CN114920777B - Jiyuan oridonin phosphorus-containing serial derivatives, preparation method and application thereof - Google Patents

Jiyuan oridonin phosphorus-containing serial derivatives, preparation method and application thereof Download PDF

Info

Publication number
CN114920777B
CN114920777B CN202210538036.XA CN202210538036A CN114920777B CN 114920777 B CN114920777 B CN 114920777B CN 202210538036 A CN202210538036 A CN 202210538036A CN 114920777 B CN114920777 B CN 114920777B
Authority
CN
China
Prior art keywords
compound
phosphorus
jiyuan
cancer
preparation
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN202210538036.XA
Other languages
Chinese (zh)
Other versions
CN114920777A (en
Inventor
可钰
贾小苹
张怡欣
肖泽霖
张建业
徐霞
刘宏民
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Zhengzhou University
Original Assignee
Zhengzhou University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Zhengzhou University filed Critical Zhengzhou University
Priority to CN202210538036.XA priority Critical patent/CN114920777B/en
Publication of CN114920777A publication Critical patent/CN114920777A/en
Application granted granted Critical
Publication of CN114920777B publication Critical patent/CN114920777B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/655—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having oxygen atoms, with or without sulfur, selenium, or tellurium atoms, as the only ring hetero atoms
    • C07F9/6552—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having oxygen atoms, with or without sulfur, selenium, or tellurium atoms, as the only ring hetero atoms the oxygen atom being part of a six-membered ring
    • C07F9/65522—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having oxygen atoms, with or without sulfur, selenium, or tellurium atoms, as the only ring hetero atoms the oxygen atom being part of a six-membered ring condensed with carbocyclic rings or carbocyclic ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/6564—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having phosphorus atoms, with or without nitrogen, oxygen, sulfur, selenium or tellurium atoms, as ring hetero atoms
    • C07F9/6571—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having phosphorus atoms, with or without nitrogen, oxygen, sulfur, selenium or tellurium atoms, as ring hetero atoms having phosphorus and oxygen atoms as the only ring hetero atoms
    • C07F9/6574—Esters of oxyacids of phosphorus
    • C07F9/65744—Esters of oxyacids of phosphorus condensed with carbocyclic or heterocyclic rings or ring systems
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Medicinal Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Steroid Compounds (AREA)

Abstract

The invention relates to the fields of natural products and pharmaceutical chemistry, and discloses a phosphorus-containing series derivative of Jiyuan oridonin, a preparation method and application thereof. The preparation method comprises the following steps: the series of derivatives are obtained by reacting Jiyuan oridonin as raw material with different phosphorus-containing reagents under the premise of not damaging active centers. The compound has good anti-tumor activity and good stability, can be used for preparing anticancer drugs, and is applied to clinically treating esophageal cancer, pancreatic cancer, gastric cancer, colon cancer, liver cancer and the like. It has the following general formula:

Description

济源冬凌草甲素含磷系列衍生物及其制备方法和用途Jiyuan Rubescensine A phosphorus-containing series derivatives and preparation method and use thereof

技术领域Technical Field

本发明涉及天然产物及药物化学领域，具体涉及济源冬凌草甲素含磷系列衍生物及其合成方法和应用。The invention relates to the field of natural products and medicinal chemistry, and in particular to a series of phosphorus-containing derivatives of Rubescensine A from Jiyuan, and a synthesis method and application thereof.

背景技术Background Art

冬凌草系唇形科香茶菜属植物，味苦、甘，性微寒。具有清热解毒，活血止痛的功效。冬凌草的化学成分研究主要涉及二萜类、挥发油、黄酮和有机酸等，其中冬凌草甲素是冬凌草中主要的抗癌有效成分，是一种无色棱柱状结晶性粉末，几乎不溶于水，微溶于乙醚、甲醇和乙醇等有机溶剂。冬凌草甲素在体外可抑制多种肿瘤细胞生长，包括胃癌、食管癌、鼻咽癌、肝癌、肺癌、膀胱癌、结肠癌、白血病和宫颈癌等癌细胞(Bu H,Liu D,Zhang G,et al.AMPK/mTOR/ULK1 Axis-Mediated Pathway Participates in Apoptosis andAutophagy Induction by Oridonin in Colon Cancer DLD-1Cells[J].Onco TargetsTher,2020,13:8533-8545；Liu Y,Song Z,Liu Y,et al.Identification of ferroptosisas a novel mechanism for antitumor activity of natural product derivative a2in gastric cancer[J].Acta Pharm Sin B,2021,11(6):1513-1525)。在现有药物中引入含磷基团可使它们的选择性得到提高，降低副作用或提高药物的生物利用度，与临床给药中未经修饰的前药相比，经过磷修饰的前药被认为具有更高的极性，并在体内提供更强的氢键。因此，设计含磷的化合物是创新药物研发的重要内容和方向之一。Rubescens is a plant of the genus Rhizoma in the family Lamiaceae. It tastes bitter and sweet and is slightly cold in nature. It has the effects of clearing away heat and detoxifying, promoting blood circulation and relieving pain. The chemical composition research of Rubescens mainly involves diterpenes, volatile oils, flavonoids and organic acids, among which Rubescensine A is the main anti-cancer active ingredient in Rubescens. It is a colorless prismatic crystalline powder, almost insoluble in water, and slightly soluble in organic solvents such as ether, methanol and ethanol. Oridonin can inhibit the growth of various tumor cells in vitro, including gastric cancer, esophageal cancer, nasopharyngeal cancer, liver cancer, lung cancer, bladder cancer, colon cancer, leukemia and cervical cancer (Bu H, Liu D, Zhang G, et al. AMPK/mTOR/ULK1 Axis-Mediated Pathway Participates in Apoptosis and Autophagy Induction by Oridonin in Colon Cancer DLD-1 Cells[J]. Onco Targets Ther, 2020, 13: 8533-8545; Liu Y, Song Z, Liu Y, et al. Identification of ferroptosis as a novel mechanism for antitumor activity of natural product derivative a2 in gastric cancer[J]. Acta Pharm Sin B, 2021, 11(6): 1513-1525). Introducing phosphorus-containing groups into existing drugs can improve their selectivity, reduce side effects or increase the bioavailability of drugs. Compared with unmodified prodrugs in clinical administration, phosphorus-modified prodrugs are believed to have higher polarity and provide stronger hydrogen bonds in vivo. Therefore, designing phosphorus-containing compounds is one of the important contents and directions of innovative drug research and development.

本发明人前期通过提取分离从济源冬凌草中得到了骨架为对映-贝壳杉烯型二萜的先导化合物--济源冬凌草甲素(Jiyuan Oridonin A，JOA)。它是一个醛式与半缩醛式的混合物，主要以半缩醛式形式存在，其中的半缩醛单元虽然是其不稳定的主要原因，但半缩醛良好的反应活性也为我们提供了可修饰结构单元。在JOA与二氯化磷酸酯试剂反应过程中，磷酸酯与其7,14-OH结合，使半缩醛结构单元转变为醛式结构，并获得抗肿瘤活性显著提高的化合物。基于此，发明人设计将不同的含磷基团引入JOA结构中，以期获得抗肿瘤活性提高的化合物。The inventors previously obtained a lead compound, Jiyuan Oridonin A (JOA), whose skeleton is an enantio-kaurene diterpene, from Jiyuan Oridonium by extraction and separation. It is a mixture of aldehyde and hemiacetal, mainly existing in the form of hemiacetal. Although the hemiacetal unit is the main reason for its instability, the good reactivity of hemiacetal also provides us with a modifiable structural unit. During the reaction of JOA with dichlorophosphate reagent, the phosphate combines with its 7,14-OH, converting the hemiacetal structural unit into an aldehyde structure, and obtaining a compound with significantly improved antitumor activity. Based on this, the inventors designed to introduce different phosphorus-containing groups into the JOA structure in order to obtain a compound with improved antitumor activity.

在JOA的结构中引入含磷基团，得到具有良好抗肿瘤活性、高生物利用度、理化性质稳定且低毒的化合物，并能尽早开发成新药，有利于我国自主知识产权的新药研发。相关内容目前尚未见相关文献报道。By introducing phosphorus-containing groups into the structure of JOA, compounds with good anti-tumor activity, high bioavailability, stable physical and chemical properties and low toxicity can be obtained, and they can be developed into new drugs as soon as possible, which is conducive to the research and development of new drugs with independent intellectual property rights in my country. There is no relevant literature report on this matter.

发明内容Summary of the invention

本发明目的在于提供济源冬凌草甲素含磷系列衍生物，提高其抗肿瘤作用及稳定性，为其在临床上的应用提供可能。The purpose of the present invention is to provide a series of phosphorus-containing derivatives of Jiyuan Rubescensine A, improve the anti-tumor effect and stability thereof, and provide the possibility for its clinical application.

本发明的另一目的在于提供其制备方法以及其在制备抗肿瘤药物的应用。Another object of the present invention is to provide a preparation method thereof and application thereof in the preparation of anti-tumor drugs.

为实现本发明目的，本发明将济源冬凌草甲素通过与不同含磷试剂反应得到系列衍生物，提高其稳定性，同时保留或增强其抗肿瘤活性。To achieve the purpose of the present invention, the present invention reacts Jiyuan Rubescensine A with different phosphorus-containing reagents to obtain a series of derivatives, thereby improving the stability thereof while retaining or enhancing its anti-tumor activity.

本发明所述济源冬凌草甲素含磷衍生物结构通式如下所示：The general structural formula of the phosphorus-containing derivative of Rubescensine A of the present invention is as follows:

其中通式I中n为1-5；代表具有不同链长的烷基；Wherein n in the general formula I is 1-5; represents an alkyl group with different chain lengths;

通式Ⅱ中n为1-5；R1为C1-5烷基；优选甲基或者乙基；In the general formula II, n is 1-5; R1 is a C1-5 alkyl group; preferably a methyl group or an ethyl group;

通式Ⅲ中R2为苯基、C1-5烷基或C1-5卤代烷基；优选苯基、乙基或氯乙基；In the general formula III, R 2 is phenyl, C1-5 alkyl or C1-5 haloalkyl; preferably phenyl, ethyl or chloroethyl;

本发明所述的济源冬凌草甲素含磷系列衍生物通式I通过如下合成路线得到：The phosphorus-containing series derivatives of Rubescensine A of the present invention of general formula I are obtained by the following synthetic route:

1、称取济源冬凌草甲素溶于四氢呋喃，在加入催化量的对甲苯磺酸后，加入不同链长的炔醇，萃取干燥后柱色谱纯化得到化合物1a-1e。1. Weigh Jiyuan Rubescensine A and dissolve it in tetrahydrofuran. After adding a catalytic amount of p-toluenesulfonic acid, add alkynols of different chain lengths. After extraction and drying, purify by column chromatography to obtain compounds 1a-1e.

2、将上述所得化合物1a-1e溶于四氢呋喃/水＝4:1中，加入无水硫酸铜与抗坏血酸钠，与叠氮三苯基膦盐反应。反应结束后萃取干燥，浓缩，柱色谱纯化得到目标衍生物(通式I)。2. Dissolve the above obtained compounds 1a-1e in tetrahydrofuran/water = 4:1, add anhydrous copper sulfate and sodium ascorbate, and react with triphenylphosphine azide. After the reaction is completed, extract and dry, concentrate, and purify by column chromatography to obtain the target derivative (general formula I).

