Betulinic acid derivative prepared from active ingredients of white birch, preparation method and application thereof

Abstract

The present invention discloses a betulinic acid derivative prepared from active ingredients of birch, and its preparation method and application. The betulinic acid derivative has a structure shown in formula (I): #imgabs0# wherein R is selected from substituted or unsubstituted C 6～12 Aryl, substituted or unsubstituted C 5～12 The present invention uses the betulic acid structure as the parent core to design and synthesize a series of betulic acid derivatives, which exhibit high α-glucosidase inhibitory activity and can be used as α-glucosidase inhibitors to treat or prevent diabetes.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical chemistry, and in particular to a betulinic acid derivative prepared from active ingredients of birch, and a preparation method and application thereof. Background Art

[0002] Type 2 diabetes mellitus (T2DM) has become one of the major chronic diseases. In particular, T2DM is characterized by hyperglycemia, which can lead to the occurrence of serious diabetic complications, such as vascular damage and cardiovascular disease. Therefore, controlling postprandial hyperglycemia has become an effective therapy for the treatment of T2DM and the prevention of diabetic complications. Carbohydrates, such as starch and dextrin in food, are the main source of blood glucose in the human body. α-glucosidase present in the gastrointestinal tract can catalyze the cleavage of α-1,4 glycosidic bonds of carbohydrates to form glucose, which is absorbed by the small intestine and then enters the blood circulation. Inhibiting the activity of intestinal α-glucosidase can effectively control postprandial hyperglycemia. Acarbose is an α-glucosidase inhibitor; however, its inhibitory effect is poor.

[0003] Therefore, it is necessary to develop an α-glucosidase inhibitor with better inhibitory effect. Summary of the Invention

[0004] The present invention aims to solve at least one of the technical problems existing in the prior art. To this end, in a first aspect, the present invention provides a betulinic acid derivative prepared from active ingredients of birch, which has better α-glucosidase inhibitory activity.

[0005] The second aspect of the present invention also provides a method for preparing betulic acid derivatives.

[0006] The third aspect of the present invention also provides a pharmaceutical composition.

[0007] The fourth aspect of the present invention also provides an α-glucosidase inhibitor.

[0008] The fifth aspect of the present invention also provides an application of betulinic acid derivatives.

[0009] According to the first aspect of the present invention, an embodiment provides a betulic acid derivative or a pharmaceutically acceptable salt thereof, wherein the betulic acid derivative has a structure shown in formula (I):

[0010]

[0011] wherein R is selected from substituted or unsubstituted C 6～12 Aryl, substituted or unsubstituted C 5～12 of heteroaryl.

[0012] The betulinic acid derivatives or pharmaceutically acceptable salts thereof according to the embodiments of the present invention have at least the following beneficial effects:

[0013] The present invention uses the betulic acid structure as the mother core to design and synthesize a series of betulic acid derivatives, which exhibit high α-glucosidase inhibitory effect and can be used as α-glucosidase inhibitors for treating or preventing diabetes.

[0014] According to some embodiments of the present invention, the R is selected from substituted or unsubstituted phenyl, the substitution is monosubstituted or polysubstituted, substituted or unsubstituted C 1～6 Alkyl, pyridyl.

[0015] According to some embodiments of the present invention, R is selected from the following groups:

[0016]

[0017] According to some embodiments of the present invention, the betulic acid derivative is selected from one of the following structures:

[0018]

[0019]

[0020] According to the second aspect of the present invention, the method for preparing the betulic acid derivatives provided in an embodiment comprises the following steps:

[0021] S1. Under anaerobic conditions, betulinic acid, a first organic solvent, chloroacetyl chloride or chloroacetic anhydride, an acid-binding agent, and DMAP are reacted, followed by post-treatment to obtain intermediate 2;

[0022] S2, under anaerobic conditions, mixing intermediate 2, a second organic solvent and sodium azide to carry out a substitution reaction to obtain intermediate 3;

[0023] S3, under anaerobic conditions, intermediate 3, The reducing agent, the copper catalyst and the third organic solvent react to obtain the betulinic acid derivative represented by formula (I);

[0024] Among them, the structural formulas of betulinic acid, intermediate 2 and intermediate 3 are as follows:

[0025]

[0026] According to some embodiments of the present invention, the first organic solvent, the second organic solvent, and the third organic solvent are independently selected from at least one of tetrahydrofuran, pyridine, acetone, DMF, and DMSO.

[0027] According to some embodiments of the present invention, in step S1, the reaction temperature is 15-35°C.

[0028] According to some embodiments of the present invention, in step S1, the reaction time is 1 to 5 hours.

[0029] According to some embodiments of the present invention, in step S1, the acid binding agent is selected from at least one of DIPEA and triethylamine.

[0030] According to some embodiments of the present invention, the copper catalyst is selected from at least one of copper sulfate pentahydrate, cuprous bromide or cuprous oxide.

[0031] According to some embodiments of the present invention, the reducing agent is selected from at least one of sodium ascorbate and DIPEA.

[0032] According to some embodiments of the present invention, in step S2, the reaction time is 8 to 16 hours.

[0033] According to some embodiments of the present invention, in step S3, the reaction time is 2 to 18 hours.

[0034] According to some embodiments of the present invention, the post-treatment step includes concentration, extraction and column chromatography.

[0035] The third aspect of the present invention provides a pharmaceutical composition comprising the above-mentioned betulic acid derivatives or pharmaceutically acceptable salts thereof; and pharmaceutically acceptable excipients.

[0036] The fourth aspect of the present invention provides an α-glucosidase inhibitor, comprising the above-mentioned betulic acid derivative or a pharmaceutically acceptable salt thereof.

[0037] A fifth aspect of the present invention provides the use of the betulic acid derivatives described above in the preparation of products for preventing and / or treating diabetes.

[0038] According to some embodiments of the present invention, the product includes at least one of a medicine or a health product.

[0039] Definitions and General Terms

[0040] "Substituted or unsubstituted C 6～12 The term "aryl" refers to an all-carbon monocyclic or fused polycyclic group having a completely conjugated π electron system and a total number of carbon atoms of 6 to 12. Optionally, at least one H in the aryl group may be substituted by a corresponding group as defined herein.

[0041] "Substituted or unsubstituted C 5～12"Heteroaryl" means a monocyclic or fused ring group of ring atoms containing one, two, three or four ring heteroatoms selected from N, O or S, the remaining ring atoms being C, and having a completely conjugated π electron system and a total of 5 to 12 carbon atoms. Optionally, at least one H in the heteroaryl group is replaced by a corresponding group as defined herein.

[0042] “C 1～6 "alkyl" means an alkyl group having a total carbon number of 1 to 6, including C 1-6 Straight chain alkyl, C 1-6 Branched alkyl and C 3-6 The cycloalkyl group may be, for example, a straight-chain alkyl group having 1, 2, 3, 4, 5 or 6 carbon atoms, a branched-chain alkyl group having 1, 2, 3, 4, 5 or 6 carbon atoms, or a cycloalkyl group having 3, 4, 5 or 6 carbon atoms, for example, a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a tert-butyl group, an n-pentyl group, an isopentyl group, a n-hexyl group, a cyclopropyl group, a methylcyclopropyl group, an ethylcyclopropyl group, a cyclopentyl group, a methylcyclopentyl group, a cyclohexyl group, and the like.

[0043] In the present invention, "substituted or unsubstituted" means that the group may or may not be further substituted by one or more groups selected from the following: 1～6 alkyl, halogen, nitro, hydroxy, alkenyl, and alkynyl.

[0044] The structural formula in the present invention appears Indicates the attachment site of a group.

[0045] The term "pharmaceutically acceptable" as used herein refers to substances that are acceptable for pharmaceutical use from a toxicological point of view and do not adversely interact with the active ingredient.

