CN116574081A

CN116574081A - Chlorogenic acid-Huang Qinsu conjugate and preparation method and application thereof

Info

Publication number: CN116574081A
Application number: CN202310830734.1A
Authority: CN
Inventors: 徐长江; 李玲; 游胜勇; 谢传奇; 徐建国
Original assignee: Institute of Applied Chemistry Jiangxi Academy of Sciences
Current assignee: Institute of Applied Chemistry Jiangxi Academy of Sciences
Priority date: 2023-07-07
Filing date: 2023-07-07
Publication date: 2023-08-11
Anticipated expiration: 2043-07-07
Also published as: CN116574081B

Abstract

The invention provides chlorogenic acid-Huang Qinsu conjugate and a preparation method and application thereof. The chlorogenic acid-Huang Qinsu conjugate has a structure shown in a general formula I or a general formula II, wherein an active carboxylic acid position in the chlorogenic acid structure is subjected to chemical modification, and is coupled with an active molecule Huang Qinsu, so that the drug effect is enhanced through the dual effects of anti-inflammatory and uric acid reduction; in order to find chlorogenic acid derivatives with strong activity and better stability, the positions of phenolic hydroxyl groups, alcoholic hydroxyl groups and the like in the structure of the chlorogenic acid derivatives are protected, so that the metabolic stability of the medicament in vivo is enhanced. The synthesis method of chlorogenic acid-Huang Qinsu conjugate is simple and efficient, and has the characteristics of increasing fat solubility and stability, changing absorption performance, improving drug effect, reducing toxicity and the like compared with the original drug.

Description

Chlorogenic acid-Huang Qinsu conjugate and preparation method and application thereof

Technical Field

The invention relates to the technical field of pharmaceutical chemistry, in particular to chlorogenic acid-Huang Qinsu conjugate and a preparation method and application thereof.

Background

The current anti-gout drugs mainly comprise three major categories of xanthine oxidase inhibitors, uric acid excretion promoting drugs and anti-inflammatory drugs. Xanthine oxidase inhibitor and uric acid excretion promoting drug are mainly used for reducing uric acid level, and anti-inflammatory drug is used for acute onset of gout, and the effect of treating acute gout is poor when the xanthine oxidase inhibitor and the uric acid excretion promoting drug are used alone. The uric acid reducing medicine and the anti-inflammatory medicine can be used in combination to exert better effect on the treatment of acute gout.

Chlorogenic acid has good anti-inflammatory activity, has good prevention and treatment effects on inflammation-related diseases, such as obvious treatment effects on cow mastitis, and can also have anti-inflammatory effect by inhibiting hyaluronidase activity. Chlorogenic acid can effectively inhibit NLRP3 inflammatory body activation and has good potential for preventing and treating acute gout. The poor stability and pharmacokinetic properties of chlorogenic acid limit its clinical application, and thus chlorogenic acid is often structurally modified to improve stability and anti-inflammatory activity, such as 1-carboxyethylated products and 3, 5-dicaffeoyl-substituted chlorogenic acid derivatives, which have improved anti-inflammatory activity over chlorogenic acid.

The flavonoid has antioxidant, antibacterial, antiviral and anticancer effects. Flavone compounds have been studied thoroughly to inhibit XOD (xanthine oxidase) and reduce serum uric acid levels, wherein Huang Qinsu, quercetin, morin and other flavone compounds have strong affinity for XOD and are good natural XODI (xanthine oxidase inhibitors). Compared with chemical small molecules such as NOSAOTAN and the like, the flavonoid XODI has good activity and small toxic and side effects, but the development and application of the flavonoid in gout treatment are limited by the poor physicochemical property and drug generation property of the flavonoid, and modification of the flavonoid by chemical modification to optimize the patentability of the flavonoid has important significance in expanding the application of the flavonoid in gout treatment.

In view of good NLRP3 inhibition activity of chlorogenic acid, application potential of chlorogenic acid in acute gout treatment and good XOD inhibition activity of flavonoid compounds, the invention designs and synthesizes chlorogenic acid-Huang Qinsu conjugate connected by different connecting chains according to a drug split principle.

Disclosure of Invention

The invention aims to provide chlorogenic acid-Huang Qinsu conjugate, a preparation method thereof and application thereof in gout treatment. The chlorogenic acid-Huang Qinsu conjugate has good XOD and NLRP3 inhibition activity and shows good gout treatment effect. The synthesis method of chlorogenic acid-Huang Qinsu conjugate is simple and efficient. Compared with the original medicine, the chlorogenic acid-Huang Qinsu conjugate provided by the invention has the characteristics of increasing fat solubility and stability, changing absorption performance, improving medicine effect, reducing toxicity and the like.

Chlorogenic acid-Huang Qinsu conjugate of the invention is a compound with a structure shown in a general formula I or a general formula II:

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wherein: r is R ₁ And R is ₂ May be the same or different, and is respectively any one of H, alkyl, aryl, heterocyclic aryl, C (O) -alkyl, C (O) -aryl, C (O) -heterocyclic aryl and C (O) -amino;

R ₃ and R is ₄ May be the same or different, and is respectively any one of H, alkyl, aryl, heterocyclic aryl, C (O) -alkyl, C (O) -aryl, C (O) -heterocyclic aryl and C (O) -amino;

In formula I: m=1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14, n=1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12;

in formula ii, m=1, 2, 3, 4, 5 or 6; n=1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12.

The invention provides a preparation method of chlorogenic acid-Huang Qinsu conjugate, which comprises the following steps:

1eq of compound 4a, compound 4b, compound 4c or compound 4d and 1eq of HOBT, 2eq of EDCI are weighed into a reaction vessel, DMF is added for dissolution, stirring reaction is carried out at room temperature for 10 min, then 1.2 eq of compound 7a is added, clear water is added after the reaction at room temperature is complete, ethyl acetate is used for extraction, the combined organic layers are sequentially washed by clear water and saturated saline, the organic layers are dried by anhydrous sodium sulfate and concentrated under reduced pressure and pass through a column to obtain pale yellow foam solid which is compound 10a, compound 10b, compound 10c or compound 10d, namely chlorogenic acid-Huang Qinsu conjugate.

The invention provides another preparation method of chlorogenic acid-Huang Qinsu conjugate, which comprises the following steps:

1eq of compound 4a and 1eq of HOBT and 2eq of EDCI are weighed into a reaction vessel, after DMF is added for dissolution, stirring reaction is carried out at room temperature, then any one of compound 7b, compound 7c, compound 7d and compound 7e is added, clear water is added after the reaction at room temperature is completed, then ethyl acetate is used for extraction, the organic layers are combined and washed sequentially by clear water and saturated saline, and the organic layers are dried by anhydrous sodium sulfate and concentrated under reduced pressure and pass through a column to obtain pale yellow foam solid which is compound 11a, compound 11b, compound 11c or compound 11d, namely chlorogenic acid-Huang Qinsu conjugate.

1eq of compound 4a and 1eq of HOBT and 2eq of EDCI are weighed into a reaction vessel, after DMF is added for dissolution, stirring reaction is carried out at room temperature, then any one of compound 9a, compound 9b, compound 9c and compound 9d is added, clear water is added after the reaction at room temperature is completed, ethyl acetate is used for extraction, the combined organic layers are sequentially washed by clear water and saturated saline, the organic layers are dried by anhydrous sodium sulfate and then concentrated under reduced pressure and pass through columns, and light yellow foamy solid which is compound 12a, compound 12b, compound 12c or compound 12d is obtained, namely chlorogenic acid-Huang Qinsu coupling compound.

The studies to which the present invention relates include, but are not limited to, the following: firstly, active carboxylic acid sites in chlorogenic acid structures are chemically modified, active molecules Huang Qinsu are coupled, and the drug effect is enhanced through the dual effects of anti-inflammatory and uric acid reduction; secondly, in order to find chlorogenic acid derivatives with strong activity and better stability, the positions of phenolic hydroxyl groups, alcoholic hydroxyl groups and the like in the structure of the chlorogenic acid derivatives are protected, so that the metabolic stability of the medicament in vivo is enhanced. Thirdly, pharmacological activity screening of chlorogenic acid-Huang Qinsu conjugate mainly comprises in-vitro and in-vivo anti-inflammatory and uric acid reducing activity test. Through analysis of structure-activity relationship, the research result of structure-activity relationship is used for further guiding chemical synthesis, and a new medicine with stronger activity and more stable chemical property can be found out and used for clinic.

The connecting chain is widely and specifically studied in the development of proteolytic targeting chimeras and antibody coupled drugs, and the connecting chain is found to have important effects on physicochemical properties, drug formation and activity of coupled molecules. The invention synthesizes a series of chlorogenic acid-Huang Qinsu conjugates containing different connecting chains and protecting groups, and systematically examines the influence of factors such as the length, the type, the rigidity and the like of the connecting chains on the coupling physical properties and the activity of chlorogenic acid-Huang Qinsu.

In another aspect, the invention is also directed to the pharmacological activity of chlorogenic acid-Huang Qinsu conjugates. Hyperuricemia is the biochemical basis for the onset of gout, and xanthine oxidase is a key enzyme in the uric acid production process. Huang Qinsu has xanthine oxidase inhibiting effect, and can reduce uric acid production and uric acid level in vivo by inhibiting xanthine oxidase activity. Thus, the inhibitory activity of chlorogenic acid-Huang Qinsu conjugate on xanthine oxidase was first measured. The synthesized chlorogenic acid-Huang Qinsu conjugate has good xanthine oxidase inhibition activity.

Meanwhile, inflammation plays a key role in the development of gout diseases, NLRP3 inflammatory corpuscles are important recognition receptors in the development process of gout inflammation, can be used as action targets in gout treatment, and can effectively inhibit the release of inflammatory factors in the development process of gout by inhibiting NLRP3 inflammatory corpuscles. Chlorogenic acid can well inhibit NLRP3 activity, and can inhibit the release of inflammatory factors and the development of inflammatory processes. Therefore, the invention subsequently tests the NLRP3 inhibitory activity of the synthesized chlorogenic acid-Huang Qinsu conjugate, and discovers that the chlorogenic acid-Huang Qinsu conjugate has better NLRP3 inhibitory activity.

The invention provides application of chlorogenic acid-Huang Qinsu conjugate in preparing medicines for preventing or treating gouty diseases, wherein the gouty diseases comprise acute gouty arthritis, chronic gouty arthritis, intermittent gouty arthritis, drug-induced liver injury, acute kidney injury or neuropathic pain and the like.

The invention provides application of chlorogenic acid-Huang Qinsu conjugate in preparing medicines for preventing or treating inflammatory infectious diseases, wherein the inflammatory infectious diseases comprise gouty arthritis, inflammatory bowel disease, cerebral (spinal) membranitis, pancreatitis, peritonitis, vasculitis (disease), glomerulonephritis, hepatitis, keratitis, cataract, senile macular degeneration and optic neuritis diseases.

The invention provides chlorogenic acid-Huang Qinsu conjugate with a novel structure through chemical modification of chlorogenic acid. The chlorogenic acid-Huang Qinsu conjugate provided by the invention has the following characteristics: firstly, the fat solubility is increased, the pro-drug enters the body in a form of pro-drug, and the pro-drug is metabolized to exert the drug effect in the acid environment of in-vivo enzyme catalysis and gastric juice after oral administration, so that the half-life period is prolonged and the drug effect is enhanced; secondly, the stability is enhanced, so that the inherent parent nucleus structure of the medicine is not damaged, and the integrity of the active site is maintained; thirdly, the curative effect of the medicine is improved through the double medicine effect.

Drawings

FIG. 1 shows the inhibitory activity of compound 10a, compound 10b, compound 10c, and compound 10d, respectively, on XOD.

FIG. 2 shows the inhibitory activity of compound 12a, compound 12b, compound 12c, and compound 12d, respectively, on XOD.

FIG. 3 is the effect of chlorogenic acid-Huang Qinsu conjugate on lipopolysaccharide-induced NO production in RAW264.7 cells.

FIG. 4 is the effect of chlorogenic acid-Huang Qinsu conjugate on lipopolysaccharide-induced survival of RAW264.7 cells.

FIG. 5 is the effect of chlorogenic acid-Huang Qinsu conjugate on lipopolysaccharide induced RAW264.7 cell morphology.

FIG. 6 is a hydrogen spectrum of compound 10 a.

FIG. 7 is a carbon spectrum of compound 10 a.

Detailed Description

For ease of reference, certain terms therein are now defined.

"alkyl" refers to an unsubstituted or substituted straight, branched, or cyclic alkyl carbon chain of up to 15 carbon atoms. Straight chain alkyl groups include methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl. Cyclic alkyl ("cycloalkyl") groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. The alkyl group may be substituted with one or more substituents. Non-limiting examples of such substituents include NH ₂ 、NO ₂ 、N(CH ₃ ) ₂ 、ONO ₂ 、F、Cl、Br、I、OH、OCH ₃ 、CO ₂ H、CO ₂ CH ₃ CN, aryl, and heteroaryl. The term "alkyl" also refers to unsubstituted or substituted straight, branched or cyclic alkyl groups containing up to 15 carbon atoms containing at least one heteroatom in the chain (e.g., nitrogen, oxygen or sulfur). The above straight-chain alkyl group includes, for example, CH ₂ CH ₂ OCH ₃ 、CH ₂ CH ₂ N(CH ₃ ) ₂ And CH (CH) ₂ CH ₂ SCH ₃ . Branched groups include, for example, CH ₂ CH(OCH ₃ )CH ₃ 、CH ₂ CH(N(CH ₃ ) ₂ )CH ₃ And CH (CH) ₂ CH(OCH ₃ )CH ₃ . The above cyclic group includes, for example, CH (CH) ₂ CH ₂ ) ₂ O、H(CH ₂ CH ₂ ) ₂ NCH ₃ And CH (CH) ₂ CH ₂ ) ₂ S, S. The above alkyl groups may be substituted with one or more substituents. Non-limiting examples of such substituents include NH ₂ 、NO ₂ 、N(CH ₃ ) ₂ 、ONO ₂ 、F、Cl、Br、I、OH、OCH ₃ 、CO ₂ H、CO ₂ CH ₃ CN, aryl, and heteroaryl.

The term "aryl" as used herein refers to an unsubstituted or substituted aromatic compound, carbocyclic group. Aryl is either a monocyclic or polycyclic fused compound. For example, phenyl is a monocyclic aryl. Naphthyl is an example of an aryl group having multiple rings fused. Aryl groups may be substituted with one or more substituents. Non-limiting examples of substituents include NH ₂ 、NO ₂ 、N(CH ₃ ) ₂ 、ONO ₂ 、F、Cl、Br、I、OH、OCH ₃ 、CO ₂ H、CO ₂ CH ₃ CN, aryl, and heteroaryl.

The chlorogenic acid-Huang Qinsu conjugate provided by the invention has the structure shown in the general formula I:

wherein: r is R ₁ And R is ₂ May be the same or different and are each H, alkyl, aryl, heteroaryl, C (O) -alkyl, C (O) -aryl, C (O) -heteroaryl, C (O) -amino; r is R ₃ And R is ₄ May be the same or different and are each H, alkyl, aryl, heteroaryl, C (O) -alkyl, C (O) -aryl, C (O) -heteroaryl, C (O) -amino; m=1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14; n=1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12.

Chlorogenic acid-Huang Qinsu conjugate of formula I is synthesized as follows:

the chlorogenic acid-Huang Qinsu conjugate provided by the invention has the structure shown in the following general formula II:

wherein: r is R ₁ And R is ₂ May be the same or different and are each H, alkyl, aryl, heteroaryl, C (O) -alkyl, C (O) -aryl, C (O) -heteroaryl, C (O) -amino; r is R ₃ And R is ₄ May be the same or different and are each H, alkyl, aryl, heteroaryl, C (O) -alkyl, C (O) -aryl, C (O) -heteroaryl, C (O) -amino; m=1, 2, 3, 4, 5, 6; n=1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12.

Chlorogenic acid-Huang Qinsu conjugate of the general formula II is synthesized as follows:

example 1

Synthesis of Compound 1: huang Qinsu (Apigenin, 10 g, 37mmol,1 eq) and potassium carbonate (10 g, 74 mmol,2 eq) were weighed into a 250 mL round bottom flask, dried DMF (100 mL) was added and dissolved, then stirred at room temperature for 15 min, then added dropwise under ice bath conditions MOM-Br was added and transferred to room temperature for reaction overnight. After TLC monitored completion of the reaction, most DMF was distilled off under reduced pressure, and after adding an equal volume of clear water to the remaining DMF (about 30 mL), pH was adjusted to neutral with 3N diluted hydrochloric acid, followed by extraction with ethyl acetate/tetrahydrofuran mixed solvent (3/1, 60 mL ×5), and the combined organic layers were washed sequentially with clear water (50 mL ×2), saturated brine (60 mL), dried over anhydrous sodium sulfate, and spin-dried over column (DCM/thf=100/1-30/1). Yellow solid 2.2. 2.2g was obtained in 19% yield. ¹ H NMR (400 MHz, DMSO) δ: 12.96 (s, 1H), 10.41 (s, 1H), 7.96 (d, J = 8.8 Hz, 2H), 6.94 (d, J = 8.8 Hz, 2H), 6.86 (s, 1H), 6.81 (d, J = 2.2 Hz, 1H), 6.44 (d, J = 2.2 Hz, 1H), 5.32 (s, 2H), 3.42 (s, 3H)。

Synthesis of compound 2 a: compound 1 (2.2 g, 7 mmol,1 eq) and potassium carbonate (1.93 g, 14 mmol,2 eq) were weighed into a 50 mL round bottom flask, dried DMF (20 mL) was added to dissolve and stirred at room temperature for 15min, then ethyl bromoacetate (0.93 mL, 8.4 mmol,1.2 eq) was added dropwise under ice bath conditions and after addition was transferred to room temperature for reaction 4-6 h. After the completion of the reaction by TLC, 20 mL clear water was added, extraction was performed with ethyl acetate (40 mL ×3), and the combined organic layers were washed with clear water (20 mL ×2) and saturated brine (20 mL) in this order, and the organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure to pass through the column (PE/ea=6/1-2/1). Yellow solid 1.76, g was obtained in 65% yield. ¹ H NMR (400 MHz, DMSO) δ: 8.00 (d, J = 8.8 Hz, 2H), 7.10 (d, J = 8.8 Hz, 2H), 6.97 (d, J = 2.0 Hz, 1H), 6.72 (s, 1H), 6.48 (d, J = 2.0 Hz, 1H), 5.33 (s, 2H), 4.91 (s, 2H), 4.18 (qd, J = 7.0, 2.4 Hz, 2H), 3.42 (s, 3H), 1.22 (td, J = 7.2, 2.8 Hz, 3H)。

Synthesis of compound 2 b: compound 1 (2.2 g, 7 mmol,1 eq) and potassium carbonate (1.93 g, 14 mmol,2 eq) were weighed into a 50 mL round bottom flask, dried DMF (20 mL) was added and stirred at room temperature for 15min, ethyl 4-bromobutyrate (1.26 mL, 8.4 mmol,1.2 eq) was added dropwise under ice bath conditions and after addition was warmed to 65 ℃ and reacted overnight. After TLC monitoring the reaction completion, 20 mL clear water was added, extracted with ethyl acetate (40 mL X3), and the combined organic layers were washed successively with clear water (20 mL X2), saturated brine (20 mL), and the organic layers were dried over anhydrous sodium sulfate and then reduced Concentrating by pressing (PE/EA=6/1-2/1). Yellow solid 1.38, g was obtained in 46% yield. ¹ H NMR (400 MHz, DMSO) δ: 12.93 (s, 1H), 10.85 (s, 1H), 8.02 (d, J = 8.8 Hz, 2H), 7.10 (d, J = 9.0 Hz, 2H), 6.87 (s, 1H), 6.51 (d, J = 2.0 Hz, 1H), 6.20 (d, J = 2.0 Hz, 1H), 5.33 (s, 2H), 4.13–4.01 (m, 4H), 3.42 (s, 3H), 2.38 (t, J = 7.2 Hz, 2H), 1.82–1.62 (m, 2H), 1.18 (t, J = 7.2 Hz, 3H)。

Synthesis of Compound 2 c: compound 1 (2.2 g, 7 mmol,1 eq) and potassium carbonate (1.93 g, 14 mmol,2 eq) were weighed into a 50 mL round bottom flask, dried DMF (20 mL) was added to dissolve and stirred at room temperature for 15 min, followed by dropwise addition of ethyl 10-bromodecanoate (2.06 mL, 8.4 mmol, 1.2 eq) under ice-bath conditions and reaction was completed at 65 ℃ overnight. After completion of the TLC monitoring reaction, 20. 20 mL clear water was added, extracted with ethyl acetate (40 mL X3), and the combined organic layers were washed successively with clear water (20 mL X2), saturated brine (20 mL), dried over anhydrous sodium sulfate and spin-dried over column (PE/EA=6/1-2/1). Yellow solid 1.86 g was obtained in 52% yield. ¹ H NMR (400 MHz, DMSO) δ: 12.93 (s, 1H), 10.85 (s, 1H), 8.02 (d, J = 8.9 Hz, 2H), 7.09 (d, J = 8.9 Hz, 2H), 6.86 (s, 1H), 6.50 (d, J = 2.0 Hz, 1H), 6.20 (d, J = 2.0 Hz, 1H), 5.32 (s, 2H), 4.13–3.98 (m, 4H), 3.43 (s, 2H), 2.26 (t, J = 7.4 Hz, 2H), 1.79 – 1.69 (m, 2H), 1.56–1.47 (m, 2H), 1.41 (d, J = 7.6 Hz, 2H), 1.26 (s, 8H), 1.21–1.14 (m, 3H)。

Synthesis of Compound 2 d: compound 1 (2.2 g, 7 mmol,1 eq) and potassium carbonate (1.93 g, 14 mmol,2 eq) were weighed into a 50 mL round bottom flask, dried DMF (20 mL) was added to dissolve and stirred at room temperature for 15 min, then 12-bromododecanoic acid ethyl ester (2.2 mL, 8.4 mmol, 1.2 eq) was added dropwise under ice-bath conditions and after addition was warmed to 65 ℃ to react overnight. After completion of TLC detection, 20 g mL of clear water was added, followed by extraction with ethyl acetate (40. 40 mL ×5), and the combined organic layers were washed successively with clear water (20. 20 mL ×2) and saturated brine (20. 20 mL), dried over anhydrous sodium sulfate, and then spin-dried (PE/EA=6/1-2/1). Yellow solid 2.24, g was obtained in 61% yield. ¹ H NMR (400 MHz, DMSO) δ: 12.93 (s, 1H), 10.85 (s, 1H), 8.01 (d, J = 8.8 Hz, 2H), 7.09 (d, J = 8.8 Hz, 2H), 6.86 (s, 1H), 6.50 (d, J = 2.0 Hz, 1H), 6.20 (d, J = 2.0 Hz, 1H), 5.33 (s, 2H), 4.13–3.97 (m, 4H), 3.43 (s, 2H), 2.26 (t, J = 7.4 Hz, 2H), 1.79–1.67 (m, 2H), 1.56–1.46 (m, 2H), 1.40 (d, J = 7.8 Hz, 2H), 1.25 (s, 10H), 1.17 (t, J = 7.0 Hz, 3H)。

Synthesis of Compound 3a Compound 2a (1.76 g, 4.6 mmol,1 eq) was dissolved in a mixture of THF/MeCN=1/1 (40 mL), and Trimethyliodosilane (TMSI) (3.9 mL, 27.6 mmol, 6.0 eq) was slowly added dropwise under ice bath conditions in Ar atmosphere, and the mixture was warmed to room temperature after the addition and reacted overnight. After TLC monitored completion of the reaction, the reaction was quenched to pale yellow or colorless by addition of saturated aqueous sodium thiosulfate, followed by extraction with EA (40 mL ×2), and the combined organic layers were washed sequentially with saturated aqueous sodium thiosulfate (40 mL), saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure and passed through a column (DCM/thf=50/1-20/1) to give a yellow solid 1.23 g in 75% yield. ¹ H NMR (400 MHz, DMSO) δ: 12.90 (s, 1H), 10.87 (s, 1H), 8.03 (d, J = 8.8 Hz, 2H), 7.11 (d, J = 8.8 Hz, 2H), 6.88 (s, 1H), 6.51 (d, J = 1.8 Hz, 1H), 6.21 (d, J = 1.8 Hz, 1H), 4.92 (s, 2H), 4.19 (q, J = 7.0 Hz, 2H), 1.23 (t, J = 7.0 Hz, 3H)。

Synthesis of Compound 3b Compound 2b (1.38 g, 3.22 mmol,1 eq) was dissolved in a mixture of THF/MeCN=1/1 (30 mL), TMSI (2.75 mL, 19.3 mmol, 6.0 eq) was slowly added dropwise under Ar ice bath conditions, and the reaction was allowed to proceed to room temperature overnight after the addition. After TLC monitored completion of the reaction, the reaction was quenched to pale yellow or colorless by addition of saturated aqueous sodium thiosulfate, followed by extraction with EA (40 mL ×2), and the combined organic layers were washed with saturated aqueous sodium thiosulfate (40 mL), saturated brine, dried over anhydrous sodium sulfate and spin-dried over column (DCM/thf=50/1-20/1) to give a yellow solid 878 mg in 71% yield. ¹ H NMR (400 MHz, DMSO) δ: 12.93 (s, 1H), 10.85 (s, 1H), 8.02 (d, J = 8.8 Hz, 2H), 7.10 (d, J = 9.0 Hz, 2H), 6.87 (s, 1H), 6.51 (d, J = 2.0 Hz, 1H), 6.20 (d, J = 2.0 Hz, 1H), 4.13– 4.01 (m, 4H), 2.38 (t, J = 7.2 Hz, 2H), 1.82–1.62 (m, 2H), 1.18 (t, J = 7.2 Hz, 3H)。

Chemical combinationSynthesis of Compound 3c Compound 2c (1.86 g, 3.64 mmol,1 eq) was dissolved in a mixture of THF/MeCN=1/1 (40 mL), TMSI (3.1 mL,21.8 mmol, 6.0 eq) was slowly added dropwise under Ar atmosphere ice bath conditions, and the reaction was allowed to warm to room temperature overnight after the addition. After TLC monitored completion of the reaction, the reaction was quenched to pale yellow or colorless by addition of saturated aqueous sodium thiosulfate solution, followed by extraction with EA (40 mL ×3), and the combined organic layers were washed with saturated aqueous sodium thiosulfate solution (40 mL), saturated brine, dried over anhydrous sodium sulfate and spin-dried over column (DCM/thf=50/1-20/1) to give 1.03g of yellow solid in 60% yield. ¹ H NMR (400 MHz, DMSO) δ 12.93 (s, 1H), 10.85 (s, 1H), 8.02 (d, J = 8.8 Hz, 2H), 7.09 (d, J = 8.8 Hz, 2H), 6.86 (s, 1H), 6.50 (d, J = 2.0 Hz, 1H), 6.20 (d, J = 2.0 Hz, 1H), 4.13– 3.98 (m, 4H), 2.26 (t, J = 7.4 Hz, 2H), 1.79–1.69 (m, 2H), 1.56–1.47 (m, 2H), 1.41 (d, J = 7.6 Hz, 2H), 1.26 (s, 8H), 1.21–1.14 (m, 3H)。

Synthesis of Compound 3d Compound 2d (2.24 g, 4.27 mmol,1 eq) was dissolved in a mixture of THF/MeCN=1/1 (40 mL), TMSI (3.7 mL, 25.6 mmol, 6.0 eq) was slowly added dropwise under Ar ice bath conditions and the reaction was allowed to proceed to room temperature overnight after the addition. After TLC monitored completion of the reaction, the reaction was quenched to pale yellow or colorless by addition of saturated aqueous sodium thiosulfate, followed by extraction with EA (40 mL ×2), and the combined organic layers were washed with saturated aqueous sodium thiosulfate (40 mL), saturated brine, dried over anhydrous sodium sulfate and spin-dried over column (DCM/thf=50/1-20/1) to give a yellow solid 854 mg in 42% yield. ¹ H NMR (400 MHz, DMSO) δ: 12.93 (s, 1H), 10.85 (s, 1H), 8.01 (d, J = 8.8 Hz, 2H), 7.09 (d, J = 8.8 Hz, 2H), 6.86 (s, 1H), 6.50 (d, J = 2.0 Hz, 1H), 6.20 (d, J = 2.0 Hz, 1H), 4.13– 3.97 (m, 4H), 2.26 (t, J = 7.4 Hz, 2H), 1.79–1.67 (m, 2H), 1.56–1.46 (m, 2H), 1.40 (d, J = 7.8 Hz, 2H), 1.25 (s, 10H), 1.17 (t, J = 7.0 Hz, 3H)。

Synthesis of Compound 4a Compound 3a (1.23 g, 3.45 mmol, 1 eq) was dissolved in THF (20 mL), followed by addition of an aqueous solution of lithium hydroxide (0.58 g, 13.8 mmol,4 eq) (5 mL) and reaction at 30℃overnight. After the completion of the reaction by the next day of TLC monitoring,THF was distilled off under reduced pressure, pH was adjusted to 2-3 with 3N diluted hydrochloric acid, followed by extraction with ethyl acetate (20 mL x 5), and the combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate and dried by spin-drying to give 767 mg as a pale yellow solid product in 68% yield. ¹ H NMR (400 MHz, DMSO) δ: 12.91 (s, 1H), 10.87 (s, 1H), 8.07 (d, J = 8.8 Hz, 2H), 7.12 (d, J = 8.8 Hz, 2H), 6.8I (d, J = 2.0 Hz, 1H), 6.51 (d, J = 2.0 Hz, 1H), 6.21 (s, 1H), 4.84 (s, 2H)。

Synthesis of Compound 4b Compound 3b (878 mg, 2.30 mmol, 1 eq) was dissolved in THF (20 mL), followed by addition of an aqueous solution of lithium hydroxide (0.36 g, 9.15 mmol,4 eq) (5 mL) and reaction at 30℃overnight. After completion of the reaction by TLC on the next day, THF was distilled off under reduced pressure, pH was adjusted to 2-3 with 3N diluted hydrochloric acid, followed by extraction with ethyl acetate (20 mL x 5), and the combined organic layers were washed with saturated brine and dried over anhydrous sodium sulfate to give a pale yellow solid product 530 mg in 65% yield. ¹ H NMR (400 MHz, DMSO) δ: 12.93 (s, 1H), 10.85 (s, 1H), 8.02 (d, J = 8.8 Hz, 2H), 7.10 (d, J = 9.0 Hz, 2H), 6.87 (s, 1H), 6.51 (d, J = 2.0 Hz, 1H), 6.20 (d, J = 2.0 Hz, 1H), 4.13–4.01 (m, 4H), 1.82–1.62 (m, 2H)。

Synthesis of Compound 4c Compound 3c (1.03 g, 2.2 mmol, 1 eq) was dissolved in THF (20 mL), followed by addition of an aqueous solution of lithium hydroxide (0.35 g, 8.8 mmol,4 eq) (5 mL) and reaction at 30℃overnight. After completion of the reaction by TLC monitoring the next day, THF was distilled off under reduced pressure, the pH was adjusted to 2-3 with 3N diluted hydrochloric acid, followed by extraction with ethyl acetate (20 mL X5), and the combined organic layers were washed with saturated brine and dried over anhydrous sodium sulfate to give 677 mg of a pale yellow solid product in 70% yield. ¹ H NMR (400 MHz, DMSO) δ: 12.93 (s, 1H), 10.85 (s, 1H), 8.02 (d, J = 8.8 Hz, 2H), 7.09 (d, J = 8.8 Hz, 2H), 6.86 (s, 1H), 6.50 (d, J = 2.0 Hz, 1H), 6.20 (d, J = 2.0 Hz, 1H), 4.13– 3.98 (m, 2H), 2.26 (t, J = 7.4 Hz, 2H), 1.79–1.69 (m, 2H), 1.56–1.47 (m, 2H), 1.41 (d, J = 7.6 Hz, 2H), 1.26 (s, 8H)。

Synthesis of Compound 4d Compound 3d (854 mg, 1.77 mmol,1 eq) was dissolved in THF (15 mL) followed by addition of hydroxideAn aqueous solution (3 mL) of lithium (0.28 g, 7.1 mmol,4 eq) was reacted overnight at 30 ℃. After completion of the reaction by TLC monitoring the next day, THF was distilled off under reduced pressure, the pH was adjusted to 2-3 with 3N diluted hydrochloric acid, followed by extraction with ethyl acetate (20 mL X5), and the combined organic layers were washed with saturated brine and dried over anhydrous sodium sulfate to give 530 mg of a pale yellow solid product in 66% yield. ¹ H NMR (400 MHz, DMSO) δ: 12.93 (s, 1H), 10.85 (s, 1H), 8.01 (d, J = 8.8 Hz, 2H), 7.09 (d, J = 8.8 Hz, 2H), 6.86 (s, 1H), 6.50 (d, J = 2.0 Hz, 1H), 6.20 (d, J = 2.0 Hz, 1H), 4.13–3.97 (m, 2H), 2.26 (t, J = 7.4 Hz, 2H), 1.79–1.67 (m, 2H), 1.56–1.46 (m, 2H), 1.40 (d, J = 7.8 Hz, 2H), 1.25 (s, 10H)。

Example 2

Synthesis of Compound 5:

chlorogenic acid (10.6 g, 30.0 mmol,1 eq) and DMAP (732.0 mg, 6.0 mmol,0.2 eq) were weighed into a 250 mL round bottom flask followed by acetic anhydride (20 mL) and pyridine (10 mL) and reacted overnight at room temperature after addition. After completion of the reaction by TLC the next day, the reaction solution was poured into an ice-water mixture 100 mL and stirred at room temperature for 30 min. Then, the pH is adjusted to 2-3 by 3N dilute hydrochloric acid, then white solid is separated out, the solid is filtered out and washed by proper amount of clear water, and the white solid of the target product is obtained after drying, namely about 12g,71 percent of yield.

Synthesis of compound 6 a: compound 5 (1.1 g, 2.0 mmol,1 eq), HOBT (0.27 g, 2.0 mmol,1 eq), EDCI (764 mg, 4.0 mmol,2 eq) were weighed into a 100 mL round bottom flask, dissolved in dichloromethane (50 mL) and stirred at room temperature for 15min. N-Boc-ethylenediamine (640 mg, 4.0 mmol,2 eq) was then added and the reaction stirred at room temperature for 2-3h. After TLC monitored completion of the reaction, dichloromethane (50 mL) was added for dilution, followed by washing with clear water (20 mL ×2), saturated brine (20 mL) in sequence, the organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure, and the crude product was purified by column chromatography (DCM/meoh=80/1-20/1) to give target product 6a as a white foamy solid 650 mg,46% yield.

Synthesis of compound 7 a: compound 6a (0.9 mmol,650 mg,1eq) was dissolved in DCM (10.35 mL), followed by TFA (45 mmol,3.45 mL, 50 eq) with a volume ratio (DCM: TFA) =3/1 and the reaction was stirred at room temperature for 2-3h. After completion of the reaction by TLC, dichloromethane was removed under reduced pressure, followed by addition of dichloromethane and repeated 3 times to give a crude product which was purified by column chromatography to give a white foamy solid 500 mg, 91% yield. ¹ H NMR (400 MHz, CDCl ₃ ) δ: 8.00 (s, 1H), 7.76 (s, 2H), 7.58 (d, J = 15.8 Hz, 1H), 7.45–7.34 (m, 2H), 7.21 (d, J = 8.2 Hz, 1H), 6.31 (d, J = 16.0 Hz, 1H), 5.61–5.43 (m, 2H), 5.10 (dd, J = 10.2, 2.8 Hz, 1H), 3.52 (s, 1H), 3.10 (d, J = 21.2 Hz, 2H), 2.92 (d, J = 33.8 Hz，1H), 2.87 (s, 1H), 2.62 (d, J = 12.8 Hz, 2H), 2.45 (d, J = 14.2 Hz, 1H), 2.29 (d, J = 1.0 Hz, 6H), 2.13 (s, 3H), 2.06 (d, J = 4.8 Hz, 3H), 1.97 (s, 4H)；C ₂₈ H ₃₄ N ₂ O ₁₃ +H ⁺ , Cal 607.2139 Found, 607.2149; +Na ⁺ , Cal 629.1959, Found, 629.1965。

Synthesis of compound 6 b: compound 5 (1.1 g, 2.0 mmol,1 eq), HOBT (0.27 g, 2.0 mmol,1 eq), EDCI (764 mg, 4.0 mmol,2 eq) were weighed into a 100 mL round bottom flask, dissolved in dichloromethane (50 mL) and stirred at room temperature for 15 min. N-Boc-1, 5-diaminopentane (808, mg, 4.0 mmol,2 eq) was then added and the reaction stirred at room temperature for 2-3h. After TLC monitored completion of the reaction, dichloromethane (50 mL) was added for dilution, followed by washing with clear water (20 mL ×2), saturated brine (20 mL) in sequence, the organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure, and the crude product was purified by column chromatography (DCM/meoh=80/1-20/1) to give 610 mg of the desired product as a white foamy solid in 41% yield.

Synthesis of compound 7 b: compound 6b (0.82 mmol,610 mg,1eq) was dissolved in DCM (9.4 mL), followed by TFA (41 mmol,3.14 mL, 50 eq) with a volume ratio (DCM: TFA) =3/1 and the reaction was stirred at room temperature for 2-3h. After TLC monitoring the completion of the reaction, the dichloromethane was dried by spin-drying, followed by addition of dichloromethane and repeated 3 times, and the crude product was purified by column chromatography to give a white color Foam solid 500 mg, 91% yield. ¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.60 (d, J = 16.0 Hz, 1H), 7.46–7.35 (m, 2H), 7.23 (d, J = 8.2 Hz, 1H), 6.33 (d, J = 16.0 Hz, 1H), 5.67–5.63 (m, 2H), 5.53 (td, J = 10.6, 4.2 Hz, 1H), 5.10 (dd, J = 10.2, 3.2 Hz, 1H), 3.71 (d, J = 8.2 Hz, 1H), 2.85 (d, J = 16.2 Hz, 1H), 2.52 (dd, J = 19.6, 6.4 Hz, 2H), 2.31 (d, J = 2.6 Hz, 6H), 2.17 (s, 3H), 2.09 (s, 3H), 1.99 (s, 3H), 1.86 (d, J = 11.6 Hz, 2H), 1.69-1.58 (m, 4H), 1.42–1.23 (m, 2H), 1.2–1.00 (m, 2H)；C ₃₁ H ₄₀ N ₂ O ₁₃ + H ⁺ , Cal 649.2609 Found, 649.2615; + Na ⁺ , 671.2428, Found, 671.2434。

Synthesis of Compound 6 c: compound 5 (1.1 g, 2.0 mmol,1 eq), HOBT (0.27 g, 2.0 mmol,1 eq), EDCI (764 mg, 4.0 mmol,2 eq) were weighed into a 100 mL round bottom flask, dissolved in dichloromethane (50 mL) and stirred at room temperature for 15min. N-Boc-1, 6-diaminohexane (864 mg, 4.0 mmol,2 eq) was then added and the reaction stirred at room temperature for 2-3h. After TLC monitored completion of the reaction, dichloromethane (50 mL) was added for dilution, followed by washing with clear water (20 mL ×2), saturated brine (20 mL) in sequence, the organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure, and the crude product was purified by column chromatography (DCM/meoh=80/1-20/1) to give the desired product 6c as a white foamy solid 1.2 g,78% yield.

Synthesis of Compound 7 c: compound 6c (1.56 mmol,1.2 g,1eq) was dissolved in DCM (18 mL) followed by TFA (78 mmol, 6.0 mL, 50.0 eq) with a volume ratio (DCM: TFA) =3/1 and the reaction was stirred at room temperature for 2-3h. After completion of the reaction by TLC, dichloromethane was removed under reduced pressure, followed by addition of dichloromethane and repeated 3 times to give a crude product which was purified by column chromatography to give 890 mg as a white foam solid in 86% yield. ¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.53 (d, J = 15.8 Hz, 1H), 7.34 (d, J = 8.6 Hz, 1H), 7.30 (s, 1H), 7.14 (d, J = 8.2 Hz, 1H), 6.61 (s, 1H), 6.26 (d, J = 16.0 Hz, 2H), 5.54 (s, 1H), 5.41 (dd, J = 13.8, 9.8 Hz, 1H), 5.04 (d, J = 9.4 Hz, 1H), 3.06 (d, J = 43.2 Hz, 2H), 2.83 (s, 2H), 2.62 (d, J = 13.8 Hz, 1H), 2.44 (dd, J = 29.2, 12.0 Hz, 2H), 2.22 (s, 6H), 2.10–2.02 (m, 3H), 1.99 (d, J = 2.8 Hz, 4H), 1.92 (s, 3H), 1.51 (s, 2H), 1.31 (s, 2H), 1.23–1.06 (m, 4H)；C ₃₂ H ₄₂ N ₂ O ₁₃ + H ⁺ , Cal 663.2765 Found, 663.2777; + Na ⁺ ,685.2585, Found,685.2598。

Synthesis of Compound 6 d: compound 5 (1.03 g, 1.83 mmol,1 eq), HOBT (0.247 g, 1.83 mmol,1 eq), EDCI (700 mg, 3.66 mmol,2 eq) was weighed into a 100 mL round bottom flask, dissolved in dichloromethane (50 mL) and stirred at room temperature for 15min. N-Boc-1, 7-diaminoheptane (842 mg, 3.66 mmol,2 eq) was then added and the reaction stirred at room temperature for 2-3h. After TLC monitored completion of the reaction, dichloromethane (50 mL) was added for dilution, followed by washing with clear water (20 mL ×2), saturated brine (20 mL) in sequence, the organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure, and the crude product was purified by column chromatography (DCM/meoh=80/1-20/1) to give 600 mg of the target product as a white foamy solid in 42% yield.

Synthesis of Compound 7 d: compound 6d (0.77 mmol,600 mg,1eq) was dissolved in DCM (8.85 mL) followed by TFA (38.5 mmol,2.95 mL, 50.0 eq) with a volume ratio (DCM: TFA) =3/1 and the reaction was stirred at room temperature for 2-3h. After completion of the reaction by TLC, dichloromethane was removed under reduced pressure, followed by addition of dichloromethane and repeated 3 times to give a crude product which was purified by column chromatography to give 417 mg as a white foamy solid in 80% yield. ¹ H NMR (400 MHz, CDCl ₃ ): δ 7.60 (d, J = 15.8 Hz, 1H), 7.45–7.33 (m, 2H), 7.22 (d, J = 8.4 Hz, 1H), 6.50 (s, 1H), 6.33 (d, J = 16.0 Hz, 1H), 5.62 (d, J = 3.2 Hz, 1H), 5.50 (td, J = 10.2, 4.8 Hz, 1H), 5.12 (dd, J = 10.0, 3.2 Hz, 1H), 3.50 (s, 4H), 3.25–3.04 (m, 2H), 2.78 (d, J = 15.2 Hz, 1H), 2.51 (dd, J = 31.2, 14.6 Hz, 2H), 2.30 (s, 6H), 2.15 (s, 3H), 2.08 (s, 3H), 1.98 (s, 4H), 1.58 (s, 2H), 1.39 (s, 2H), 1.32–1.13 (m, 6H)；C ₃₃ H ₄₄ N ₂ O ₁₃ + H ⁺ , Cal 677.2922 Found, 677.2930.+ Na ⁺ ,699.2741, Found,699.2751。

Synthesis of compound 6 e: compound 5 (0.79 g, 1.4 mmol,1 eq), HOBT (0.19 g, 1.4 mmol,1 eq), EDCI (535 mg, 2.8 mmol,2 eq) were weighed into a 100 mL round bottom flask, dissolved in dichloromethane (50 mL) and stirred at room temperature for 15min. N-Boc-1, 12-diaminododecane (840 mg, 2.8 mmol,2 eq) was then added and the reaction stirred at room temperature for 2-3h. After TLC monitored completion of the reaction, dichloromethane (50 mL) was added for dilution, followed by washing with clear water (20 mL ×2), saturated brine (20 mL) in sequence, the organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure, and the crude product was purified by column chromatography (DCM/meoh=80/1-20/1) to give 550 mg of the target product as a white foamy solid in 46% yield.

Synthesis of compound 7 e: compound 6e (0.65 mmol,550 mg,1.0 eq) was dissolved in DCM, followed by TFA (32.5 mmol,2.4 mL, 50 eq) with a volume ratio (DCM: TFA) =3/1 and the reaction was stirred at room temperature for 2-3h. After completion of the reaction by TLC, dichloromethane was removed under reduced pressure, followed by addition of dichloromethane and repeated 3 times to give a crude product which was purified by column chromatography to give 400 mg as a white foam in 83% yield. ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.03 (s, 1H), 7.44–7.34 (m, 2H), 7.22 (d, J = 8.4 Hz, 1H), 6.33 (d, J = 16.0 Hz, 1H), 6.17 (t, J = 5.6 Hz, 1H), 5.62 (d, J = 3.4 Hz, 1H), 5.52 (td, J = 10.7, 4.5 Hz, 1H), 5.11 (dd, J = 10.2, 3.6 Hz, 1H), 3.78 (s, 4H), 3.19 (d, J = 6.8 Hz, 2H), 2.82 (s, 1H), 2.57–2.46 (m, 2H), 2.30 (s, 6H), 2.16 (s, 3H), 2.08 (s, 3H), 1.98 (s, 4H), 1.60 (s, 2H), 1.43 (s, 2H), 1.22 (s, 16H)；C ₃₈ H ₅₄ N ₂ O ₁₃ + H ⁺ , Cal 747.3704 Found, 747.3706; + Na ⁺ ,769.3524, Found,769.3511。

Example 3

Synthesis of compound 8 a: compound 5 (2.0 g, 3.55 mmol,1 eq), HOBT (0.48 g, 3.55 mmol,1 eq), EDCI (1.36 g, 7.10 mmol,2 eq) were weighed into a 100 mL round bottom flask, dissolved in dichloromethane (50 mL) and stirred at room temperature for 15 min. Followed by addition of t-butoxycarbonyl-imino-polyethylene glycol-amino (Boc-NH-PEG-NH) ₂ ) (1.5. 1.5 mL, 7.1 mmol,2 eq) and stirred at room temperature for 2-3h. TLC monitoringAfter completion of the reaction, dichloromethane (50 mL) was added for dilution, followed by washing with clear water (20 mL ×2), saturated brine (20 mL), drying over anhydrous sodium sulfate and spin-drying, and the crude product was purified by column chromatography (DCM/meoh=80/1-20/1) to give the desired product as a white foamy solid 1.58g in 59% yield.

Synthesis of compound 9 a: compound 8a (1.58 g, 2.1 mmol,1 eq) was dissolved in DCM (22 mL), followed by TFA (7.4 mL, 0.105mol, 50.0 eq) with a volume ratio (DCM: TFA) =3/1 and the reaction was stirred at room temperature for 2-3h. After the completion of the reaction, the dichloromethane was removed under reduced pressure and the operation was repeated 3 times, and the obtained crude product was purified by column chromatography to give the objective product as a white foam solid 1.12. 1.12 g in 82% yield. ¹ H NMR (400 MHz, CDCl ₃ ) δ: 8.16 (s, 3H), 7.61 (d, J = 15.8 Hz, 1H), 7.49–7.37 (m, 2H), 7.22 (d, J = 8.4 Hz, 1H), 6.34 (d, J = 16.0 Hz, 1H), 5.60 (d, J = 3.4 Hz, 1H), 5.53 (td, J = 10.6, 4.6 Hz, 1H), 5.16 (dd, J = 10.2, 3.6 Hz, 1H), 3.71 (d, J = 4.6 Hz, 2H), 3.54 (d, J = 4.6 Hz, 2H), 3.48–3.35 (m, 2H), 3.21 (s, 2H), 2.81 (d, J = 15.8 Hz, 1H), 2.61 (dd, J = 20.8, 7.8 Hz, 2H), 2.30 (d, J = 2.2 Hz, 6H), 2.21 (s, 3H), 2.08 (s, 3H), 1.99 (d, J = 5.8 Hz, 4H)；C ₃₀ H ₃₈ N ₂ O ₁₄ + H ⁺ , Cal 651.2401 Found, 651.2411.+ Na ⁺ ,673.2221, Found, 673.2230。

Synthesis of compound 8 b: compound 5 (2.0 g, 3.55 mmol,1 eq), HOBT (0.48 g, 3.55 mmol,1 eq), EDCI (1.36 g, 7.10 mmol,2 eq) were weighed into a 100 mL round bottom flask, dissolved in dichloromethane (50 mL) and stirred at room temperature for 15min. Followed by the addition of tert-butyl 2- (2- (2-aminoethoxy) ethoxy) ethylcarbamate (Boc-NH-PEG 2-NH) ₂ ) (1.25 mL, 7.1 mmol,2 eq) and allowed to react for 2-3h at room temperature with stirring. After TLC monitored completion of the reaction, dichloromethane (50 mL) was added for dilution, followed by washing with clear water (20 mL ×2), saturated brine (20 mL), dried over anhydrous sodium sulfate and spin-dried, the crude product was column chromatographed (DCM/meoh=80/1-20/1) to give the desired product as a white foamy solid 1.6 g, 59% yield.

Synthesis of compound 9 b: compound 8b (1).6g, 2.01 mmol,1 eq) was dissolved in DCM (22 mL) and TFA (7.4 mL, 0.10mol, 50.0 eq) was then added, wherein the volume ratio (DCM: TFA) =3/1 and the reaction stirred at room temperature for 2-3h. After the completion of the reaction, the dichloromethane was removed under reduced pressure and the operation was repeated 3 times, and the obtained crude product was purified by column chromatography to give the objective product as a white foam solid 1.12. 1.12 g in 80% yield. C (C) ₃₂ H ₄₂ N ₂ O ₁₅ + H ⁺ , Cal 695.2663 Found, 695.2672.+ Na ⁺ ,717.2483, Found,717.2491。

Synthesis of Compound 8 c: compound 5 (1.0 g, 1.77 mmol,1 eq), HOBT (0.25 g, 1.77 mmol,1 eq), EDCI (0.7 g, 3.55 mmol,2 eq) was weighed into a 50 mL round bottom flask, dissolved in dichloromethane (25 mL) and stirred at room temperature for 15min. Followed by the addition of 13-amino-5, 8, 11-trioxa-2-azatridecanoic acid 1, 1-dimethylethyl ester (Boc-NH-PEG 3-NH) ₂ ) (1.0 g, 3.55 mmol,2 eq) and stirred at room temperature for reaction 2-3h. After TLC monitored completion of the reaction, dichloromethane (25 mL) was added for dilution, followed by washing with clear water (10 mL ×2), saturated brine (10 mL), dried over anhydrous sodium sulfate and spin-dried, the crude product was column chromatographed (DCM/meoh=80/1-20/1) to give the desired product as a white foamy solid 0.8g in 45% yield.

Synthesis of compound 9 c: compound 8c (0.8 g, 0.95 mmol,1 eq) was dissolved in DCM (10.5 mL) followed by TFA (3.5 mL, 47.5 mmol, 50.0 eq) with a volume ratio (DCM: TFA) =3/1 and the reaction was stirred at room temperature for 2-3h. After the completion of the reaction by TLC, dichloromethane was removed under reduced pressure and the procedure was repeated 3 times, the crude product obtained was purified by column chromatography to give the target product as a white foam solid 595 mg in 85% yield. ¹ H NMR (400 MHz, CDCl ₃ )δ: 8.23 (s, 2H), 8.02 (s, 1H), 7.62 (d, J = 16.0 Hz, 1H), 7.45–7.37 (m, 2H), 7.23 (d, J = 8.4 Hz, 1H), 6.34 (d, J = 16.0 Hz, 1H), 5.68–5.44 (m, 2H), 5.13 (dt, J = 11.6, 5.8 Hz, 1H), 3.81 (s, 2H), 3.69–3.52 (m, 6H), 3.51–3.32 (m, 2H), 3.24 (s, 2H), 2.82 (t, J = 12.6 Hz, 1H), 2.69–2.50 (m, 6H), 2.30 (d, J = 2.6 Hz, 6H), 2.21 (s, 3H), 2.09 (s, 3H), 2.04–1.95 (m, 4H)；C ₃₄ H ₄₆ N ₂ O ₁₆ + H ⁺ , Cal 739.2926 Found, 739.2930; + Na ⁺ ,761.2745, Found,761.2750。

Synthesis of Compound 8 d: compound 5 (1.0 g, 1.77 mmol,1 eq), HOBT (0.25 g, 1.77 mmol,1 eq), EDCI (0.7 g, 3.55 mmol,2 eq) was weighed into a 50 mL round bottom flask, dissolved in dichloromethane (25 mL) and stirred at room temperature for 15min. Subsequent addition of O- (2-aminoethyl) -O' - [2- (tert-butoxycarbonyl-amino) ethyl group]Tetraethylene glycol (Boc-NH-PEG 4-NH) ₂ ) (1.5. 1.5 mL, 7.1 mmol,2 eq) and stirred at room temperature for 2-3h. After TLC monitored completion of the reaction, dichloromethane (25 mL) was added for dilution, followed by washing with clear water (10 mL ×2), saturated brine (10 mL), dried over anhydrous sodium sulfate and spin-dried, the crude product was column chromatographed (DCM/meoh=80/1-20/1) to afford the desired product 8c as a white foamy solid 900 mg, 58% yield.

Synthesis of Compound 9 d: compound 8d (900 mg, 1.02mmol, 1 eq) was dissolved in DCM (11.4 mL) followed by TFA (3.8 mL, 51 mmol, 50.0 eq) with a volume ratio (DCM: TFA) =3/1 and the reaction was stirred at room temperature for 2-3h. After the completion of the reaction by TLC, dichloromethane was removed under reduced pressure, and the operation was repeated 3 times, and the obtained crude product was purified by column chromatography to obtain the objective product 9c as a white foam solid 654 mg in 82% yield. ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.16 (s, 3H), 7.60 (d, J = 16.0 Hz, 1H), 7.45–7.35 (m, 2H), 7.23 (d, J = 8.4 Hz, 1H), 6.33 (d, J = 16.0 Hz,1H), 5.61 (d, J = 3.6 Hz, 1H), 5.53 (d, J = 4.2 Hz, 1H), 5.12 (dd, J = 10.2, 3.6 Hz, 1H), 3.87–3.78 (m,2H), 3.68 (d, J = 3.6 Hz, 2H), 3.60 (dd, J = 15.2, 9.2 Hz, 8H), 3.51–3.33 (m, 3H), 3.20 (s, 2H), 2.82 (d, J = 16.2 Hz, 1H), 2.63 (d, J = 13.2 Hz, 1H), 2.52 (dd, J = 16.2, 3.2 Hz, 1H), 2.31 (d, J = 2.8 Hz, 6H), 2.21 (s, 3H), 2.09 (s, 3H), 2.04–1.94 (m, 4H); C ₃₆ H ₅₀ N ₂ O ₁₇ + H ⁺ , Cal 783.3188, Found, 783.3190。

Example 4

Compound 10aAnd (3) synthesis: compound 4a (65.6 mg, 0.2 mmol, 1.0 eq) and HOBT (27 mg, 0.2 mmol, 1.0 eq) were weighed out, EDCI (76.4 mg, 0.4 mmol, 1.0 eq) was dissolved in a 25mL round bottom flask, DMF (5 mL) was added and reacted at room temperature with stirring for 10 min, followed by compound 7a (163 mg, 0.24 mmol, 1.2 eq) and reacted at room temperature for 2-3h. After TLC monitored the reaction was complete, clear water 5mL was added, followed by extraction with ethyl acetate (15 mL ×5), and the combined organic layers were washed sequentially with clear water (15 mL ×2), saturated brine (10 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure through the column (DCM/meoh=100/1-20/1). This gave 112.9. 112.9 mg as a pale yellow foam solid in 56.0% yield. The hydrogen spectrum of compound 10a is shown in fig. 6, and the carbon spectrum of compound 10a is shown in fig. 7. ¹ HNMR (400 MHz, CDCl ₃ ) δ: 12.75 (s, 1H), 9.34 (s, 1H), 7.60-7.57 (m, 3H), 7.39-7.36 (m, 4H), 7.20 (d, J = 6.8 Hz, 1H), 6.80 (d, J = 5.8 Hz, 2H), 6.31 (t, J = 11.6 Hz, 4H), 5.67 (s, 1H), 5.57 (s, 1H), 5.17 (d, J = 9.2 Hz, 1H), 4.39 (s, 2H), 3.59-3.45 (m, 4H), 2.86 (d, J = 13.8 Hz, 1H), 2.75-2.48 (m, 3H), 2.30 (s, 6H), 2.22 (s, 3H), 2.10 (s, 3H), 1.99 (s, 3H); ¹³ CNMR (101 MHz, CDCl ₃ ) δ: 182.0, 171.3, 170.4, 170.3, 167.0, 169.4, 168.2, 168.0, 165.9, 163.7, 163.0, 161.7, 159.7, 157.6, 144.1, 143.8, 142.5, 132.8, 127.9, 126.6, 124.2, 124.0, 122.9, 118.2, 114.9, 104.7, 99.4, 94.5, 81.1, 71.8, 68.2, 67.3, 66.9, 40.7, 38.8, 38.2, 30.7, 21.8, 21.1, 20.7, 20.6, 20.58; HRMS: C ₄₅ H ₄₄ N ₂ O ₁₉ + H ⁺ , Cal 917.2617; Found, 917.2607; C ₄₅ H ₄₄ N ₂ O ₁₉ + Na ⁺ , Cal 939.2436 Found, 939.2413。

Example 5

Synthesis of Compound 10 b: compound 4b (71.2 mg, 0.2 mmol, 1.0 eq) and HOBT (27. mg, 0.2 mmol, 1.0 eq) were weighed into a 25mL round bottom flask, and after dissolution in DMF (5 mL) was added, the reaction was stirred at room temperature for 10 min, followed byCompound 7a (163 mg, 0.24 mmol, 1.2 eq) was added later and reacted at room temperature for 2-3h. After TLC monitored the reaction was complete, clear water 5mL was added, followed by extraction with ethyl acetate (15 mL ×5), and the combined organic layers were washed sequentially with clear water (15 mL ×2), saturated brine (10 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure through the column (DCM/meoh=100/1-20/1). This gave 88.3. 88.3 mg as a pale yellow foam solid in 48.0% yield. ¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.65–7.51 (m, 3H), 7.37–7.35 (m, 3H), 7.25–7.13 (m, 1H), 6.77 (s, 2H), 6.49–6.21 (m, 3H), 5.64–5.55 (m, 3H), 5.13 (s, 1H), 3.96 (s, 2H), 3.76 (s, 2H), 3.43–3.31 (m, 3H), 2.80 (s, 1H), 2.62–2.42( m, 3H), 2.30 (s, 6H), 2.23 (s, 3H), 2.10 (s, 3H), 1.99 (s, 5H), 1.87 (s, 3H); ¹³ C NMR (101 MHz, CDCl ₃ ): δ 182.2, 174.6, 172.4, 172.3, 171.4, 170.3, 170.2, 169.9, 168.2, 168.0, 165.7, 163.8, 161.9, 157.7, 144.0, 143.8, 142.5, 132.8, 127.8, 126.6, 124.0, 122.9, 118.3, 114.7, 107.9, 106.6, 106.4, 106.0, 104.7, 103.4, 99.5, 94.4, 81.0, 77.4, 71.8, 68.1, 68.0, 67.7, 29.2, 25.6, 23.9, 22.9, 21.8, 21.0, 20.7, 20.6, 20.6; HRMS: C ₄₇ H ₄₈ N ₂ O ₁₉ +H ⁺ ,Cal 945.2924, Found 945.2912; C ₄₇ H ₄₈ N ₂ O ₁₉ +Na ⁺ Cal 967.2743 Found 967.2750。

Example 6

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Synthesis of Compound 10 c: compound 4c (88 mg, 0.2 mmol, 1.0 eq) and HOBT (27 mg, 0.2 mmol, 1.0 eq) were weighed out, EDCI (76.4 mg, 0.4 mmol, 1.0 eq) was dissolved in a 25mL round bottom flask, DMF (5 mL) was added and stirred at room temperature for 10 min, followed by compound 7a (163 mg, 0.24 mmol, 1.2 eq) and reacted at room temperature for 2-3h. After TLC monitoring the reaction was completed, clear water 5 was added thereto mL followed by extraction with ethyl acetate (15 mL X5), and the combined organic layers were washed successively with clear water (15 mL X2), saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure and passed through a column (DCM/MeO) H=100/1-20/1). 70.0. 70.0 mg as pale yellow foam solid was obtained in 31.3% yield. ¹ H NMR (400 MHz, CDCl ₃ ): δ 7.65 (d, J = 8.2 Hz, 2H), 7.50 (d, J = 15.8 Hz, 1H), 7.30–7.27 (m, 3H), 7.12 (d, J = 8.2 Hz,1H), 6.81 (d, J = 8.0 Hz, 2H), 6.57 (s, 1H), 6.40 (d, J = 16.2 Hz, 2H), 6.31–6.17 (m, 2H), 5.63–5.38 (m, 1H), 5.06 (dd, J = 9.8, 2.8 Hz, 1H), 3.85 (s, 2H), 3.31–3.21 (m, 3H), 2.73 (d, J = 15.0 Hz, 1H), 2.50 (dd, J = 38.6, 12.6 Hz, 2H), 2.32–2.17 (m, 7H), 2.12 (s, 4H), 2.01 (s, 3H), 1.90 (s, 3H), 1.66–1.64 (m, 2H), 1.53–1.51 (m, 2H), 1.36–1.10 (m, 12H); ¹³ C NMR (101 MHz, CDCl ₃ ) δ: 182.4, 177.0, 175.6, 171.5, 170.2, 170.0, 168.2, 168.0, 165.7, 164.2, 163.9, 162.2, 162.0, 157.9, 144.0, 143.8, 142.5, 132.8, 128.0, 126.6, 124.0, 123.0, 122.9, 118.3, 114.9, 104.7, 103.5, 99.6, 94.5, 80.9, 71.8, 68.3, 68.1, 67.1, 40.7, 39.1, 38.0, 36.4, 34.0, 30.9, 29.4, 29.2, 29.0, 25.9, 25.6, 24.9, 21.7, 21.0, 20.7, 20.6, 20.56; HRMS: C ₅₃ H ₆₀ N ₂ O ₁₉ + H ⁺ , Cal 1029.3863 Found, 1029.3867; C ₅₃ H ₆₀ N ₂ O ₁₉ + Na ⁺ , 1051.3682, Found 1051.3687。

Example 7

Synthesis of Compound 10 d: compound 4d (91 mg, 0.2 mmol, 1.0 eq) and HOBT (27 mg, 0.2 mmol, 1.0 eq) were weighed out, EDCI (76.4 mg, 0.4 mmol, 1.0 eq) was dissolved in a 25mL round bottom flask, DMF (5 mL) was added and stirred at room temperature for 10 min, followed by compound 7a (163 mg, 0.24 mmol, 1.2 eq) and reacted at room temperature for 2-3h. After TLC monitored the reaction was complete, clear water 5mL was added, followed by extraction with ethyl acetate (15 mL ×5), and the combined organic layers were washed sequentially with clear water (15 mL ×2), saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure and passed through a column (DCM/meoh=100/1-20/1) to give 80.0 mg as a pale yellow foamy solid with a yield of 35.2%. ¹ HNMR (400 MHz, CDCl ₃ )δ: 7.65 (d, J = 8.0 Hz, 2H), 7.51 (d, J = 15.8 Hz, 1H), 7.31-7.23 (m, 3H), 7.13 (d, J = 8.2 Hz, 1H), 6.81 (d, J = 8.2 Hz, 2H), 6.51 (s, 1H), 6.40 (d, J = 15.8 Hz, 2H), 6.32-6.18 (m, 2H), 5.56-5.45 (m, 2H), 5.14-4.94 (m, 1H), 3.84 (s, 2H), 3.33-3.22 (m, 3H), 2.86 (d, J = 30.5 Hz, 1H), 2.73 (d, J = 15.8 Hz, 1H), 2.50 (dd, J = 40.2, 13.1 Hz, 2H), 2.22 (s, 7H), 2.13 (s, 4H), 2.01 (s, 3H), 1.91 (s, 3H), 1.64 (s, 2H), 1.53 (d, J = 5.2 Hz, 2H), 1.40-1.11 (m, 12H); ¹³ C NMR (101 MHz, CDCl ₃ ) δ: 182.4, 175.5, 171.3, 170.2, 169.9, 168.1, 168.0, 165.6, 164.1, 162.9, 162.0, 157.9, 144.0, 143.8, 142.5, 132.9, 128.0, 126.6, 124.0, 122.9, 118.3, 114.9, 104.8, 103.7, 99.7, 94.4, 81.0, 71.8, 68.1, 67.0, 60.4, 40.8, 39.2, 38.1, 36.7, 36.4, 30.9, 29.2, 25.9, 25.6, 21.8, 21.0, 20.7, 20.6, 20.56; HRMS: C ₅₄ H ₆₂ N ₂ O ₁₉ + H ⁺ , Cal 1043.4020 Found, 1043.4021。

Example 8

Synthesis of Compound 11 a: compound 4a (98.4 mg, 0.3 mmol, 1.0 eq) and HOBT (40.5 mg, 0.3 mmol, 1.0 eq) were weighed out, EDCI (114.6 mg, 0.6 mmol, 2.0 eq) was dissolved in 25mL round bottom flask, DMF (5 mL) was added and stirred at room temperature for 10 min, followed by compound 7b (232.8 mg, 0.36 mmol, 1.2 eq) and reacted at room temperature for 2-3h. After TLC monitored the reaction was complete, clear water 5mL was added, followed by extraction with ethyl acetate (15 mL ×5), and the combined organic layers were washed sequentially with clear water (15 mL ×2), saturated brine (10 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure through the column (DCM/meoh=100/1-20/1). 110. 110 mg as a pale yellow foamy solid was obtained in 37.5% yield. ¹ H NMR (400 MHz, CDCl ₃ ) δ: 12.71 (s, 1H), 9.53 (s, 1H), 7.68 (d, J = 8.2 Hz, 2H), 7.57 (d, J = 15.8 Hz, 1H), 7.40–7.32 (m, 2H), 7.23–7.19 (m, 1H), 7.04–6.82 (m, 3H), 6.59 (s, 1H), 6.39–6.29 (m, 4H), 5.63 (d, J = 2.8 Hz, 1H), 5.54 (td, J = 10.4, 4.6 Hz, 1H), 5.13 (dd, J = 10.2, 2.8 Hz, 1H), 4.49 (s, 2H), 3.34–3.26 (m, 2H), 3.19–3.15 (m, 1H), 2.98–2.83 (m, 1H), 2.66–2.45 (m, 3H), 2.30 (s, 6H), 2.18 (s, 3H), 2.08 (s, 3H), 2.01–1.95 (m, 4H), 1.63–1.45 (m, 4H), 1.34–1.28 (m, 2H); ¹³ C NMR (101 MHz, CDCl ₃ ) δ: 182.2, 170.6, 170.2, 169.9, 169.7, 168.2, 168.1, 165.8, 163.9, 163.2, 161.9, 159.9, 157.7, 144.1, 143.8, 142.5, 132.8, 128.0, 126.6, 124.6, 124.0, 122.9, 118.3, 115.1, 104.7, 104.1, 99.5, 98.7, 94.5, 81.1, 71.7, 68.1, 67.2, 50.6, 39.5, 38.9, 38.2, 30.8, 28.9, 28.7, 23.8, 21.7, 21.0, 20.7, 20.6, 20.57; HRMS: C ₄₈ H ₅₀ N ₂ O ₁₉ + H ⁺ , Cal 959.3081 Found, 959.3016; C ₄₈ H ₅₀ N ₂ O ₁₉ + Na ⁺ Cal 981.2900, Found 981.2802。

Example 9

Synthesis of Compound 11 b: compound 4a (98.4 mg, 0.3 mmol, 1.0 eq) and HOBT (40.5 mg, 0.3 mmol, 1.0 eq) were weighed out, EDCI (114.6 mg, 0.6 mmol, 2.0 eq) was dissolved in 25mL round bottom flask, DMF (5 mL) was added and reacted at room temperature with stirring for 10 min, followed by compound 7c (237.6 mg, 0.36 mmol, 1.2 eq) and 2-3h at room temperature. After TLC monitored the reaction was complete, clear water 5mL was added, followed by extraction with ethyl acetate (15 mL ×5), and the combined organic layers were washed sequentially with clear water (15 mL ×2), saturated brine (10 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure through the column (DCM/meoh=100/1-20/1). 70 mg as a pale yellow foam solid was obtained in 23.6% yield. ¹ H NMR (400 MHz, CDCl ₃ ) δ: 12.61 (s, 1H), 9.35 (s, 1H), 7.62 (d, J = 8.2 Hz, 2H), 7.51 (d, J = 15.8 Hz, 1H), 7.31– 7.28 (m, 2H), 7.13 (d, J = 8.2 Hz, 1H), 6.83 (d, J = 7.6 Hz, 3H), 6.44 (s, 1H), 6.35–6.32 (m, 2H), 6.26–6.22 (m, 2H), 5.56 (d, J = 2.8 Hz, 1H), 5.46 (td, J = 10.4, 4.6 Hz, 1H), 5.05 (dd, J = 10.4, 2.8 Hz, 1H), 4.42 (s, 2H), 3.26–3.16 (m, 2H), 3.10–3.05 (m, 1H), 2.90–2.76 (m, 1H), 2.55–2.43 (m, 3H), 2.22 (s, 6H), 2.10 (s, 3H), 2.01 (s, 3H), 1.98–1.85 (m, 4H), 1.80–1.77 (m, 2H), 1.46–1.39 (m, 4H), 1.22–1.17 (m, 2H); ¹³ C NMR (101 MHz, CDCl ₃ ): δ 182.2, 170.5, 170.2, 169.9, 169.7, 168.2, 168.1, 167.9, 144.1, 143.8, 142.5, 132.8, 128.1, 126.6, 124.7, 124.0, 122.9, 118.3, 115.1, 107.9, 104.8, 104.2, 99.6, 94.4, 81.2, 71.7, 68.1, 68.0, 67.2, 39.4, 38.8, 38.3, 30.7, 29.2, 29.1, 25.8, 25.6, 21.7, 21.0, 20.7, 20.6, 20.58; HRMS: C ₄₉ H ₅₂ N ₂ O ₁₉ + Na ⁺ Cal 995.3056, Found 995.3063。

Example 10

Synthesis of Compound 11 c: compound 4a (65.0 mg, 0.2 mmol, 1.0 eq) and HOBT (27.0 mg, 0.2 mmol, 1.0 eq) were weighed out, EDCI (76.0 mg, 0.4 mmol, 2.0 eq) was dissolved in 25mL round bottom flask, DMF (5 mL) was added and stirred at room temperature for 10 min, followed by compound 7d (162.2 mg, 0.24 mmol, 1.2 eq) and reacted at room temperature for 2-3h. After TLC monitored the reaction was complete, clear water 5mL was added, followed by extraction with ethyl acetate (15 mL ×5), and the combined organic layers were washed sequentially with clear water (15 mL ×2), saturated brine (10 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure through the column (DCM/meoh=100/1-20/1). 100. 100 mg as a pale yellow foamy solid was obtained in a yield of 50.2%. ¹ H NMR (400 MHz, CDCl ₃ ) δ: 12.69 (s, 1H), 9.34 (s, 1H), 7.74 (d, J = 8.6 Hz, 2H), 7.59 (d, J = 16.0 Hz, 1H), 7.42–7.33 (m, 2H), 7.21 (d, J = 8.2 Hz, 1H), 6.94 (d, J = 8.6 Hz, 2H), 6.80 (s, 1H), 6.45–6.43 (m, 2H), 6.40-6.37 (m, 1H), 6.36–6.28 (m, 2H), 5.64 (d, J = 3.2 Hz, 1H), 5.57 (ddd, J = 19.6, 14.8, 3.6 Hz, 2H), 5.13 (dd, J = 10.2, 3.2 Hz, 1H), 4.52 (s, 2H), 3.41–3.08 (m, 4H), 2.93–2.80 (m, 1H), 2.67–2.44 (m, 2H), 2.34–2.3 (m, 8H), 2.18 (s, 3H), 2.08 (s, 3H), 2.05–1.91 (m, 4H), 1.58–1.39 (m, 4H), 1.26 (s, 6H); ¹³ C NMR (101 MHz, CDCl ₃ ): δ 182.3, 170.4, 170.2, 169.9, 169.6, 168.2, 168.0, 167.9, 165.8, 163.9, 163.2, 162.0, 159.9, 157.7, 144.1, 143.8, 142.5, 132.8, 128.1, 126.6, 124.8, 124.0, 122.9, 118.3, 115.1, 104.8, 104.3, 99.7, 94.4, 81.2, 71.7, 68.1, 67.3, 67.2, 53.5, 39.8, 39.1, 38.3, 30.7, 29.2, 29.2, 28.6, 26.6, 21.7, 21.0, 20.7, 20.6, 20.58; HRMS: C ₅₀ H ₅₄ N ₂ O ₁₉ + H ⁺ , Cal 987.3394 Found, 987.3395; C ₅₀ H ₅₄ N ₂ O ₁₉ + Na ⁺ Cal 1009.3213, Found 1009.3211。

Example 11

Synthesis of Compound 11 d: compound 4a (65.0 mg, 0.2 mmol, 1.0 eq) and HOBT (27.0 mg, 0.2 mmol, 1.0 eq) were weighed out, EDCI (76.0 mg, 0.4 mmol, 2.0 eq) was dissolved in 25mL round bottom flask, DMF (5 mL) was added and stirred at room temperature for 10 min, followed by compound 7e (180.0 mg, 0.24 mmol, 1.2 eq) and reacted at room temperature for 2-3h. After TLC monitored the reaction was complete, clear water 5mL was added, followed by extraction with ethyl acetate (15 mL ×5), and the combined organic layers were washed sequentially with clear water (15 mL ×2), saturated brine (10 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure through the column (DCM/meoh=100/1-20/1). 110. 110 mg as a pale yellow foamy solid was obtained in a yield of 51.2%. ¹ H NMR (400 MHz, CDCl ₃ ): δ 7.73 (d, J = 7.8 Hz, 2H), 7.52 (d, J = 16.0 Hz, 1H), 7.33–7.29 (m, 2H), 7.15 (d, J = 8.2 Hz, 1H), 6.92 (d, J = 8.3 Hz, 2H), 6.56-6.53 (m, 1H), 6.42 (d, J = 20.4 Hz, 2H), 6.33–6.16 (m, 3H), 5.62–5.41 (m, 2H), 5.06 (dd, J = 10.2, 3.6 Hz, 1H), 4.49 (s, 2H), 3.27-3.26 (m, 2H), 3.13-3.12 (m, 2H),2.89–2.73 (m, 1H), 2.55-2.44 (m, 2H), 2.23 (d, J = 2.8 Hz, 6H), 2.12 (s, 3H), 2.02 (s, 3H), 1.98–1.87 (m, 5H), 1.43–1.35 (m, 4H), 1.18–1.05 (m, 16H); ¹³ C NMR (101 MHz, CDCl ₃ ): δ 182.3, 170.3, 170.1, 169.8, 169.5, 168.1, 167.9, 167.8, 167.0, 163. 9, 163.3, 162.1, 159.9, 157.8, 144.2, 143.9, 142.5, 132.8, 128.2, 126.6, 125.0, 124.0, 122.9, 118.2, 115.2, 104.9, 104.4, 99.7, 94.5, 81.3, 71.7, 68.1, 67.3, 39.9, 39.2, 38.4, 30.7, 29.7, 29.5, 29.5, 29.3, 29.3, 29.2, 29.17, 26.8, 21.7, 21.0, 20.7, 20.6, 20.55; HRMS: C ₅₅ H ₆₄ N ₂ O ₁₉ + H ⁺ , Cal 1057.4182 Found, 1057.4187。

Example 12

Synthesis of compound 12 a: compound 4a (49.1 mg, 0.15 mmol, 1.0 eq) and HOBT (20.0 mg, 0.15 mmol, 1.0 eq) were weighed out, EDCI (57.3 mg, 0.3 mmol, 2.0 eq) was dissolved in 25mL round bottom flask, DMF (5 mL) was added and reacted at room temperature with stirring for 10 min, followed by compound 9a (117.0 mg, 0.18 mmol, 1.2 eq) and reacted at room temperature for 2-3h. After TLC monitored the reaction was complete, clear water 5mL was added, followed by extraction with ethyl acetate (15 mL ×5), and the combined organic layers were washed sequentially with clear water (15 mL ×2), saturated brine (10 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure through the column (DCM/meoh=100/1-20/1). Obtained as a pale yellow foamy solid 65 mg in 44.2% yield. ¹ HNMR (400 MHz, CDCl ₃ ): δ 12.73 (s, 1H), 7.66 (d, J = 8.2 Hz, 2H), 7.57 (d, J = 15.8 Hz, 1H), 7.38-7.36 (m, 3H), 7.21 (d, J = 8.2 Hz, 1H), 6.99 (s, 1H), 6.89 (d, J = 8.2 Hz, 2H), 6.39-6.30 (m, 4H), 5.74-5.49 (m, 2H), 5.15 (d, J = 7.4 Hz, 1H), 4.55 (s, 2H), 3.579-3.55 (m, 7H), 3.31 (s, 1H), 2.89 (d, J = 15.8 Hz, 1H), 2.61 (dd, J = 41.0, 13.6 Hz, 2H), 2.30 (s, 6H), 2.18 (s, 3H), 2.08 (s, 3H), 2.05- 1.94 (m, 4H); ¹³ C NMR (100 MHz, CDCl ₃ ) δ: 182.2, 170.7, 170.2, 169.9, 169.7, 168.3, 168.2, 168.1, 163.9, 163.0, 161.7, 160.0, 157.7, 144.3, 143.9, 142.5, 132.7, 128.0, 126.7, 124.4, 124.1, 123.0, 118.1, 115.1, 104.8, 104.1, 99.6, 94.6, 81.1, 71.6, 69.7, 69.4, 68.1, 67.5, 67.3, 39.7, 39.1, 38.4, 30.7, 21.6, 21.0, 20.7, 20.64, 20.60; HRMS: C ₄₇ H ₄₈ N ₂ O ₂₀ + H ⁺ , Cal: 961.2897 Found, 961.2881; C ₄₇ H ₄₈ N ₂ O ₂₀ + Na ⁺ , Cal 983.2693 Found, 983.2697。

Example 13

Synthesis of compound 12 b: compound 4a (65.0 mg, 0.2 mmol, 1.0 eq) and HOBT (27.0 mg, 0.2 mmol, 1.0 eq) were weighed out, EDCI (76.0 mg, 0.4 mmol, 2.0 eq) was dissolved in a 25mL round bottom flask, DMF (5 mL) was added and reacted at room temperature with stirring for 10 min, followed by compound 9b (161.0 mg, 0.24 mmol, 1.2 eq) at room temperature for 2-3h. After TLC monitored the reaction was complete, clear water 5mL was added, followed by extraction with ethyl acetate (15 mL ×5), and the combined organic layers were washed sequentially with clear water (15 mL ×2), saturated brine (10 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure through the column (DCM/meoh=100/1-20/1). Light yellow foamy solid 104 mg was obtained in 52.0% yield. ¹ HNMR (400 MHz, CDCl ₃ ): δ 12.73 (s, 1H), 9.12 (s, 1H), 7.72 (d, J = 8.0 Hz, 2H), 7.57 (d, J = 15.9 Hz, 1H), 7.38-7.35 (m, 2H), 7.24-7.15 (m, 2H), 6.91 (d, J = 7.8 Hz, 2H), 6.74 (s, 1H), 6.44-6.30 (m, 4H), 5.64-5.54 (m, 2H), 5.13 (d, J = 7.6 Hz, 1H), 4.52 (s, 2H), 3.62-3.52 (m, 12H), 2.87 (d, J = 15.8 Hz, 1H), 2.58 (dd, J = 40.0, 13.4 Hz, 3H), 2.30 (s, 6H), 2.19 (s, 3H), 2.09 (s, 3H), 2.00 (s, 3H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 182.2, 170.6, 170.2, 169.9, 169.6, 168.2, 168.1, 168.0, 165.8, 163.6, 163.1, 162.0, 159.8, 157.7, 144.1, 143.8, 142.5, 132.8, 128.0, 126.6, 124.7, 124.0, 122.9, 118.2, 115.1, 104.9, 104.2, 99.5, 94.4, 81.1, 71.7, 70.2, 70.1, 69.5, 68.1, 67.2, 39.5, 38.9, 38.2, 30.8, 21.6, 21.0, 20.7, 20.64, 20.60; HRMS: C ₄₉ H ₅₂ N ₂ O ₂₁ + H ⁺ , Cal 1005.3135 Found, 1005.3134; C ₄₉ H ₅₂ N ₂ O ₂₁ + Na ⁺ , Cal 1027.2955, Found, 1027.2954。

Example 14

Synthesis of Compound 12 c: compound 4a (49.1 mg, 0.15 mmol, 1.0 eq) and HOBT (20.0 mg, 0.15 mmol, 1.0 eq) were weighed out, EDCI (57.3 mg, 0.3 mmol, 2.0 eq) was dissolved in 25mL round bottom flask, DMF (5 mL) was added and stirred at room temperature for 10 min, followed by compound 9c (124.8 mg, 0.18 mmol, 1.2 eq) and reacted at room temperature for 2-3h. After TLC monitored the reaction was complete, clear water 5mL was added, followed by extraction with ethyl acetate (15 mL ×5), and the combined organic layers were washed sequentially with clear water (15 mL ×2), saturated brine (10 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure through the column (DCM/meoh=100/1-20/1). 67. 67 mg as a pale yellow foamy solid was obtained in 43.7% yield. ¹ HNMR (400 MHz, CDCl ₃ ): δ 12.72 (s, 1H), 9.14 (s, 1H), 7.73 (d, J = 7.6 Hz, 2H), 7.58 (d, J = 16.0 Hz, 1H), 7.39-7.35 (m, 2H), 7.25-7.11 (m, 2H), 6.92 (d, J = 7.4 Hz, 2H), 6.78 (s, 1H), 6.52-6.23 (m, 4H), 5.73-5.47 (m, 2H), 5.12 (d, J = 7.4 Hz, 1H), 4.53 (s, 2H), 3.73 -3.43 (m, 15H), 3.37 (s, 1H), 2.86 (d, J = 15.8 Hz, 1H), 2.57 (dd, J = 41.0, 14.2 Hz, 3H), 2.31 (s, 6H), 2.18 (s, 3H), 2.08 (s, 3H), 1.99 (s, 3H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 182.2, 170.6, 170.2, 169.8, 169.6, 168.2, 168.0, 167.9, 165.8, 163.6, 163.1, 162.1, 159.8, 157.7, 144.0, 143.8, 142.5, 132.8, 128.0, 126.6, 124.7, 124.0, 122.9, 118.3, 115.1, 104.9, 104.3, 99.6, 94.3, 81.0, 71.8, 70.4, 70.2, 70.1, 69.6, 68.1, 67.2, 67.1, 39.6, 39.0, 38.2, 30.8, 21.6, 21.0, 20.7, 20.64, 20.6; HRMS: C ₅₁ H ₅₆ N ₂ O ₂₂ + Na ⁺ , Cal 1071.3217 Found, 1071.3214。

Example 15

Synthesis of Compound 12 d: compound 4a (49.1 mg, 0.15 mmol, 1.0 eq) and HOBT (20.0 mg, 0.15 mm) were weighed outol, 1.0 eq), EDCI (57.3 mg, 0.3 mmol, 2.0 eq) was dissolved in a 25mL round bottom flask, DMF (5 mL) was added and the reaction was stirred at room temperature for 10 min, followed by addition of compound 9d (132.0 mg, 0.18 mmol, 1.2 eq) and reaction at room temperature for 2-3h. After TLC monitored the reaction was complete, clear water 5mL was added, followed by extraction with ethyl acetate (15 mL ×5), and the combined organic layers were washed sequentially with clear water (15 mL ×2), saturated brine (10 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure through the column (DCM/meoh=100/1-20/1). Obtained as a pale yellow foam 48, mg in 30.0% yield. ¹ H NMR (400 MHz, CDCl ₃ ): δ 12.72 (s, 1H), 9.12 (s, 1H), 7.76 (d, J = 5.0 Hz, 2H), 7.58 (d, J = 15.8 Hz, 1H), 7.36 (s, 2H), 7.23 (s, 2H), 6.95 (d, J = 4.8 Hz, 2H), 6.85 (s, 1H), 6.55–6.23 (m, 4H), 5.64-5.57 (m, 2H), 5.11 (d, J = 9.8 Hz, 1H), 4.54 (s, 2H), 3.72–3.45 (m, 19H), 3.36 (s, 1H), 2.86 (d, J = 15.2 Hz, 1H), 2.68-2.48 (m, 3H), 2.31 (s, 6H), 2.19 (s, 3H), 2.09 (s, 3H), 1.99 (s, 3H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 182.2, 170.6, 170.2, 169.8, 169.6, 168.2, 168.0, 167.8, 165.7, 163.6, 163.2, 162.1, 159.9, 157.7, 144.0, 143.8, 142.5, 132.8, 128.1, 126.6, 124.8, 124.0, 123.3, 122.9, 118.3, 115.2, 105.0, 104.4, 99.6, 94.3, 81.0, 71.8, 70.4, 70.2, 70.15, 69.7, 68.1, 67.3, 67.1, 39.6, 39.0, 38.3, 30.8, 21.6, 21.0, 20.7, 20.64, 20.60; HRMS: C ₅₃ H ₆₀ N ₂ O ₂₃ + Na ⁺ , Cal 1115.3479; Found, 1115.3481。

Determination of XOD inhibitory Activity of chlorogenic acid-Huang Qinsu conjugate: to the reaction solution containing 67 mM phosphate buffer (ph=7.4) with XOD (activity of 20 nM, 5 mU/mL), chlorogenic acid-Huang Qinsu (CA) series compound and positive control allopurinol were added at different concentrations (6.25, 12.5, 25.0, 50, 100 μmol/L), respectively, and no CA group was added as a control. The mixture was pre-incubated at 37℃for 15 min, 50 mM xanthine was added to the mixture, immediately after which the absorbance was measured at 295 nm, 1 time per minute for 30 min, and the percent (%) inhibition was calculated according to the following equation, whereby the drug concentration required for 50% inhibition was calculated as IC ₅₀ Values. Test compounds and allopurinol were dissolved in DMSO, so DMSO was used as a negative control.

Inhibition% = [ (control reaction rate-experimental reaction rate)/(control reaction rate) ]x100

The test results are shown in fig. 1 and 2: compounds 10c, 10d were found to exhibit good inhibitory activity against XOD, with more optimal compound 10c: IC ₅₀ = 30.742 μM，10d: IC ₅₀ = 22.425 μm, which has slightly better inhibitory activity than allopurinol: IC (integrated circuit) ₅₀ = 40.547 μM。

Determination of anti-inflammatory Activity of chlorogenic acid-Huang Qinsu conjugate: RAW264.7 cells were seeded in 96-well plates at a concentration of 5X 10 at 100. Mu.L/well ⁴ Well, culturing at 37 ℃ under constant temperature and humidity overnight; treatment with the test compound (concentration 40. Mu.g/mL, 50. Mu.L/well) 2 h and LPS (concentration 4. Mu.g/mL, 50. Mu.L/well) 24 h(experiments are divided into a control group, a 1% DMSO negative control group and an experimental group (10 a-10d, 12a-12 d). Wherein the control group is normal cultured cells, and neither compound nor LPS is added for stimulation, the DMSO negative control group, namely a model group, is not added with compound and then is added with LPS for stimulation, and the experimental group is added with LPS for stimulation after the compound is added for culture; 100. Mu.L of supernatant was collected in another 1 new 96-well plate, 100. Mu.L of formulated reagent for qualitative test of urinary nitrite (griess) A/B (v/v=1:1) was added to each well, and the mixture was shaken at room temperature for 10 min, absorbance was measured at 540 and nm, and NO production level was analyzed according to standard curve regression equation; MTT (5 mg/mL, 10. Mu.L/well) was added to the remaining wells and incubated at 37℃in the absence of light for 4 h; the supernatant was discarded, DMSO was added at 200. Mu.L/well, and the absorbance (A) was measured at 492 nm by shaking at room temperature for 10 min.

The cell viability calculation formula is: survival (%) = (experimental group a-blank group a)/(control group a-blank group) 100 in order to evaluate the anti-inflammatory activity of the synthesized Huang Qinsu-chlorogenic acid conjugate, the effect of the compound on NO production, cell survival and cell morphology of RAW264.7 cells was tested.

* Indicating significant differences compared to the control group, # indicates significant differences compared to the model group, the numbers represent the degree of difference significance, # p <0.01, # p <0.001, # p <0.0001 compared to the control group, # p <0.05, # p <0.01, # p <0.001, # p <0.0001 compared to the model group.

Effect of compound on NO formation: compounds 10a, 10b, 10c, 10d at a concentration of 10. Mu.g/mL have good inhibitory activity against lipopolysaccharide-induced NO production in RAW264.7 cells, with compound 10b exhibiting the strongest inhibitory effect. Other compounds had less effect on lipopolysaccharide-induced NO production by RAW264.7 cells (fig. 3).

Effect of compounds on cell viability: compounds 10b, 10c, 10d and 12b at a concentration of 10 μg/mL had less effect on the lipopolysaccharide-induced decrease in RAW264.7 cell viability (percentage calculated as compared to the control group); while compounds 10a, 12a may protect cells from lipopolysaccharide-induced decrease in cell viability; while compounds 12b, 12c have an accelerating effect on lipopolysaccharide-induced decrease in RAW264.7 cell viability (fig. 4).

Effect of compounds on cell morphology: compounds 10a, 10b, 10c, 10d and 12b at a concentration of 10 μg/mL had less effect on lipopolysaccharide-induced changes in RAW264.7 cell morphology; compound 12c has an enhancing effect on lipopolysaccharide-induced changes in RAW264.7 cell morphology (fig. 5). In FIG. 5, C represents a control group, and L represents an LPS-lipopolysaccharide group.

From the above, the chlorogenic acid-Huang Qinsu conjugate can be used for preparing medicines for preventing or treating gouty diseases, wherein the gouty diseases comprise acute gouty arthritis, chronic gouty arthritis, intermittent gouty arthritis, drug-induced liver injury, acute kidney injury or neuropathic pain and the like.

Chlorogenic acid-Huang Qinsu conjugate of the invention can be used for preparing medicines for preventing or treating inflammatory infectious diseases, wherein the inflammatory infectious diseases comprise gouty arthritis, inflammatory bowel disease, cerebral (spinal) membranitis, pancreatitis, peritonitis, vasculitis (disease), glomerulonephritis, hepatitis, keratitis, cataract, senile macular degeneration and optic neuritis diseases.

The preferred embodiments of the invention disclosed above are intended only to assist in the explanation of the invention. The preferred embodiments are not exhaustive or to limit the invention to the precise form disclosed. Obviously, many modifications and variations are possible in light of the above teaching. The embodiments were chosen and described in order to best explain the principles of the invention and the practical application, to thereby enable others skilled in the art to best understand and utilize the invention. The invention is limited only by the claims and the full scope and equivalents thereof.

Claims

1. Chlorogenic acid-Huang Qinsu conjugate is characterized by having a structure shown in a general formula I or a general formula II:

，/>，

wherein: r is R ₁ And R is ₂ Respectively is any one of H, alkyl, aryl, heterocyclic aryl, C (O) -alkyl, C (O) -aryl, C (O) -heterocyclic aryl and C (O) -amino;

R ₃ and R is ₄ Respectively is any one of H, alkyl, aryl, heterocyclic aryl, C (O) -alkyl, C (O) -aryl, C (O) -heterocyclic aryl and C (O) -amino;

in formula ii, m=1, 2, 3, 4, 5 or 6; n=1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12.

2. A method for preparing chlorogenic acid-Huang Qinsu conjugate according to claim 1, comprising the following steps:

3. The method for preparing chlorogenic acid-Huang Qinsu conjugate according to claim 2, wherein the synthesis of compound 7a is as follows:

synthesis of Compound 5: 1eq chlorogenic acid and 0.2 eq DMAP are weighed into a reaction vessel, acetic anhydride 20 mL and pyridine 10 mL are added, after the reaction is completed, the reaction solution is poured into an ice-water mixture and stirred at room temperature, then the pH is regulated to 2-3 by 3N dilute hydrochloric acid, white solid is separated out, the solid is filtered out, and is washed by a proper amount of clear water, and the white solid of the target product is obtained after drying, namely the compound 5;

synthesis of compound 6 a: 1eq of compound 5,1eq of HOBT and 2eq of EDCI are weighed into a reaction vessel, after being dissolved by adding dichloromethane 50 mL, stirring at room temperature, after being added with 2eq of N-tert-butoxycarbonyl-ethylenediamine, stirring at room temperature for reaction, after the reaction is completed, adding dichloromethane 50 mL for dilution, then washing with clear water and saturated saline in sequence, drying an organic layer by anhydrous sodium sulfate, concentrating under reduced pressure, and obtaining a target product of compound 6a by column chromatography;

synthesis of compound 7 a: 1eq of compound 6a was dissolved in DCM, followed by addition of 50eq of TFA, volume ratio DCM: TFA=3/1, stirring reaction at room temperature, after completion of the reaction, dichloromethane was removed under reduced pressure, followed by addition of dichloromethane and re-spinning dry, the crude product obtained was purified by column chromatography to give compound 7a as a white foam solid.

4. A method for preparing chlorogenic acid-Huang Qinsu conjugate according to claim 1, which comprises the following steps:

5. The method for preparing chlorogenic acid-Huang Qinsu conjugate according to claim 4, wherein the step of synthesizing compound 7b, compound 7c, compound 7d or compound 7e is as follows:

synthesis of Compound 5: 1eq chlorogenic acid and 0.2 eq DMAP are weighed into a reaction vessel, acetic anhydride 20 mL and pyridine 10 mL are added, after the reaction is completed, the reaction solution is poured into an ice-water mixture and stirred at room temperature, then the pH is adjusted to 2-3 by 3N dilute hydrochloric acid, white solid is separated out, the solid is filtered out, and is washed by a proper amount of clear water, and the white solid of the target product is obtained after drying, namely the compound 5;

Synthesis of compound 6b, compound 6c, compound 6d or compound 6 e: 1eq of compound 5,1eq HOBT and 2eq EDCI are weighed into a reaction vessel, after methylene dichloride 50 mL is added for dissolution, stirring is carried out at room temperature, then 2eq N-tert-butoxycarbonyl-1, 5-diaminopentane, N-tert-butoxycarbonyl-1, 6-diaminohexane, N-tert-butoxycarbonyl-1, 7-diaminoheptane and N-tert-butoxycarbonyl-1, 12-diaminododecane are added, after the reaction is completed, methylene dichloride is added for dilution, then clear water and saturated saline water are sequentially used for washing, the organic layer is dried by anhydrous sodium sulfate and then concentrated under reduced pressure, and the crude product is subjected to column chromatography to obtain a target product of compound 6b, compound 6c, compound 6d or compound 6e;

synthesis of compound 7b, compound 7c, compound 7d or compound 7 e: 1eq of compound 6b, compound 6c, compound 6d or compound 6e was dissolved in DCM, 50eq of TFA was then added, the volume ratio of DCM: TFA=3/1, the reaction was stirred at room temperature, after completion of the reaction, the dichloromethane was removed under reduced pressure, then dichloromethane was added and again dried by spinning, and the crude product obtained was purified by column chromatography to give compound 7b, compound 7c, compound 7d or compound 7e as a white foamy solid.

6. A method for preparing chlorogenic acid-Huang Qinsu conjugate according to claim 1, comprising the following steps:

7. The method for producing chlorogenic acid-Huang Qinsu conjugate according to claim 6, wherein the step of synthesizing compound 9a, compound 9b, compound 9c or compound 9d is as follows:

synthesis of compound 8a, compound 8b, compound 8c or compound 8 d: 1eq of compound 5,1eq HOBT, 2eq EDCI is called into a reaction vessel, after methylene chloride is added for dissolution, stirring is carried out at room temperature, then any one of 2eq tert-butoxycarbonyl-imino-polyethylene glycol-amino, 2- (2- (2-amino ethoxy) ethyl carbamic acid tert-butyl ester, 3-amino-5, 8, 11-trioxa-2-aza-tridecanoic acid 1, 1-dimethyl ethyl ester and O- (2-amino ethyl) -O' - [2- (tert-butoxycarbonyl-amino) ethyl ] tetraethylene glycol is added, after the reaction is completed, methylene chloride is added for dilution, and then washing and drying are carried out, and the crude product is subjected to column chromatography to obtain compound 8a, compound 8b, compound 8c or compound 8d;

Synthesis of compound 9a, compound 9b, compound 9c or compound 9 d: 1eq of compound 8a, compound 8b, compound 8c or compound 8d was dissolved in DCM, followed by 50.0 eq of TFA, wherein the volume ratio DCM: TFA=3/1, the reaction was stirred at room temperature, after completion of the reaction, the dichloromethane was removed under reduced pressure, and the crude product obtained was purified by column chromatography to give compound 9a, compound 9b, compound 9c or compound 9d.

8. Use of chlorogenic acid-Huang Qinsu conjugate according to claim 1 in the preparation of a medicament for preventing or treating gouty diseases.

9. Use of chlorogenic acid-Huang Qinsu conjugate according to claim 1 for the preparation of a medicament for preventing or treating inflammatory infectious diseases.