本发明所述的济源冬凌草甲素含磷系列衍生物通式Ⅱ通过如下合成路线得到：The phosphorus-containing series derivatives of Rubescensine A of the present invention are obtained by the following synthetic route:

1、称取济源冬凌草甲素溶于四氢呋喃，在加入催化量的对甲苯磺酸后，加入不同链长的溴醇，萃取干燥后柱色谱纯化得到化合物2a-2e。1. Weigh out Jiyuan Rubescensine A and dissolve it in tetrahydrofuran. After adding a catalytic amount of p-toluenesulfonic acid, add bromohydrins of different chain lengths. After extraction and drying, purify by column chromatography to obtain compounds 2a-2e.

2、将上述所得化合物2a-2e溶于乙醇中，加入叠氮化钠，在80℃下反应回流，反应结束后萃取干燥，浓缩，柱色谱纯化得到化合物3a-3e。2. The above-obtained compounds 2a-2e were dissolved in ethanol, sodium azide was added, and the mixture was refluxed at 80°C. After the reaction was completed, the mixture was extracted, dried, concentrated, and purified by column chromatography to obtain compounds 3a-3e.

3、将上述所得化合物3a-3e溶于二氯甲烷，加入亚磷酸三甲酯或者亚磷酸三乙酯，再进行水解，反应结束后萃取干燥，浓缩，柱色谱纯化得到目标衍生物(通式Ⅱ)。3. The above-obtained compounds 3a-3e are dissolved in dichloromethane, trimethyl phosphite or triethyl phosphite is added, and then hydrolyzed. After the reaction is completed, the compounds are extracted, dried, concentrated, and purified by column chromatography to obtain the target derivatives (Formula II).

本发明所述的济源冬凌草甲素含磷系列衍生物通式Ⅲ通过如下合成路线得到：The phosphorus-containing series derivatives of Rubescensine A of the present invention of general formula III are obtained by the following synthetic route:

称取济源冬凌草甲素溶于四氢呋喃，加入三乙胺，与不同的二氯化磷酸酯反应，反应结束后萃取干燥，浓缩，柱色谱纯化得到目标衍生物(通式Ⅲ)。Weigh Jiyuan Rubescensine A and dissolve it in tetrahydrofuran, add triethylamine, and react with different dichlorophosphates. After the reaction is completed, extract and dry, concentrate, and purify by column chromatography to obtain the target derivative (general formula III).

本发明创新点及优点：本发明选用济源冬凌草甲素(JOA)作为母核，通过引入三苯基膦盐、磷酰胺以及磷酸酯，得到所需的含磷衍生物，该类化合物具有抗癌活性，可用于制备抗肿瘤药物，应用于临床治疗食管癌、胰腺癌、结肠癌、胃癌、肝癌等，具有很好的开发前景。Innovation and advantages of the present invention: The present invention uses Jiyuan Rubescensine A (JOA) as the mother nucleus, and obtains the required phosphorus-containing derivatives by introducing triphenylphosphine salts, phosphoramides and phosphates. This type of compound has anti-cancer activity and can be used to prepare anti-tumor drugs. It is used in the clinical treatment of esophageal cancer, pancreatic cancer, colon cancer, gastric cancer, liver cancer, etc., and has a good development prospect.

具体实施方式DETAILED DESCRIPTION

通过以下具体实例进一步举例说明本发明，但应注意本发明的范围并不受这些实施例的任何限制。The present invention is further illustrated by the following specific examples, but it should be noted that the scope of the present invention is not limited by these examples.

实施例1：Embodiment 1:

化合物Ⅰ-1的制备Preparation of Compound Ⅰ-1

称取100mg化合物1a，溶于THF/H2O＝4/1的溶液中，加入117mg叠氮三苯基膦盐，加入39mg抗坏血酸钠，再加入16mg无水硫酸铜，反应6-7h，旋干，乙酸乙酯与饱和食盐水萃取，干燥浓缩，柱色谱纯化，产率为87％。1H NMR(400MHz,DMSO-d6)δ7.91(d,J＝1.8Hz,2H),7.89(s,2H),7.83(d,J＝1.3Hz,1H),7.81(d,J＝1.6Hz,2H),7.78(q,J＝2.7Hz,8H),5.78(s,1H),5.35(s,1H),5.11(s,1H),5.06(d,J＝2.8Hz,1H),4.43(t,J＝6.9Hz,2H),4.37-4.32(m,2H),4.19(d,J＝7.1Hz,1H),3.97(dd,J＝4.1,1.4Hz,1H),3.93(dd,J＝9.6,6.9Hz,1H),3.69(s,2H),3.57-3.52(m,1H),2.86(td,J＝6.9,2.6Hz,2H),2.74(d,J＝9.1Hz,1H),2.65(t,J＝12.6Hz,1H),2.01(t,J＝7.2Hz,3H),1.90(d,J＝8.8Hz,1H),1.57-1.49(m,3H),1.41(d,J＝2.4Hz,1H),1.38(s,1H),1.28(s,2H),1.27(s,2H),1.24(s,2H),1.19(d,J＝5.6Hz,1H),1.15(d,J＝9.1Hz,2H),0.90(s,3H),0.81(s,3H).13C NMR(101MHz,DMSO-d6)δ206.28,152.83,144.47,135.41,135.38,134.09(*2),133.99(*2),130.77(*2),130.64(*2),122.89,119.24(*2),118.39(*2),116.45,99.76,69.99,67.30,65.65,63.54,57.77,56.64,55.39,48.84,48.34,42.97,42.28,40.86,39.76,34.15,33.15,30.83,30.66,30.50,26.74,24.76,22.56,21.19,20.34,19.83,19.23,18.19.HR-MS(ESI):计算值C46H55N3O5P+[M+H]+:761.3952,实测值761.3953.Weigh 100 mg of compound 1a, dissolve in a solution of THF/H 2 O=4/1, add 117 mg of triphenylphosphine azido salt, add 39 mg of sodium ascorbate, and then add 16 mg of anhydrous copper sulfate, react for 6-7 hours, spin dry, extract with ethyl acetate and saturated brine, dry and concentrate, and purify by column chromatography. The yield is 87%. 1 H NMR (400 MHz, DMSO-d 6 )δ7.91(d,J＝1.8Hz,2H),7.89(s,2H),7.83(d,J＝1.3Hz,1H),7.81(d,J＝1.6Hz,2H),7.78(q,J＝2.7Hz,8H),5.78(s,1H),5.35(s,1H),5.11(s,1H),5.06 (d,J＝2.8Hz,1H),4.43(t,J＝6.9Hz,2H),4.37-4.32(m,2H),4.19(d,J＝7.1Hz,1H),3.97(dd,J＝4.1,1.4Hz,1H),3.93(dd,J＝9.6,6.9Hz,1H),3.69(s,2H), 3 .57-3.52(m,1H),2.86(td,J＝6.9,2.6Hz,2H),2.74(d,J＝9.1Hz,1H),2.65(t,J＝12.6Hz,1H),2.01(t,J＝7.2Hz,3H),1.90(d,J＝8.8Hz,1H),1.57-1.49( m,3H),1.41(d,J＝2.4Hz,1H),1.38(s,1H),1.28(s,2H),1.27(s,2H),1.24(s,2H),1.19(d,J＝5.6Hz,1H),1.15(d,J＝9.1Hz,2H),0.90(s,3H),0.81(s ,3H). 13 C NMR (101MHz, DMSO-d 6 ) δ206.28,152.83,144.47,135.41,135.38,134.09(*2),133.99(*2),130.77(*2),130.64(*2),122.89,119.24(*2),118. 39(*2),116.45,99.76,69.99,67.30,65.65,63.54, 57.77,56.64,55.39,48.84,48.34,42.97,42.28,40.86,39.76,34.15,33.15,30.83,30.66,30.50,26.74,24.76,22.56,21.19,20.34,19.83,19.23,18.19. HR-MS (ESI): calculated for C 46 H 55 N 3 O 5 P + [M+H] + : 761.3952, found 761.3953.

实施例2：Embodiment 2:

化合物Ⅰ-2的制备Preparation of Compound Ⅰ-2

用100mg化合物1b代替化合物1a，其他操作同实施例1，得白色固体，产率为74％。1H NMR(400MHz,DMSO-d6)δ7.91(dh,J＝5.8,1.6Hz,3H),7.85(s,1H),7.83(d,J＝1.4Hz,1H),7.81(d,J＝1.6Hz,2H),7.79-7.76(m,8H),5.80(s,1H),5.38(d,J＝1.3Hz,1H),5.16(d,J＝2.8Hz,1H),5.13-5.11(m,1H),4.57(t,J＝2.0Hz,1H),4.54-4.46(m,1H),4.41(t,J＝6.7Hz,2H),4.26(d,J＝7.3Hz,1H),4.00-3.98(m,1H),3.78(dt,J＝9.5,6.2Hz,1H),3.72-3.64(m,2H),3.35-3.31(m,1H),2.83(d,J＝9.0Hz,1H),2.66(t,J＝7.7Hz,3H),2.63-2.57(m,1H),2.02(t,J＝7.2Hz,3H),1.82(t,J＝7.4Hz,2H),1.57(dd,J＝9.9,4.2Hz,1H),1.49(d,J＝7.7Hz,2H),1.37(d,J＝8.5Hz,3H),1.31-1.29(m,1H),1.24(s,1H),1.22-1.18(m,2H),1.10(dd,J＝8.2,6.0Hz,1H),0.92(s,3H),0.82(s,3H).13C NMR(101MHz,DMSO-d6)δ206.26,152.77,146.74,135.41,135.38,134.08(*3),133.98(*3),130.76(*3),130.64(*3),122.42,119.22,118.37,116.57,99.77,70.08,67.23,65.59,63.72,57.80,56.74,48.89,48.35,43.01,42.39,40.89,40.59,39.89,34.17,33.17,30.77,30.60,29.91,24.80,22.43,21.23,20.33,19.82,19.22,18.29.HR-MS(ESI):计算值C47H57N3O5P+[M+H]+:774.4030,实测值774.4026.100 mg of compound 1b was used to replace compound 1a. Other operations were the same as in Example 1 to obtain a white solid with a yield of 74%. 1 H NMR (400 MHz, DMSO-d 6 ) δ7.91 (dh, J=5.8,1.6 Hz, 3H), 7.85 (s, 1H), 7.83 (d, J=1.4 Hz, 1H), 7.81 (d, J=1.6 Hz, 2H), 7.79-7.76 (m, 8H), 5.80 (s, 1H), 5.38 (d, J=1.3 Hz, 1H), 5.16 (d, J=2.8 Hz, 1H),5.13-5.11(m,1H),4.57(t,J＝2.0Hz,1H),4.54-4.46(m,1H),4.41(t,J＝6.7Hz,2H),4.26(d,J＝7.3Hz,1H),4.00-3.98(m,1H),3.78(dt,J＝9.5,6 .2Hz,1H),3.7 2-3.64(m,2H),3.35-3.31(m,1H),2.83(d,J＝9.0Hz,1H),2.66(t,J＝7.7Hz,3H),2.63-2.57(m,1H),2.02(t,J＝7.2Hz,3H),1.82(t,J＝7.4Hz,2H),1.57 (dd,J＝9.9,4 .2Hz,1H),1.49(d,J＝7.7Hz,2H),1.37(d,J＝8.5Hz,3H),1.31-1.29(m,1H),1.24(s,1H),1.22-1.18(m,2H),1.10(dd,J＝8.2,6.0Hz,1H),0.92(s,3H),0 .82(s,3H). 13 C NMR(101MHz,DMSO-d 6 )δ206.26,152.77,146.74,135.41,135.38,134.08(*3),133.98(*3),130.76(*3),130.64(*3),122.42,119.22,118.37,116.57,99.77,70.08,6 7.23,65.59,63.72,5 7.80,56.74,48.89,48.35,43.01,42.39,40.89,40.59,39.89,34.17,33.17,30.77,30.60,29.91,24.80,22.43,21.23,20.33,19.82,19.22,18.29. HR-MS (ESI): calculated for C 47 H 57 N 3 O 5 P + [M+H] + : 774.4030, found 774.4026.