[0046] Pharmaceutically acceptable excipients used in the present invention include any solvent, solid excipient, diluent, binder, disintegrant, or other liquid excipient, dispersant, flavoring agent or suspending agent, surfactant, isotonic agent, thickener, emulsifier, preservative, solid binder, glidant or lubricant, etc., suitable for the specific target dosage form. As described in the following documents: In Remington: The Science and Practice of Pharmacy, 21st edition, 2005, ed. D. B. Troy, Lippincott Williams & Wilkins, Philadelphia, and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and J. C. Boylan, 1988-1999, Marcel Dekker, New York, the contents of these documents are summarized, indicating that different excipients can be used in the preparation of pharmaceutically acceptable compositions and their known preparation methods. Except to the extent that any conventional excipients are incompatible with the compounds of the present invention, for example by producing any adverse biological effects or interacting in a deleterious manner with any other component of the pharmaceutically acceptable composition, their use is contemplated by the present invention.

[0047] Examples of pharmaceutically acceptable excipients include, but are not limited to, ion exchangers; aluminum; aluminum stearate; lecithin; serum proteins, such as human serum albumin; buffer substances, such as phosphates; glycine; sorbic acid; potassium sorbate; partial glyceride mixtures of saturated vegetable fatty acids; water; salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts; colloidal silicon; magnesium trisilicate; polyvinylpyrrolidone; polyacrylates; waxes; polyethylene-polyoxypropylene-blocking polymers; lanolin; sugars, such as lactose, glucose, and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as carboxymethyl cellulose. sodium cellulose, ethylcellulose and cellulose acetate; gum powder; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycol compounds such as propylene glycol and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethanol; phosphate buffered solution; and other nontoxic suitable lubricants such as sodium lauryl sulfate and magnesium stearate; coloring agents; release agents; coatings; sweeteners; flavoring agents; fragrances; preservatives and antioxidants.

[0048] The pharmaceutical compositions of the compounds of the present invention can be administered in unit dosage forms. The dosage forms can be liquid or solid. Liquid dosage forms can be true solutions, colloids, microparticles, or suspensions. Other dosage forms include tablets, capsules, dripping pills, aerosols, pills, powders, solutions, suspensions, emulsions, granules, suppositories, and lyophilized powder injections.

[0049] Tablets and capsules for oral administration may contain excipients such as binders, such as syrup, gum arabic, sorbitol, tragacanth, or polyvinyl pyrrolidone; fillers, such as lactose, sucrose, corn starch, calcium phosphate, sorbitol, glycine; lubricants, such as magnesium stearate, talc, polyethylene glycol, silica; disintegrants, such as potato starch; or acceptable wetting agents, such as sodium lauryl sulfate. Tablets may be coated by methods known in the pharmaceutical industry.

[0050] Oral liquids can be prepared as suspensions, solutions, emulsions, syrups, or elixirs of hydrated oils, or as dry products, supplemented with water or other suitable media before use. Such liquid preparations may contain conventional additives such as suspending agents, sorbitol, methyl cellulose, glucose syrup, gelatin, hydroxyethyl cellulose, carboxymethyl cellulose, aluminum stearate gel, hydrogenated edible oils and fats, emulsifiers such as lecithin, sorbitan monooleate, and gum arabic; or non-aqueous vehicles (which may include edible oils) such as almond oil, fats such as glycerol, ethylene glycol, or ethanol; and preservatives such as methyl or propyl parahydroxybenzoate or sorbic acid. Flavorings or coloring agents may be added as needed.

[0051] Other features and advantages of the present invention will be set forth in the description which follows, and in part will be obvious from the description, or may be learned by practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0052] The above and / or additional aspects and advantages of the present invention will become apparent and readily understood from the following description of the embodiments with reference to the accompanying drawings, in which:

[0053] Figure 1 A graph showing the half-maximal inhibitory concentration of a betulic acid derivative Z13 as an α-glucosidase inhibitor for α-glucosidase in vitro is provided for Example 2 of the present invention;

[0054] Figure 2 This is a diagram showing the enzyme kinetics of the betulinic acid derivative Z13 used as an α-glucosidase inhibitor against α-glucosidase in vitro in Example 3 of the present invention;

[0055] Figure 3 This is a diagram showing the substrate kinetics of the betulic acid derivative Z13 used as an α-glucosidase inhibitor in vitro for α-glucosidase in Example 4 of the present invention. DETAILED DESCRIPTION

[0056] The following are specific embodiments of the present invention, and the technical solutions of the present invention are further described in conjunction with the embodiments, but the present invention is not limited to these embodiments.

[0057] Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

[0058] Example 1

[0059] Example 1 provides a betulic acid derivative, the reaction equation and preparation formula of which are as follows:

[0060]

[0061] S1. Betulinic acid (1 eq) was dissolved in 400 μl THF. Chloroacetyl chloride (2.8 eq), DIPEA (1.7 eq), and DMAP (1.7 eq) were added in sequence under anaerobic conditions. The mixture was reacted at room temperature for 2 h. After monitoring the reaction, the excess solvent was removed by rotary evaporation. The reaction solution was extracted with ethyl acetate (3 × 15 mL). The organic phase was collected and washed with water. Intermediate 2 was obtained by column chromatography.

[0062] S2. Dissolve intermediate 2 (1 eq) and sodium azide (5 eq) in 2 mL of DMF and react overnight at room temperature by nucleophilic substitution under anaerobic conditions. After monitoring the reaction, extract the reaction solution with ethyl acetate (3 × 15 mL) and collect the organic phase. Wash the organic phase with water and obtain intermediate 3 by column chromatography.

[0063] S3. Dissolve the intermediate 3 (1 eq) in DMSO and add the R-containing acetylene compound in sequence. Sodium ascorbate (1 eq) and copper sulfate pentahydrate (1.01 eq) were reacted at room temperature under anaerobic conditions for 2-18 hours. The reaction solution was extracted with ethyl acetate (3 x 15 mL), and the organic phase was collected and washed with water. The organic phase was purified by column chromatography to obtain betulinic acid derivatives.

[0064] The general structural formula (Z1-Z26) of the prepared betulinic acid derivatives is:

[0065]

[0066] Wherein, the structural formula of the R substituent is shown in Table 1:

[0067] Table 1

[0068]

[0069]

[0070] The structures of betulinic acid derivatives Z1 to Z26 were characterized by NMR and melting point. The properties and NMR characterization results of each compound are shown below:

[0071]

[0072] (Z1).Yield 87.8%mp:215.7.-216.3℃; 1 H NMR(500MHz,Chloroform-d)δ7.92(s,1H),7.84(t,J=1.9Hz,1H),7.72(dt,J=7.6,1.5Hz,1H),7.36(t,J=7.8Hz,1H),7.31(dt, J=8.1,1.6Hz,1H),5.21(d,J=2.3Hz,2H),4.73(d,J=2.3Hz,1H),4.62–4.56(m,2H),2.99(td,J=10.6,4.8Hz,1H),2.26(dt,J=13 .0,3.3Hz,1H),2.18(td,J=12.3,3.6Hz,1H),1.96(q,J=7.5Hz,2H),1.68(s,8H),1.61(d,J=11.3Hz,2H),1.52–1.45(m,3H),1. 44–1.33(m,7H),1.31–1.23(m,3H),1.17(dt,J=13.4,3.2Hz,1H),0.96(s,3H),0.92(s,3H),0.83(d,J=3.3Hz,6H),0.75(s,3H). 13 C NMR(126MHz,Chloroform-d)δ181.90,166.03,150.52,147.07,134.96,132.22,13 0.30,128.43,126.03,124.02,121.51,109.88,84.18,56.48,55.43,51.38,50.47 ,49.33,47.04,42.55,40.79,38.47,38.37,38.02,37.19,37.17,34.28,32.26,30.67,29.79,28.15,25.51,23.73,20.98,19.47,18.22,16.48,16.26,16.12,14.79.

