was allowed to proceed at room temperature for 24 h. After the reaction was completed by TLC monitoring, water was added to quench the reaction. The mixture was extracted with petroleum ether, dried over anhydrous Na2SO4, concentrated, and then subjected to silica gel column chromatography [V(PE):V(EA) = 20:1], finally yielding 82 mg of a green oily substance, BCA, with a yield of 70%. 1H NMR spectrum: 1 ¹H NMR (400MHz, CDCl₃) δ: 8.25(s, 2H), 7.43(s, 2H), 7.39(d, J = 10.4Hz, 2H), 7.04(d, J = 10.5Hz, 2H), 2.91–2.85(m, 10H), 2.70(s, 6H), 1.37(t, J = 7.9Hz, 6H); ¹H NMR spectrum: 13C10 NMR (100MHz, CDCl3) δ: 145.88, 143.70, 137.60, 136.83, 136.60, 135.95, 134.68, 125.69, 123.34, 115.38, 34.10, 24.32, 17.59, 12.05; High-resolution mass spectrometry: HRMS (ESI) m / z: [M+H] + calcd for C 28 H 30 367.2420, found367.2422.

[0266] The results showed that not only in step A, the yield of brominated product 5a (76%) was higher than that of iodinated product 5b (55%), but also in the Suzuki coupling reaction in step B, the conversion of brominated product 5a to BCA was higher than that of iodinated product 5b. Specific yields are shown in the table below:

[0267]

[0268] a The yields of 5a and 5b generated from 4′, respectively; b The yields of BCA produced from 5a and 5b via the Suzuki coupling reaction are given. c The overall yield is obtained by multiplying the yields from steps A and B.

[0269] Example 12: Synthesis of compounds GABI-1 to GABI-7

[0270]

[0271] Specific steps: Take 2a (83.2 mg, 0.3 mmol), 6′, 7′, 8′ or 9′ (0.45 mmol), Pd2(dba)3 (13.7 mg, 0.015 mmol), t Bu-Xphos (23.8 mg, 0.03 mmol) and NaO t Bu (40.4 mg, 0.42 mmol) was added to a 25 mL Schlenk tube, and the tube was purged with N2 three times. Then, 3 mL of redistilled toluene was added under N2 atmosphere, and the reaction was carried out at 110 °C for 24 h. The reaction was monitored by TLC [V(PE):V(EA) = 1:1] to ensure completion. After the reaction solution cooled to room temperature, it was poured into water and extracted three times with ethyl acetate. The organic phases were combined and dried over anhydrous Na2SO4. After the solvent was evaporated, the solution was separated by silica gel column chromatography [V(PE):V(EA):V(Et3N) = 200:200:1].

[0272] 80.2 mg of the blue solid product GABI-1 was obtained, with a yield of 85%; mp 141–142 °C; 1H NMR spectrum: 1 ¹H NMR (400MHz, CD₂Cl₂) δ: 8.37 (s, 1H), 8.20 (s, 1H), 7.88–7.82 (m, 1H), 7.60–7.53 (m, 2H), 7.38–7.33 (m, 3H), 7.23 (d, J = 10.6 Hz, 1H), 3.21–3.13 (m, 1H), 2.88 (s, 3H), 2.59 (s, 3H), 1.41 (d, J = 6.9 Hz, 6H); ¹H NMR spectrum: 13 C NMR (100MHz, CD₂Cl₂) δ: 145.80, 144.31, 143.80, 142.81, 140.34, 136.94, 136.37, 135.86, 135.02, 134.80, 127.40, 123.85, 122.94, 120.55, 117.29, 111.59, 108.22, 38.78, 24.88, 24.46, 10.65; Infrared spectrum: IR (KBr) ν: 3450, 3054, 2960, 29 23,2863,1897,1776,1725,1710,1609,1556,1531,1496,1456,1403,1390,1373,1299,1270,1217,1154,1142,1121 ,1098,1070,1051,1004,968,931,885,864,849,815,789,779,765,746,719,667,636,628,570,546,476,432,404cm -1 High-resolution mass spectrometry: HRMS (ESI) m / z: [M+H] + calcd for C 22 H 22 N2 315.1856, found 315.1856.

[0273] 38.9 mg of the blue solid product GABI-2 was obtained, with a yield of 39%; mp 115–117 °C; 1H NMR spectrum: 1¹H NMR (500MHz, CD₂Cl₂) δ: 8.38 (d, J = 2.0Hz, 1H), 8.27 (s, 1H), 7.61–7.54 (m, 2H), 7.48 (dd, J = 8.9, 4.7Hz, 1H), 7.32 (s, 1H), 7.24 (d, J = 10.6Hz, 1H), 7.11 (td, J = 9.1, 2.5Hz, 1H), 3.17 (hept, J = 6.9Hz, 1H), 2.88 (s, 3H), 2.57 (s, 3H), 1.41 (d, J = 6.9Hz, 6H); ¹H NMR spectrum: 13 C NMR (125MHz, CD₂Cl₂) δ: 160.0 (d, J = 237.6 Hz), 146.2, 145.3, 144.2 (d, J = 12.5 Hz), 143.0, 139.7, 136.9, 136.4, 136.2, 135.1, 131.6, 127.6, 117.3, 112.3 (d, J = 7.2 Hz), 112.2 (d, J = 8.7 Hz), 108.1, 106.0 (d, J = 24.4 Hz), 38.8, 24.9, 24.5, 10.6; fluorine NMR spectrum: 19 F NMR (376MHz, CD₂Cl₂) δ: -108.10; Infrared spectrum: IR (KBr) ν: 3434, 2961, 2923, 2863, 1859, 1731, 1622, 1607, 1591, 1563, 1530, 1497, 1439, 1391, 1373, 1360, 1309, 1297, 1260, 1216, 1129, 1115, 1095, 1051, 1003, 970, 951, 868, 851, 828, 817, 803, 782, 750, 720, 704, 669, 645, 630, 613, 558, 483, 439 cm⁻¹ -1 High-resolution mass spectrometry: HRMS (ESI) m / z: [M+H] + calcd forC 22 H 21 FN2 333.1762, found 333.1761.