实施例3：Embodiment 3:

化合物Ⅰ-3的制备Preparation of Compound Ⅰ-3

用100mg化合物1c代替化合物1a，其他操作同实施例1，得白色固体，产率为81％。1H NMR(400MHz,Chloroform-d)δ7.78(s,6H),7.76(d,J＝2.6Hz,3H),7.70(s,4H),5.93(s,1H),5.30(d,J＝1.4Hz,1H),5.18(s,1H),4.89-4.82(m,3H),4.77(t,J＝4.5Hz,1H),4.61(d,J＝6.0Hz,2H),4.17-4.09(m,3H),3.89-3.85(m,2H),3.83(d,J＝6.6Hz,2H),3.79(q,J＝4.5Hz,2H),3.72(d,J＝3.5Hz,1H),3.52-3.46(m,2H),3.40(dd,J＝9.8,5.1Hz,2H),3.08(td,J＝6.9,4.2Hz,1H),2.98(dd,J＝15.0,9.3Hz,2H),2.74(d,J＝6.6Hz,3H),2.32(d,J＝10.6Hz,3H),1.34(t,J＝5.8Hz,3H),1.17(d,J＝7.0Hz,3H),0.96(s,3H),0.84(s,3H).13CNMR(101MHz,DMSO-d6)δ206.28,152.83,144.47,135.41,135.38,134.09(*2),133.99(*2),130.77(*2),130.64(*2),122.89,119.24(*2),118.39(*2),116.45,99.53,84.87,71.77,70.15,67.11,65.55,63.58,52.31，57.80,56.79,48.92,43.05,42.43,40.89,40.38，39.84,34.17,33.17,30.52,28.93,25.41,24.78,21.22,20.20 18.28,17.93.HR-MS(ESI):计算值C48H59N3O5P+[M+H]+:788.4187,实测值788.4167.100 mg of compound 1c was used to replace compound 1a. Other operations were the same as in Example 1 to obtain a white solid with a yield of 81%. 1 H NMR (400 MHz, Chloroform-d) δ7.78 (s, 6H), 7.76 (d, J = 2.6 Hz, 3H), 7.70 (s, 4H), 5.93 (s, 1H), 5.30 (d, J = 1.4 Hz, 1H), 5.18 (s, 1H), 4.89-4.82 (m, 3H), 4.77 (t, J = 4.5 Hz, 1H), 4.61 (d, J = 6.0 Hz, 2H), 4.17-4.09 (m, 3H), 3.89-3.85 (m, 2H), 3.83 (d, J = 6.6 Hz, 2H), 3.79 (q, J＝4.5Hz,2H),3.72(d,J＝3.5Hz,1H),3.52-3.46(m,2H),3.40(dd,J＝9.8,5.1Hz,2H),3.08(td,J＝6.9,4.2Hz,1H),2.98(dd,J＝15.0,9.3Hz,2H),2.74(d,J＝ 6.6Hz,3H),2.32(d,J＝10.6Hz,3H),1.34(t,J＝5.8Hz,3H),1.17(d,J＝7.0Hz,3H),0.96(s,3H),0.84(s,3H). 13 CNMR(101MHz,DMSO-d 6 )δ206.28,152.83,144.47,135.41,135.38,134.09(*2),133.99(*2),130.77(*2),130.64(*2),122.89,119.24(*2),118.39(*2),116.45,99.53, 84.87,7 1.77, 70.15, 67.11, 65.55, 63.58, 52.31, 57.80, 56.79, 48.92, 43.05, 42.43, 40.89, 40.38, 39.84, 34.17, 33.17, 30.52, 28.93, 25.41, 24.78, 21.22, 20.20 18.28, 17.93. HR-MS (ESI): calculated for C 48 H 59 N 3 O 5 P + [M+H] + : 788.4187, found 788.4167.

实施例4：Embodiment 4:

化合物Ⅰ-4的制备Preparation of Compound Ⅰ-4

用100mg化合物1d代替化合物1a，其他操作同实施例1，得白色固体，产率为78％。1H NMR(400MHz,Chloroform-d)δ7.80(s,2H),7.78(s,4H),7.75(s,2H),7.71(s,3H),7.69(s,4H),5.95(s,1H),5.31(d,J＝2.7Hz,1H),5.18-5.16(m,1H),4.82(s,1H),4.81-4.75(m,1H),4.59(t,J＝6.7Hz,2H),4.13-4.10(m,1H),3.89-3.85(m,1H),3.80(q,J＝8.3Hz,2H),3.33-3.26(m,1H),3.00(d,J＝9.1Hz,1H),2.79-2.74(m,2H),2.68(d,J＝9.9Hz,3H),2.55(s,3H),2.37-2.27(m,3H),2.04-1.98(m,1H),1.77-1.71(m,1H),1.68(t,J＝3.1Hz,1H),1.60(s,2H),1.57(d,J＝9.5Hz,3H),1.43(d,J＝5.1Hz,2H),1.37(d,J＝4.9Hz,3H),1.34-1.29(m,2H),1.17(t,J＝7.0Hz,1H),0.95(s,3H),0.83(d,J＝2.5Hz,3H).13C NMR(101MHz,CDCl3)δ206.04,151.66,135.03,135.00(*3),133.67(*3),133.57,130.54(*3),130.42(*3),118.50,117.64,116.91,100.05,77.29,70.68,67.85,66.18,64.78,57.90,56.56,48.49,48.40,42.59,41.75,40.67,39.96,33.97,32.86,30.58,30.04,29.87,29.11,28.71,25.75,25.53,24.70,21.77,21.26,21.00,20.92,19.14,18.25,15.17.HR-MS(ESI):计算值C49H61N3O5P+[M+H]+:802.4343,实测值802.4332.100 mg of compound 1d was used to replace compound 1a, and other operations were the same as in Example 1 to obtain a white solid with a yield of 78%. NMR(400MHz,Chloroform-d)δ7.80(s,2H),7.78(s,4H),7.75(s,2H),7.71(s,3H),7.69(s,4H),5.95(s,1H),5.31(d,J＝2.7Hz,1H),5.18-5.16(m,1H),4.82(s ,1H),4.81-4.75(m,1H),4.59(t,J＝6.7Hz,2H),4.13-4.10(m,1H),3.89-3.85(m,1H),3.80(q,J＝8.3Hz,2H),3.33-3.26(m,1H),3.00(d,J＝9.1H z,1H),2.79-2.74(m,2H),2.68(d,J＝9.9Hz,3H),2.55(s,3H),2.37-2.27(m,3H),2.04-1.98(m,1H),1.77-1.71(m,1H),1.68(t,J＝3.1Hz,1H),1.60( s,2H),1.57(d,J＝9.5Hz,3H),1.43(d,J＝5.1Hz,2H),1.37(d,J＝4.9Hz,3H),1.34-1.29(m,2H),1.17(t,J＝7.0Hz,1H),0.95(s,3H),0.83(d,J＝2.5Hz,3H ). 13C NMR(101MHz, CDCl3)δ206.04,151.66,135.03,135.00(*3),133.67(*3),133.57,130.54(*3),130.42(*3),118.50,117.64,116.91,100.05,77.29,70. 68,67.85,66.18,64.78,57.90,56 .56,48.49,48.40,42.59,41.75,40.67,39.96,33.97,32.86,30.58,30.04,29.87,29.11,28.71,25.75,25.53,24.70,21.77,21.26,21.00,20.92,19.14,18.25,15.17. HR-MS (ESI): calculated for C 49 H 61 N 3 O 5 P + [M+H] + :802.4343, found 802.4332.

实施例5：Embodiment 5:

化合物Ⅰ-5的制备Preparation of Compound Ⅰ-5

用100mg化合物1e代替化合物1a，其他操作同实施例1，得白色固体，产率为83％。1H NMR(400MHz,Chloroform-d)δ7.81(s,3H),7.76(d,J＝1.9Hz,3H),7.70(d,J＝3.5Hz,3H),7.67(dd,J＝8.1,3.2Hz,6H),5.94(s,1H),5.32(s,1H),5.18(s,1H),4.83(s,1H),4.81-4.77(m,1H),4.58(d,J＝6.9Hz,3H),4.12-4.09(m,1H),3.84-3.83(m,1H),3.82-3.79(m,2H),3.49(qd,J＝7.0,4.7Hz,1H),3.29(dt,J＝9.3,6.0Hz,2H),3.02(d,J＝9.5Hz,1H),2.83-2.79(m,1H),2.77(d,J＝2.2Hz,1H),2.66(d,J＝5.7Hz,2H),2.61(t,J＝7.7Hz,2H),2.35-2.30(m,3H),2.09-2.04(m,1H),1.69-1.65(m,3H),1.59(d,J＝2.3Hz,3H),1.44(d,J＝5.6Hz,3H),1.41(s,3H),1.33(d,J＝4.0Hz,2H),1.17(t,J＝7.0Hz,1H),0.96(s,3H),0.84(s,3H).13C NMR(101MHz,CDCl3)δ205.97,151.59,135.01,134.99,133.67(*3),133.57(*3),130.53(*3),130.41(*3),120.36，118.49,117.64,116.93,100.19,77.28,70.87,68.49,66.22,64.81,57.99,57.94,56.56,48.44,42.44,41.72,40.65,39.98,39.52,33.98,32.85,30.56,30.04,29.87,28.92,28.04,25.77,25.18,24.70,21.80,21.30,21.02,19.16,19.13,18.29,15.37.HR-MS(ESI):计算值C50H63N3O5P+[M+H]+:816.4500,实测值816.4499.100 mg of compound 1e was used to replace compound 1a, and the other operations were the same as in Example 1 to obtain a white solid with a yield of 83%. NMR(400MHz,Chloroform-d)δ7.81(s,3H),7.76(d,J＝1.9Hz,3H),7.70(d,J＝3.5Hz,3H),7.67(dd,J＝8.1,3.2Hz,6H),5.94(s,1H),5.32(s,1H),5.18(s,1H),4.83 (s,1H),4.81-4.77(m,1H),4.58(d,J＝6.9Hz,3H),4.12-4.09(m,1H),3.84-3.83(m,1H),3.82-3.79(m,2H),3.49(qd,J＝7.0,4.7Hz,1H),3.29(dt,J＝9 . 3,6.0Hz,2H),3.02(d,J＝9.5Hz,1H),2.83-2.79(m,1H),2.77(d,J＝2.2Hz,1H),2.66(d,J＝5.7Hz,2H),2.61(t,J＝7.7Hz,2H),2.35-2.30(m,3H),2.09-2 .04(m,1H),1.69-1.65(m,3H),1.59(d,J＝2.3Hz,3H),1.44(d,J＝5.6Hz,3H),1.41(s,3H),1.33(d,J＝4.0Hz,2H),1.17(t,J＝7.0Hz,1H),0.96(s,3H),0. 84(s,3H). 13 C NMR(101MHz, CDCl3)δ205.97,151.59,135.01,134.99,133.67(*3),133.57(*3),130.53(*3),130.41(*3),120.36,118.49,117.64,116.93,100.19,77 .28,70.87,68.49,66.22,64.81,57.99,57 .94,56.56,48.44,42.44,41.72,40.65,39.98,39.52,33.98,32.85,30.56,30.04,29.87,28.92,28.04,25.77,25.18,24.70,21.80,21.30,21.02,19.16,19.13,18.29,15.37. HR-MS (ESI): calculated for C 50 H 63 N 3 O 5 P + [M+H] + :816.4500, found 816.4499.