[0073]

[0074] (Z2).Yield 38.9%mp:234.3-234.4℃; 1H NMR(500MHz,Chloroform-d)δ7.87(s,1H),7.78–7.75(m,2H),7.47–7.44(m,2H),5.20(d,J=2.9Hz,2H),4.73(d,J=2.3Hz,1H),4.62–4.56(m,2H),3.00(td,J=10.8,4.9Hz,1H),2.27(dt,J=13.0,3.4Hz,1H),2.18(td,J=12.3,3.6Hz,1H),1.98(dq,J=11.3,7.4Hz,2H),1.72–1.66(m,7H),1.59(d,J=11.4Hz,1H),1.53–1.45(m,3H),1.42(ddd,J=11.9,5.6,3.3Hz,3H),1.35(s,7H),1.30–1.23(m,3H),0.96(s,3H),0.92(s,3H),0.83(d,J=3.4Hz,6H),0.74(s,3H). 13 C NMR(126MHz,Chloroform-d)δ181.90,166.16,151.55,150.50,148.34,127.62,125.92,125.72,120.81。

[0075]

[0076] (Z3).Yield 88.8%mp:213.7-214.6℃; 1H NMR(500MHz,Chloroform-d)δ8.36(s,1H),8.11(dd,J=7.8,1.8Hz,1H),7.66(dd,J=8.1,1.2Hz,1H),7.42(td,J=7.5,1.2Hz,1H),7.21(td,J=7.7,1.8Hz,1H),5.24(d,J=2.0Hz,2H),4.73(d,J=2.3Hz,1H),4.61–4.56(m,2H),2.99(td,J=10.7,4.8Hz,1H),2.26(dt,J=12.9,3.3Hz,1H),2.20–2.15(m,1H),2.01–1.94(m,2H),1.73–1.66(m,7H),1.60(t,J=11.3Hz,2H),1.52–1.46(m,3H),1.45–1.31(m,7H),1.30–1.22(m,3H),1.19–1.14(m,1H),0.96(s,3H),0.91(s,3H),0.83(d,J=5.4Hz,6H),0.73(s,3H). 13 C NMR(126MHz,Chloroform-d)δ181.90,166.04,150.49,145.87,133.68,131.16,130.83,129.62,127.87,124.52,121.38,109.89,84.10,56.48,55.44,51.44,50.47,49.33,47.04,42.55,40.79,38.48,38.37,38.01,37.19,37.16,34.28,32.25,30.66,29.78,28.17,25.51,23.72,20.97,19.46,18.21,16.51,16.26,16.11,14.78。

[0077]

[0078] (Z4).Yield 78.9%mp:241.01-242.0℃; 1H NMR(500MHz,Chloroform-d)δ8.00(t,J=1.8Hz,1H),7.92(s,1H),7.77(dt,J=7.7,1.4Hz,1H),7.49–7.46(m,1H),7.30(t,J=7.9Hz,1H),5.21(d,J=2.2Hz,2H),4.73(d,J=2.3Hz,1H),4.61–4.56(m,2H),3.03–2.97(m,1H),2.26(dt,J=12.9,3.3Hz,1H),2.18(td,J=12.3,3.6Hz,1H),1.99–1.93(m,2H),1.68(s,7H),1.61(d,J=11.4Hz,3H),1.52–1.46(m,3H),1.42–1.35(m,6H),1.32–1.23(m,3H),1.18(dd,J=13.6,3.3Hz,1H),0.96(s,3H),0.92(s,3H),0.83(d,J=3.1Hz,6H),0.75(s,3H). 13 C NMR(126MHz,Chloroform-d)δ181.92,166.02,150.49,146.93,132.44,131.37,130.57,128.93,124.50,123.11,121.52,109.89,84.19,56.48,55.44,51.41,50.47,49.33,47.04,42.56,40.79,38.47,38.37,38.03,37.20,37.17,34.28,32.25,30.66,29.79,28.16,25.51,23.74,20.98,19.47,18.22,16.49,16.26,16.12,14.79。

[0079]

[0080] (Z5).Yield 69.6%mp:222.1-222.9℃; 1H NMR(500MHz,Chloroform-d)δ7.90(s,1H),7.70(d,J=1.9Hz,1H),7.60(d,J=7.8Hz,1H),7.32(t,J=7.6Hz,1H),7.16(d,J=7.5Hz,1H),5.26–5.16(m,2H),4.73(d,J=2.2Hz,1H),4.62–4.56(m,2H),3.00(td,J=10.7,4.8Hz,1H),2.40(s,3H),2.27(dt,J=12.9,3.3Hz,1H),2.20–2.15(m,1H),2.00–1.93(m,2H),1.73–1.66(m,7H),1.60(t,J=11.3Hz,2H),1.54–1.46(m,3H),1.43–1.35(m,6H),1.30–1.24(m,5H),0.96(s,3H),0.91(s,3H),0.83(d,J=4.2Hz,6H),0.74(s,3H). 13 C NMR(126MHz,Chloroform-d)δ181.91,166.13,150.50,148.42,138.71,130.24,129.25,128.89,126.69,123.09,121.11,109.89,84.07,56.48,55.44,51.40,50.48,49.33,47.04,42.55,40.79,38.47,38.38,38.02,37.19,37.18,34.28,32.25,30.66,29.79,28.15,25.52,23.73,21.58,20.98,19.47,18.22,16.48,16.26,16.11,14.79。

[0081]

[0082] (Z6).Yield 57.2%mp:201.1-202.0℃; 1H NMR(500MHz,Chloroform-d)δ7.89(s,1H),7.73(d,J=7.7Hz,2H),7.24(d,J=7.3Hz,2H),5.20(d,J=2.7Hz,2H),4.73(d,J=2.3Hz,1H),4.61–4.54(m,2H),3.00(td,J=10.8,4.8Hz,1H),2.65(s,1H),2.38(s,3H),2.27(dt,J=12.9,3.3Hz,1H),2.18(td,J=12.3,3.6Hz,1H),1.97(dq,J=12.4,7.6Hz,2H),1.71–1.66(m,6H),1.60(t,J=11.3Hz,1H),1.53–1.45(m,3H),1.44–1.35(m,6H),1.32–1.24(m,5H),1.17(dt,J=13.4,3.2Hz,1H),0.96(s,3H),0.91(s,3H),0.83(d,J=3.6Hz,6H),0.74(s,3H).13C NMR(126MHz,Chloroform-d)δ181.78,166.12,150.52,138.37,129.69,127.55,125.90,109.87,84.06,56.47,55.44,51.44,50.48,49.33,47.04,42.56,40.80,38.47,38.37,38.02,37.19,37.17,34.28,32.26,31.72,30.67,29.79,28.14,25.52,23.72,22.79,21.45,20.98,19.47,18.22,16.47,16.26,16.11,14.79,14.27。

[0083]

[0084] (Z7).Yield 88.6%mp:223.4-224.3℃; 1H NMR(500MHz,Chloroform-d)δ7.92(s,1H),7.84(t,J=1.9Hz,1H),7.72(dt,J=7.5,1.4Hz,1H),7.36(t,J=7.8Hz,1H),7.31(ddd,J=8.0,2.1,1.2Hz,1H),5.21(d,J=2.2Hz,2H),4.73(d,J=2.3Hz,1H),4.61–4.56(m,2H),2.99(td,J=10.8,4.8Hz,1H),2.63(s,1H),2.26(dt,J=12.9,3.3Hz,1H),2.18(td,J=12.3,3.6Hz,1H),1.99–1.93(m,2H),1.68(s,7H),1.58(s,2H),1.52–1.46(m,3H),1.43–1.34(m,7H),1.28–1.25(m,2H),1.23–1.14(m,1H),0.96(s,3H),0.92(s,3H),0.83(d,J=3.2Hz,6H),0.75(s,3H). 13 C NMR(126MHz,Chloroform-d)δ181.99,166.11,163.87,161.90,150.50,147.49,127.77,127.71,126.69,120.93,116.09,115.92,109.88,84.12,56.48,55.43,51.43,50.47,49.33,47.04,42.55,40.79,38.47,38.37,38.02,37.19,34.27,32.25,30.67,29.79,28.16,25.51,23.74,22.79,20.97,19.46,18.22,16.48,16.26,16.12,14.79。