[0274] 57.8 mg of the blue solid product GABI-3 was obtained, with a yield of 58%; mp 91–93 °C; 1H NMR spectrum: 1¹H NMR (500MHz, CD₂Cl₂) δ: 8.38 (d, J = 2.0 Hz, 1H), 8.25 (s, 1H), 7.84 (dd, J = 8.9, 4.8 Hz, 1H), 7.59 (dd, J = 10.7, 2.0 Hz, 1H), 7.31 (s, 1H), 7.27–7.21 (m, 2H), 7.11 (td, J = 9.2, 2.5 Hz, 1H), 3.17 (hept, J = 6.9 Hz, 1H), 2.88 (s, 3H), 2.58 (s, 3H), 1.41 (d, J = 6.9 Hz, 6H); ¹H NMR spectrum: 13 C NMR (125MHz, CD₂Cl₂) δ: 160.7 (d, J = 239.8 Hz), 146.3, 144.6, 143.0, 140.2, 139.5, 136.9, 136.4, 136.3, 135.2, 135.1, 127.6, 121.2 (d, J = 10.0 Hz), 117.3, 111.5 (d, J = 25.8 Hz), 107.9, 98.3 (d, J = 27.7 Hz), 38.8, 24.9, 24.5, 10.6; fluorine NMR spectrum: 19 F NMR (376MHz, CD₂Cl₂) δ: -10⁵.72; Infrared spectrum: IR (KBr) ν: 3458, 3106, 2956, 2922, 2865, 1862, 1715, 1617, 1595, 1557, 1530, 1497, 1401, 1389, 1371, 1347, 1312, 1263, 1212, 1172, 1111, 1086, 1071, 1050, 1000, 968, 952, 936, 900, 842, 818, 808, 771, 749, 720, 665, 627, 613, 564, 538, 440 cm⁻¹ -1 High-resolution mass spectrometry: HRMS (ESI) m / z: [M+H] + calcd for C 22 H 21 FN2 333.1762,found333.1761.

[0275] 40.7 mg of the blue solid product GABI-4 was obtained, with a yield of 39%; mp 141–143 °C; 1H NMR spectrum: 1¹H NMR (500MHz, CD₂Cl₂) δ: 8.37 (d, J = 2.0 Hz, 1H), 8.24 (s, 1H), 7.86 (d, J = 2.0 Hz, 1H), 7.58 (dd, J = 10.6, 1.9 Hz, 1H), 7.47 (d, J = 8.6 Hz, 1H), 7.35–7.29 (m, 2H), 7.24 (d, J = 10.6 Hz, 1H), 3.17 (hept, J = 6.9 Hz, 1H), 2.87 (s, 3H), 2.56 (s, 3H), 1.41 (d, J = 6.9 Hz, 6H); ¹H NMR spectrum: 13 C NMR (125MHz, CD₂Cl₂) δ: 146.3, 145.1, 144.9, 143.0, 139.6, 136.9, 136.4, 136.2, 135.2, 133.7, 128.5, 127.6, 124.3, 120.2, 117.4, 112.6, 108.1, 38.8, 24.9, 24.5, 10.6; Infrared spectrum: IR (KBr) ν: 2955, 2920 ,2863,1873,1726,1605,1562,1530,1495,1461,1431,1392,1372,1306,1294,1240,1215,1099,1 072,1052,1001,968,938,910,870,819,805,780,732,719,691,666,637,624,597,555,485,422cm -1 High-resolution mass spectrometry: HRMS (ESI) m / z: [M+H] + calcdfor C 22 H 21 ClN2349.1464, found 349.1464.

[0276] 53.2 mg of the blue solid product GABI-5 was obtained, with a yield of 51%; mp 58–59 °C; 1H NMR spectrum: 1 ¹H NMR (500MHz, CD₂Cl₂) δ: 8.38 (d, J = 2.0 Hz, 1H), 8.21 (s, 1H), 7.80 (d, J = 8.6 Hz, 1H), 7.59 (dd, J = 10.6, 2.0 Hz, 1H), 7.52 (d, J = 2.0 Hz, 1H), 7.34–7.30 (m, 2H), 7.25 (d, J = 10.6 Hz, 1H), 3.17 (hept, J = 6.9 Hz, 1H), 2.88 (s, 3H), 2.56 (s, 3H), 1.41 (d, J = 6.9 Hz, 6H); ¹H NMR spectrum: 13C NMR (125MHz, CD₂Cl₂) δ: 146.4, 144.7, 143.0, 142.5, 139.4, 136.8, 136.4, 136.3, 135.7, 135.2, 129.7, 127.6, 123.7, 121.4, 117.5, 111.7, 108.1, 38.8, 24.9, 24.5, 10.6; Infrared spectrum: IR (KBr) ν: 3076, 2 957,2865,1712,1611,1530,1494,1451,1401,1388,1371,1332,1303,1275,1262,1210,1137,11 08,1086,1051,999,967,907,849,811,781,749,731,694,666,632,599,560,538,499,428,405cm -1 High-resolution mass spectrometry: HRMS (ESI) m / z: [M+H] + calcd forC 22 H 21 ClN2 349.1464, found 349.1465.

[0277] 47.5 mg of the blue solid product GABI-6 was obtained, with a yield of 46%; mp 53–55 °C; 1H NMR spectrum: 1 ¹H NMR (500MHz, CD₂Cl₂) δ: 8.38 (d, J = 2.0 Hz, 1H), 8.23 ​​(s, 1H), 7.81 (d, J = 8.8 Hz, 1H), 7.58 (dd, J = 10.6, 2.0 Hz, 1H), 7.34 (s, 1H), 7.24 (d, J = 10.6 Hz, 1H), 7.02–6.94 (m, 2H), 3.79 (s, 3H), 3.17 (hept, J = 6.8 Hz, 1H), 2.88 (s, 3H), 2.57 (s, 3H), 1.41 (d, J = 6.9 Hz, 6H); ¹³C NMR spectrum: 13C NMR (125MHz, CD₂Cl₂) δ: 158.0, 146.2, 143.3, 143.0, 139.8, 136.9, 136.3, 136.2, 135.6, 135.2, 127.5, 120.6, 117.7, 113.1, 108.3, 94.9, 56.2, 38.8, 24.9, 24.5, 10.6; Infrared spectrum: IR (KBr)ν:2958,2866,2833,1717,1618,1588,1558,1532,1493,1456,1402,1388,1372,13 45,1308,1274,1224,1089,1050,1027,969,944,821,769,735,710,668,633,565,419cm -1 High-resolution mass spectrometry: HRMS (ESI) m / z: [M+H] + calcd for C 23 H 24 N2O 345.1960,found 345.1960.

[0278] 43.4 mg of the blue solid product GABI-7 was obtained, with a yield of 42%; mp 139–141 °C; 1H NMR spectrum: 1 ¹H NMR (500MHz, CD₂Cl₂) δ: 8.40–8.31 (m, 2H), 7.58 (dd, J = 10.7, 2.0 Hz, 1H), 7.49 (s, 1H), 7.43 (d, J = 8.9 Hz, 1H), 7.32 (s, 1H), 7.24 (d, J = 10.6 Hz, 1H), 7.00 (dd, J = 8.9, 2.4 Hz, 1H), 3.87 (s, 3H), 3.17 (hept, J = 6.9 Hz, 1H), 2.87 (s, 3H), 2.57 (s, 3H), 1.40 (d, J = 6.9 Hz, 6H); ¹H NMR spectrum: 13C NMR (125MHz, CD₂Cl₂) δ: 157.4, 146.3, 144.2, 143.4, 143.0, 139.6, 136.9, 136.3, 136.2, 135.2, 129.3, 127.6, 117.3, 114.5, 112.4, 108.1, 102.0, 56.2, 38.8, 24.9, 24.5, 10.6; Infrared spectrum: IR (KBr) ν: 3416, 3104, 2960, 1619, 15 93,1555,1532,1488,1452,1400,1385,1372,1305,1275,1235,1215,1197,1184,1151,1136,1102,1086,104 9,1031,995,967,946,892,852,840,827,817,806,785,752,720,669,650,634,607,566,544,477,437,403cm -1 High-resolution mass spectrometry: HRMS (ESI) m / z: [M+H] + calcd for C 23 H 24 N2O 345.1960, found345.1960.

[0279] Example 13: Synthesis of compound CABI-1

[0280]

[0281] Specific steps: Take CA (39.3 mg, 0.15 mmol), 6′ (26.6 mg, 0.225 mmol), Pd2(dba)3 (6.9 mg, 0.0075 mmol), t Bu-Xphos (11.9 mg, 0.015 mmol) and NaO t Bu (20.2 mg, 0.225 mmol) was added to a 25 mL Schlenk tube, and the tube was purged with N2 three times. Then, 3 mL of redistilled toluene was added under N2 atmosphere, and the reaction was carried out at 110 °C for 24 h. The reaction was monitored by TLC [V(PE):V(EA) = 1:1] to ensure complete reaction. After the reaction solution cooled to room temperature, it was poured into water and extracted three times with ethyl acetate. The organic phases were combined and dried over anhydrous Na2SO4. After the solvent was evaporated, silica gel column chromatography [V(PE):V(EA):V(Et3N) = 200:200:1] was used to separate 27.6 mg of the blue compound CABI-1, with a yield of 61%. mp 160–161 °C; 1H NMR spectrum: 1¹H NMR (500MHz, CD₂Cl₂) δ: 8.34 (d, J = 1.9 Hz, 1H), 8.19 (s, 1H), 7.89–7.82 (m, 1H), 7.59–7.51 (m, 2H), 7.38–7.32 (m, 3H), 7.20 (d, J = 10.5 Hz, 1H), 2.92 (q, J = 7.6 Hz, 2H), 2.87 (s, 3H), 2.58 (s, 3H), 1.38 (t, J = 7.6 Hz, 3H); ¹H NMR spectrum: 13 C NMR (125MHz, CD₂Cl₂) δ: 145.8, 144.3, 143.8, 140.4, 138.7, 137.4, 136.9, 136.5, 136.2, 135.0, 127.3, 123.9, 123.0, 120.5, 117.1, 111.6, 108.3, 34.2, 24.5, 17.5, 10.6; Infrared spectrum: IR(KBr) v: 344 4,3087,3047,2960,2928,2866,1611,1556,1532,1496,1455,1401,1370,1320,1302,1271,121 7,1146,1127,1087,1060,1006,962,933,920,865,816,798,782,766,749,716,669,626,434cm -1 High-resolution mass spectrometry: HRMS (ESI) m / z: [M+H] + calcd for C 21 H 20 N2 301.1698, found 301.1698.

[0282] Example 14: Synthesis of compounds AzBI-1 to AzBI-7

[0283]

[0284] Specific steps: Take Az (83.2 mg, 0.3 mmol), 6′, 7′, 8′ or 9′ (0.45 mmol), Pd2(dba)3 (13.7 mg, 0.015 mmol), t Bu-Xphos (23.8 mg, 0.03 mmol) and NaO tBu (40.4 mg, 0.42 mmol) was added to a 25 mL Schlenk tube, and the tube was purged with N2 three times. Then, 3 mL of redistilled toluene was added under N2 atmosphere, and the reaction was carried out at 110 °C for 24 h. The reaction was monitored by TLC [V(PE):V(EA) = 1:1] to ensure completion. After the reaction solution cooled to room temperature, it was poured into water and extracted three times with ethyl acetate. The organic phases were combined and dried over anhydrous Na2SO4. After the solvent was evaporated, the solution was separated by silica gel column chromatography [V(PE):V(EA):V(Et3N) = 200:200:1].

[0285] 55.7 mg of the purple solid product AzBI-1 was obtained, with a yield of 76%; mp 128–129 °C; 1H NMR spectrum: 1 ¹H NMR (500MHz, CD₂Cl₂) δ: 8.48 (s, 1H), 8.43 (d, J = 9.3Hz, 2H), 8.05 (d, J = 8.1Hz, 1H), 7.85 (d, J = 7.9Hz, 1H), 7.68 (t, J = 9.9Hz, 1H), 7.61 (s, 2H), 7.45 (t, J = 7.6Hz, 1H), 7.41–7.33 (m, 3H); ¹H NMR spectrum: 13 C NMR (125MHz, CD₂Cl₂) δ: 145.1, 143.5, 143.0, 140.4, 136.9, 136.4, 133.4, 125.5, 124.4, 123.5, 120.9, 112.0, 107.4; Infrared spectrum: IR (KBr) ν: 3126, 3051, 3016, 1689, 16 09,1570,1538,1513,1500,1452,1405,1297,1282,1259,1224,1216,1189,1155 ,1133,1020,947,930,899,885,853,841,787,726,643,615,584,519,472,423cm -1 High-resolution mass spectrometry: HRMS (ESI) m / z: [M+H] + calcd for C 17 H 12 N2 244.1000, found 244.1001.

[0286] 28.3 mg of the purple solid product AzBI-2 was obtained, with a yield of 36%; 1H NMR spectrum: 1¹H NMR (500MHz, CD₂Cl₂) δ: 8.50 (s, 1H), 8.43 (d, J = 9.1 Hz, 2H), 7.97 (dd, J = 8.9, 4.6 Hz, 1H), 7.69 (t, J = 9.9 Hz, 1H), 7.57 (s, 2H), 7.53 (dd, J = 9.2, 2.5 Hz, 1H), 7.38 (t, J = 9.8 Hz, 2H), 7.20 (td, J = 9.1, 2.5 Hz, 1H); ¹H NMR spectrum: 13 C NMR (125MHz, CD₂Cl₂) δ: 160.1 (d, J = 238.3 Hz), 145.6 (d, J = 12.7 Hz), 144.4, 143.1, 140.4, 137.3, 136.7, 130.1, 125.6, 112.6 (d, J = 14.5 Hz), 112.4, 107.4, 106.6 (d, J = 24.0 Hz); fluorine NMR spectrum: 19 F NMR (376MHz, CD₂Cl₂) δ: -107.53; Infrared spectrum: IR (KBr) ν: 3125, 3029, 1710, 1617, 1597, 1574, 1538, 1511, 1496, 1473, 1440, 1404, 1372, 1310, 1284, 1230, 1209, 1193, 1144, 1100, 1016, 953, 909, 899, 886, 864, 821, 788, 734, 644, 631, 584, 544, 481, 444, 430, 404 cm⁻¹ -1 High-resolution mass spectrometry: HRMS (ESI) m / z: [M+H] + calcd forC 17 H 11 FN2 263.0979, found 263.0978.

[0287] 37.0 mg of the purple solid product AzBI-3 was obtained, with a yield of 47%; mp 160–162 °C; 1H NMR spectrum: 1 ¹H NMR (500MHz, CD₂Cl₂) δ: 8.49–8.39 (m, 3H), 7.80 (dd, J = 8.8, 4.9 Hz, 1H), 7.75–7.66 (m, 2H), 7.55 (s, 2H), 7.39 (t, J = 9.8 Hz, 2H), 7.14 (td, J = 9.2, 2.5 Hz, 1H); ¹H NMR spectrum: 13CNMR (125MHz, CD₂Cl₂) δ: 160.9 (d, J = 239.9 Hz), 143.7 (d, J = 3.1 Hz), 143.0, 141.5, 140.4, 137.4, 136.9, 133.6 (d, J = 13.4 Hz), 125.7, 121.7 (d, J = 10.2 Hz), 111.7 (d, J = 25.0 Hz), 107.4, 98.9 (d, J = 28.8 Hz); fluorine NMR spectrum: 19 F NMR (376MHz, CD₂Cl₂) δ: -105.00; Infrared spectrum: IR (KBr) ν: 3122, 3033, 1688, 1615, 1597, 1575, 1538, 1509, 1472, 1456, 1409, 1343, 1295, 1273, 1225, 1195, 1167, 1117, 1096, 1012, 959, 951, 899, 866, 846, 817, 784, 772, 736, 701, 643, 625, 615, 607, 585, 570, 489, 455, 434, 417 cm⁻¹ -1 High-resolution mass spectrometry: HRMS (ESI) m / z: [M+H] + calcd for C 17 H 11 FN2 263.0979, found 263.0978.

[0288] 27.5 mg of the purple solid product AzBI-4 was obtained, with a yield of 33%; 1H NMR spectrum: 1 ¹H NMR (500MHz, CD₂Cl₂) δ: 8.48 (s, 1H), 8.44 (d, J = 9.2Hz, 2H), 7.96 (d, J = 8.7Hz, 1H), 7.84 (d, J = 1.9Hz, 1H), 7.70 (t, J = 9.9Hz, 1H), 7.57 (s, 2H), 7.44–7.35 (m, 3H); ¹H NMR spectrum: 13C NMR (125MHz, CD₂Cl₂) δ: 146.0, 144.3, 143.0, 140.4, 137.4, 136.9, 132.2, 128.9, 125.7, 124.8, 120.7, 112.8, 107.5; Infrared spectrum: IR (KBr) ν: 3066, 3020, 1609, 1573 ,1537,1518,1457,1426,1403,1372,1342,1305,1281,1212,1194,1117,1057, 1020,951,911,898,861,850,798,725,653,629,606,588,581,476,429,411cm -1 High-resolution mass spectrometry: HRMS (ESI) m / z: [M+H] + calcdfor C 17 H 11 ClN2 279.0684, found 279.0682.

[0289] 36.7 mg of the purple solid product AzBI-5 was obtained, with a yield of 44%; mp 156–158 °C; 1H NMR spectrum: 1 ¹H NMR (500MHz, CD₂Cl₂) δ: 8.50–8.40 (m, 3H), 8.04 (d, J = 2.0Hz, 1H), 7.78 (d, J = 8.6Hz, 1H), 7.71 (t, J = 9.9Hz, 1H), 7.57 (s, 2H), 7.43–7.33 (m, 3H); ¹H NMR spectrum: 13 C NMR (125MHz, CD₂Cl₂) δ: 143.8, 143.7, 142.8, 140.4, 137.5, 136.9, 134.1, 130.2, 125.7, 124.1, 121.8, 112.1, 107.5; Infrared spectrum: IR (KBr) ν: 3090, 3018, 1612, 1594, 1577, 154 1,1518,1453,1404,1373,1335,1312,1286,1269,1224,1195,1138,1108,1066,101 8,936,912,900,847,831,800,750,718,687,654,646,617,598,582,554,482,419cm -1 High-resolution mass spectrometry: HRMS (ESI) m / z: [M+H] + calcd for C 17 H 11ClN2 279.0684,found 279.0683.