实施例6：Embodiment 6:

化合物Ⅱ-1的制备Preparation of Compound II-1

将100mg化合物3a溶于二氯甲烷中，加入89mg亚磷酸三甲酯，加入0.5mL水，反应24h后处理，柱色谱纯化，得白色固体，产率为56％。1H NMR(400MHz,DMSO-d6)δ5.79(s,1H),5.38(d,J＝1.2Hz,1H),5.13-5.11(m,1H),5.07(d,J＝2.8Hz,1H),4.87(dt,J＝11.6,6.8Hz,1H),4.54(t,J＝2.0Hz,1H),4.47(q,J＝8.4Hz,1H),4.17(d,J＝7.1Hz,1H),4.00-3.97(m,1H),3.69(dt,J＝10.2,6.3Hz,1H),3.58(d,J＝1.0Hz,3H),3.56(d,J＝1.0Hz,3H),3.01-2.90(m,2H),2.81(d,J＝9.1Hz,1H),2.71-2.63(m,1H),2.63-2.56(m,1H),2.04(d,J＝12.9Hz,1H),1.57(ddd,J＝13.7,6.4,4.3Hz,1H),1.43(d,J＝12.7Hz,1H),1.36(s,1H),1.32(dd,J＝9.8,5.4Hz,1H),1.29-1.26(m,1H),1.26-1.24(m,1H),1.22(d,J＝8.3Hz,1H),1.19(d,J＝2.8Hz,1H),1.16(s,1H),1.11-1.04(m,1H),0.93(s,3H),0.83(s,3H).13C NMR(101MHz,DMSO-d6)δ206.21,152.84,116.45,100.20,69.99,68.79,65.68,63.69,57.79,56.65,52.85,52.79,48.79,43.01,42.24,41.26,40.87,39.97,34.16,33.16,30.52,24.77,21.22,18.28.HR-MS(ESI):计算值C24H38NO8P[M+Na]+:522.2227,实测值522.2223.100 mg of compound 3a was dissolved in dichloromethane, 89 mg of trimethyl phosphite and 0.5 mL of water were added, and the mixture was reacted for 24 h and then treated and purified by column chromatography to obtain a white solid with a yield of 56%. 1 H NMR (400 MHz, DMSO-d 6 )δ5.79(s,1H),5.38(d,J＝1.2Hz,1H),5.13-5.11(m,1H),5.07(d,J＝2.8Hz,1H),4.87(dt,J＝11.6,6.8Hz,1H),4.54(t,J＝2.0Hz,1H),4.47(q,J＝8.4Hz,1H ),4.17(d,J＝7.1Hz,1H),4.00-3.97(m,1H),3.69(dt,J＝10.2,6.3Hz,1H),3.58(d,J＝1.0Hz,3H),3.56(d,J＝1.0Hz,3H),3.01-2.90(m,2H),2.81(d,J＝9 .1Hz, 1H),2.71-2.63(m,1H),2.63-2.56(m,1H),2.04(d,J＝12.9Hz,1H),1.57(ddd,J＝13.7,6.4,4.3Hz,1H),1.43(d,J＝12.7Hz,1H),1.36(s,1H),1.32(dd,J＝ 9.8,5.4Hz,1H),1.29-1.26(m,1H),1.26-1.24(m,1H),1.22(d,J＝8.3Hz,1H),1.19(d,J＝2.8Hz,1H),1.16(s,1H),1.11-1.04(m,1H),0.93(s,3H),0. 83(s,3H). 13 C NMR (101 MHz, DMSO-d 6 ) δ 206.21, 152.84, 116.45, 100.20, 69.99, 68.79, 65.68, 63.69, 57.79, 56.65, 52.85, 52.79, 48.79, 43.01, 42.24, 41.26, 40.87, 39.97, 34.16, 33.16, 30.52, 24.77, 21.22, 18.28. HR-MS (ESI): calculated for C 24 H 38 NO 8 P [M+Na] + : 522.2227, found 522.2223.

实施例7：Embodiment 7:

化合物Ⅱ-2的制备Preparation of Compound II-2

将100mg化合物3a溶于二氯甲烷中，加入亚磷酸三乙酯，其他操作同实施例6，产率为48％。1H NMR(400MHz,DMSO-d6)δ5.79(s,1H),5.37(d,J＝1.2Hz,1H),5.11(d,J＝1.1Hz,1H),5.07(d,J＝2.8Hz,1H),4.78-4.73(m,1H),4.54(t,J＝2.0Hz,1H),4.51-4.44(m,1H),4.17(d,J＝7.1Hz,1H),3.99(q,J＝1.4Hz,1H),3.98(d,J＝1.2Hz,1H),3.93(d,J＝1.5Hz,2H),3.91(t,J＝1.2Hz,2H),3.88(dd,J＝7.2,1.6Hz,2H),3.71-3.66(m,1H),2.95(tt,J＝11.3,8.4Hz,3H),2.80(d,J＝9.1Hz,1H),2.67-2.62(m,1H),2.62-2.56(m,1H),2.07-1.98(m,2H),1.37-1.34(m,2H),1.29(d,J＝6.9Hz,1H),1.25(d,J＝1.3Hz,1H),1.23(s,1H),1.22(d,J＝0.8Hz,6H),1.21-1.20(m,3H),0.92(s,3H),0.82(s,3H).13C NMR(101MHz,DMSO-d6)δ206.21,152.83,116.46,100.27,69.93,68.95,65.68,63.70,61.75,61.70,57.79,56.64,48.76,43.00,42.23,41.28,40.85,39.66,34.16,33.15,30.52,24.77,21.07,18.28,16.61,16.54.HR-MS(ESI):计算值C24H38NO8P[M+Na]+:550.2540,实测值550.2535.100 mg of compound 3a was dissolved in dichloromethane, triethyl phosphite was added, and the other operations were the same as those in Example 6. The yield was 48%. 1 H NMR (400 MHz, DMSO-d 6 )δ5.79(s,1H),5.37(d,J＝1.2Hz,1H),5.11(d,J＝1.1Hz,1H),5.07(d,J＝2.8Hz,1H),4.78-4.73(m,1H),4.54(t,J＝2.0Hz,1H),4.51-4.44(m,1H),4.17( d,J＝7.1Hz,1H),3.99(q,J＝1.4Hz,1H),3.98(d,J＝1.2Hz,1H),3.93(d,J＝1.5Hz,2H),3.91(t,J＝1.2Hz,2H),3.88(dd,J＝7.2,1.6Hz ,2H),3.71-3.66(m,1H),2.95(tt,J＝11.3,8.4Hz,3H),2.80(d,J＝9.1Hz,1H),2.67-2.62(m,1H),2.62-2.56(m,1H),2.07-1.98(m,2H),1.37-1.34(m ,2H),1.29(d,J＝6.9Hz,1H),1.25(d,J＝1.3Hz,1H),1.23(s,1H),1.22(d,J＝0.8Hz,6H),1.21-1.20(m,3H),0.92(s,3H),0.82(s,3H). 13 C NMR (101 MHz, DMSO-d 6 ) δ 206.21, 152.83, 116.46, 100.27, 69.93, 68.95, 65.68, 63.70, 61.75, 61.70, 57.79, 56.64, 48.76, 43.00, 42.23, 41.28, 40.85, 39.66, 34.16, 33.15, 30.52, 24.77, 21.07, 18.28, 16.61, 16.54. HR-MS (ESI): calculated for C 24 H 38 NO 8 P [M+Na] + : 550.2540, found 550.2535.

实施例8：Embodiment 8:

化合物Ⅱ-3的制备Preparation of Compound II-3

以化合物3b代替化合物3a，其他操作同实施例6，产率为59％。1H NMR(400MHz,DMSO-d6)δ5.80(s,1H),5.38(d,J＝1.2Hz,1H),5.12-5.09(m,2H),4.96(dt,J＝11.6,6.6Hz,1H),4.53(dd,J＝2.8,1.5Hz,1H),4.44(p,J＝8.3Hz,1H),4.19(d,J＝7.1Hz,1H),3.99-3.96(m,1H),3.76(dt,J＝9.6,6.0Hz,1H),3.58(d,J＝1.3Hz,3H),3.55(d,J＝1.3Hz,3H),2.90-2.82(m,2H),2.80(d,J＝9.5Hz,1H),2.70-2.63(m,1H),2.63-2.56(m,1H),2.02-1.95(m,1H),1.68(p,J＝6.5Hz,2H),1.57(ddd,J＝13.7,6.3,4.3Hz,1H),1.43(d,J＝13.0Hz,2H),1.37(q,J＝3.2Hz,2H),1.35-1.31(m,1H),1.29(d,J＝5.9Hz,1H),1.20(t,J＝3.1Hz,1H),1.17(d,J＝6.4Hz,1H),1.08(ddd,J＝15.8,11.6,3.8Hz,1H),0.93(s,3H),0.82(s,3H).13C NMR(101MHz,DMSO-d6)δ206.27,152.78,116.50,99.63,70.13,65.57,65.32,63.64,57.79,56.81,52.77,52.71,48.93,43.06,42.34,40.90,39.84,38.50,34.17,33.16,32.25,30.52,24.78,21.20,18.25.HR-MS(ESI):计算值C25H40NO8P[M+Na]+:536.2384,实测值536.2376.Compound 3b was used instead of compound 3a, and other operations were the same as in Example 6. The yield was 59%. 1 H NMR (400 MHz, DMSO-d 6 )δ5.80(s,1H),5.38(d,J＝1.2Hz,1H),5.12-5.09(m,2H),4.96(dt,J＝11.6,6.6Hz,1H),4.53(dd,J＝2.8,1.5Hz,1H),4.44(p,J＝8.3Hz,1H),4.19(d,J＝7. 1Hz,1H),3.99-3.96(m,1H),3.76(dt,J＝9.6,6.0Hz,1H),3.58(d,J＝1.3Hz,3H),3.55(d,J＝1.3Hz,3H),2.90-2.82(m,2H),2.80(d,J＝9.5Hz,1H),2.70-2 . 63(m,1H),2.63-2.56(m,1H),2.02-1.95(m,1H),1.68(p,J＝6.5Hz,2H),1.57(ddd,J＝13.7,6.3,4.3Hz,1H),1.43(d,J＝13.0Hz,2H),1.37(q,J＝3.2Hz,2 H),1.35-1.31(m,1H),1.29(d,J＝5.9Hz,1H),1.20(t,J＝3.1Hz,1H),1.17(d,J＝6.4Hz,1H),1.08(ddd,J＝15.8,11.6,3.8Hz,1H),0.93(s,3H),0.82(s,3H ). 13 C NMR (101 MHz, DMSO-d 6 ) δ 206.27, 152.78, 116.50, 99.63, 70.13, 65.57, 65.32, 63.64, 57.79, 56.81, 52.77, 52.71, 48.93, 43.06, 42.34, 40.90, 39.84, 38.50, 34.17, 33.16, 32.25, 30.52, 24.78, 21.20, 18.25. HR-MS (ESI): calculated for C 25 H 40 NO 8 P [M+Na] + : 536.2384, found 536.2376.