[0085]

[0086] (Z8).Yield 96.4%mp:220.9-221.0℃; 1H NMR(500MHz,Chloroform-d)δ8.31(td,J=7.6,1.9Hz,1H),8.07(d,J=3.5Hz,1H),7.32(tdd,J=7.5,5.2,1.9Hz,1H),7.27(d,J=1.3Hz,1H),7.14(ddd,J=11.1,8.2,1.3Hz,1H),5.27–5.18(m,2H),4.73(d,J=2.2Hz,1H),4.61–4.56(m,2H),2.99(td,J=10.7,4.8Hz,1H),2.27(dt,J=12.9,3.4Hz,1H),2.17(td,J=12.3,3.6Hz,1H),1.98(dq,J=12.2,9.1,7.8Hz,2H),1.72–1.66(m,7H),1.64–1.57(m,2H),1.53–1.45(m,3H),1.40(ddd,J=21.7,12.3,4.2Hz,6H),1.30–1.24(m,4H),1.17(dt,J=13.4,3.2Hz,1H),0.96(s,3H),0.91(s,3H),0.83(d,J=5.3Hz,6H),0.73(s,3H). 13 C NMR(126MHz,Chloroform-d)δ182.03,166.04,160.40,158.43,150.49,141.74,141.72,129.63,129.56,128.05,128.03,124.78,124.75,124.30,124.20,118.53,118.42,115.90,115.73,109.90,84.10,56.49,55.45,51.39,50.48,49.34,47.05,42.56,40.80,38.48,38.38,38.01,37.19,34.29,32.26,30.67,29.79,28.14,25.52,23.71,20.98,19.47,18.22,16.45,16.26,16.11,14.79。

[0087]

[0088] (Z9).Yield 43.7%mp:227.8.1-228.6℃; 1H NMR(500MHz,Chloroform-d)δ8.05(dd,J=7.9,1.5Hz,1H),7.95(s,1H),7.83(dd,J=8.2,1.3Hz,1H),7.67(td,J=7.7,1.3Hz,1H),7.51(td,J=7.8,1.4Hz,1H),5.22(s,2H),4.73(d,J=2.3Hz,1H),4.61–4.55(m,2H),2.99(td,J=10.7,4.8Hz,1H),2.26(dt,J=12.8,3.3Hz,1H),2.17(td,J=12.3,3.6Hz,1H),1.97(p,J=8.9,7.9Hz,2H),1.72–1.64(m,7H),1.60(t,J=11.4Hz,1H),1.54–1.45(m,3H),1.43–1.31(m,7H),1.30–1.22(m,4H),1.17(dt,J=13.4,3.3Hz,1H),0.96(s,3H),0.91(s,3H),0.83(s,6H),0.72(s,3H). 13 C NMR(126MHz,Chloroform-d)δ182.09,165.84,150.49,148.41,142.50,132.71,131.34,129.23,124.63,124.57,124.19,109.89,84.22,56.49,55.44,51.41,50.47,49.33,47.05,42.55,40.79,38.48,38.37,38.00,37.19,37.17,34.28,32.25,30.66,29.79,28.15,25.51,23.69,20.97,19.46,18.21,16.43,16.24,16.11,14.79 . 。

[0089]

[0090] (Z10).Yield 77.5%mp:245.1-245.5℃;25-2F 1H NMR(500MHz,Chloroform-d)δ7.93(s,1H),7.37(h,J=4.9Hz,2H),6.78(tt,J=8.9,2.3Hz,1H),5.22(d,J=2.0Hz,2H),4.73(d,J=2.3Hz,1H),4.62–4.57(m,2H),2.99(td,J=10.7,4.7Hz,1H),2.26(dt,J=13.0,3.4Hz,1H),2.19(dt,J=12.2,6.1Hz,1H),1.97(q,J=8.0Hz,2H),1.68(s,7H),1.60(t,J=11.3Hz,1H),1.54–1.45(m,3H),1.44–1.32(m,7H),1.26(dd,J=13.7,9.6Hz,4H),1.18–1.14(m,1H),0.96(s,3H),0.92(s,3H),0.83(d,J=3.2Hz,6H),0.75(s,3H). 13 C NMR(126MHz,Chloroform-d)δ165.94,164.58,164.47,162.60,162.50,150.51,146.42,133.59,121.85,109.89,108.91,108.86,108.75,108.69,103.86,103.66,103.46,84.26,56.52,55.43,51.38,50.48,49.34,47.06,42.57,40.80,38.48,38.37,38.03,37.20,34.28,32.29,30.69,29.84,29.80,28.16,25.52,23.74,21.00,19.47,18.23,16.48,16.27,16.15,14.79。

[0091]

[0092] (Z11).Yield 58.6%mp:235.2-236.4℃; 1H NMR(500MHz,DMSO-d6)δ12.07(s,1H),8.39(s,1H),7.11–7.04(m,2H),6.92(d,J=7.5Hz,1H),6.52(dd,J=8.1,2.3Hz,1H),5.48(d,J=17.5Hz,1H),5.36(d,J=17.5Hz,1H),5.19(s,2H),4.68(d,J=2.5Hz,1H),4.55(s,1H),4.46(t,J=8.1Hz,1H),2.98–2.90(m,1H),2.21(t,J=12.0Hz,1H),2.11(d,J=9.3Hz,1H),1.83–1.74(m,2H),1.64(s,4H),1.61–1.56(m,3H),1.51(t,J=10.9Hz,1H),1.45–1.24(m,12H),1.21–1.12(m,1H),1.08(d,J=9.5Hz,1H),0.97–0.95(m,1H),0.93(s,3H),0.86(s,3H),0.79(d,J=5.4Hz,6H),0.67(s,3H). 13 C NMR(126MHz,DMSO-d6)δ177.26,166.90,150.33,149.09,147.02,131.03,129.40,122.34,113.63,112.98,110.47,109.69,82.07,55.42,54.41,50.65,49.58,48.51,46.64,42.04,40.24,37.60,37.50,36.59,36.34,33.68,31.69,30.09,29.22,27.55,25.03,23.22,20.47,18.96,17.67,16.13,15.86,15.69,14.37。

[0093]

[0094] (Z12).Yield 50.4%mp:213.2-213.6℃; 1H NMR(500MHz,Chloroform-d)δ7.54(s,1H),5.14(d,J=2.6Hz,2H),4.73(d,J=2.3Hz,1H),4.60(t,J=2.0Hz,1H),4.54(dd,J=11.4,4.8Hz,1H),3.95(t,J=5.9Hz,2H),2.99(t,J=5.9Hz,2H),2.27(dt,J=12.9,3.3Hz,1H),2.17(td,J=12.2,3.6Hz,1H),1.96(q,J=8.4Hz,2H),1.72–1.64(m,7H),1.60(t,J=11.4Hz,2H),1.53–1.44(m,3H),1.43–1.31(m,7H),1.29–1.21(m,3H),1.17(dt,J=13.5,3.3Hz,1H),1.02(ddd,J=19.4,14.0,6.1Hz,1H),0.96(s,3H),0.91(s,3H),0.81(d,J=7.8Hz,6H),0.71(s,3H). 13 C NMR(126MHz,Chloroform-d)δ181.81,166.17,150.54,145.98,123.20,109.85,84.00,61.71,56.48,55.40,51.29,50.44,49.34,47.05,42.54,40.77,38.46,38.33,37.98,37.18,37.16,34.25,32.27,30.67,29.80,28.75,28.11,25.49,23.68,20.96,19.47,18.22,16.41,16.26,16.16,14.78 . 。

[0095]