[0290] 30.4 mg of the purple solid product AzBI-6 was obtained, with a yield of 37%; mp 132–134 °C; 1H NMR spectrum: 1 ¹H NMR (500MHz, CD₂Cl₂) δ: 8.43 (d, J = 9.1 Hz, 2H), 8.39 (s, 1H), 7.74 (d, J = 8.9 Hz, 1H), 7.68 (t, J = 9.9 Hz, 1H), 7.57 (s, 2H), 7.49 (d, J = 2.4 Hz, 1H), 7.37 (t, J = 9.8 Hz, 2H), 7.01 (dd, J = 8.8, 2.4 Hz, 1H), 3.92 (s, 3H); ¹H NMR spectrum: 13 C NMR (125MHz, CD₂Cl₂) δ: 158.1, 143.5, 142.3, 140.4, 139.2, 137.1, 136.6, 134.1, 125.6, 121.2, 112.5, 107.5, 95.8, 56.3; Infrared spectrum: IR (KBr) ν: 3093, 3081, 3004, 2956, 2836, 1975, 1854, 1793, 1625, 1590, 1574,1539,1517,1494,1479,1467,1453,1439,1409,1375,1349,1319,1287,1276,1230,1216,120 4,1174,1130,1112,1035,930,898,817,809,789,765,744,732,701,642,622,583,467,441,420cm -1 High-resolution mass spectrometry: HRMS (ESI) m / z: [M+H] + calcd for C 18 H 14 N2O 275.1179, found 275.1178.

[0291] 36.2 mg of the purple solid product AzBI-7 was obtained, with a yield of 44%; mp 118–119 °C; 1H NMR spectrum: 1¹H NMR (500MHz, CD₂Cl₂) δ: 8.45 (s, 1H), 8.41 (d, J = 9.2Hz, 2H), 7.92 (d, J = 8.9Hz, 1H), 7.67 (t, J = 9.9Hz, 1H), 7.57 (s, 2H), 7.40–7.32 (m, 3H), 7.07 (dd, J = 8.9, 2.5Hz, 1H), 3.90 (s, 3H); ¹H NMR spectrum: 13 C NMR (125MHz, CD₂Cl₂) δ: 157.2, 145.9, 143.6, 143.2, 140.5, 136.9, 136.4, 127.9, 125.6, 114.2, 112.4, 107.2, 103.0, 56.1; Infrared spectrum: IR (KBr) ν: 3300, 3115, 3075, 3010, 2979, 2943, 2843, 1702, 1671, 161 5,1593,1574,1539,1521,1483,1441,1411,1388,1379,1354,1313,1297,1278,1239,1197,1157, 1106,1016,945,926,900,871,824,808,790,752,727,696,646,621,598,586,565,508,474,433cm -1 High-resolution mass spectrometry: HRMS (ESI) m / z: [M+H] + calcd for C 18 H 14 N2O 275.1179, found 275.1179.

[0292] Example of effect 1: Stability

[0293] The final product BGA obtained in Example 3 remained a blackish-green crystalline form after being stored at room temperature in the dark for one year, and its purity showed no significant change after NMR analysis. In contrast, GA stored under the same conditions had a melting point of around 33°C, and even after multiple purification and storage processes, it eventually turned into a green oily substance, with no product observed on TLC, and NMR analysis confirmed significant deterioration with disordered peak shapes.

[0294] The final product A obtained in Example 4 2-2 After being stored at room temperature away from light for one year, it remained a blue-purple solid with a melting point above 320°C. TLC showed no obvious impurities, while GAS-Na stored under the same conditions appeared as a green oily substance, and TLC showed complete deterioration.

[0295] The final product CAS-Na obtained in Example 9 remained a blue solid after being stored at room temperature in the dark for one year, and its purity showed no significant change during NMR analysis. Because the product is easily decomposed by heat, its melting point was not detected. In contrast, CA stored under the same conditions showed a significant color change within one week, and TLC revealed partial deterioration, requiring storage in a low-temperature (2-8°C) N2 atmosphere.

[0296] The final products GABI-1 to GABI-7 obtained in Example 12 were stored in the dark at a low temperature of 2 to 8°C for a period of time. The color did not change significantly and remained a blue solid. TLC monitoring showed no obvious impurity generation, and NMR analysis also showed no significant change in purity.

[0297] The final product CABI-1 obtained in Example 13 was stored in the dark at a low temperature of 2-8°C for a period of time. The color did not change significantly and it remained a blue solid. TLC monitoring showed no obvious impurity generation, and NMR analysis also showed no significant change in purity.

[0298] The final products AzBI-1 to AzBI-7 obtained in Example 14 were stored at a low temperature of 2 to 8°C in the dark for a period of time. The color did not change significantly and remained a purple solid. TLC monitoring showed no obvious impurity generation, and NMR analysis also showed no significant change in purity.

[0299] Example 2: In vitro antioxidant assay (FRAP method)

[0300] (1) Experimental Principle

[0301] The FRAP (Ferric reducing antioxidant power) method, proposed by Benzie and Strain in 1996, is a sensitive and accurate test for total antioxidant capacity (Brahmi, F. et al. Antioxidants. 2018, 7, 184.). Under acidic conditions, antioxidants can reduce Ferric-tri-pyridyl-tri-azine (F... 3+ -TPTZ) is reduced to blue Fe 2+ -TPTZ (maximum absorption wavelength 593 nm), and finally, the blue Fe was measured at 593 nm. 2+ The total antioxidant capacity of a sample can be obtained by measuring the absorbance of -TPTZ.

[0302] (2) Experimental materials

[0303] Preparation of test sample solutions: Four accurately weighed compounds were thoroughly dissolved in dimethyl sulfoxide to prepare 0.4258 mM GA, 0.2726 mM BGA, 0.5342 mM GAS-Na, and 0.2345 mM A, respectively.2-2 Prepare the solutions as needed. Preparation of standard solutions: Weigh 0.02507 g of FeSO4·7H2O provided by the Total Antioxidant Capacity Assay Kit (Beyotime), add 901.8 μL of dimethyl sulfoxide to form a 100 mM solution, and dilute to concentrations of 50, 10, 5, 2.5, 1.5, and 0.75 mM respectively, for later use. Preparation of FRAP working solution: For every 100 assays, mix 15 mL of TPTZ diluent with 1.5 mL of TPTZ solution, homogenize, and then add 1.5 mL of assay buffer to form the FRAP working solution. Use within 1-2 hours after preparation.

[0304] (3) Experimental methods

[0305] Determination of the standard curve: Pipette 180 μL of FRAP working solution into each well of a 96-well plate. There are six concentrations of standard solution, with six replicates for each concentration. A blank control group and a positive control group are also included. Add 5 μL of the corresponding drug concentration to each well. Add 5 μL of distilled water to the blank control group and 5 μL of 1 mM Trolox (water-soluble vitamin E) solution to the positive control group. Mix gently and incubate at 37°C for 3 min, then measure the absorbance at 593 nm. Sample determination: Follow the same method as the standard curve determination, with six replicates. This is used to detect the absorbance of the sample solution itself at 593 nm at different concentrations. Finally, substitute the absorbance values ​​of the experimental groups into the standard curve to calculate the corresponding total antioxidant capacity. Relative antioxidant capacity = total antioxidant capacity / sample concentration.

[0306] (4) Experimental Results

[0307] Table 1. Compounds GA, BGA, GAS-Na, and A determined by FRAP method. 2-2 in vitro antioxidant capacity

[0308]

[0309] From Table 1 and Figure 1 The results showed that the relative antioxidant capacity of the dimer compound BGA of the present invention was significantly improved compared with GA alone (P<0.01). This indicates that guaiac is an effective antioxidant fragment, and its direct dimerization or even oligomerization at the 2-position does not affect its antioxidant capacity.

[0310] Example 3: In vitro antioxidant test (DPPH free radical scavenging method)

[0311] (1) Experimental Principle

[0312] DPPH (1,1-diphenyl-2-picrylhydrazyl) is a stable nitrogen-centered free radical. Its ethanol solution is purple, with a maximum absorption wavelength of 517 nm. When a free radical scavenger is added to the DPPH solution, its lone pair electrons are paired, the solution color lightens, turning yellow or pale yellow, and the absorbance at 517 nm decreases. The degree of change is linearly related to the degree of free radical scavenging.

[0313] (2) Experimental materials

[0314] Solution preparation: Prepare a 0.2 mM DPPH stock solution with anhydrous ethanol, and then prepare 0.4 mM solutions of GA, BGA, GAS-Na, and A with anhydrous ethanol or a mixture of anhydrous ethanol and dimethyl sulfoxide. 2-2 Store at 0–4°C away from light.

[0315] (3) Experimental methods

[0316] Add 100 μL of DPPH solution and 100 μL of different compound solutions to a 96-well plate, mix well, and incubate at room temperature in the dark for 0.5 h. Then, measure the OD value at 517 nm using a microplate reader, and record it as A. s Subsequently, the change in OD value at 517 nm was tracked after 3 hours. Similarly, 100 μL of compound solution and 100 μL of anhydrous ethanol were added to a 96-well plate, mixed well, and the OD value was measured after standing at room temperature for a period of time, and recorded as A0; 100 μL of DPPH solution and 100 μL of anhydrous ethanol were added to a 96-well plate, mixed well, and the OD value was measured after standing at room temperature for a period of time, and recorded as A. c Each compound was configured with 3 duplicate wells. According to the formula: Scavenging rate (%) = [1 - (A)] s -A0) / A c ]×100%, calculate the DPPH scavenging rate of the compound.

[0317] (4) Experimental Results

[0318] Table 2. Scavenging rates of compounds on DPPH free radicals at different reaction times.

[0319]

[0320]

[0321] Oxidative stress caused by excessive free radicals in the human body can lead to acute and chronic inflammation. In 1997, Kourounakis found that when the concentration of GA was twice that of DPPH, the clearance rate reached 50% after 3 hours of co-treatment (Kourounakis, AP; Rekka, EA; Kourounakis, PNJPharm.Pharmacol.1997,49,938.). This experiment studied compounds GA, BGA, GAS-Na, and A. 2-2 The changes in the scavenging effect of four compounds after co-treatment with DPPH for 0.5 h and 3 h under the same experimental conditions were studied, with GA as a positive control.

[0322] The experimental results are shown in Table 2. The DPPH scavenging rate of all compounds was significantly increased after 3 hours of treatment compared to 0.5 hours. BGA exhibited the highest DPPH scavenging rate (84.21%) after 3 hours of treatment. 2-2 It also showed good performance in DPPH· clearance (58.02%), while the DPPH· clearance rate of GA was 32.20% at this time. Figure 2 It can be known that BGA and A 2-2 All four compounds showed significant improvements in free radical scavenging compared to GA (P<0.01 and P<0.0001). Overall, at 0.5 h and 3 h, the scavenging rates of the four compounds for DPPH·, from highest to lowest, were: BGA > A. 2-2 The free radical scavenging abilities of the GA and GAS-Na compounds were significantly higher than those of the monomer and its sodium sulfonate derivatives, with the monomer and dimer of *Guaiacomonas guaiac* showing slightly higher scavenging abilities than their corresponding sodium sulfonate derivatives. While the scavenging effects of the four compounds on DPPH· showed some delay, the dimers BGA and A... 2-2 The scavenging effect on DPPH free radicals was good even after 3 hours of reaction.

[0323] The experimental results show that, at the same concentration, the dimer compounds BGA and A... 2-2 The free radical scavenging rates of the compounds are all more than twice that of their corresponding monomers. The antioxidant capacity of the compounds of this invention exhibits a certain synergistic effect compared to their corresponding monomers.

[0324] Example 4: Griess method for detecting the inhibitory effect of the compound on LPS-induced RAW 264.7 mouse macrophage inflammation model.

[0325] (1) Experimental Principle

[0326] Lipopolysaccharide (LPS) is a major component of the outer membrane of pathogenic Gram-negative bacteria and can stimulate macrophages to produce pro-inflammatory factors. Nitric oxide (NO) is a biological messenger molecule whose production depends on inducible nitric oxide synthase (iNOS) and plays a crucial role in inflammatory responses. NO is readily oxidized to NO2 in vivo or in aqueous solution. - Under acidic conditions, NO2 - It undergoes a diazo reaction with the diazonium salt sulfanilamide to form a diazo compound, which further couples with naphthylvinyldiamine in Griess' reagent. The product exhibits a maximum absorption peak at 540 nm. Therefore, the absorbance at this point can be used to determine the NO production status in the cell model.

[0327] (2) Experimental materials

[0328] Mouse mononuclear macrophage RAW 264.7 cell line, test compound, lipopolysaccharide (LPS), DMEM high glucose medium, fetal bovine serum, and nitric oxide (NO) detection kit.

[0329] (3) Experimental methods

[0330] Mouse monocyte / macrophage RAW 264.7 cell line was cultured in a high-glucose DMEM culture medium containing 10% FBS at 37°C in a 5% CO2 incubator. The cells exhibited logarithmic growth, and the medium was changed every 24 hours. Cells were passaged when they reached 80%–90% confluence. After passage 3 and achieving stable cell counts, they were sporulated at 5 × 10⁻⁶ cells / year. 4 Inoculate 96-well plates at a density of [number] cells / mL and incubate for 24 h. Add prepared compounds of different concentrations and co-incubate for 1 h. After incubation, add LPS (100 ng / well) to the drug-treated group and the model control group, and add complete culture medium to the blank group. Continue incubation for 24 h, collect the supernatant, and follow the NO kit instructions. Measure the absorbance at 540 nm and calculate the NO content and inhibition rate.

[0331] (4) Experimental Results

[0332] This experiment is divided into two parts. The first part investigates compounds GA, BGA, GAS-Na, and A. 2-2The inhibition rate of NO in a macrophage inflammation model was investigated, with a model group (LPS stimulation only, no drug administration) and a control group (no LPS stimulation and no drug administration). NO concentration was calculated based on a pre-established linear relationship (standard curve) between NO concentration and OD value. The second part of the experiment investigated the inhibition rate of compounds CABI-1, GABI-1~GABI-7, and AzBI-1~AzBI-7 on NO in a macrophage inflammation model, with a model group (LPS stimulation only, no drug administration) as a control. The NO inhibition rate (%) was calculated as [1 - (OD value / model group)] × 100%.

[0333] Table 3 shows the inhibition rate of compounds on NO production in a macrophage inflammation model.

[0334]

[0335] The experimental results in the first part are shown in Table 3 and... Figure 3 As shown, the results indicate that A 2-2 It can significantly inhibit the production of NO inflammatory factors, and this inhibitory effect is concentration-dependent. Adding 1 μg / mL of A... 2-2 The NO concentration was significantly reduced afterward (NO inhibition rate 19.5%), and the inhibition rates reached 46.3% and 93.0% when the administered concentration was increased to 5 μg / mL and 25 μg / mL, respectively. The inhibitory effects of the other three compounds, GAS-Na, BGA, and GA, did not show a significant concentration dependence, and their inhibitory effects at different concentrations were all inferior to those of A. 2-2 In 2001, Guarrera et al. found that commercially available Veralga, containing GAS-Na, was... The anti-inflammatory activity of the gel is comparable to that of OTC hydrocortisone cream (Guarrera, M.; Turbino, L.; Rebora, AJ Eur. Acad. Dermatol. Venereol. 2001, 15, 486.). Therefore, A 2-2 It is a potentially highly effective anti-inflammatory substance that is superior to the commercially available active ingredient GAS-Na.

[0336] Table 4 shows the inhibition rate of compounds on NO production in a macrophage inflammation model.

[0337]

[0338]

[0339]

[0340] "-" indicates that the data was measured.

[0341] The experimental results in Part II are shown in Table 4 and... Figure 4 As shown, Figure 4 The inhibition rate of the target compound on LPS-induced NO production in macrophages was demonstrated using GAS-Na as a positive control. A higher inhibition rate indicates a better anti-inflammatory effect of the compound. Specifically, the inhibition rates (%) of GABI-1 to GABI-5 were calculated as follows: [1 - (OD value / model group 1)] × 100%; the inhibition rates (%) of CABI-1, GABI-6 to GABI-7, and GAS-Na were calculated as follows: [1 - (OD value / model group 2)] × 100%; the inhibition rates (%) of AzBI-1 to AzBI-5 were calculated as follows: [1 - (OD value / model group 3)] × 100%; and the inhibition rates (%) of AzBI-6 to AzBI-7 were calculated as follows: [1 - (OD value / model group 4)] × 100%. The results showed that at a drug concentration of 1 μM, among compounds GABI-1 to GABI-7, except for GABI-6, whose NO inhibition rate (2.00%) was lower than that of the positive control GAS-Na (5.40%), the NO inhibition rates of the other azurite derivatives were all higher than those of GAS-Na; while the NO inhibition rates of compounds AzBI-1 to AzBI-7 were all lower than those of GAS-Na (5.40%). At a drug concentration of 10 μM, GABI-1 had the highest NO inhibition rate (21.77%), and GABI-2 (19.97%) and GABI-6 (20.25%) also showed NO inhibition rates approximately twice as high as those of GAS-Na (10.16%), demonstrating good anti-inflammatory activity; among compounds AzBI-1 to AzBI-7, only AzBI-1 had a NO inhibition rate (11.75%) superior to that of GAS-Na. When the drug concentration was 20 μM, GABI-1, GABI-2, GABI-3, GABI-5, GABI-6, GABI-7 and AzBI-7 showed better NO inhibition rates than GAS-Na (23.07%), among which GABI-6 (35.85%) and GABI-7 (37.16%) were significantly better than GAS-Na.

[0342] Example 5: Effect of the compound on the viability of RAW 264.7 macrophages determined by CCK-8 assay.

[0343] (1) Experimental materials

[0344] Mouse mononuclear macrophage RAW 264.7 cell line, test compound, CCK-8 assay kit, DMEM high glucose medium, fetal bovine serum.

[0345] (2) Experimental methods

[0346] Macrophages in the logarithmic growth phase were scraped with a disposable spatula and dispersed into single cells using a pipette. The appropriate number of cells were counted and added to the appropriate culture medium at a concentration of 5 × 10⁻⁶.4 The solution was seeded at a density of 100 μL / mL into 96-well plates and incubated for 24 h. Compound A was added to each of the drug-treated groups. 2-2 GAS-Na, BGA, and GA were administered at concentrations of 1, 5, and 25 μg / mL, with 5 replicates per group. The blank control group received the corresponding volume of cell culture medium and was incubated for 24 h. The top layer of culture medium in the 96-well plate was discarded, and 100 μL of 10% CCK-8 serum-free DMEM solution was added. The plate was incubated at 37°C in a 5% CO2 cell culture incubator for 45 min. The OD value at 450 nm was measured using a microplate reader, and cell viability was calculated. Cell viability (%) = (OD value of the drug-treated group / OD value of the blank control group) × 100.

[0347] (3) Experimental Results

[0348] Table 5. Effects of compounds on the viability of RAW 264.7 macrophages.

[0349]

[0350] This experiment is divided into three parts. The first part studies compounds GA, BGA, GAS-Na, and A. 2-2 The experiments investigated the cytotoxicity of compounds CA, BCA, and CAS-Na in the first part of the study, and the experiments investigated the cytotoxicity of compounds GABI-1 to AZBI-7 in the third part. Table 8 shows that after co-culturing macrophages with different concentrations of the test compounds for 24 hours, the cell viability was approximately 100%, indicating that none of the four compounds had significant cytotoxicity to macrophages and therefore exhibited high safety.

[0351] Table 6. Effects of compounds on the viability of RAW 264.7 macrophages.

[0352]

[0353]

[0354] Table 6 shows that at concentrations of 1 μg / mL, 5 μg / mL, and 25 μg / mL, these six compounds did not exhibit significant cytotoxicity against macrophages. Furthermore, except for compound CA, which showed a cell survival rate of 88% at 25 μg / mL, the cell survival rates of the other compounds were all above 90% at all concentrations.

[0355] Table 7. Effects of compounds on the viability of RAW 264.7 macrophages.

[0356]

[0357]

[0358] Table 7 shows that, except for AzBI-6, the cell viability of the other compounds was above 80% at concentrations of 1 μM, 10 μM, and 20 μM. Overall, the cytotoxicity of compounds GABI-1 to GABI-7 was significantly lower than that of compounds AzBI-1 to AzBI-7. Except for GABI-7, the cell viability of compounds GABI-1 to GABI-6 was above 90% at all administered concentrations. Cell viability (%) of compounds GABI-1 to GABI-6 = (OD value of administered group / OD value of blank group 1) × 100%; Cell viability (%) of compounds CABI-1, AzBI-1, and AzBI-2 = (OD value of administered group / OD value of blank group 2) × 100%; Cell viability (%) of compounds AzBI-3 to AzBI-7 = (OD value of administered group / OD value of blank group 3) × 100%.

[0359] Therefore, the results of Examples 1 to 5 show that the dimer compounds and benzimidazole compounds of the present invention have good stability, antioxidant activity, anti-inflammatory activity and high safety. In particular, the sodium sulfonate salts of azurite dimer compounds have excellent antioxidant and anti-inflammatory properties and have strong development and application prospects. They are expected to become active pharmaceutical ingredients for neuroprotection, anti-radiation, anti-allergy, anti-inflammatory, anti-ulcer and wound healing.

Claims

1. A compound as shown in Formula I: YLZ Formula I; in, The definitions of each substituent in Formula I are those in group (a). Group (a): Y is ; Z is ; R 1a and R 3a Independently H or Cl- 12 alkyl; R 1g and R 3g Independently -SO3Q; Q is an alkali metal; R 1b R 1c R 1d R 1e R 1f R 3b R 3c R 3d R 3e and R 3f Independently H or Cl- 12 alkyl; L means it does not exist.

2. The compound of formula I as claimed in claim 1, characterized in that, The compound shown in Formula I satisfies one or more of the following conditions: (1) The alkali metal is Li, Na or K; (2) R 1a and R 3a In the context, C1- 12 The alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl; (3) R 1b R 1c R 1d R 1e R 1f R 3b R 3c R 3d R 3e and R 3f In the context, C1- 12 The alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl.

3. The compound of formula I as described in claim 2, characterized in that, The compound shown in Formula I satisfies one or two of the following conditions: (1) The alkali metal mentioned is Na; (2) R 1b R 1c R 1d R 1e R 1f R 3b R 3c R 3d R 3e and R 3f In the context, C1- 12 The alkyl group is independently methyl, ethyl, or isopropyl.

4. The compound of formula I as claimed in claim 1, characterized in that, R 1d and R 3d For H.

5. The compound of formula I as claimed in claim 1, characterized in that, The compound shown in Formula I satisfies one or two of the following conditions: (1) for or ; (2) for or .

6. The compound of formula I as claimed in claim 1, characterized in that, The compound represented by Formula I is any one of the following compounds: 。 7. A method for preparing the compound of Formula I as described in any one of claims 1-6, characterized in that, The method for preparing the compound as shown in Formula I is Method 3: When R in Y and Z 1g and R 3g Independently -SO3Q, R 1a and R 3a Independently for C1- 12 When alkyl, the preparation method of the compound as shown in Formula I is Method 3: Method 3: It includes the following steps: reacting an alkali metal base and compound V in a solvent to obtain the compound shown in Formula I: ； The compound V is compound V-1 of the following formula: ； In compound V-1, R 5g and R 7g Independently -SO3H, R 5a and R 7a Independently for C1- 12 Alkyl; R 1b R 1c R 1d R 1e R 1f R 3b R 3c R 3d R 3e and R 3f The definitions are the same as those described in any one of claims 1-6.

8. A compound V-1: ； In compound V-1, R 5g and R 7g Independently -SO3H, R 5a and R 7a Independently for C1- 12 Alkyl; R 1b R 1c R 1d R 1e R 1f R 3b R 3c R 3d R 3e and R 3f The definitions are as described in any one of claims 1-6.

9. The compound according to claim 8, characterized in that, The compound V-1 is the following compound: 。 10. A pharmaceutical composition, characterized in that, It comprises a compound as shown in Formula I as described in any one of claims 1-6 and a pharmaceutically acceptable carrier.

11. The use of a compound as shown in Formula I as claimed in any one of claims 1-6 or a pharmaceutical composition as claimed in claim 10 in the preparation of an anti-inflammatory drug.

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