实施例9：Embodiment 9:

化合物Ⅱ-4的制备Preparation of Compound II-4

以化合物3b代替化合物3a，加入亚磷酸三乙酯，其他操作同实施例6，产率为61％。1HNMR(400MHz,DMSO-d6)δ5.79(s,1H),5.37(t,J＝1.1Hz,1H),5.10(d,J＝3.1Hz,2H),4.85(dt,J＝11.1,6.6Hz,1H),4.52(dd,J＝3.1,1.5Hz,1H),4.43(p,J＝8.4Hz,1H),4.17(d,J＝7.2Hz,1H),3.97(dd,J＝4.2,1.4Hz,1H),3.93-3.87(m,4H),3.75(dt,J＝9.7,6.1Hz,1H),2.89-2.81(m,2H),2.78(d,J＝9.2Hz,1H),2.67-2.61(m,1H),2.58(t,J＝9.0Hz,1H),1.98(d,J＝12.6Hz,1H),1.66(q,J＝6.7Hz,2H),1.57(dd,J＝12.6,7.1Hz,1H),1.43(d,J＝13.0Hz,2H),1.36(d,J＝3.5Hz,2H),1.32(d,J＝9.9Hz,1H),1.29(d,J＝2.8Hz,1H),1.24-1.20(m,6H),1.19(d,J＝4.2Hz,1H),1.17(s,1H),1.13-1.02(m,2H),0.92(s,3H),0.82(s,3H).13C NMR(101MHz,DMSO-d6)δ206.28,152.77,116.51,99.65,70.12,65.57,63.64,61.61,61.56,57.80,56.83,48.93,43.07,42.34,40.90,39.83,38.59,34.17,33.16,32.32,30.52,28.35,24.78,21.19,18.26,16.62,16.55.HR-MS(ESI):计算值C27H44NO8P[M+Na]+:564.2697,实测值564.2688.Compound 3b was used instead of compound 3a, triethyl phosphite was added, and other operations were the same as those in Example 6. The yield was 61%. 1 HNMR (400 MHz, DMSO-d 6 )δ5.79(s,1H),5.37(t,J＝1.1Hz,1H),5.10(d,J＝3.1Hz,2H),4.85(dt,J＝11.1,6.6Hz,1H),4.52(dd,J＝3.1,1.5Hz,1H),4.43(p,J＝8.4Hz,1H),4.17(d,J＝ 7.2Hz,1H),3.97(dd,J＝4.2,1.4Hz,1H),3.93-3.87(m,4H),3.75(dt,J＝9.7,6.1Hz,1H),2.89-2.81(m,2H),2.78(d,J＝9.2Hz,1H),2.67-2.61(m ,1H),2.58(t,J＝9.0Hz,1H),1.98(d,J＝12.6Hz,1H),1.66(q,J＝6.7Hz,2H),1.57(dd,J＝12.6,7.1Hz,1H),1.43(d,J＝13.0Hz,2H),1.36(d,J＝3.5Hz,2H),1. 32(d,J＝9.9Hz,1H),1.29(d,J＝2.8Hz,1H),1.24-1.20(m,6H),1.19(d,J＝4.2Hz,1H),1.17(s,1H),1.13-1.02(m,2H),0.92(s,3H),0.82(s,3H). 13 C NMR (101 MHz, DMSO-d 6 ) δ 206.28, 152.77, 116.51, 99.65, 70.12, 65.57, 63.64, 61.61, 61.56, 57.80, 56.83, 48.93, 43.07, 42.34, 40.90, 39.83, 38.59, 34.17, 33.16, 32.32, 30.52, 28.35, 24.78, 21.19, 18.26, 16.62, 16.55. HR-MS (ESI): calculated for C 27 H 44 NO 8 P [M+Na] + : 564.2697, found 564.2688.

实施例10：Embodiment 10:

化合物Ⅱ-5的制备Preparation of Compound II-5

以化合物3c代替化合物3a，其他操作同实施例6，产率为52％。1H NMR(400MHz,Chloroform-d)δ6.02(s,1H),5.39(d,J＝1.0Hz,1H),5.19(dd,J＝8.4,1.2Hz,2H),4.80(d,J＝1.5Hz,1H),4.79-4.68(m,2H),4.62(q,J＝8.8Hz,1H),4.16-4.04(m,2H),3.75(d,J＝2.1Hz,2H),3.72(s,3H),3.69(s,3H),3.51(tdd,J＝8.0,4.3,1.7Hz,1H),3.33(dt,J＝10.1,6.3Hz,2H),2.99-2.94(m,2H),2.94-2.89(m,3H),2.88(dd,J＝4.5,2.1Hz,1H),2.85(d,J＝3.1Hz,1H),2.85-2.82(m,1H),2.81-2.79(m,1H),2.79-2.76(m,1H),1.47(d,J＝3.1Hz,2H),1.40(s,1H),1.29(s,1H),1.25(s,1H),0.97(s,3H),0.87(s,3H).13C NMR(101MHz,CDCl3)δ206.21,152.84,116.45,100.20,69.99,68.79,65.68,63.69,57.79,56.65,52.85,52.79,48.79,43.01,42.24,41.26,40.87,39.97,34.16,33.16,30.52,28.56,28.13,24.77,21.22,18.28.HR-MS(ESI):计算值C26H42NO8P[M+Na]+:550.2540,实测值550.2537.Compound 3c was used to replace compound 3a. Other operations were the same as in Example 6. The yield was 52%. 1 H NMR (400 MHz, Chloroform-d) δ6.02 (s, 1H), 5.39 (d, J = 1.0 Hz, 1H), 5.19 (dd, J = 8.4, 1.2 Hz, 2H), 4.80 (d, J = 1.5 Hz, 1H), 4.79-4.68 (m, 2H), 4.62 (q, J = 8.8 Hz, 1H), 4.16-4.04 (m, 2H), 3.75 (d, J = 2.1 Hz, 2H), 3.72 (s, 3H), 3.69 (s, 3H), 3.51 (tdd, J = 8.0, 4.3, 1.7 Hz, 1H), 3 .33(dt,J＝10.1,6.3Hz,2H),2.99-2.94(m,2H),2.94-2.89(m,3H),2.88(dd,J＝4.5,2.1Hz,1H),2.85(d,J＝3.1Hz,1H),2.85-2.82(m,1H),2.81-2.79( m,1H),2.79-2.76(m,1H),1.47(d,J＝3.1Hz,2H),1.40(s,1H),1.29(s,1H),1.25(s,1H),0.97(s,3H),0.87(s,3H). 13 C NMR (101 MHz, CDCl3) δ 206.21, 152.84, 116.45, 100.20, 69.99, 68.79, 65.68, 63.69, 57.79, 56.65, 52.85, 52.79, 48.79, 43.01, 42.24, 41.26, 40.87, 39.97, 34.16, 33.16, 30.52, 28.56, 28.13, 24.77, 21.22, 18.28. HR-MS (ESI): calculated for C 26 H 42 NO 8 P [M + Na] + : 550.2540, found 550.2537.

实施例11：Embodiment 11:

化合物Ⅱ-6的制备Preparation of Compound II-6

以化合物3c代替化合物3a，加入亚磷酸三乙酯，其他操作同实施例6，产率为54％。1HNMR(400MHz,DMSO-d6)δ5.79(s,1H),5.38(t,J＝1.1Hz,1H),5.10(d,J＝3.1Hz,2H),4.85(dt,J＝11.1,6.6Hz,1H),4.52(dd,J＝3.1,1.5Hz,1H),4.43(p,J＝8.4Hz,1H),4.17(d,J＝7.2Hz,1H),3.97(dd,J＝4.2,1.4Hz,1H),3.93-3.87(m,4H),3.75(dt,J＝9.7,6.1Hz,1H),2.89-2.81(m,2H),2.78(d,J＝9.2Hz,1H),2.67-2.61(m,1H),2.58(t,J＝9.0Hz,1H),1.98(d,J＝12.6Hz,1H),1.66(q,J＝6.7Hz,2H),1.57(dd,J＝12.6,7.1Hz,1H),1.43(d,J＝13.0Hz,2H),1.36(d,J＝3.5Hz,2H),1.32(d,J＝9.9Hz,1H),1.29(d,J＝2.8Hz,1H),1.24-1.20(m,6H),1.19(d,J＝4.2Hz,1H),1.17(s,1H),1.13-1.02(m,2H),0.92(s,3H),0.82(s,3H).13C NMR(101MHz,DMSO-d6)δ206.30,152.75,116.51,99.65,70.12,65.57,63.64,62.61,62.56,57.80,56.83,48.93,43.07,42.34,40.90,39.83,38.59,34.17,33.16,32.32,30.52,28.17,24.78,18.26,16.62,16.55.HR-MS(ESI):计算值C28H46NO8P[M+Na]+:578.2853,实测值578.2848.Compound 3c was used instead of compound 3a, triethyl phosphite was added, and the other operations were the same as those in Example 6. The yield was 54%. 1 HNMR (400 MHz, DMSO-d 6 )δ5.79(s,1H),5.38(t,J＝1.1Hz,1H),5.10(d,J＝3.1Hz,2H),4.85(dt,J＝11.1,6.6Hz,1H),4.52(dd,J＝3.1,1.5Hz,1H),4.43(p,J＝8.4Hz,1H),4.17(d,J＝ 7.2Hz,1H),3.97(dd,J＝4.2,1.4Hz,1H),3.93-3.87(m,4H),3.75(dt,J＝9.7,6.1Hz,1H),2.89-2.81(m,2H),2.78(d,J＝9.2Hz,1H),2.67-2.61(m ,1H),2.58(t,J＝9.0Hz,1H),1.98(d,J＝12.6Hz,1H),1.66(q,J＝6.7Hz,2H),1.57(dd,J＝12.6,7.1Hz,1H),1.43(d,J＝13.0Hz,2H),1.36(d,J＝3.5Hz,2H),1. 32(d,J＝9.9Hz,1H),1.29(d,J＝2.8Hz,1H),1.24-1.20(m,6H),1.19(d,J＝4.2Hz,1H),1.17(s,1H),1.13-1.02(m,2H),0.92(s,3H),0.82(s,3H). 13 C NMR (101 MHz, DMSO-d 6 ) δ 206.30, 152.75, 116.51, 99.65, 70.12, 65.57, 63.64, 62.61, 62.56, 57.80, 56.83, 48.93, 43.07, 42.34, 40.90, 39.83, 38.59, 34.17, 33.16, 32.32, 30.52, 28.17, 24.78, 18.26, 16.62, 16.55. HR-MS (ESI): calculated for C 28 H 46 NO 8 P [M+Na] + : 578.2853, found 578.2848.