[0096] (Z13).Yield 85.6%mp:248.9-249.9℃; 1H NMR(500MHz,Chloroform-d)δ7.57(d,J=1.7Hz,1H),5.27–5.00(m,4H),4.73(d,J=2.3Hz,1H),4.59(d,J=1.9Hz,1H),4.51(dd,J=11.7,4.6Hz,1H),3.06–2.98(m,1H),2.26(ddd,J=32.8,10.6,3.6Hz,2H),1.98(qd,J=10.2,8.1,5.9Hz,2H),1.71–1.62(m,7H),1.63–1.51(m,7H),1.46(qd,J=9.6,9.0,5.3Hz,1H),1.37(dd,J=13.0,3.3Hz,2H),1.33(d,J=4.6Hz,4H),1.29–1.22(m,4H),1.19–1.11(m,1H),1.08–0.99(m,1H),0.95–0.94(m,3H),0.89–0.86(m,3H),0.77(dd,J=12.6,3.7Hz,6H),0.55(d,J=12.5Hz,3H). 13 C NMR(126MHz,Chloroform-d)δ181.32,165.72,152.91,150.72,121.78,109.75,84.01,62.45,62.18,56.50,55.26,51.65,50.28,49.35,46.97,42.47,40.72,38.23,37.91,37.16,34.09,32.35,31.57,30.69,30.31,29.78,28.03,25.42,23.67,23.01,22.66,20.91,19.52,18.42,16.36,16.22,14.72 . 。

[0097]

[0098] (Z14).Yield 79.9%mp:210.4.1-210.7℃; 1H NMR(500MHz,Chloroform-d)δ8.32(d,J=6.4Hz,1H),8.24(dd,J=7.8,1.8Hz,1H),7.45(dd,J=8.1,1.3Hz,1H),7.38(td,J=7.6,1.4Hz,1H),7.29(td,J=7.7,1.7Hz,1H),5.28–5.21(m,2H),4.73(d,J=2.3Hz,1H),4.60(q,J=1.7Hz,1H),4.59–4.55(m,1H),2.99(td,J=10.6,4.7Hz,1H),2.26(dt,J=13.1,3.4Hz,1H),2.17(td,J=12.3,3.6Hz,1H),2.03–1.92(m,2H),1.75–1.65(m,7H),1.60(t,J=11.3Hz,2H),1.55–1.44(m,4H),1.44–1.34(m,7H),1.32–1.23(m,5H),1.17(dt,J=13.4,3.3Hz,1H),0.96(s,3H),0.91(s,3H),0.83(d,J=5.4Hz,6H),0.72(s,3H). 13 C NMR(126MHz,Chloroform-d)δ181.93,166.05,150.50,144.53,131.42,130.33,130.06,129.30,129.15,127.33,124.73,109.89,84.10,56.48,55.44,51.44,50.47,49.33,47.04,42.55,40.79,38.47,38.37,38.00,37.19,34.28,32.25,30.66,29.79,28.15,25.51,23.71,20.97,19.47,18.21,16.48,16.26,16.11,14.79,14.27。

[0099]

[0100] (Z15).Yield 90%mp:205.2-206.6℃; 1H NMR(500MHz,Chloroform-d)δ7.90(s,1H),7.80–7.75(m,2H),7.42–7.38(m,2H),5.21(d,J=2.7Hz,2H),4.73(d,J=2.2Hz,1H),4.60(d,J=3.1Hz,1H),4.59–4.55(m,1H),3.00(td,J=10.7,4.8Hz,1H),2.27(dt,J=13.0,3.5Hz,1H),2.18(td,J=12.3,3.6Hz,1H),2.02–1.93(m,2H),1.68(s,7H),1.60(t,J=11.4Hz,2H),1.53–1.44(m,3H),1.39(dtd,J=16.3,13.1,11.3,4.2Hz,7H),1.31–1.24(m,3H),1.17(dt,J=13.4,3.3Hz,1H),0.96(s,3H),0.92(s,3H),0.83(d,J=2.9Hz,6H),0.75(s,3H). 13 CNMR(126MHz,Chloroform-d)δ181.88,166.07,150.50,147.32,134.20,129.22,128.99,127.21,121.19,109.89,84.17,56.48,55.44,51.37,50.48,49.34,47.05,42.56,40.80,38.48,38.37,38.03,37.20,37.17,34.28,32.25,30.67,29.79,28.15,25.51,23.74,20.98,19.47,18.22,16.48,16.26,16.13,14.79。

[0101]

[0102] (Z16).Yield 78.5%mp:253.6-254.4℃; 1H NMR(500MHz,Chloroform-d)δ7.79(s,1H),7.75(dt,J=5.8,2.9Hz,1H),7.28(d,J=3.2Hz,3H),5.23(d,J=2.3Hz,2H),4.73(d,J=2.2Hz,1H),4.63–4.56(m,2H),2.99(td,J=10.7,4.9Hz,1H),2.47(s,3H),2.26(dt,J=13.1,3.3Hz,1H),2.18(td,J=12.2,3.6Hz,1H),1.97(dd,J=10.1,5.6Hz,2H),1.71(s,7H),1.66–1.57(m,3H),1.49(t,J=12.2Hz,3H),1.45–1.33(m,7H),1.27(d,J=9.5Hz,2H),1.18(dd,J=13.9,3.5Hz,1H),0.96(s,3H),0.92(s,3H),0.83(d,J=2.8Hz,6H),0.73(s,3H). 13 C NMR(126MHz,Chloroform-d)δ181.34,168.09,162.06,151.35,150.64,144.78,139.66,132.86,131.57,130.03,128.37,126.77,109.81,83.36,56.47,55.51,50.49,49.36,47.04,42.55,40.80,38.44,38.03,37.20,35.87,34.31,32.31,31.72,30.70,29.82,28.01,25.55,23.62,22.79,20.98,20.02,19.48,18.27,16.50,16.28,16.12,14.78,14.27。

[0103]

[0104] (Z17).Yield 40%mp:227.8-228.4℃; 1H NMR(500MHz,Chloroform-d)δ8.05(dd,J=7.9,1.5Hz,1H),7.95(s,1H),7.83(dd,J=8.2,1.3Hz,1H),7.67(td,J=7.7,1.3Hz,1H),7.51(td,J=7.8,1.4Hz,1H),5.22(s,2H),4.73(d,J=2.3Hz,1H),4.61–4.55(m,2H),2.99(td,J=10.7,4.8Hz,1H),2.26(dt,J=12.8,3.3Hz,1H),2.17(td,J=12.3,3.6Hz,1H),1.97(p,J=8.9,7.9Hz,2H),1.72–1.64(m,7H),1.60(t,J=11.4Hz,1H),1.54–1.45(m,3H),1.43–1.31(m,7H),1.30–1.22(m,4H),1.17(dt,J=13.4,3.3Hz,1H),0.96(s,3H),0.91(s,3H),0.83(s,6H),0.72(s,3H). 13 C NMR(126MHz,Chloroform-d)δ182.09,165.84,150.49,148.41,142.50,132.71,131.34,129.23,124.63,124.57,124.19,109.89,84.22,56.49,55.44,51.41,50.47,49.33,47.05,42.55,40.79,38.48,38.37,38.00,37.19,37.17,34.28,32.25,30.66,29.79,28.15,25.51,23.69,20.97,19.46,18.21,16.43,16.24,16.11,14.79。

[0105]

[0106] (Z18).Yield 79.8%mp:228.1-228.9℃; 1H NMR(500MHz,Chloroform-d)δ8.63(t,J=1.9Hz,1H),8.25(dt,J=7.8,1.4Hz,1H),8.21–8.18(m,1H),8.08(s,1H),7.62(t,J=8.0Hz,1H),5.25(d,J=2.0Hz,2H),4.73(d,J=2.2Hz,1H),4.62–4.57(m,2H),2.99(td,J=10.7,4.7Hz,1H),2.27(dt,J=13.0,3.4Hz,1H),2.17(td,J=12.3,3.6Hz,1H),2.01–1.93(m,2H),1.69(d,J=4.3Hz,7H),1.61(d,J=11.3Hz,1H),1.49(td,J=12.1,10.0,3.3Hz,3H),1.44–1.32(m,7H),1.30–1.23(m,3H),1.17(dt,J=13.4,3.2Hz,1H),1.04(td,J=12.9,4.9Hz,1H),0.96(s,3H),0.91(s,3H),0.84(d,J=7.2Hz,6H),0.76(s,3H). 13 C NMR(126MHz,Chloroform-d)δ182.17,165.95,150.48,148.80,146.19,132.27,131.70,130.12,123.04,122.08,120.72,109.90,84.30,56.49,55.43,51.42,50.46,49.33,47.05,42.55,40.79,38.48,38.36,38.04,37.20,37.16,34.26,32.25,30.66,29.79,28.16,25.50,23.75,20.97,19.46,18.21,16.48,16.26,16.14,14.79。

[0107]

[0108] (Z19).Yield 73.9%mp:234.3-234.4℃; 1H NMR(500MHz,Chloroform-d)δ8.60(dt,J=4.9,1.4Hz,1H),8.28(s,1H),8.19(dd,J=7.9,1.1Hz,1H),7.79(td,J=7.7,1.8Hz,1H),7.26–7.22(m,1H),5.21(s,2H),4.73(d,J=2.3Hz,1H),4.61–4.56(m,2H),3.00(td,J=10.8,4.9Hz,1H),2.27(dt,J=12.9,3.3Hz,1H),2.21–2.15(m,1H),2.00–1.93(m,2H),1.69(d,J=17.4Hz,7H),1.60(t,J=11.3Hz,2H),1.53–1.46(m,3H),1.44–1.32(m,7H),1.29–1.24(m,2H),1.17(dt,J=13.3,3.3Hz,1H),1.04(ddt,J=20.4,12.3,6.7Hz,1H),0.96(s,3H),0.91(s,3H),0.82(d,J=6.5Hz,6H),0.72(s,3H). 13 C NMR(126MHz,Chloroform-d)δ181.47,165.90,150.54,149.37,148.61,137.31,123.74,123.20,120.61,109.88,84.05,56.47,55.43,51.44,50.48,49.34,47.06,42.56,40.80,38.46,38.38,38.02,37.19,34.29,32.29,31.57,30.69,30.32,29.80,28.14,25.53,23.71,20.99,19.47,18.23,16.46,16.26,16.11,14.79。

[0109]

[0110] (Z20).Yield 89.7%mp:215.2-216.0℃; 1H NMR(500MHz,Chloroform-d)δ7.91(s,1H),7.86–7.82(m,2H),7.43(dd,J=8.4,7.0Hz,2H),7.37–7.33(m,1H),5.21(d,J=3.1Hz,2H),4.73(d,J=2.3Hz,1H),4.60(q,J=1.9Hz,1H),4.59–4.55(m,1H),3.00(td,J=10.7,4.8Hz,1H),2.27(dt,J=13.0,3.4Hz,1H),2.18(td,J=12.3,3.6Hz,1H),1.97(q,J=8.0Hz,2H),1.73–1.67(m,7H),1.61(t,J=11.4Hz,2H),1.50(td,J=11.7,9.9,5.9Hz,3H),1.44–1.35(m,6H),1.30–1.23(m,4H),1.17(dt,J=13.4,3.2Hz,1H),0.96(s,3H),0.92(s,3H),0.83(d,J=4.2Hz,6H),0.74(s,3H). 13 C NMR(126MHz,Chloroform-d)δ182.01,166.13,150.49,148.36,130.45,128.99,128.44,125.98,121.10,109.89,84.07,56.49,55.44,51.37,50.47,49.33,47.04,42.55,40.79,38.48,38.37,38.02,37.19,37.17,34.28,32.25,31.72,30.66,29.79,28.14,25.51,23.72,22.79,20.97,19.46,18.21,16.47,16.26,16.11,14.79,14.26。

[0111]

[0112] (Z21).Yield 55.9%mp:258.1-259.0℃; 1H NMR(500MHz,Chloroform-d)δ9.04–9.00(m,1H),8.60(d,J=5.5Hz,1H),8.28(dt,J=8.0,1.9Hz,1H),8.04(s,1H),7.42(dd,J=8.0,4.9Hz,1H),5.24(d,J=2.8Hz,2H),4.73(d,J=2.2Hz,1H),4.61–4.56(m,2H),3.01(ddd,J=14.5,9.6,4.2Hz,1H),2.31–2.18(m,2H),1.97(q,J=8.1Hz,2H),1.68(s,7H),1.64–1.57(m,2H),1.52–1.45(m,3H),1.44–1.35(m,7H),1.33(s,2H),1.31–1.22(m,7H),1.17(dt,J=13.5,3.3Hz,1H),1.09–0.97(m,2H),0.96(s,3H),0.92(s,3H),0.83(d,J=6.0Hz,6H),0.74(s,3H). 13 C NMR(126MHz,Chloroform-d)δ181.12,165.93,150.64,148.83,146.59,145.03,133.82,126.99,124.20,121.64,119.21,109.81,84.28,56.46,55.44,51.43,50.50,49.34,47.08,42.57,40.80,38.45,38.37,38.02,37.23,37.19,34.28,32.35,31.57,30.71,30.31,29.82,28.15,25.54,23.74,22.79,21.00,19.48,18.23,16.48,16.27,16.14,14.79,14.26。

[0113]

[0114] (Z22).Yield 87.6%mp:249.2.-250.2℃; 1H NMR(500MHz,Chloroform-d)δ8.00(s,1H),7.96(d,J=8.1Hz,2H),7.69(d,J=8.1Hz,2H),5.23(d,J=2.5Hz,2H),4.73(d,J=2.3Hz,1H),4.61–4.57(m,2H),2.99(td,J=10.8,4.9Hz,1H),2.28–2.24(m,1H),2.21–2.14(m,1H),2.02–1.93(m,2H),1.71(d,J=3.3Hz,7H),1.60(t,J=11.4Hz,2H),1.49(td,J=9.8,4.8Hz,3H),1.44–1.33(m,7H),1.31–1.22(m,3H),1.18(dt,J=13.3,3.2Hz,1H),0.96(s,3H),0.92(s,3H),0.84(d,J=4.7Hz,6H),0.75(s,3H). 13 C NMR(126MHz,Chloroform-d)δ181.73,166.00,150.50,146.97,133.86,130.43,130.17,126.12,126.05,126.02,121.96,109.89,84.27,56.47,55.44,51.46,50.49,49.33,47.04,42.56,40.80,38.47,38.37,38.03,37.20,37.17,34.28,32.25,30.66,29.79,28.16,25.51,23.75,20.98,19.46,18.22,16.49,16.26,16.11,14.79,14.05。

[0115]

[0116] (Z23).Yield 86.6%mp:210.9-212.1℃; 1H NMR(500MHz,Chloroform-d)δ7.83(s,1H),7.40(dd,J=3.6,1.1Hz,1H),7.31(dd,J=5.1,1.2Hz,1H),7.08(dd,J=5.1,3.5Hz,1H),5.19(d,J=3.4Hz,2H),4.73(d,J=2.2Hz,1H),4.62–4.55(m,2H),3.00(td,J=10.7,4.8Hz,1H),2.27(dt,J=13.1,3.4Hz,1H),2.17(td,J=12.3,3.6Hz,1H),2.03–1.93(m,2H),1.72–1.65(m,7H),1.62(d,J=11.4Hz,1H),1.53–1.44(m,3H),1.42–1.35(m,6H),1.30–1.22(m,5H),1.17(dt,J=13.4,3.2Hz,1H),0.96(s,3H),0.92(s,3H),0.83(d,J=2.4Hz,6H),0.75(s,3H).13C NMR(126MHz,Chloroform-d)δ182.01,166.00,150.50,143.40,132.70,127.78,125.37,124.55,120.61,109.89,84.15,56.48,55.43,51.34,50.46,49.33,47.04,42.55,40.79,38.47,38.37,38.02,37.19,34.27,32.25,30.66,29.79,28.15,25.51,23.72,22.79,20.97,19.46,18.21,16.49,16.26,16.12,14.79,14.27。

[0117]

[0118] (Z24).Yield 59.8%mp:248.9.1-249.9℃; 1H NMR(500MHz,Chloroform-d)δ7.62(s,1H),5.23–5.08(m,2H),4.82(d,J=13.4Hz,1H),4.76(s,1H),4.73(d,J=2.1Hz,1H),4.61–4.58(m,1H),4.51(dd,J=11.7,4.6Hz,1H),3.02(td,J=10.7,4.9Hz,1H),2.31–2.19(m,2H),2.01–1.95(m,2H),1.64(d,J=3.8Hz,7H),1.57(t,J=11.3Hz,3H),1.42(dt,J=25.7,12.0Hz,7H),1.33(d,J=3.5Hz,2H),1.26(d,J=6.1Hz,3H),1.17–1.13(m,1H),0.95(s,3H),0.89(s,3H),0.78(s,3H),0.75(s,3H),0.53(s,3H). 13 C NMR(126MHz,Chloroform-d)δ165.65,150.71,123.37,109.72,84.00,60.55,56.53,55.64,55.24,51.59,50.24,49.37,46.98,42.45,40.89,40.70,38.18,37.89,37.16,37.09,34.06,32.40,30.70,29.82,29.77,28.00,25.40,23.66,20.91,19.51,18.39,16.32,16.28,16.20,14.70。

[0119]

[0120] (Z25).Yield 14.5%mp:238.1-239.9℃; 1H NMR(500MHz,Chloroform-d)δ8.25(s,1H),5.22(s,2H),4.73(d,J=2.3Hz,1H),4.60(dd,J=3.6,1.9Hz,1H),4.59–4.55(m,1H),3.96(s,3H),2.99(td,J=10.7,4.7Hz,1H),2.26(dt,J=12.9,3.3Hz,1H),2.17(td,J=12.3,3.6Hz,1H),1.96(q,J=8.3Hz,2H),1.72–1.63(m,7H),1.60(s,1H),1.49(td,J=12.9,12.0,3.0Hz,3H),1.44–1.33(m,7H),1.31–1.22(m,3H),1.17(dt,J=13.4,3.2Hz,1H),1.06–0.97(m,1H),0.96(s,3H),0.92(s,3H),0.82(d,J=3.7Hz,6H),0.73(s,3H). 13 CNMR(126MHz,Chloroform-d)δ182.02,166.05,150.49,147.27,134.26,129.24,128.89,127.24,121.25,109.90,84.19,56.49,55.44,51.41,50.47,49.34,47.05,42.56,40.79,38.48,38.37,38.03,37.20,34.28,32.25,31.73,30.67,29.79,28.16,25.51,23.74,22.79,20.98,19.47,18.22,16.48,16.26,16.13,14.79,14.27。

[0121]

[0122] (Z26).Yield 87.5%mp:268.0-268.1℃; 1H NMR(500MHz,Chloroform-d)δ7.68(s,1H),5.19–5.12(m,2H),4.89(d,J=12.5Hz,1H),4.76–4.74(m,1H),4.73(d,J=2.3Hz,1H),4.69(d,J=12.5Hz,1H ),4.61–4.59(m,1H),4.55(dd,J=11.4,4.8Hz,1H),3.90(ddd,J=11.5,8.4, 2.9Hz,1H),3.59–3.52(m,1H),3.03–2.96(m,1H),2.27(dt,J=13.0,3.4Hz, 1H),2.18(td,J=12.2,3.6Hz,1H),1.97(dq,J=11.7,9.0,7.8Hz,2H),1.85 –1.79(m,1H),1.72–1.65(m,7H),1.63–1.57(m,4H),1.56–1.45(m,6H),1.3 8(dtd,J=15.8,13.1,11.2,4.6Hz,7H),1.31–1.22(m,3H),1.17(dt,J=13.5 ,3.3Hz,1H),0.96(s,3H),0.92(s,3H),0.82(d,J=5.2Hz,6H),0.72(s,3H). 13 C NMR(126MHz,Chloroform-d)δ181.68,166.12,150.51,145.81,124.07,109. 88,98.18,83.99,62.35,60.53,56.47,55.43,51.25,50.47,49.33,47.03,42 .55,40.79,38.46,38.36,37.99,37.19,34.28,32.25,30.66,30.56,29.79,28.12,25.50,23.70,20.97,19.47,19.40,18.22,16.45,16.26,16.12,14.78.

[0123] Example 2: α-glucosidase inhibitory activity test of betulinic acid derivatives

[0124] 1. Preparation of reagents and standard solutions

[0125] (1) 100 mM phosphate buffer (PBS, pH 6.8): Weigh a certain amount of potassium dihydrogen phosphate and disodium hydrogen phosphate and dissolve them in ultrapure water to dissolve the dilution reagent.

[0126] (2) Preparation of α-glucosidase solution: Add an appropriate amount of 100 mM PBS to an enzyme with an activity of 100 U to a working concentration of 0.05 U / mL, and freeze in aliquots.

[0127] (3) Substrate preparation: Accurately weigh an appropriate amount of 4-nitrophenyl-D-pyranoglucoside (PNPG), dissolve it in 100 mM PBS solution, and prepare a substrate working solution with a concentration of 0.25 mM. Vortex mix thoroughly and prepare it fresh before each experiment.

[0128] Preparation of test drugs: Accurately weigh an appropriate amount of the drug to be tested, dissolve it in DMSO to prepare a 10 mM stock solution, and store it at -20°C in the dark. Before the experiment, dilute it with DMSO to the desired concentration (0-200 μM), with a DMSO content of 5%.

[0129] 2. Experimental steps

[0130] (1) 10 μL of α-glucosidase with a working concentration of 0.05 U / mL, 130 μL of 100 mM phosphate buffer (pH 6.8), and 10 μL of compounds of different concentrations (betulinic acid derivatives 1 to 13 prepared in Example 1) were added to a 96-well plate in sequence. The blank control group was replaced with 10 μL of 5% DMSO to replace 10 μL of the compound. Acarbose was used as a positive control. Four replicate wells were set up in parallel for each group. The enzyme reaction system was placed on a microplate reader and incubated at 37°C for 10 min.

[0131] (2) Subsequently, 50 μL of substrate PNPG was added to the enzyme reaction system to initiate the enzyme reaction. The microplate was placed on a microplate reader and incubated at 37°C for 10 min. During the incubation process, the time was evenly distributed three times. The readings were read once at a wavelength of 405 nm during each time period. The readings were recorded as OD1, OD2, and OD3.

[0132] (3) The α-glucosidase inhibitory activity of the test compound was calculated according to the following formula:

[0133] Inhibition rate (%) = [(OD3-OD)-(OD1-OD)] / OD3-OD×100%

[0134] Where OD represents the absorbance value of the blank control group. Data processing: MS Excel was used to analyze and process the data, and Origin 9.1 was used to calculate the half-maximal inhibitory concentration (IC 50 ), IC 50 It represents the concentration of the test compound required to inhibit the activity of α-glucosidase by 50% under the experimental conditions.

[0135] 3. Results Analysis

[0136] The α-glucosidase inhibitory activity of the synthesized compounds was evaluated by in vitro enzymatic assay, and the results are shown in Table 2:

[0137] Table 2. Evaluation of the in vitro inhibitory activity of the screened compounds against α-glucosidase

[0138]

[0139]

[0140] The IC value of the positive control drug acarbose 50 The derivative Z13 was screened out and showed the strongest α-glucosidase inhibitory activity, IC 50 The value is 2.83±0.09μM, which is 216 times that of acarbose. Figure 1 The results showed that this small molecule compound exhibited strong binding affinity when interacting with α-glucosidase. Therefore, it can be inferred that the betulinic acid skeleton structure plays a very important role in the good α-glucosidase inhibitory activity of this type of compound. Betulinic acid derivatives can be used as α-glucosidase inhibitors to treat or prevent diabetes.

[0141] Example 3: Enzyme kinetics experiment

[0142] The α-glucosidase inhibitory activity of the synthesized active compounds was evaluated using in vitro enzyme kinetics experiments:

[0143] 1. Preparation of reagents and standard solutions

[0144] (1) 100 mM phosphate buffer (PBS, pH 6.8): Weigh a certain amount of potassium dihydrogen phosphate and disodium hydrogen phosphate and dissolve them in ultrapure water to dissolve the dilution reagent.

[0145] (2) Preparation of α-glucosidase solution: Add an appropriate amount of 100 mM PBS to an enzyme with an activity of 100 U to prepare working concentrations of 0.0375 U / mL, 0.05 U / mL, 0.0625 U / mL, and 0.075 U / mL, respectively, and freeze in aliquots.

[0146] (3) Substrate preparation: Accurately weigh an appropriate amount of 4-nitrophenyl-D-pyranoglucoside (PNPG), dissolve it in 100 mM PBS solution, and prepare a substrate working solution with a concentration of 0.25 mM. Vortex mix thoroughly and prepare it fresh before each experiment.

[0147] Preparation of test drugs: Accurately weigh an appropriate amount of the drug to be tested, dissolve it in DMSO to prepare a 10 mM stock solution, and store it at -20°C in the dark. Before the experiment, dilute it with DMSO to the desired concentration (0-200 μM), with a DMSO content of 5%.

[0148] 2. Experimental steps

[0149] (1) 10 μL of α-glucosidase with concentrations of 0.0375 U / mL, 0.05 U / mL, 0.0625 U / mL, and 0.075 U / mL, 130 μL of 100 mM phosphate buffer (pH 6.8), and 10 μL of compounds of different concentrations (the betulinic acid compound 13 prepared in Example 1) were added to a 96-well plate in sequence. The blank control group was replaced with 10 μL of DMSO with a content equal to 5% to replace 10 μL of the compound. Acarbose was used as a positive control. Four replicate wells were set up in parallel for each group. The enzyme reaction system was placed on a microplate reader and incubated at 37°C for 10 min.

[0150] (2) Subsequently, 50 μL of 0.25 mM PNPG was added to the enzyme reaction system to initiate the enzyme reaction. The microplate was placed on a microplate reader and incubated at 37°C for 15 min. During the incubation process, the sample was read three times at a wavelength of 405 nm. The readings were recorded as OD1, OD2, and OD3.

[0151] (3) Data processing: MS Excel was used to analyze and process the data. The reaction rate of the enzyme reaction system was ΔOD / min.

[0152] 3. Results Analysis

[0153] The results of enzyme kinetic inhibition type evaluation test are as follows Figure 2 As shown. Figure 2 It can be seen that the inhibitor binds to the enzyme through non-covalent bonds to inhibit the activity of the enzyme, which is a reversible inhibition.

[0154] Example 4: Substrate kinetics experiment

[0155] In vitro substrate kinetics experiments were used to evaluate the α-glucosidase inhibitory activity of the synthesized active compounds:

[0156] 1. Preparation of reagents and standard solutions

[0157] (1) 100 mM phosphate buffer (PBS, pH 6.8): Weigh a certain amount of potassium dihydrogen phosphate and disodium hydrogen phosphate and dissolve them in ultrapure water to dissolve the dilution reagent.

[0158] (2) Preparation of α-glucosidase solution: Add an appropriate amount of 100 mM PBS to an enzyme with an activity of 100 U to a working concentration of 0.05 U / mL, and freeze in aliquots.

[0159] (3) Substrate preparation: Accurately weigh an appropriate amount of 4-nitrophenyl-D-pyranoglucoside (PNPG), dissolve it in 100 mM PBS solution, and prepare substrate working solutions with concentrations of (0.25 mM, 0.5 mM, 0.75 mM, 1 mM). Vortex mix thoroughly and prepare freshly before each experiment.

[0160] Preparation of test drugs: Accurately weigh an appropriate amount of the drug to be tested, dissolve it in DMSO to prepare a 10 mM stock solution, and store it at -20°C in the dark. Before the experiment, dilute it with DMSO to the desired concentration (0-200 μM), with a DMSO content of 5%.

[0161] 2. Experimental steps

[0162] (1) 10 μL of 0.5 U / mL α-glucosidase, 130 μL of 100 mM phosphate buffer (pH 6.8), and 10 μL of compounds of different concentrations (the betulinic acid compound 13 prepared in Example 1) were added to a 96-well plate in sequence. The blank control group was replaced with 10 μL of 5% DMSO to replace 10 μL of the compound. Acarbose was used as a positive control. Four replicate wells were set up in parallel for each group. The enzyme reaction system was placed on a microplate reader and incubated at 37°C for 10 min.

[0163] (2) Subsequently, 50 μL of substrate PNPG of different concentrations was added to the enzyme reaction system to initiate the enzyme reaction. The microplate was placed on a microplate reader and incubated at 37°C for 10 min. During the incubation process, the time was evenly distributed three times, and the reading was taken once at a wavelength of 405 nm during each time period. The readings were recorded as OD1, OD2, and OD3.

[0164] (3) Data processing: MS Excel was used to analyze and process the data. The reaction rate of the enzyme reaction system was ΔOD / min.

[0165] 3. Results Analysis

[0166] The experimental results are as follows Figure 3 Shown is a mixed inhibitor.

[0167] In summary, the present invention provides a series of betulinic acid derivatives Z1 to Z26, and determines their activity against α-glucosidase. The results show that most of the synthesized derivatives exhibit significant inhibitory activity against α-glucosidase, especially derivative Z13, which exhibits the strongest α-glucosidase inhibitory effect, with IC 50The value is 2.83±0.09μM, which is 216 times that of acarbose, and can be used as an α-glucosidase inhibitor to treat or prevent diabetes.

[0168] The above is a detailed description of the embodiments of the present invention, but the present invention is not limited to the above embodiments. Various changes can be made within the knowledge of ordinary technicians in the relevant technical field without departing from the scope of the present invention.

Claims

1. A betulic acid derivative or a pharmaceutically acceptable salt thereof, characterized in that: The betulinic acid derivatives have a structure shown in formula (I): ； Wherein, the R is selected from the following groups: 。 2. The betulic acid derivative or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The betulic acid derivatives are selected from the following structures: 。 3. The method for preparing betulic acid derivatives according to claim 1 or 2, characterized in that: The steps include: S1. Under anaerobic conditions, betulinic acid, a first organic solvent, chloroacetyl chloride or chloroacetic anhydride, an acid-binding agent, and DMAP are reacted, followed by post-treatment to obtain intermediate 2; S2, under anaerobic conditions, mixing intermediate 2, a second organic solvent and sodium azide to carry out a substitution reaction to obtain intermediate 3; S3, under anaerobic conditions, intermediate 3, , a reducing agent, a copper catalyst and a third organic solvent to react to obtain a betulinic acid derivative represented by formula (I); Among them, the structural formulas of betulinic acid, intermediate 2 and intermediate 3 are as follows: 。 4. The method for preparing betulic acid derivatives according to claim 3, characterized in that: The first organic solvent, the second organic solvent and the third organic solvent are independently selected from at least one of tetrahydrofuran, pyridine, acetone, DMF and DMSO.

5. The method for preparing betulic acid derivatives according to claim 3, characterized in that: In step S1, the acid binding agent is selected from at least one of DIPEA and triethylamine.

6. A pharmaceutical composition, characterized in that The invention comprises the betulic acid derivative or a pharmaceutically acceptable salt thereof according to claim 1 or 2; and a pharmaceutically acceptable excipient.

7. An α-glucosidase inhibitor, characterized in that The invention comprises the betulic acid derivatives or pharmaceutically acceptable salts thereof according to claim 1 or 2.

8. Use of the betulic acid derivatives according to claim 1 or 2 in the preparation of medicaments for preventing and / or treating diabetes.

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