实施例12：Embodiment 12:

化合物Ⅱ-7的制备Preparation of Compound II-7

以化合物3d代替化合物3a，其他操作同实施例6，产率为71％。1H NMR(400MHz,Chloroform-d)δ6.03-6.02(m,1H),5.40(d,J＝1.0Hz,1H),5.22(d,J＝1.3Hz,1H),4.85(d,J＝1.5Hz,1H),4.76(q,J＝8.7Hz,1H),4.07(dd,J＝4.3,1.5Hz,1H),3.88-3.81(m,1H),3.80-3.69(m,1H),3.58(d,J＝1.3Hz,3H),3.55(d,J＝1.3Hz,3H),3.42(dt,J＝9.7,7.0Hz,1H),2.97(dt,J＝9.5,1.3Hz,1H),2.83-2.76(m,2H),2.06-2.01(m,1H),1.72(dd,J＝14.2,6.2,4.3Hz,1H),1.51(d,J＝3.5Hz,1H),1.48(q,J＝2.4Hz,2H),1.43-1.41(m,2H),1.41-1.39(m,1H),1.37(d,J＝5.9Hz,1H),1.34(dd,J＝6.8,2.3Hz,2H),1.25(s,1H),1.24(s,1H),1.23(d,J＝2.9Hz,2H),1.21(d,J＝3.1Hz,1H),1.19-1.10(m,2H),0.97(s,3H),0.87(s,3H).13CNMR(101MHz,CDCl3)δ205.35,150.86,117.46,99.79,77.24,71.40,66.35,65.17,63.94,58.15,56.85,52.79,52.43,48.45,42.07,41.46,40.55,39.84,33.98,32.80,30.45,29.69,24.58,20.93,18.23,15.47,15.43.HR-MS(ESI):计算值C27H44NO8P[M+Na]+:564.2697,实测值564.2692.Compound 3d was used instead of compound 3a, and other operations were the same as in Example 6. The yield was 71%. NMR(400MHz,Chloroform-d)δ6.03-6.02(m,1H),5.40(d,J＝1.0Hz,1H),5.22(d,J＝1.3Hz,1H),4.85(d,J＝1.5Hz,1H),4.76(q,J＝8.7Hz,1H),4.07(dd,J＝4.3,1.5Hz, 1H),3.88-3.81(m,1H),3.80-3.69(m,1H),3.58(d,J＝1.3Hz,3H),3.55(d,J＝1.3Hz,3H),3.42(dt,J＝9.7,7.0Hz,1H),2.97(dt,J＝9.5,1.3Hz,1H),2.83- 2 .76(m,2H),2.06-2.01(m,1H),1.72(dd,J＝14.2,6.2,4.3Hz,1H),1.51(d,J＝3.5Hz,1H),1.48(q,J＝2.4Hz,2H),1.43-1.41(m,2H),1.41-1.39(m,1H), 1.37(d,J＝5.9Hz,1H),1.34(dd,J＝6.8,2.3Hz,2H),1.25(s,1H),1.24(s,1H),1.23(d,J＝2.9Hz,2H),1.21(d,J＝3.1Hz,1H),1.19-1.10(m,2H),0.97(s, 3H),0.87(s,3H). 13 C NMR (101 MHz, CDCl3) δ 205.35, 150.86, 117.46, 99.79, 77.24, 71.40, 66.35, 65.17, 63.94, 58.15, 56.85, 52.79, 52.43, 48.45, 42.07, 41.46, 40.55, 39.84, 33.98, 32.80, 30.45, 29.69, 24.58, 20.93, 18.23, 15.47, 15.43. HR-MS (ESI): calculated for C 27 H 44 NO 8 P [M + Na] + : 564.2697, found 564.2692.

实施例13：Embodiment 13:

化合物Ⅱ-8的制备Preparation of Compound II-8

以化合物3d代替化合物3a，加入亚磷酸三乙酯，其他操作同实施例6，产率为62％。1HNMR(400MHz,Chloroform-d)δ5.40(d,J＝1.0Hz,1H),5.22(d,J＝1.2Hz,1H),4.85(d,J＝1.5Hz,1H),4.76(q,J＝8.7Hz,1H),4.13-4.10(m,1H),4.10-4.03(m,2H),3.94-3.89(m,4H),3.87-3.82(m,1H),3.75-3.69(m,1H),3.55-3.46(m,1H),3.46-3.41(m,1H),2.97(dt,J＝9.6,1.3Hz,1H),2.84-2.77(m,2H),2.06-1.98(m,2H),1.74-1.68(m,1H),1.51(d,J＝2.3Hz,1H),1.49-1.46(m,3H),1.45(d,J＝0.9Hz,1H),1.43-1.39(m,3H),1.37(d,J＝2.5Hz,1H),1.36-1.33(m,3H),1.32(d,J＝1.0Hz,1H),1.30(d,J＝2.3Hz,1H),1.28(d,J＝5.3Hz,1H),1.26(s,1H),1.24(d,J＝7.0Hz,3H),1.22-1.19(m,3H),0.97(s,3H),0.87(s,3H).13C NMR(101MHz,CDCl3)δ206.15,148.35,116.62,99.36,77.25,73.12,66.94,64.24,63.75,58.08,57.29,52.80,52.43,50.57,48.88,42.08,41.47,40.55,39.84,33.98,32.80,30.45,29.73,27.21,24.58,20.93,18.23,16.16,16.09.HR-MS(ESI):计算值C29H48NO8P[M+Na]+:592.3010,实测值592.2993.Compound 3d was used instead of compound 3a, triethyl phosphite was added, and the other operations were the same as those in Example 6. The yield was 62%. HNMR(400MHz,Chloroform-d)δ5.40(d,J＝1.0Hz,1H),5.22(d,J＝1.2Hz,1H),4.85(d,J＝1.5Hz,1H),4.76(q,J＝8.7Hz,1H),4.13-4.10(m,1H),4.10-4.03(m,2H),3 .94-3.89(m,4H),3.87-3.82(m,1H),3.75-3.69(m,1H),3.55-3.46(m,1H),3.46-3.41(m,1H),2.97(dt,J＝9.6,1.3Hz,1H),2.84-2.77(m,2H),2.06 -1.98(m,2H),1.74-1.68(m,1H),1.51(d,J＝2.3Hz,1H),1.49-1.46(m,3H),1.45(d,J＝0.9Hz,1H),1.43-1.39(m,3H),1.37(d,J＝2.5Hz,1H),1.36-1.3 3(m,3H),1.32(d,J＝1.0Hz,1H),1.30(d,J＝2.3Hz,1H),1.28(d,J＝5.3Hz,1H),1.26(s,1H),1.24(d,J＝7.0Hz,3H),1.22-1.19(m,3H),0.97(s,3H),0.8 7(s,3H). 13 C NMR (101 MHz, CDCl3) δ 206.15, 148.35, 116.62, 99.36, 77.25, 73.12, 66.94, 64.24, 63.75, 58.08, 57.29, 52.80, 52.43, 50.57, 48.88, 42.08, 41.47, 40.55, 39.84, 33.98, 32.80, 30.45, 29.73, 27.21, 24.58, 20.93, 18.23, 16.16, 16.09. HR-MS (ESI): calculated for C 29 H 48 NO 8 P [M+Na] + : 592.3010, found 592.2993.

实施例14：Embodiment 14:

化合物Ⅱ-9的制备Preparation of Compound II-9

以化合物3e代替化合物3a，其他操作同实施例6，产率为65％。1H NMR(400MHz,Chloroform-d)δ6.04-6.02(m,1H),5.41-5.39(m,1H),5.21(dd,J＝6.0,1.3Hz,1H),4.84(dd,J＝21.9,1.4Hz,1H),4.76(dd,J＝9.7,8.5Hz,1H),4.26-4.15(m,1H),4.09-4.04(m,4H),3.85(dd,J＝9.6,7.1,5.4Hz,1H),3.75-3.63(m,1H),3.43(dq,J＝9.5,7.0Hz,1H),3.33(dt,J＝9.5,6.1Hz,1H),3.00(d,J＝8.3Hz,1H),2.96-2.90(m,2H),2.84-2.74(m,3H),1.77-1.73(m,1H),1.73-1.68(m,1H),1.48(d,J＝3.2Hz,2H),1.38(s,2H),1.37(d,J＝1.6Hz,1H),1.32(d,J＝1.5Hz,6H),1.29(d,J＝1.8Hz,1H),1.25(d,J＝3.3Hz,2H),1.24-1.20(m,2H),0.97(s,3H),0.87(s,3H).13C NMR(101MHz,CDCl3))δ206.25,152.79,116.50,99.59,70.12,67.52,65.54,63.61,57.82,56.77,52.73,52.35,51.03,48.91,43.03,42.43,40.90,39.85,34.17,33.17,30.54,29.65,28.74,26.26,25.79,24.79,21.21,18.28.HR-MS(ESI):计算值C28H46NO8P[M+Na]+:578.2853,实测值578.2849.Compound 3e was used to replace compound 3a. Other operations were the same as in Example 6. The yield was 65%. 1 H NMR (400 MHz, Chloroform-d) δ 6.04-6.02 (m, 1H), 5.41-5.39 (m, 1H), 5.21 (dd, J = 6.0, 1.3 Hz, 1H), 4.84 (dd, J = 21.9, 1.4 Hz, 1H), 4.76 (dd, J = 9.7, 8.5 Hz, 1H), 4.26-4.15 (m, 1H), 4.09-4.04 (m, 4H), 3.85 (dd, J = 9.6, 7.1, 5.4 Hz, 1H), 3.75-3.63 (m, 1H), 3.43 (dq, J = 9.5, 7.0 Hz, 1H), 3.33 (dt, J = 9.5,6.1Hz,1H),3.00(d,J＝8.3Hz,1H),2.96-2.90(m,2H),2.84-2.74(m,3H),1.77-1.73(m,1H),1.73-1.68(m,1H),1.48(d,J＝3.2Hz,2H),1.38(s,2H ),1.37(d,J＝1.6Hz,1H),1.32(d,J＝1.5Hz,6H),1.29(d,J＝1.8Hz,1H),1.25(d,J＝3.3Hz,2H),1.24-1.20(m,2H),0.97(s,3H),0.87(s,3H). 13 C NMR (101 MHz, CDCl3) δ 206.25, 152.79, 116.50, 99.59, 70.12, 67.52, 65.54, 63.61, 57.82 , 56.77, 52.73, 52.35, 51.03, 48.91, 43.03, 42.43, 40.90, 39.85, 34.17, 33.17, 30.54, 29.65, 28.74, 26.26, 25.79, 24.79, 21.21, 18.28. HR-MS (ESI): calculated for C 28 H 46 NO 8 P [M+Na] + : 578.2853, found 578.2849.

实施例15：Embodiment 15:

化合物Ⅱ-10的制备Preparation of Compound II-10

以化合物3e代替化合物3a，加入亚磷酸三乙酯，其他操作同实施例6，产率为68％。1HNMR(400MHz,DMSO-d6)δ5.79(s,1H),5.37(d,J＝1.2Hz,1H),5.11(d,J＝1.1Hz,1H),5.07(d,J＝2.8Hz,1H),4.81-4.66(m,2H),4.54(t,J＝2.0Hz,1H),4.51-4.45(m,1H),4.17(d,J＝7.1Hz,1H),3.98(dt,J＝4.0,1.4Hz,2H),3.93-3.89(m,4H),3.70-3.65(m,1H),3.01-2.91(m,3H),2.80(d,J＝9.1Hz,1H),2.70-2.63(m,1H),2.62-2.55(m,1H),2.48-2.38(m,1H),2.08-1.98(m,2H),1.56(ddd,J＝13.8,6.5,4.4Hz,2H),1.45-1.40(m,2H),1.38-1.34(m,3H),1.26(d,J＝1.5Hz,1H),1.24(s,2H),1.23(s,1H),1.22(d,J＝0.8Hz,6H),1.21-1.20(m,3H),0.92(s,3H),0.82(s,3H).13C NMR(101MHz,DMSO-d6)δ206.23,152.80,116.51,99.58,70.12,67.52,65.54,63.61,57.82,56.77,52.73,52.35,51.03,48.91,43.03,42.43,40.90,39.85,34.17,33.17,30.54,29.65,28.74,26.26,25.79,24.79,21.21,18.28.16.63,16.54HR-MS(ESI):计算值C30H50NO8P[M+Na]+:606.3166,实测值606.3147.Compound 3e was used instead of compound 3a, triethyl phosphite was added, and the other operations were the same as those in Example 6. The yield was 68%. 1 HNMR (400 MHz, DMSO-d 6 )δ5.79(s,1H),5.37(d,J＝1.2Hz,1H),5.11(d,J＝1.1Hz,1H),5.07(d,J＝2.8Hz,1H),4.81-4.66(m,2H),4.54(t,J＝2.0Hz,1H),4.51-4.45(m,1H),4.17( d,J＝7.1Hz,1H),3.98(dt,J＝4.0,1.4Hz,2H),3.93-3.89(m,4H),3.70-3.65(m,1H),3.01-2.91(m,3H),2.80(d,J＝9.1Hz, 1H),2.70-2.63(m,1H),2.62-2.55(m,1H),2.48-2.38(m,1H),2.08-1.98(m,2H),1.56(ddd,J＝13.8,6.5,4.4Hz,2H),1.45-1.40(m,2H),1.38-1.34 (m,3H),1.26(d,J＝1.5Hz,1H),1.24(s,2H),1.23(s,1H),1.22(d,J＝0.8Hz,6H),1.21-1.20(m,3H),0.92(s,3H),0.82(s,3H). 13 C NMR (101 MHz, DMSO-d 6 ) δ 206.23, 152.80, 116.51, 99.58, 70.12, 67.52, 65.54, 63.61, 57.82, 56.77, 52.73, 52.35, 51.03, 48.91, 43.03, 42.43, 40.90, 39.85, 34.17, 33.17, 30.54, 29.65, 28.74, 26.26, 25.79, 24.79, 21.21, 18.28. 16.63, 16.54 HR-MS (ESI): calcd. for C 30 H 50 NO 8 P [M+Na] + :606.3166, measured value 606.3147.

实施例16：Embodiment 16:

化合物Ⅲ-1的制备Preparation of compound III-1

称取200mg JOA溶解于超干THF中，冰浴条件下加入145mg二氯化磷酸苯酯，加入116mg的三乙胺，反应5-6h，旋干，用乙酸乙酯和饱和碳酸氢钠萃取，干燥浓缩，柱色谱纯化，得白色固体，产率为74％。1H NMR(400MHz,DMSO-d6)δ10.25(s,1H),7.32(t,J＝7.8Hz,2H),7.20(t,J＝7.3Hz,1H),6.99(d,J＝8.0Hz,2H),5.64(s,1H),5.33(s,1H),5.13(s,1H),4.91(ddd,J＝21.0,12.4,4.7Hz,1H),4.79(d,J＝2.3Hz,1H),3.89-3.82(m,1H),3.33(d,J＝10.3Hz,2H),3.18(s,1H),2.65-2.56(m,1H),2.43-2.32(m,2H),1.91-1.85(m,1H),1.79-1.76(m,1H),1.68-1.61(m,1H),1.48(dd,J＝13.2,2.2Hz,1H),1.44-1.39(m,1H),1.34(d,J＝11.3Hz,1H),1.22(s,1H),1.19(s,1H),0.97(s,3H),0.71(s,3H).13C NMR(101MHz,DMSO-d6)δ206.31,200.04,149.88,145.84,129.60,125.30,119.90,116.27,82.36,79.69,61.95,61.68,54.50,51.62,50.21,42.68,40.42,36.68,33.24,33.09,31.16,25.89,20.69,18.52.HR-MS(ESI):计算值C26H30O7P[M+H]+:485.1724,实测值485.1714.Weigh 200 mg of JOA and dissolve it in ultra-dry THF. Add 145 mg of phenyl dichlorophosphate and 116 mg of triethylamine under ice bath conditions. React for 5-6 hours, spin dry, extract with ethyl acetate and saturated sodium bicarbonate, dry and concentrate, and purify by column chromatography to obtain a white solid with a yield of 74%. 1 H NMR (400 MHz, DMSO-d 6 )δ10.25(s,1H),7.32(t,J＝7.8Hz,2H),7.20(t,J＝7.3Hz,1H),6.99(d,J＝8.0Hz,2H),5.64(s,1H),5.33(s,1H),5.13(s,1H),4.91(ddd,J＝21.0,12.4,4 .7Hz,1H),4.79(d,J＝2.3Hz,1H),3.89-3.82(m,1H),3.33(d,J＝10.3Hz,2H),3.1 8(s,1H),2.65-2.56(m,1H),2.43-2.32(m,2H),1.91-1.85(m,1H),1.79-1.76(m,1H),1.68-1.61(m,1H),1.48(dd,J＝13.2,2.2Hz,1H),1.44-1.39(m ,1H),1.34(d,J=11.3Hz,1H),1.22(s,1H),1.19(s,1H),0.97(s,3H),0.71(s,3H). 13 C NMR (101 MHz, DMSO-d 6 ) δ 206.31, 200.04, 149.88, 145.84, 129.60, 125.30, 119.90, 116.27, 82.36, 79.69, 61.95, 61.68, 54.50, 51.62, 50.21, 42.68, 40.42, 36.68, 33.24, 33.09, 31.16, 25.89, 20.69, 18.52. HR-MS (ESI): calculated for C 26 H 30 O 7 P [M+H] + : 485.1724, found 485.1714.

实施例17：Embodiment 17:

化合物Ⅲ-2的制备Preparation of compound III-2

以二氯化磷酸乙酯替代二氯化磷酸苯酯，其他操作同实施例16，得白色固体，产率为72％。1H NMR(400MHz,DMSO-d6)δ10.19(s,1H),5.82(s,1H),5.40(s,1H),4.75(s,1H),4.75-4.67(m,1H),3.94-3.89(m,2H),3.86(d,J＝4.1Hz,1H),3.10(s,1H),2.69(t,J＝13.1Hz,1H),2.39(d,J＝12.8Hz,1H),2.14(ddd,J＝13.6,5.2,2.4Hz,1H),1.92-1.85(m,1H),1.72(d,J＝1.6Hz,1H),1.64-1.59(m,1H),1.45-1.40(m,2H),1.33(t,J＝11.7Hz,2H),1.24(d,J＝7.0Hz,3H),1.23(d,J＝4.7Hz,1H),1.19(s,1H),1.18-1.11(m,1H),0.95(s,3H),0.69(s,3H).13C NMR(101MHz,DMSO-d6)δ205.73,200.90,146.51,115.73,80.51,77.81,61.69,61.54,54.24,51.69,50.33,42.54,40.40,39.46,36.48,33.19,32.86,31.12,24.64,20.56,18.50,16.32.HR-MS(ESI):计算值C22H30O7P[M+H]+:437.1724,实测值437.1716.Ethyl dichlorophosphate was used to replace phenyl dichlorophosphate. Other operations were the same as in Example 16 to obtain a white solid with a yield of 72%. 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.19 (s, 1H), 5.82 (s, 1H), 5.40 (s, 1H), 4.75 (s, 1H), 4.75-4.67 (m, 1H), 3.94-3.89 (m, 2H), 3.86 (d, J = 4.1 Hz, 1H), 3.10 (s, 1H), 2.69 (t, J = 13.1 Hz, 1H), 2.39 (d, J = 12.8 Hz, 1H), 2.14 (ddd, J = 13.6, 5.2, 2.4 Hz, 1 H),1.92-1.85(m,1H),1.72(d,J＝1.6Hz,1H),1.64-1.59(m,1H),1.45-1.40(m,2H),1.33(t,J＝11.7Hz,2H),1.24(d,J＝7.0Hz,3H),1.23(d,J＝4.7Hz,1H) ,1.19(s,1H),1.18-1.11(m,1H),0.95(s,3H),0.69(s,3H). 13 C NMR (101 MHz, DMSO-d 6 ) δ 205.73, 200.90, 146.51, 115.73, 80.51, 77.81, 61.69, 61.54, 54.24, 51.69, 50.33, 42.54, 40.40, 39.46, 36.48, 33.19, 32.86, 31.12, 24.64, 20.56, 18.50, 16.32. HR-MS (ESI): calculated for C 22 H 30 O 7 P [M+H] + : 437.1724, found 437.1716.

实施例18：Embodiment 18:

化合物Ⅲ-3的制备Preparation of compound III-3

以二氯化磷酸氯乙酯替代二氯化磷酸苯酯，其他操作同实施例16，得白色固体，产率为67％。1H NMR(400MHz,DMSO-d6)δ10.20(s,1H),5.82(s,1H),5.40(s,1H),4.76(d,J＝1.8Hz,1H),4.76-4.67(m,1H),3.98(dt,J＝4.0,1.4Hz,2H),3.86(d,J＝4.1Hz,1H),3.70-3.65(t,2H),3.10(s,1H),2.70(q,J＝13.2Hz,1H),2.39(d,J＝13.0Hz,1H),2.14(ddd,J＝13.5,5.1,2.2Hz,1H),1.92-1.85(m,1H),1.72(d,J＝1.6Hz,1H),1.62(ddd,J＝14.6,4.5,2.7Hz,1H),1.44-1.39(m,2H),1.34(d,J＝11.3Hz,1H),1.28(d,J＝15.6Hz,1H),1.23(t,J＝3.5Hz,1H),1.19(s,1H),0.95(s,3H),0.69(s,3H).13C NMR(101MHz,DMSO-d6)δ206.27,201.42,147.03,116.25,81.11,78.41,62.19,62.04,55.38,54.75,52.20,50.85,43.05,42.97,40.92,39.86,36.99,33.71,31.64,25.16,21.06,19.01.3.98(dt,J＝4.0,1.4Hz,2H).HR-MS(ESI):计算值C22H29ClO7P[M+H]+:471.1334,实测值471.1323.Substituting dichloroethyl phosphate for dichlorophenyl phosphate, the other operations were the same as in Example 16 to obtain a white solid with a yield of 67%. 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.20 (s, 1H), 5.82 (s, 1H), 5.40 (s, 1H), 4.76 (d, J = 1.8 Hz, 1H), 4.76-4.67 (m, 1H), 3.98 (dt, J = 4.0, 1.4 Hz, 2H), 3.86 (d, J = 4.1 Hz, 1H), 3.70-3.65 (t, 2H), 3.10 (s, 1H), 2.70 (q, J = 13.2 Hz, 1H), 2.39 (d, J = 13.0 Hz, 1H), 2.14 (ddd, J = 13.5,5.1,2.2Hz,1H),1.92-1.85(m,1H),1.72(d,J＝1.6Hz,1H),1.62(ddd,J＝14.6,4.5,2.7Hz,1H),1.44-1.39(m,2H),1.34(d,J＝11.3Hz,1H),1.28(d ,J＝15.6Hz,1H),1.23(t,J＝3.5Hz,1H),1.19(s,1H),0.95(s,3H),0.69(s,3H). 13 C NMR (101 MHz, DMSO-d 6 ) δ 206.27, 201.42, 147.03, 116.25, 81.11, 78.41, 62.19, 62.04, 55.38, 54.75, 52.20, 50.85, 43.05, 42.97, 40.92, 39.86, 36.99, 33.71, 31.64, 25.16, 21.06, 19.01.3.98 (dt, J=4.0, 1.4 Hz, 2H). HR-MS (ESI): calculated for C 22 H 29 ClO 7 P [M+H] + : 471.1334, found 471.1323.

实施例19：本发明合成的济源冬凌草甲素含磷系列衍生物对人食管癌细胞Eca-109，人胃癌细胞MGC-803，人胰腺癌细胞PANC-1、SW-1990，人直肠癌细胞HT-29，人肝癌细胞HepG-2的抗肿瘤活性。Example 19: Antitumor activity of the phosphorus-containing series of derivatives of Rubescensine A synthesized by the present invention against human esophageal cancer cells Eca-109, human gastric cancer cells MGC-803, human pancreatic cancer cells PANC-1, SW-1990, human rectal cancer cells HT-29, and human liver cancer cells HepG-2.

实验方法：以下百分含量若没有特别说明均为质量百分含量Experimental method: The following percentages are by mass unless otherwise specified.

以济源冬凌草甲素和冬凌草甲素为阳性对照，选用以上六种癌细胞测定所合成的化合物72h抗增殖活性：将对数生长期的细胞用含0.25％ EDTA的胰酶消化之后配制成浓度为1-10×104个/mL的细胞悬液，按照1000-10000个细胞/孔接种于96孔板中，每孔快速顶壁加入100μL细胞悬液，接种完毕后，将96孔板放置在体积百分含量5％ CO2的37℃湿度培养箱中。24h后弃干净板中上清液，每孔加入含有不同药物浓度的培养液200μL，每个化合物按照一定浓度梯度设置三个平行孔，对照组加空白的培养液200μL，再放入体积百分含量5％CO2的37℃湿度培养箱中培养72h。每孔加入100μL 30％ TCA溶液于4℃下固定60min；用去离子水洗3-4次，风干；每孔加入0.4％ SRB 100μL室温染色30min；用1％的乙酸洗3次，风干；每孔加入150μL的Tris base后用平板摇床摇15min,待摇匀后，使用酶标仪测定吸光密度值(OD),检测波长540nm。以溶剂对照处理细胞为对照组，按下述公式计算化合物对细胞的抑制率，并按中效方程计算半数抑制浓度IC50：抑制率(％)＝(对照组OD均值-给药组OD均值)/对照组OD均值*100％With Jiyuan Rubescensine A and Rubescensine A as positive controls, the above six cancer cells were selected to determine the 72h antiproliferative activity of the synthesized compounds: the cells in the logarithmic growth phase were digested with trypsin containing 0.25% EDTA and then prepared into a cell suspension with a concentration of 1-10×10 4 /mL, and inoculated in a 96-well plate at 1000-10000 cells/well, and 100μL of cell suspension was quickly added to the top wall of each well. After inoculation, the 96-well plate was placed in a 37°C humidity incubator with a volume percentage of 5% CO 2. After 24h, the supernatant in the clean plate was discarded, and 200μL of culture solution containing different drug concentrations was added to each well. Three parallel wells were set for each compound according to a certain concentration gradient, and 200μL of blank culture solution was added to the control group, and then placed in a 37°C humidity incubator with a volume percentage of 5% CO 2 for 72h. Add 100 μL of 30% TCA solution to each well and fix at 4°C for 60 min; wash with deionized water 3-4 times and air dry; add 100 μL of 0.4% SRB to each well and stain at room temperature for 30 min; wash with 1% acetic acid 3 times and air dry; add 150 μL of Tris base to each well and shake on a flatbed shaker for 15 min. After shaking well, use an enzyme marker to measure the absorbance density (OD) at a wavelength of 540 nm. The solvent control treated cells were used as the control group, and the inhibition rate of the compound on the cells was calculated according to the following formula, and the half-maximal inhibitory concentration IC 50 was calculated according to the median effect equation: Inhibition rate (%) = (control group OD mean - drug group OD mean) / control group OD mean * 100%

济源冬凌草甲素含磷衍生物抗肿瘤活性Antitumor activity of phosphorus-containing derivatives of Rubescensine A from Jiyuan

ORI为冬凌草甲素；JOA为济源冬凌草甲素。ORI is Rubescensine A; JOA is Jiyuan Rubescensine A.

上述实验结果表明：本发明所述化合物具有较良好的体外抗肿瘤活性，可用于制备抗肿瘤药物，应用于临床治疗食管癌、胃癌、直肠癌、胰腺癌、肝癌等。以本发明化合物作为活性成分用于制备新的抗癌药物，具有潜在的应用价值。The above experimental results show that the compounds of the present invention have good in vitro antitumor activity and can be used to prepare antitumor drugs for clinical treatment of esophageal cancer, gastric cancer, rectal cancer, pancreatic cancer, liver cancer, etc. The compounds of the present invention are used as active ingredients to prepare new anticancer drugs, which have potential application value.

药物稳定性试验Drug stability testing

根据《中国药典》2020年版药物稳定性试验指导原则，对抗肿瘤活性最好的化合物Ⅲ-1进行稳定性试验，结果如下表所示：According to the guidelines for drug stability testing of the 2020 edition of the Chinese Pharmacopoeia, compound III-1 with the best anti-tumor activity was subjected to a stability test. The results are shown in the following table:

化合物Ⅲ-1影响因素试验(1批供试品)Compound III-1 influencing factor test (1 batch of test products)

化合物Ⅲ-1加速试验和长期试验(3批供试品)Accelerated test and long-term test of compound III-1 (3 batches of test products)

Claims (4)

1.济源冬凌草甲素含磷系列衍生物，其特征在于，具有通式所示结构：1. Jiyuan oridonin A series of phosphorus-containing derivatives is characterized by having a structure shown by the general formula: 其中n为1-5；R1为C1-5烷基；R2为苯基、C1-5烷基或C1-5卤代烷基。wherein n is 1-5; R 1 is C1-5 alkyl; R 2 is phenyl, C1-5 alkyl or C1-5 haloalkyl. 2.如权利要求1所述的济源冬凌草甲素含磷系列衍生物，其特征在于，n为1-5；R1为甲基或者乙基；R2为苯基、乙基或氯乙基。2. Jiyuan oridonin A phosphorus-containing series derivatives as claimed in claim 1, characterized in that n is 1-5; R 1 is methyl or ethyl; R 2 is phenyl, ethyl or chlorine Ethyl. 3.如权利要求1所述的济源冬凌草甲素含磷系列衍生物，其特征在于，选自如下化合物：3. Jiyuan oridonin A phosphorus-containing series derivatives as claimed in claim 1, characterized in that it is selected from the following compounds: 。 . 4.如权利要求1-3任一所述的济源冬凌草甲素含磷系列衍生物在制备药物中的应用，其特征在于，以其为活性成分，制备治疗食管癌、胰腺癌、胃癌、结肠癌或肝癌药物。4. Application of Jiyuan oridonin A phosphorus-containing series derivatives in the preparation of medicines as described in any one of claims 1 to 3, characterized in that it is used as an active ingredient to prepare and treat esophageal cancer, pancreatic cancer, and gastric cancer. , colon cancer or liver cancer drugs.
CN202210538036.XA 2022-05-17 2022-05-17 Jiyuan oridonin phosphorus-containing serial derivatives, preparation method and application thereof Active CN114920777B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202210538036.XA CN114920777B (en) 2022-05-17 2022-05-17 Jiyuan oridonin phosphorus-containing serial derivatives, preparation method and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202210538036.XA CN114920777B (en) 2022-05-17 2022-05-17 Jiyuan oridonin phosphorus-containing serial derivatives, preparation method and application thereof

Publications (2)

Publication Number Publication Date
CN114920777A CN114920777A (en) 2022-08-19
CN114920777B true CN114920777B (en) 2023-11-17

Family

ID=82809353

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202210538036.XA Active CN114920777B (en) 2022-05-17 2022-05-17 Jiyuan oridonin phosphorus-containing serial derivatives, preparation method and application thereof

Country Status (1)

Country Link
CN (1) CN114920777B (en)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106749305A (en) * 2016-11-16 2017-05-31 中国药科大学 Oridonin derivative, the Preparation Method And The Use of A- rings transformation
CN108299458A (en) * 2017-12-28 2018-07-20 青岛海洋生物医药研究院股份有限公司 Oridonin derivative and its preparation method and application
CN112979671A (en) * 2021-03-05 2021-06-18 郑州大学 Oridonin-naphthalimide fluorescent probe as well as preparation method and application thereof

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106749305A (en) * 2016-11-16 2017-05-31 中国药科大学 Oridonin derivative, the Preparation Method And The Use of A- rings transformation
CN108299458A (en) * 2017-12-28 2018-07-20 青岛海洋生物医药研究院股份有限公司 Oridonin derivative and its preparation method and application
CN112979671A (en) * 2021-03-05 2021-06-18 郑州大学 Oridonin-naphthalimide fluorescent probe as well as preparation method and application thereof

Also Published As

Publication number Publication date
CN114920777A (en) 2022-08-19

Similar Documents

Publication Publication Date Title
KR20150046315A (en) Tenofovir prodrug and pharmaceutical uses thereof
JP6679624B2 (en) Tenofovir monobenzyl ester phosphamide prodrug, process for its preparation and use
CN104860993B (en) A kind of chromocor compound prodrug and application thereof
KR20080108322A (en) Method for preparing HIV protease inhibitor
CN102286047A (en) 2'-deoxidized-2'-fluorin-4'-triazole substituted-beta-D cytidine analogue as well as preparation method and application thereof
CN103275051B (en) A kind of 7,3 ', 4 '-trihydroxyflavone derivative and preparing the application in Hepatoma therapy medicine
CN102603836A (en) Schisandrin C simplifier, schisandrin analogue, preparation method and applications thereof
CN111655710B (en) Gemcitabine phosphorus-containing prodrugs
CN114380864B (en) A dihydroartemisinin derivative, preparation method, pharmaceutical composition and its application in the preparation of antitumor drugs
JP6890132B2 (en) Uridine phosphoramides for antivirals, their preparation methods and their use in medicine
CN114920777B (en) Jiyuan oridonin phosphorus-containing serial derivatives, preparation method and application thereof
AU618536B2 (en) Novel 3',4'-dinitrogen substituted epipodophyllotoxin glucoside derivatives
CN106317030B (en) A kind of 4- indyl coumarin derivative and its preparation method and application
CN104987356A (en) Ursolic acid-glycolysis inhibitor DCA conjugate and application thereof
CN110903289B (en) Purine-aminomethyl-pyridone derivatives and their preparation methods and uses
CN108658957B (en) A substituted chromenol ester compound and its application in the preparation of anticancer drugs
CN109867708B (en) Preparation method and application of glycyrrhetinic acid series derivatives (TNGA-X) with anti-tumor effect
CN114213435B (en) Preparation and application of bis- (10-deoxydihydroartemisinin) -phloroglucinol derivative
CN116102608B (en) A stigmasterol triazole derivative and its preparation method and application
CN103919770A (en) Application of curcumin analog S5 containing thiapyrone structure in preparation of anti-inflammation drugs
KR102378151B1 (en) New derivatives of β-apopicropodophyllin
CN106496271A (en) The preparation method and purposes of quinolizidine kind alkaloid derivant
CN113549106A (en) A kind of compretin derivative and its preparation method and application
CN112645959A (en) Nitric ester NO donor type hematoporphyrin derivative and preparation method and application thereof
CN103739597A (en) 14-deoxy-14,15-didehydro andrographolide derivatives and medicine composition and application thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant