WO2020098658A1

WO2020098658A1 - 20-substituted camptothecin derivative, preparation method therefor and application thereof

Info

Publication number: WO2020098658A1
Application number: PCT/CN2019/117646
Authority: WO
Inventors: 江涛; 张海霖; 吴贯召; 张逸轩; 李静; 戚欣
Original assignee: 中国海洋大学
Priority date: 2018-11-12
Filing date: 2019-11-12
Publication date: 2020-05-22
Also published as: CN111171041B; CN111171041A

Abstract

The present invention provides a compound having a structure shown in formula (I), an isomer thereof, a pharmaceutically acceptable salt thereof, a preparation method therefor, and a pharmaceutical composition comprising said compound. Provided is an application of the present compound and pharmaceutical composition thereof in preparation of a drug for preventing and/or treating a cancer. The present compound has a good ex-vivo cytotoxic activity and a good in-vivo antitumor effect. The preparation method for the compound is easy and raw materials are easy to obtain. Therefore, the present compound has a wide medicinal prospect. (I)

Description

20-position substituted camptothecin derivatives and preparation method and application thereof

Technical field

The invention belongs to the technical field of medicine, and specifically relates to a new type of camptothecin derivative and a preparation method and application thereof.

Background technique

Camptothecin derivatives are known as one of the three major discoveries of anti-cancer drugs in the 1990s, showing broad-spectrum anti-tumor activity and great research and application value. The early camptothecin compounds formed a ternary complex with Top I and DNA to block DNA replication and transcription, and had the disadvantages of low selectivity and toxic side effects. With the development of science and technology, molecular targeted anti-tumor drugs have become a research hotspot in recent years. Such drugs can selectively act on signal transduction pathways related to tumor cell differentiation and proliferation. -Methylenedioxycamptothecin is the starting compound, which is structurally modified to develop more efficient and less toxic antitumor candidate compounds.

Summary of the invention

The present invention provides a camptothecin derivative and its preparation method and application in anti-tumor, which can solve the shortcomings of low selectivity and large toxic and side effects existing in the prior art, and can be used to prepare drugs for preventing or treating tumors.

In order to solve the above technical problems, on the one hand, the present invention provides a compound having the structure shown in formula (I), its isomer and a pharmaceutically acceptable salt:

Where X is-(CH ₂ ) _n -,-(CH ₂ ) _n O-,-(CH ₂ ) _n NH-,-(CH ₂ ) _n CONH-,-(CH ₂ ) _n NHCO (CH ₂ ) _m- , (CH ₂ ) _n- (CH = CH) _p- (CH ₂ ) _m -,-(CH ₂ ) _n- (CH = CH) _p- (CH ₂ ) _m O-, -O (CH ₂ ) _n SS (CH ₂ ) _m O- or -O (CH ₂ ) _n SS (CH ₂ ) _m OCOY-;

Y is - (CH ₂₎ _n - or - (CH ₂₎ _n CONH-;

n and m are each independently 0 or a positive integer, preferably, n and m are each independently a positive integer, and more preferably, n and m are each independently 1, 2, 3, 4, 5, 6, 7 , 8, 9 or 10;

p is a positive integer, preferably, p is 1 or 2;

R is selected from any one of a hydrogen atom, a substituted or unsubstituted sulfonamide group, a substituted or unsubstituted base, a substituted or unsubstituted saturated nitrogen-containing heterocyclic ring, and a substituted or unsubstituted alkyl group; preferably Wherein R is any one of a substituted or unsubstituted sulfonamide group, a substituted or unsubstituted base group, a substituted or unsubstituted saturated nitrogen-containing heterocyclic ring, and a substituted or unsubstituted alkyl group.

When the group is a substituted group, the substituent is selected from hydrogen atom, halogen atom, hydroxyl group, nitro group, amino group, alkyl group, substituted alkyl group, alkoxy group, substituted alkoxy group, aromatic Any one of a group, a substituted aryl group, a heteroaryl group and a substituted heteroaryl group.

In a preferred embodiment, X in formula (I) is-(CH ₂ ) _n NH-, n is 1, R is hydrogen or alkyl, preferably, the compound of formula (I) is compound 2 or compound 3 .

In a preferred embodiment, in formula (I), X is-(CH ₂ ) _n NH-, n is 1, R is a substituted or unsubstituted sulfonamide group, and R has the structure shown in formula (II) :

among them,

R ₁ is alkyl, aryl, substituted aryl, heteroaryl or substituted heteroaryl; preferably R ₁ is C1-C4 alkyl, more preferably R ₁ is methyl, ethyl, n-propyl Or n-butyl; likewise preferably R ₁ can be phenyl, substituted phenyl, naphthyl, thiophene or pyridine, more preferably R ₁ is para-fluorine substitution, chlorine substitution, nitro substitution, methyl substitution or methyl Oxygen substituted phenyl;

R ₂ is phenyl or substituted phenyl, preferably para-fluorine substituted or methoxy substituted phenyl;

Particularly preferably, the compound of formula (I) is w-1, w-2, w-3, w-4, w-5, w-6, w-7, w-8, w-9, w-10, w-11, w-12, w-13, w-14, w-15, w-16, w-17, w-18, w-19, w-20, w-21, w-22, w- 23 or w-24.

The preparation method of such compounds includes the following steps:

1) 10,11-Methylenedioxycamptothecin undergoes an acylation reaction with Boc-protected glycine under the catalysis of a condensing agent, and the resulting product is deprotected by trifluoroacetic acid to obtain a 20-glycine modified 10,11-Methylenedioxycamptothecin;

2) The 20,10-glycine-modified 10,11-methylenedioxycamptothecin is catalyzed by cuprous iodide with one-pot reaction of benzenesulfonyl azide and alkynyl group containing different substituents to obtain the compound.

In another preferred embodiment of formula (I) wherein X is _{_{- (CH 2) n NHCO (}} CH 2) m -, n and m are both 1, R is a substituted or unsubstituted uracil, and R It has the structure shown in formula (III):

Wherein R ₃ is H, F or Cl.

In another preferred example, said R, X, Y, R ₁ , R ₂ and R ₃ are corresponding groups in each specific compound in the examples.

Particularly preferably, the compound of formula (I) is w-25, w-26 or w-27.

The preparation method of such compounds includes the following steps:

2) The above-mentioned 10,11-methylenedioxycamptothecin modified with glycine at position 20 is reacted with uracil acetic acid modified with different substituents under the catalysis of a condensing agent to obtain the compound.

In another preferred embodiment of formula (I) wherein X is _{_{- (CH 2) n NHCO (}} CH 2) m -, n is 1, m is 0, R is a substituted or unsubstituted saturated nitrogen-containing Ring; Preferably, the compound of formula (I) is w-28, w-29, w-30, w-31, w-32 or w-33.

The preparation method of such compounds includes the following steps:

2) The 10,11-methylenedioxycamptothecin modified with glycine at position 20 is obtained as described above, and acylation reaction with carbonyldiimidazole and heterocyclic amine is carried out under the catalysis of triethylamine to obtain the compound.

In another preferred embodiment of formula (I) wherein X is - (CH ₂₎ _n - or _{_{- (CH 2) n CONH-,}} n is 2, R is a substituted or unsubstituted alkyl group; preferably The compound of formula (I) is z-1-9 or z-2-5.

The preparation method of such compounds includes the following steps:

10,11-Methylenedioxycamptothecin, catalyzed by a condensing agent, undergoes an acylation reaction with 2S- (4′-butyric acid amide) -3R-nonadecanol whose hydroxyl group is protected by TBDMS, and the resulting product is The TBDMS protecting group is removed under trifluoroacetic acid conditions to obtain compound z-1-9; 10,11-methylenedioxycamptothecin under the catalysis of a condensing agent and (4E, 8E, 12E, 16E) -4, 8,12,16,2021-pentamethyl-4,8,12,16,20-pentaene-docosapentaenoic acid undergoes an acylation reaction to give compound z-2-5.

In another preferred embodiment, in formula (I), X is -O (CH ₂ ) _n SS (CH ₂ ) _m OCOY-, Y is-(CH ₂ ) _n -or-(CH ₂ ) _n CONH -; N and m are both 2, R is substituted or unsubstituted alkyl; preferably, the compound of formula (I) is z-3-3, z-4-2, z-5-1, z-6- 1. z-7-1 or z-8-1.

The preparation method of such compounds includes the following steps:

10,11-Methylenedioxycamptothecin reacted with phosgene and bis (2-hydroxyethyl) disulfide under DMAP catalysis to obtain 10,11- containing disulfide bond and carbonate modification at position 20 Methylenedioxycamptothecin, an acylation reaction between this compound and 4-aminooxobutanoic acid with different substitutions on the amine group under the catalysis of DMAP, DIC and EDCI to obtain the compound.

In yet another preferred embodiment, in formula (I), X is -CH ₂ NH- or -OCH ₂ CH ₂ SSCH ₂ CH ₂ O-, and R is H.

Another aspect of the present invention also provides a pharmaceutical composition comprising a therapeutically effective dose of the above compound, its isomer or pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier.

In still another aspect of the present invention, there is provided the use of the above compound, its isomer or pharmaceutically acceptable salt or the above pharmaceutical composition in the preparation of a medicament for preventing and / or treating cancer. Preferably, the cancer is lung cancer or colon cancer.

The selectivity of the camptothecin derivatives of the present invention to tumor tissue is higher than that of traditional camptothecin compounds, and the antitumor activity is significantly improved compared with the parent compound, while maintaining less toxicity, and can be used for preparation of prevention or Drugs used to treat tumors. The camptothecin derivative of the present invention has clear structural characteristics, is convenient to synthesize, simple and quick to purify, and has good biological activity. Therefore, such compounds have broad prospects in preventing or treating tumors.

The mode of administration of the compound of the present invention, its isomer, its pharmaceutically acceptable salt, or the pharmaceutical composition containing the compound is not particularly limited. Representative modes of administration include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), etc. The compounds of the present invention can be administered alone or in combination with other pharmaceutically acceptable compounds. The same or similar administration methods as dasatinib can be selected, including (but not limited to): oral, transdermal, intravenous, intramuscular, local administration, etc.

It should be understood that, within the scope of the present invention, the above technical features of the present invention and the technical features specifically described in the following (eg, embodiments) can be combined with each other, thereby forming a new or preferred technical solution. Due to space limitations, I will not repeat them here.

BRIEF DESCRIPTION

Figure 1. Effect of intragastric administration of compound w-31 on the growth of transplanted tumors in A549 mice.

Figure 2. Effect of intragastric administration of compound w-31 on body weight of A549 transplanted mice.

Figure 3. Effect of intragastric administration of compound z-1-9 on the growth of transplanted tumors in A549 mice.

Figure 4. Effect of intragastric administration of compound z-1-9 on the body weight of A549 transplanted mice.

Figure 5. Effect of intragastric administration of compound z-3-3 on the growth of transplanted tumors in A549 mice.

Figure 6. Effect of intragastric administration of compound z-3-3 on body weight of A549 transplanted mice.

detailed description

The methods and techniques of the present invention are generally performed according to conventional methods known in the art, unless otherwise stated. Nomenclature and experimental methods and techniques related to biology, pharmacology, and medical and medicinal chemistry described herein are known and commonly used in the art. Standard techniques are used for chemical synthesis methods, chemical analysis methods, pharmaceutical preparation methods, preparations and administration methods, and patient treatment methods.

Unless otherwise stated, the scientific and technical terms used herein shall have the meanings generally understood by those of ordinary skill in the art. However, the following terms have the following definitions:

The term "isomer" includes conformational isomers, optical isomers (such as enantiomers and diastereomers) and geometric isomers (such as cis-trans isomers). These isomers or combinations thereof can be used as racemic mixtures (racemates), individual enantiomers, individual diastereomers, mixtures of diastereomers, cis or trans Isomers exist.

The term "pharmaceutically acceptable salt" refers to any pharmaceutically acceptable salt of the compound of formula (I), preferably to the acid addition salt of the compound. Examples of preferred pharmaceutically acceptable salts are acid addition salts of pharmaceutically acceptable inorganic or organic acids, such as hydrohalic acid, sulfuric acid, phosphoric acid, or aliphatic or aromatic carboxylic acids or sulfonic acids, such as acetic acid, succinic acid, Lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, niacin, methanesulfonic acid, p-toluenesulfonic acid or naphthalenesulfonic acid. Examples of other preferred pharmaceutically acceptable salts are alkali metal salts (sodium or potassium), alkaline earth metal salts (calcium or magnesium) or from ammonia or such as C1-C7 alkylamines, cyclohexylamine, triethanolamine, ethylenediamine Or tris (hydroxymethyl) aminomethane, a pharmaceutically acceptable organic amine-derived ammonium salt.

The term "sulfonamide group" refers to a collective name of compound groups having a structure of -SO ₂ N-, and particularly preferably refers to a collective name of chemical groups having a structure of the following formula (II),

Wherein R1 is alkyl, aryl, substituted aryl, heteroaryl or substituted heteroaryl, and R2 is aryl or substituted aryl.

The term "base" includes substituted or unsubstituted natural bases and artificial bases; preferably, substituted or unsubstituted natural bases, including but not limited to substituted or unsubstituted adenine, substituted Or unsubstituted guanine, substituted or unsubstituted uracil, substituted or unsubstituted thymine and substituted or unsubstituted cytosine; more preferably substituted or unsubstituted uracil, such as 5- Fluorouracil.

The term "saturated nitrogen-containing heterocyclic ring" is preferably a 4-6 membered nitrogen-containing heterocyclic ring, including but not limited to substituted or unsubstituted tetrahydropyrrole, substituted or unsubstituted morpholine, substituted or unsubstituted piperidine, substituted Or unsubstituted piperazine and substituted or unsubstituted thiomorpholine and so on.

The term "alkyl" refers to any linear or branched, substituted or unsubstituted saturated hydrocarbon group, wherein the hydrocarbon group having 1 to 10 carbon atoms includes but is not limited to methyl, ethyl, n-propyl, isopropyl, cyclic Propyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, 2,4,4-trimethylpentyl, cyclopentyl , N-hexyl, isohexyl, cyclohexyl, n-heptyl, cycloheptyl, n-octyl, 2-ethylhexyl, cyclooctyl, n-nonyl, cyclononyl or n-decyl. Other long-chain alkyl groups with more carbon atoms include but are not limited to squalene, nonadecanol and the like. Typically, "alkyl" refers to C1-20 alkyl, preferably C1-10 alkyl.

The term "halogen atom" refers to any radio-stable atom in column 7 of the periodic table, ie fluorine, chlorine, bromine or iodine, preferably fluorine and chlorine.

The terms "aryl", "substituted aryl", "heteroaryl" and "substituted heteroaryl" refer to aromatic hydrocarbon rings, preferably having 5, 6 or 7 atoms, most preferably having 6 atoms Narration. "Heteroaryl" and "substituted heteroaryl" refer to aromatic hydrocarbon rings having at least one heteroatom (eg, oxygen, sulfur, or nitrogen atom) and at least one carbon atom in the ring. The "aryl" includes, but is not limited to, substituted or unsubstituted the following groups: phenyl, o-methylphenyl, m-tolyl, xylyl, naphthyl, or anthracenyl. Typically, "aryl" refers to C6-20 aryl, and "heteroaryl" refers to C2-18 heteroaryl, preferably C3-15 heteroaryl.

The term "substituted" refers to any group in which at least one hydrogen atom is replaced by a substituent selected from halogen atoms, alkyl groups, substituted alkyl groups, alkoxy groups, substituted alkoxy groups, cycloalkyl groups , Substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyacylamino, cyano, hydroxyl, carboxyl , Carboxyalkyl, keto, thioketo, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocycle Group, heterocyclyloxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -SO _2- Alkyl, -SO ₂ -substituted alkyl, -SO ₂ -aryl and -SO ₂ -heteroaryl.

The term "therapeutically effective dose" refers to any amount of the drug as described below. When used alone or in combination with another therapeutic agent, the amount of the drug can promote the regression of the disease, which manifests as a reduction in the severity of the symptoms of the disease 2. Increase the frequency and duration of the disease-free symptom period, or prevent the disorder or disability caused by the disease. The "therapeutically effective dose" of the medicine of the present invention also includes the "prophylactically effective dose". The "prophylactically effective dose" is any amount of the drug as described below, when the amount of the drug is administered alone or in combination with another therapeutic agent When there is a risk of developing a disease or a subject suffering from a disease recurrence, the occurrence or recurrence of the disease can be suppressed.

As will be apparent to those skilled in the art, the effective in vivo dosage and specific mode of administration will vary according to the type, weight and age of the mammal being treated, the specific compounds used and the specific purpose of using these compounds. Those skilled in the art can determine the effective dose level (ie, the dose level necessary to achieve the desired effect) according to conventional pharmacological methods. Generally, the human clinical application of the product starts at a lower dose level and then continuously increases the dose level until the desired effect is achieved. Alternatively, acceptable in vitro studies can be used by existing pharmacological methods to establish useful doses and routes of administration of the compositions identified by this method.

The "pharmaceutical composition" of the present invention can be made into liquid preparation forms such as tablets, capsules, powders, granules, lozenges, suppositories, oral liquids or sterile parenteral suspensions, as well as large or small volume injections, lyophilized powders, etc. Injection form. The drugs in the above dosage forms can be prepared according to conventional methods in the pharmaceutical field.

The "pharmaceutically acceptable carrier" in the present invention includes conventional diluents, fillers, binders, wetting agents, absorption enhancers, surfactants, adsorption carriers, lubricants and the like in the pharmaceutical field.

The term "cancer" refers to a large class of diseases characterized by the uncontrolled growth of abnormal cells in the body. Uncontrolled cell division and growth Division and growth result in the formation of malignant tumors or cells that invade adjacent tissues and can also be transferred to the distal part of the body through the lymphatic system or blood flow. In the present invention, another equivalent description of "treatment of cancer" is "treatment of tumor" or "anti-cancer" or "anti-tumor".

Cancer is an uncontrolled condition that prevents the growth of cells that interfere with the normal functioning of body organs and systems. A subject with cancer is a subject with objectively measurable cancer cells present in the body of the subject. Subjects at risk of developing cancer are subjects prone to developing cancer (eg based on family history, genetic predisposition), subjects exposed to radiation or other cancer-causing agents.

The compounds of the present invention and their pharmaceutical compositions can be used to prevent or treat various cancers. Examples of such cancers include breast cancer, prostate cancer, lung cancer, ovarian cancer, cervical cancer, skin cancer, melanoma, colon cancer, stomach cancer, liver cancer, esophageal cancer, kidney cancer, throat cancer, thyroid cancer, pancreatic cancer, testis Cancer, brain cancer, bone cancer and blood cancer (such as leukemia, chronic lymphocytic leukemia), etc. Other cancers include but are not limited to basal cell carcinoma, biliary tract cancer, bladder cancer, bone cancer, brain and central nervous system (CNS) cancer, cervical cancer, choriocarcinoma, colorectal cancer, connective tissue cancer, digestive system cancer, uterus Endometrial cancer, esophageal cancer, eye cancer, head and neck cancer, intraepithelial tumor, laryngeal cancer, lung cancer (small cell, large cell), lymphoma (including Hodgkin lymphoma and non-Hodgkin lymphoma), melanoma , Neuroblastoma, oral cancer (such as lips, tongue, mouth and pharynx), retinoblastoma, rhabdomyosarcoma, respiratory cancer, sarcoma, uterine cancer, urinary system cancer, and other cancers and sarcomas. The compounds of the present invention and compositions thereof are preferably useful for colon cancer and / or lung cancer.

The present invention also provides a method for preventing and / or treating cancer, the method comprising administering to a patient a therapeutically effective dose of a compound having the structure represented by formula (I), an isomer thereof, or a pharmaceutical thereof as described above An acceptable salt, or a pharmaceutical composition containing the compound. Preferably, the cancer is lung cancer or colon cancer.

Some examples of the compounds of the present invention include the compounds shown in Table A (especially the compounds other than Nos. 2 and 3):

Table A

The compounds of the present invention can generally be obtained by multi-step reaction synthesis using 10,11-methylenedioxycamptothecin (FL118) unless otherwise specified.

The 10,11-methylenedioxycamptothecin can be purchased from the market, or can be prepared by the following method: using piperaldehyde as the starting material, after two steps of nitrification with concentrated nitric acid and hydrogen palladium-carbon reduction 6-Amino-3,4-methylenedioxybenzaldehyde was obtained, and then 5 ′ (S) -1,5-dioxo- (5′-ethyl-5′-hydroxy-2′H, 5′H, 6′H-6-oxopyran)-[3 ′, 4 ′, f] -Δ ^{6 (8)} -tetrahydroindolizine under the catalysis of p-toluenesulfonic acid, Fried In the rand condensation reaction, the resulting reactant is purified by silica gel column chromatography to obtain 10,11-methylenedioxycamptothecin.

The present invention will be further described below in conjunction with specific embodiments. It should be understood that these examples are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods without specific conditions in the following examples generally follow the conventional conditions or the conditions recommended by the manufacturer. Unless stated otherwise, percentages and parts are percentages by weight and parts by weight.

Example 1 Preparation of a camptothecin derivative substituted with a 20-position sulfonamidino group

The preparation of the camptothecin derivative substituted with the 20-position sulfonamidino group of the present invention can be carried out by the following general formula.

Reaction conditions: (a) DMAP, DIC, EDCI, DCM; (b) CF ₃ COOD, CH ₂ Cl ₂ .; (C) R ₁ SO ₂ N ₃ , alkyne, Et ₃ N, CuI, DCM.

In summary, the preparation of the 20-position sulfonamidino substituted camptothecin derivative of the present invention includes the following steps:

1) 10,11-Methylenedioxycamptothecin (Compound 1, also known as FL118) undergoes an acylation reaction with Boc-protected glycine under the catalysis of a condensing agent, and the resulting product is deprotected under trifluoroacetic acid Group to obtain 10,11-methylenedioxycamptothecin modified with glycine at position 20;

2) This 20-position glycine-modified 10,11-methylenedioxycamptothecin is catalyzed by cuprous iodide with one-pot reaction of benzenesulfonyl azide and alkynyl group containing different substituents to obtain the 20-position Camptothecin derivatives substituted with sulfonamidinyl.

The specific synthesis method is as follows:

1. Preparation of 10,11-Methylenedioxy-20 (S) -O- (Boc-Glycine) Camptothecin (Compound 2)

Add 20 (S) -10,11-methylenedioxycamptothecin (200mg, 0.5mmol) to a 100mL eggplant-shaped bottle, dissolve with anhydrous dichloromethane (45mL), and then add EDCI (0.84g, 4.4mmol), DMAP (124mg, 1.02mmol), Boc-glycine (105mg, 0.6mmol), under nitrogen protection at room temperature for 12h. After the reaction was detected by TLC, it was cooled to room temperature, diluted with dichloromethane (100 mL), washed with 0.1 M dilute hydrochloric acid, distilled water, and saturated saline three times, and concentrated by rotary evaporation to obtain a yellow oily liquid. Silica gel column chromatography (dichloromethane : Acetone = 8: 1) Separately pure, 0.26g of brown solid was obtained with a yield of 95%, namely compound 2. mp 241-242 ° C; HRMS (ESI): C ₂₈ H ₂₇ N ₃ O ₉ calculated value 550.5360, found value 550.1806.

¹ H NMR (500 MHz, DMSO) δ 8.34 (s, 1H), 7.36 (s, 2H), 7.02 (d, J = 70.1 Hz, 1H), 6.24 (s, 2H), 5.45 (s, 2H), 5.08 (d, J = 25.9 Hz, 2H), 3.87 (dd, J = 54.0, 17.3 Hz, 2H), 2.10 (s, 2H), 1.32 (d, 6H), 0.94 (d, 6H).

¹³ C NMR (125MHz, DMSO) δ169.98,167.64,157.03,156.29,151.82,149.97,149.09,146.81,146.63,145.88,130.60,128.58,125.99,118.23,104.79,103.46,103.07,95.30,79.08,76.70,66.61, 50.46,42.34,41.06,30.69,28.20,23.63,7.93.

2. Preparation of 10,11-methylenedioxy-20 (S) -O-glycine camptothecin (Compound 3)

Compound 2 (200 mg, 0.36 mmol) was added to a 100 mL eggplant-shaped bottle, dissolved with anhydrous dichloromethane (45 mL), and then CF ₃ COOH (6 mL) was added, and the reaction was carried out at room temperature for 12 h under nitrogen protection. After TLC detected the reaction was complete, it was concentrated by rotary evaporation to obtain a yellow oily liquid, which was purified by silica gel column chromatography (dichloromethane: acetone = 8: 1) to obtain 0.15 g of brown solid with a yield of 95%, that is, compound 3. mp> 250 ℃; HRMS (ESI): C ₂₃ H ₁₉ N ₃ O ₇ calculated value 450.4190, found value 450.1288.

¹ H NMR (500 MHz, DMSO) δ 8.44 (s, 1H), 7.45 (s, 1H), 7.40 (s, 1H), 7.16 (s, 1H), 6.25 (s, 2H), 5.48 (d, J = 16.5 Hz, 2H), 5.16 (d, J = 20.9 Hz, 2H), 4.26 (d, J = 17.9 Hz, 1H), 4.06 (d, J = 17.9 Hz, 1H), 2.21–2.09 (m, 2H ), 0.98–0.89 (m, 3H).

¹³ C NMR (125MHz, DMSO) δ167.34,167.15,156.99,151.97,149.97,149.22,146.89,146.86,145.20,130.85,128.80,126.18,118.33,104.77,103.62,103.12,95.17,78.01,66.75,50.61,30.57, 23.63, 7.93.

3. Preparation of compound w-1

Compound 3 (30 mg, 0.07 mmol) was added to a 50 mL eggplant-shaped bottle, dissolved with anhydrous dichloromethane (25 mL), and then 2 drops of triethylamine was added, and the reaction was stirred for 10 min until the reaction solution was clear. Under the protection of nitrogen, phenylacetylene (7.32 μL, 0.07 mmol), 4-methylbenzenesulfonyl azide (19 mg, 0.084 mmol), and cuprous iodide (1.2 mg, 0.007 mmol) were added successively, and the reaction was carried out at room temperature for 12 h under nitrogen protection. After the reaction was detected by TLC, saturated ammonium chloride solution (3 mL) was added, and stirring was continued for 30 min. Dichloromethane (50 mL) was added for dilution, and the organic layer was washed three times with distilled water and saturated brine in this order, and the organic layer was dried over anhydrous MgSO ₄ . Concentrated by rotary evaporation to obtain a yellow oily liquid, which was purified by silica gel column chromatography (dichloromethane: acetone = 8: 1) to obtain 38 mg of a yellow solid with a yield of 76%, namely compound w-1. mp 151-152 ° C; HRMS (ESI): calculated value C ₃₈ H ₃₂ N ₄ O ₉ S calculated value 721.7530, measured value 721.1970.

¹ H NMR (500 MHz, DMSO) δ 8.43 (s, 1H), 7.53 (d, J = 8.0 Hz, 1H), 7.48 (s, 1H), 7.43 (d, J = 7.4 Hz, 1H), 7.39 ( s, 1H), 7.27 (d, J = 7.3Hz, 2H), 7.22–7.14 (m, 3H), 7.03 (d, J = 7.9Hz, 2H), 6.97 (s, 1H), 6.26 (d, J = 1.8Hz, 2H), 5.48 (s, 2H), 5.18 (s, 2H), 4.35 (dd, J = 17.9, 5.7Hz, 1H), 4.26 (dd, J = 17.8, 5.8Hz, 1H), 4.09 (s, 2H), 2.36 (s, 1H), 2.21 (s, 2H), 2.10 (dt, J = 13.9, 6.9 Hz, 2H), 0.84 (t, J = 7.2 Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ168.19,167.41,167.13,156.95,151.73,150.11,149.12,146.97,145.15,141.82,141.22,135.23,130.51,129.31,129.22,128.73,127.10,126.16,126.09,118.53,105.07, 103.54,103.03,94.63,77.09,66.82,50.61,42.84,38.29,30.79,21.45,21.25,7.91.

4. Preparation of compound w-2

The preparation and purification methods are the same as compound w-1, and 4-fluorobenzenesulfonyl azide is used instead of 4-methylbenzenesulfonyl azide to obtain 41 mg of yellow powdery solid, with a yield of 81%, namely compound w-2. mp 156-157 ° C; HRMS (ESI): C ₃₇ H ₂₉ FN ₄ O ₉ S calculated value 725.7164, found value 725.1712.

¹ H NMR (500 MHz, DMSO) δ 9.17 (t, J = 5.8 Hz, 1H), 8.43 (s, 1H), 7.66 (dd, J = 8.6, 5.3 Hz, 2H), 7.48 (s, 1H), 7.38 (s, 1H), 7.25 (d, J = 7.4Hz, 2H), 7.21–7.14 (m, 3H), 7.01 (t, J = 8.8Hz, 2H), 6.97 (s, 1H), 6.26 (d , J = 3.5 Hz, 2H), 5.49 (s, 2H), 5.18 (s, 2H), 4.38 (dd, J = 17.9, 5.7 Hz, 1H), 4.29 (dd, J = 17.9, 5.9 Hz, 1H) , 4.10 (s, 2H), 2.10 (dq, J = 13.0, 6.5Hz, 2H), 0.84 (t, J = 7.4Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ168.15,167.44,167.41,162.79,156.95,151.75,150.10,149.13,146.98,146.93,145.14,140.30,134.99,130.55,129.24,129.02,128.94,128.78,128.74,127.15,126.11, 118.54,115.78,115.61,105.02,103.56,103.04,94.59,77.14,66.82,50.63,42.94,38.27,30.77,7.92.

5. Preparation of compound w-3

The preparation and purification methods are the same as compound w-1. 4-chlorobenzenesulfonyl azide is used instead of 4-methylbenzenesulfonyl azide to obtain 39 mg of a yellow powdery solid with a yield of 75%, ie compound w-3. mp 160-161 ℃; HRMS (ESI): C ₃₇ H ₂₉ ClN ₄ O ₉ S calculated value 741.1680, found value 741.1416.

¹ H NMR (500 MHz, DMSO) δ 8.42 (s, 1H), 7.58 (d, J = 8.6 Hz, 2H), 7.47 (s, 1H), 7.38 (s, 1H), 7.23 (dd, J = 7.5 , 6.0Hz, 3H), 7.16 (dt, J = 20.0, 6.9Hz, 3H), 7.00 (d, J = 8.4Hz, 1H), 6.97 (s, 1H), 6.26 (d, J = 2.7Hz, 2H ), 5.49 (s, 2H), 5.17 (s, 2H), 4.39 (dd, J = 17.9, 5.7Hz, 1H), 4.30 (dd, J = 17.9, 5.9Hz, 1H), 4.10 (s, 2H) , 2.10 (dq, J = 14.2, 7.1 Hz, 2H), 0.84 (t, J = 7.4 Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ168.14,167.57,167.46,156.95,151.75,150.08,149.13,146.98,146.93,145.14,142.72,136.50,134.93,130.53,129.70,129.23,128.74,128.58,127.98,127.93,127.14, 126.10,118.52,105.03,103.56,103.03,94.59,77.16,66.82,50.62,42.97,38.29,30.75,7.92.

6. Preparation of compound w-4

The preparation and purification methods are the same as compound w-1, and 4-nitrobenzenesulfonyl azide is used instead of 4-methylbenzenesulfonyl azide to obtain 42 mg of yellow powdery solid, with a yield of 81%, namely compound w-4. mp 166-167 ° C; HRMS (ESI): C ₃₇ H ₂₉ N ₅ O ₁₁ S calculated value 752.7230, found value 752.1655.

¹ H NMR (500 MHz, DMSO) δ 8.40 (d, J = 8.9 Hz, 1H), 8.14 (d, J = 8.8 Hz, 1H), 7.92 (d, J = 8.7 Hz, 1H), 7.74 (d, J = 8.7Hz, 1H), 7.44 (s, 1H), 7.33 (s, 1H), 7.29–7.24 (m, 1H), 7.22 (d, J = 7.2Hz, 2H), 7.15 (t, J = 7.0 Hz, 2H), 7.12–7.08 (m, 1H), 6.97 (s, 1H), 6.25 (s, 2H), 5.49 (s, 2H), 5.13 (t, J = 13.2Hz, 2H), 4.45 (dd , J = 18.0, 5.7Hz, 1H), 4.36 (dd, J = 17.9, 5.9Hz, 1H), 4.12 (d, J = 6.8Hz, 1H), 3.58 (s, 1H), 2.12 (td, J = 13.8, 7.0Hz, 2H), 0.86 (t, J = 7.3Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ170.38,168.16,168.10,167.49,156.94,151.75,150.68,150.03,149.19,149.12,148.95,146.96,146.88,145.14,134.65,130.48,129.75,129.65,129.22,128.74,127.45, 127.19,124.88,123.87,118.52,104.95,103.52,103.04,94.56,77.25,66.83,50.59,43.09,42.50,38.37,30.73,7.94.

7. Preparation of compound w-5

The preparation and purification methods are the same as compound w-1, and β-naphthalenesulfonyl azide is used instead of 4-methylbenzenesulfonyl azide to obtain 39 mg of a yellow powdery solid with a yield of 73%, namely compound w-5. mp 165-166 ° C; HRMS (ESI): C ₄₁ H ₃₂ N ₄ O ₉ S calculated value 757.7860, found value 757.1960.

¹ H NMR (500 MHz, DMSO) δ 9.21 (t, J = 5.9 Hz, 1H), 8.30 (s, 1H), 8.09 (s, 1H), 7.75 (s, 1H), 7.68 (d, J = 8.2 Hz, 1H), 7.59–7.53 (m, 2H), 7.49–7.44 (m, 1H), 7.42 (s, 1H), 7.32 (d, J = 13.3Hz, 1H), 7.28 (d, J = 7.6Hz , 2H), 7.12 (t, J = 7.6Hz, 2H), 7.04 (t, J = 7.3Hz, 1H), 6.92 (s, 1H), 6.25 (d, J = 9.4Hz, 2H), 5.49 (t , J = 10.0 Hz, 2H), 5.07 (d, J = 19.1 Hz, 1H), 4.91 (d, J = 19.1 Hz, 1H), 4.45 (dd, J = 17.8, 5.9 Hz, 1H), 4.31 (dd , J = 17.8, 5.9 Hz, 1H), 4.15 (s, 2H), 2.06 (td, J = 14.7, 7.2 Hz, 2H), 0.81 (t, J = 7.4 Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ168.31,167.66,167.49,156.88,151.58,149.80,149.04,146.90,146.87,145.33,140.89,135.07,133.86,131.73,130.24,129.29,129.05,128.63,128.48,128.37,127.86, 127.19,127.08,125.98,125.89,122.47,118.25,105.15,103.46,102.98,94.56,77.25,66.76,50.47,43.03,38.24,30.64,7.92.

8. Preparation of compound w-6

The preparation and purification methods are the same as compound w-1. Ethylsulfonyl azide is used instead of 4-methylbenzenesulfonyl azide to obtain 35 mg of light yellow powdery solid with a yield of 75%, namely compound w-6. mp 161-162 ℃; HRMS (ESI): C ₃₃ H ₃₀ N ₄ O ₉ S calculated value 659.6820, found value 659.1810.

¹ H NMR (500 MHz, DMSO) δ 8.43 (d, J = 18.1 Hz, 1H), 7.48 (d, J = 12.4 Hz, 2H), 7.38 (d, J = 7.3 Hz, 2H), 7.32–7.26 ( m, 2H), 7.12 (s, 1H), 7.06 (s, 1H), 7.03 (s, 1H), 6.26 (d, J = 4.9Hz, 2H), 5.50 (t, J = 7.7Hz, 2H), 5.20 (d, J = 11.7 Hz, 2H), 3.98 (s, 1H), 3.91 (d, J = 16.3 Hz, 1H), 3.58–3.51 (m, 1H), 3.48 (s, 1H), 2.19–2.10 (m, 2H), 1.21 (d, J = 2.3 Hz, 3H), 1.11 (t, J = 7.1 Hz, 2H), 0.95-0.89 (m, 3H).

¹³ C NMR (125MHz, DMSO) δ168.21,167.42,166.77,156.94,151.87,150.07,149.19,146.93,145.28,135.56,130.64,129.44,128.85,128.78,127.21,126.13,118.52,104.98,103.58,103.08,94.40, 77.17,76.97,66.80,50.63,49.03,47.25,42.73,38.48,30.86,8.60,8.43,7.95.

9. Preparation of compound w-7

The preparation and purification methods are the same as compound w-1, and 3-pyridinesulfonyl azide is used instead of 4-methylbenzenesulfonyl azide to obtain 35 mg of yellow powdery solid with a yield of 70%, namely compound w-7. mp 156-157 ℃; HRMS (ESI): C ₃₆ H ₂₉ N ₅ O ₉ S calculated value 708.7140, found value 708.1756.

¹ H NMR (500 MHz, DMSO) δ 9.37 (t, J = 5.8 Hz, 1H), 8.40 (s, 1H), 7.88 (d, J = 7.9 Hz, 1H), 7.45 (d, J = 5.6 Hz, 1H), 7.38 (s, 1H), 7.25 (d, J = 7.4 Hz, 2H), 7.15 (dt, J = 23.8, 7.2 Hz, 5H), 6.95 (s, 1H), 6.25 (d, J = 3.6 Hz, 2H), 5.48 (s, 2H), 5.17 (s, 2H), 4.43 (dd, J = 17.9, 5.8 Hz, 1H), 4.30 (dd, J = 17.9, 5.8 Hz, 1H), 4.13 (s , 2H), 2.10 (dt, J = 14.0, 6.9Hz, 2H), 0.85 (t, J = 7.4Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ168.05,167.78,167.39,156.94,152.18,151.71,150.08,149.10,147.01,146.89,146.45,145.19,134.79,133.81,130.47,129.23,128.76,127.24,126.06,123.68,118.42, 105.04,103.52,103.02,94.49,77.18,66.79,50.60,43.06,41.78,38.38,30.75,26.89,7.94.

10. Preparation of compound w-8

The preparation and purification methods are the same as compound w-1. Methanesulfonyl azide is used instead of 4-methylbenzenesulfonyl azide to obtain 33 mg of a yellow powdery solid with a yield of 73%, namely compound w-8. mp 175-176 ° C; HRMS (ESI): calculated value of C ₃₂ H ₂₈ N ₄ O ₉ S 645.6550, measured value 645.1644.

¹ H NMR (500 MHz, DMSO) δ 8.82 (t, J = 5.6 Hz, 1H), 8.44 (s, 1H), 7.49 (s, 1H), 7.41 (s, 1H), 7.35 (d, J = 7.5 Hz, 2H), 7.26 (t, J = 7.5 Hz, 3H), 7.21–7.17 (m, 1H), 6.95 (s, 1H), 6.26 (s, 2H), 5.47 (s, 2H), 5.18 (t , J = 11.0 Hz, 2H), 4.29 (dd, J = 18.0, 5.9 Hz, 1H), 4.15 (dd, J = 17.9, 5.8 Hz, 1H), 4.05 (dd, J = 31.7, 14.7 Hz, 2H) , 2.80 (s, 3H), 2.10 (dt, J = 23.2, 7.1 Hz, 2H), 0.87 (t, J = 7.3 Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ168.20,167.44,166.53,156.96,151.87,150.07,149.19,146.91,146.87,145.47,135.44,130.63,129.47,128.88,128.81,127.24,126.15,118.36,104.98,103.58,103.06, 94.42,77.09,66.71,50.65,43.55,42.83,38.38,30.70,8.01.

11. Preparation of compound w-9

The preparation and purification methods are the same as compound w-1. Propylsulfonyl azide is used instead of 4-methylbenzenesulfonyl azide to obtain 35 mg of yellow powdery solid with a yield of 75%, ie compound w-9. mp 143-144 ° C; HRMS (ESI): C ₃₄ H ₃₂ N ₄ O ₉ S calculated value 673.7090, found value 673.1955.

¹ H NMR (500 MHz, DMSO) δ 8.42 (s, 1H), 7.78 (t, J = 6.1 Hz, 1H), 7.47 (s, 2H), 7.42 (s, 1H), 7.39–7.31 (m, 2H ), 7.32–7.20 (m, 2H), 7.04 (s, 1H), 6.27 (d, J = 4.2 Hz, 2H), 5.47 (d, J = 7.5 Hz, 2H), 5.17 (s, 2H), 4.20 (dd, J = 18.9, 6.2Hz, 1H), 4.07–3.95 (m, 1H), 2.95–2.85 (m, 2H), 2.18–2.07 (m, 2H), 1.61 (dd, J = 14.8, 7.3Hz , 2H), 0.92 (t, J = 7.2Hz, 3H), 0.88-0.83 (m, 2H), 0.78 (t, J = 7.3Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ169.59,167.62,156.96,151.86,150.12,149.16,146.94,146.87,145.54,130.62,129.66,129.44,129.01,128.82,128.78,126.11,118.30,105.00,103.56,103.08,94.76, 77.19,66.69,54.47,50.58,43.90,30.55,17.49,17.33,17.16,13.49,12.93,7.99.

12. Preparation of compound w-10

The preparation and purification methods are the same as compound w-1, butylsulfonyl azide is used instead of 4-methylbenzenesulfonyl azide to obtain 37 mg of yellow powdery solid, with a yield of 76%, namely compound w-10. mp 128-129 ° C; HRMS (ESI): calculated value of C ₃₅ H ₃₄ N ₄ O ₉ S 687.7360, found value 687.2113.

¹ H NMR (500 MHz, DMSO) δ 8.84 (t, J = 5.8 Hz, 1H), 8.45 (s, 1H), 7.50 (s, 1H), 7.43 (s, 1H), 7.35 (t, J = 7.3 Hz, 2H), 7.26 (t, J = 7.5 Hz, 2H), 7.18 (t, J = 7.4 Hz, 1H), 6.96 (s, 1H), 6.27 (d, J = 1.6 Hz, 2H), 5.47 ( s, 2H), 5.20 (s, 2H), 4.30 (dd, J = 17.8, 5.9 Hz, 1H), 4.17 (dd, J = 17.9, 5.8 Hz, 1H), 4.06 (t, J = 6.8 Hz, 2H ), 2.76 (ddd, J = 20.9, 13.5, 8.9 Hz, 2H), 2.10 (td, J = 13.9, 6.9 Hz, 2H), 1.44-1.37 (m, 2H), 1.03 (dt, J = 14.6, 7.3 Hz, 2H), 0.86 (dd, J = 14.3, 6.9Hz, 3H), 0.61 (t, J = 7.3Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ168.22,167.36,166.68,156.95,151.82,150.11,149.17, 146.93,145.33,135.59,130.64,129.44,128.78,127.21,126.17,118.49,105.04,103.59,103.07,94.44,76.98, 66.75,54.29,50.64,42.79,38.33,30.81,25.61,21.16,13.80,7.96.

13. Preparation of compound w-11

The preparation and purification method is the same as compound w-1, using methylsulfonyl azide and p-fluorophenylacetylene instead of 4-methylbenzenesulfonyl azide and phenylacetylene to obtain 34 mg of yellow powdery solid, yield 73%, that is compound w -11. mp 174-175 ° C; HRMS (ESI): C ₃₂ H ₂₇ FN ₄ O ₉ S calculated value 663.6454, found value 663.1552.

¹ H NMR (500 MHz, DMSO) δ 8.83 (t, J = 5.7 Hz, 1H), 8.43 (s, 1H), 7.48 (s, 1H), 7.42–7.37 (m, 3H), 7.10 (t, J = 8.8 Hz, 2H), 6.96 (s, 1H), 6.27 (s, 2H), 5.48 (s, 2H), 5.23–5.13 (m, 2H), 4.30 (dd, J = 17.9, 5.8 Hz, 1H) , 4.15 (dd, J = 17.9, 5.7 Hz, 1H), 4.04 (dd, J = 28.9, 14.7 Hz, 2H), 2.84 (s, 3H), 2.15–2.08 (m, 2H), 0.88 (t, J = 7.3Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ168.19,167.44,166.43,162.63,160.70,156.97,151.89,150.05,149.19,146.91,145.48,131.50,131.43,130.64,128.86,126.15,118.36,115.65,115.48,104.97,103.57, 103.07,94.42,77.11,66.72,50.64,43.61,42.82,37.62,36.24,30.69,8.00.

14. Preparation of Compound w-12

Substituting p-methoxybenzenesulfonyl azide for 4-methylbenzenesulfonyl azide, the preparation and purification methods are the same as compound w-1, to obtain a yellow powdery solid 40mg, yield 78%, that is compound w-12. mp 139-141 ° C; HRMS (ESI): C ₃₈ H ₃₂ N ₄ O ₁₀ S calculated value 737.7520, found value 737.1908.

¹ H NMR (500 MHz, DMSO) δ 8.97 (t, J = 5.6 Hz, 1H), 8.39 (s, 1H), 7.53 (t, J = 9.3 Hz, 2H), 7.45 (s, 1H), 7.38 ( s, 1H), 7.25 (d, J = 7.4 Hz, 2H), 7.19 (t, J = 7.1 Hz, 2H), 7.16–7.12 (m, 1H), 6.96 (s, 1H), 6.70 (d, J = 8.7 Hz, 2H), 6.25 (d, J = 2.2 Hz, 2H), 5.47 (s, 2H), 5.14 (s, 2H), 4.34 (dd, J = 17.9, 5.6 Hz, 1H), 4.24 (dd , J = 17.9, 5.8 Hz, 1H), 4.08 (s, 1H), 3.82 (s, 1H), 3.66 (d, J = 18.4 Hz, 3H), 2.14–2.04 (m, 2H), 0.83 (t, J = 7.3Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ168.23,167.42,166.91,161.69,156.93,151.72,149.10,146.94,146.92,145.16,135.97,135.24,130.49,129.28,129.11,128.81,128.72,128.18,127.10,126.05,118.51, 114.25,113.82,105.04,103.50,103.03,94.63,77.07,66.81,56.07,55.81,50.57,42.82,41.77,38.16,30.77,26.89,7.92.

15. Preparation of compound w-13

Using 2-thiophenesulfonyl azide and p-fluorophenylacetylene instead of 4-methylbenzenesulfonyl azide and phenylacetylene, the preparation and purification methods are the same as compound w-1, to obtain a yellow powdery solid 37mg, yield 73%, that is Compound w-13. mp 178-179 ° C; HRMS (ESI): C ₃₅ H ₂₇ FN ₄ O ₉ S ₂ calculated value 731.7384, found value 731.1271.

¹ H NMR (500 MHz, DMSO) δ 9.24 (t, J = 5.7 Hz, 1 H), 8.41 (s, 1 H), 7.55 (d, J = 4.8 Hz, 1 H), 7.47 (d, J = 10.9 Hz, 1H), 7.39 (s, 1H), 7.31 (t, J = 5.7 Hz, 3H), 7.04 (t, J = 8.8 Hz, 2H), 6.97 (d, J = 12.1 Hz, 1H), 6.83-6.79 ( m, 1H), 6.25 (d, J = 2.5 Hz, 2H), 5.47 (s, 2H), 5.17 (s, 2H), 4.42 (dd, J = 17.9, 5.8 Hz, 1H), 4.29 (dd, J = 17.9, 5.8 Hz, 1H), 4.08 (s, 2H), 2.10 (tt, J = 14.1, 7.1 Hz, 2H), 0.84 (t, J = 7.3 Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ168.05,167.35,167.23,156.93,151.75,150.08,149.13,146.97,146.93,145.38,145.06,131.36,131.29,131.23,131.19,131.17,130.53,130.19,128.76,127.14,126.08, 118.56,115.61,115.44,105.07,103.53,103.04,94.68,77.14,66.84,50.60,42.88,37.46,30.84,7.90.

16. Preparation of compound w-14

Using p-methoxybenzenesulfonyl azide and p-fluorophenylacetylene instead of 4-methylbenzenesulfonyl azide and phenylacetylene, the preparation and purification method is the same as compound w-1, powdery solid 40mg, yield 75%, namely Compound w-14. mp 221-222 ° C; HRMS (ESI): C ₃₈ H ₃₁ FN ₄ O ₁₀ S calculated value 755.7424, found value 755.1798.

¹ H NMR (500 MHz, DMSO) δ 9.07 (t, J = 5.7 Hz, 1H), 8.41 (s, 1H), 7.52 (d, J = 8.1 Hz, 2H), 7.46 (s, 1H), 7.38 ( s, 1H), 7.29 (dd, J = 8.1, 5.8 Hz, 2H), 7.02 (dd, J = 9.8, 8.3 Hz, 4H), 6.97 (s, 1H), 6.26 (d, J = 2.8 Hz, 2H ), 5.50 (d, J = 14.0 Hz, 2H), 5.16 (s, 2H), 4.36 (dd, J = 17.9, 5.7 Hz, 1H), 4.26 (dd, J = 17.9, 5.8 Hz, 1H), 4.07 (s, 2H), 2.21 (s, 3H), 2.10 (dt, J = 19.5, 6.5 Hz, 2H), 0.84 (t, J = 7.4 Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ168.21,167.42,167.06,162.56,160.63,156.95,151.74,150.08,149.12,146.96,146.93,145.15,141.85,141.15,131.29,131.23,130.51,129.20,128.72,126.13,126.08, 118.54,115.54,115.37,105.05,103.52,103.05,94.61,77.11,66.82,50.60,42.83,37.48,30.78,21.24,7.91.

17. Preparation of compound w-15

Using 4-chlorobenzenesulfonyl azide and p-fluorophenylacetylene instead of 4-methylbenzenesulfonyl azide and phenylacetylene, the preparation and purification methods are the same as compound w-1, to obtain a yellow powdery solid 41mg, yield 77%, that is Compound w-15. mp 165-167 ° C; HRMS (ESI): C ₃₇ H ₂₈ ClFN ₄ O ₉ S calculated value 759.1584, found value 759.1310.

¹ H NMR (500 MHz, DMSO) δ 9.27 (t, J = 5.8 Hz, 1H), 8.43 (s, 1H), 7.58 (d, J = 8.5 Hz, 2H), 7.48 (s, 1H), 7.38 ( s, 1H), 7.29–7.21 (m, 4H), 7.01 (t, J = 8.8Hz, 2H), 6.97 (s, 1H), 6.26 (d, J = 2.9Hz, 2H), 5.49 (s, 2H ), 5.19 (s, 2H), 4.40 (dd, J = 18.0, 5.8 Hz, 1H), 4.30 (dd, J = 17.8, 5.9 Hz, 1H), 4.08 (s, 2H), 2.15-2.08 (m, 2H), 0.85 (t, J = 7.4Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ168.16,167.51,167.47,160.66,156.95,151.75,150.07,149.14,146.99,146.93,145.13,142.68,136.52,131.25,131.19,130.96,130.54,128.76,128.73,127.94,126.12, 118.53,115.57,115.40,105.04,103.56,103.03,94.58,77.19,66.83,50.64,42.97,37.51,30.75,7.91.

18. Preparation of compound w-16

Using p-toluenesulfonyl azide and p-methoxyphenylacetylene instead of 4-methylbenzenesulfonyl azide and phenylacetylene, the preparation and purification method is the same as compound w-1, to obtain a yellow powdery solid 39mg, yield 75%, That is compound w-16. mp 158-160 ℃; HRMS (ESI): C ₃₉ H ₃₄ N ₄ O ₁₀ S calculated value 751.7790, found value 751.2076.

¹ H NMR (500 MHz, DMSO) δ 8.88 (t, J = 5.8 Hz, 1H), 8.39 (s, 1H), 7.55 (t, J = 8.2 Hz, 2H), 7.44 (s, 1H), 7.38 ( s, 1H), 7.18 (d, J = 8.5 Hz, 2H), 7.04 (d, J = 8.1 Hz, 2H), 6.97 (s, 1H), 6.75 (d, J = 8.6 Hz, 2H), 6.26 ( d, J = 2.7 Hz, 2H), 5.48 (s, 2H), 5.14 (s, 2H), 4.34 (dd, J = 17.9, 5.8 Hz, 1H), 4.24 (dd, J = 17.9, 5.9 Hz, 1H ), 4.00 (s, 2H), 3.64 (s, 3H), 2.22 (s, 3H), 2.15-2.06 (m, 2H), 0.85 (t, J = 7.4Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ168.22,167.67,167.41,158.59,156.94,151.73,150.07,149.11,146.95,146.93,145.19,141.75,141.30,130.58,130.49,129.20,128.69,126.82,126.15,126.06,118.52, 114.19,105.05,103.50,103.04,94.63,77.08,66.81,55.41,50.58,42.82,37.50,30.79,21.25,7.92.

19. Preparation of compound w-17

Using 4-chlorobenzenesulfonyl azide and p-methoxyphenylacetylene instead of 4-methylbenzenesulfonyl azide and phenylacetylene, the preparation and purification methods are the same as compound w-1, to obtain 39 mg of yellow powdery solid, yield 73% , That is compound w-17. mp 145-146 ° C; HRMS (ESI): C ₃₈ H ₃₁ ClN ₄ O ₁₀ S calculated value 771.1940, found value 771.1504.

¹ H NMR (500 MHz, DMSO) δ 9.13 (t, J = 5.8 Hz, 1H), 8.42 (s, 1H), 7.56 (d, J = 8.5 Hz, 2H), 7.47 (s, 1H), 7.38 ( s, 1H), 7.22 (d, J = 8.4 Hz, 2H), 7.14 (d, J = 8.5 Hz, 2H), 6.97 (s, 1H), 6.72 (d, J = 8.5 Hz, 2H), 6.26 ( d, J = 2.7 Hz, 2H), 5.49 (s, 2H), 5.18 (s, 2H), 4.38 (dd, J = 17.9, 5.7 Hz, 1H), 4.28 (dd, J = 18.0, 5.9 Hz, 1H ), 4.01 (s, 2H), 3.64 (s, 3H), 2.11 (dq, J = 14.1, 7.1 Hz, 2H), 0.85 (t, J = 7.4 Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ168.17,168.12,167.47,158.61,156.95,151.74,150.08,149.13,146.99,146.93,145.17,142.79,136.38,130.53,130.48,128.76,128.68,127.94,126.45,126.11,118.51, 114.17,105.05,103.56,103.03,94.59,77.16,66.82,55.41,50.63,42.95,37.48,30.75,7.93.

20. Preparation of compound w-18

Using 4-nitrobenzenesulfonyl azide and p-fluorophenylacetylene instead of 4-methylbenzenesulfonyl azide and phenylacetylene, the preparation and purification methods are the same as compound w-1, to obtain a yellow powdery solid 41mg, yield 76%, That is compound w-18. mp 217-218 ° C; HRMS (ESI): C ₃₇ H ₂₈ FN ₅ O ₁₁ S calculated value 770.7134, found value 770.1548.

¹ H NMR (500 MHz, DMSO) δ 9.50 (t, J = 5.8 Hz, 1H), 8.40 (s, 1H), 7.94 (d, J = 8.6 Hz, 2H), 7.75 (d, J = 8.6 Hz, 2H), 7.46 (s, 1H), 7.33 (s, 1H), 7.25 (dd, J = 8.2, 5.6Hz, 2H), 7.00-6.95 (m, 3H), 6.26 (s, 2H), 5.50 (s , 2H), 5.20–5.09 (m, 2H), 4.46 (dd, J = 17.9, 5.7 Hz, 1H), 4.35 (dd, J = 17.9, 5.9 Hz, 1H), 4.08 (t, J = 11.3 Hz, 2H), 2.16-2.09 (m, 2H), 0.86 (t, J = 7.3Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ168.12,167.50,162.61,160.68,156.95,151.76,150.03,149.16,148.95,146.97,146.89,145.13,131.27,130.73,130.49,128.69,127.43,126.07,123.87,118.53,115.59, 115.42,104.96,103.53,103.04,94.56,77.27,66.83,56.48,50.60,43.09,37.61,30.72,19.01,7.93.

21. Preparation of Compound w-19

Using p-fluorobenzenesulfonyl azide and p-fluorophenylacetylene instead of 4-methylbenzenesulfonyl azide and phenylacetylene, the preparation and purification method is the same as compound w-1, to obtain 43mg of yellow powdery solid, yield 82%, namely Compound w-19. mp 163-164 ° C; HRMS (ESI): C ₃₇ H ₂₈ F ₂ N ₄ O ₉ S calculated value 743.7068, found value 743.1610.

¹ H NMR (500 MHz, DMSO) δ 9.21 (t, J = 5.8 Hz, 1H), 8.42 (s, 1H), 7.66 (dd, J = 8.5, 5.4 Hz, 2H), 7.47 (s, 1H), 7.38 (s, 1H), 7.28 (dd, J = 8.2, 5.7Hz, 2H), 7.02 (dd, J = 8.7, 7.1Hz, 4H), 6.97 (s, 1H), 6.26 (d, J = 3.6Hz , 2H), 5.49 (s, 2H), 5.17 (s, 2H), 4.38 (dd, J = 17.9, 5.7Hz, 1H), 4.29 (dd, J = 17.9, 5.9Hz, 1H), 4.08 (s, 2H), 2.15-2.08 (m, 2H), 0.85 (t, J = 7.4Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ168.17,167.44,167.34,164.79,162.80,162.57,160.63,156.95,151.75,150.07,149.13,146.98,145.14,140.26,131.19,131.04,130.54,128.91,128.75,126.10,118.54, 115.77,115.56,115.39,105.01,103.54,103.03,94.57,77.16,66.82,56.48,50.61,42.93,37.49,30.77,19.01,7.91.

22. Preparation of compound w-20

Using 4-nitrobenzenesulfonyl azide and p-methoxyphenylacetylene instead of 4-methylbenzenesulfonyl azide and phenylacetylene, the preparation and purification methods are the same as compound w-1, to obtain a yellow powdery solid 41mg, yield 75 %, That is compound w-20. mp 195-196 ° C; HRMS (ESI): calculated value of C ₃₈ H ₃₁ N ₅ O ₁₂ S 782.7490, found value 782.1746.

¹ H NMR (500 MHz, DMSO) δ 9.41 (t, J = 5.7 Hz, 1H), 8.39 (s, 1H), 7.89 (d, J = 8.6 Hz, 2H), 7.70 (d, J = 8.6 Hz, 2H), 7.45 (s, 1H), 7.33 (s, 1H), 7.10 (d, J = 8.5Hz, 2H), 6.97 (s, 1H), 6.66 (d, J = 8.5Hz, 2H), 6.25 ( s, 2H), 5.50 (s, 2H), 5.13 (t, J = 14.5 Hz, 2H), 4.45 (dd, J = 17.9, 5.7 Hz, 1H), 4.34 (dd, J = 17.9, 5.9 Hz, 1H ), 4.02 (q, J = 14.6 Hz, 2H), 3.59 (s, 3H), 2.13 (tt, J = 13.9, 7.0 Hz, 2H), 0.87 (t, J = 7.3 Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ168.76,168.15,167.52,158.63,156.95,151.74,150.02,149.26,149.12,148.81,146.97,146.88,145.19,130.49,128.69,127.38,126.10,126.06,123.75,118.50,114.13, 104.98,103.52,103.03,94.57,77.27,66.82,55.34,50.60,43.07,37.53,30.71,7.95.

23. Preparation of compound w-21

Substituting p-fluorobenzenesulfonyl azide and p-methoxyphenylacetylene for 4-methylbenzenesulfonyl azide and phenylacetylene, the preparation and purification methods are the same as compound w-1, to obtain a yellow powdery solid 41mg, yield 78% , That is compound w-21. mp 159-160 ℃; HRMS (ESI): C ₃₈ H ₃₁ FN ₄ O ₁₀ S calculated value 755.7424, found value 755.1798.

¹ H NMR (500 MHz, DMSO) δ 9.04 (t, J = 5.8 Hz, 1H), 8.41 (s, 1H), 7.66 (dd, J = 8.6, 5.4 Hz, 2H), 7.46 (s, 1H), 7.38 (s, 1H), 7.16 (d, J = 8.5 Hz, 2H), 7.01 (t, J = 8.8 Hz, 2H), 6.97 (s, 1H), 6.74 (d, J = 8.6 Hz, 2H), 6.26 (d, J = 3.4 Hz, 2H), 5.49 (s, 2H), 5.16 (s, 2H), 4.36 (dd, J = 17.9, 5.7 Hz, 1H), 4.27 (dd, J = 17.9, 5.9 Hz , 1H), 4.02 (s, 2H), 3.64 (s, 3H), 2.15-2.08 (m, 2H), 0.85 (t, J = 7.4Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ168.17,167.94,167.44,162.75,158.60,156.95,151.75,150.06,149.12,146.96,145.17,140.41,130.51,128.99,128.91,128.73,126.56,126.09,118.52,115.74,115.56, 114.20,105.01,103.53,103.03,94.59,77.12,66.81,56.48,55.41,50.60,42.92,37.49,30.78,19.01,7.92.

24. Preparation of compound w-22

Using β-naphthalenesulfonyl azide and p-methoxyphenylacetylene instead of 4-methylbenzenesulfonyl azide and phenylacetylene, the preparation and purification method is the same as compound w-1, to obtain 38mg of yellow powdery solid, yield 70% , That is compound w-22. mp 169-171 ℃; HRMS (ESI): C ₄₂ H ₃₄ FN ₄ O ₁₀ S calculated value 787.8120, found value 787.2058.

¹ H NMR (500 MHz, DMSO) δ 9.13 (t, J = 5.7 Hz, 1H), 8.29 (s, 1H), 8.00 (s, 1H), 7.74 (d, J = 8.2 Hz, 1H), 7.64 ( d, J = 8.2Hz, 1H), 7.58-7.50 (m, 3H), 7.46 (t, J = 7.4Hz, 1H), 7.41 (s, 1H), 7.33 (s, 1H), 7.16 (d, J = 8.5 Hz, 2H), 6.92 (s, 1H), 6.61 (d, J = 8.5 Hz, 2H), 6.25 (d, J = 9.1 Hz, 2H), 5.49 (t, J = 10.1 Hz, 2H), 5.06 (d, J = 19.0 Hz, 1H), 4.89 (d, J = 19.0 Hz, 1H), 4.45 (dd, J = 17.8, 5.9 Hz, 1H), 4.30 (dd, J = 17.7, 5.8 Hz, 1H) ), 4.09-4.01 (m, 2H), 3.49 (s, 3H), 2.08 (tt, J = 14.4, 7.2 Hz, 2H), 0.83 (t, J = 7.3 Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ168.35,168.20,167.50,158.41,156.89,151.56,149.78,149.03,146.90,146.86,145.38,140.85,133.79,131.69,130.50,130.21,129.00,128.47,128.35,128.27,127.81, 127.11,126.49,125.96,125.83,122.44,118.21,114.02,105.17,103.45,102.97,94.57,77.27,66.75,55.21,50.47,42.98,37.37,30.62,7.94.

25. Preparation of compound w-23

Using β-naphthalenesulfonyl azide and p-fluorophenylacetylene instead of 4-methylbenzenesulfonyl azide and phenylacetylene, the preparation and purification method is the same as compound w-1, to obtain a yellow powdery solid 39mg, yield 72%, namely Compound w-23. mp 170-171 ° C; HRMS (ESI): C ₄₁ H ₃₁ FN ₄ O ₉ S calculated value 775.7764, found value 775.1896.

¹ H NMR (500 MHz, DMSO) δ 9.28 (t, J = 5.9 Hz, 1H), 8.28 (s, 1H), 8.03 (s, 1H), 7.74 (t, J = 9.8 Hz, 1H), 7.65 ( d, J = 8.2Hz, 1H), 7.58–7.51 (m, 3H), 7.50–7.43 (m, 1H), 7.41 (s, 1H), 7.32 (d, J = 7.7Hz, 1H), 7.29 (dd , J = 8.4, 5.6 Hz, 2H), 6.92–6.85 (m, 3H), 6.26 (t, J = 7.1 Hz, 2H), 5.49 (t, J = 10.3 Hz, 2H), 5.06 (d, J = 19.1Hz, 1H), 4.89 (d, J = 19.1Hz, 1H), 4.47 (dd, J = 17.8, 5.9Hz, 1H), 4.32 (dd, J = 17.8, 5.9Hz, 1H), 4.16-4.07 ( m, 2H), 2.08 (tt, J = 14.4, 7.2 Hz, 2H), 0.83 (t, J = 7.4 Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ 168.34,167.60,167.50,162.43,160.50,156.88,151.57,149.76,149.04,146.86,145.34,140.75,133.81,131.66,131.27,131.20,131.02,130.21,128.96,128.46,128.42, 128.35,127.84,127.19,125.96,125.83,122.38,118.23,115.42,115.25,105.15,103.44,102.97,94.54,77.30,66.75,50.46,43.01,37.39,30.62,7.92.

26. Preparation of compound w-24

Using 3-pyridinesulfonyl azide and p-fluorophenylacetylene instead of 4-methylbenzenesulfonyl azide and phenylacetylene, the preparation and purification methods are the same as compound w-1, to obtain a yellow powdery solid 39mg, yield 77%, that is Compound w-24. mp 173-175 ℃; HRMS (ESI): C ₃₆ H ₂₈ FN ₅ O ₉ S calculated value 726.7044, found value 726.1655.

¹ H NMR (500 MHz, DMSO) δ 9.39 (t, J = 5.8 Hz, 1H), 8.65 (s, 1H), 8.46 (s, 1H), 8.42 (s, 1H), 7.88 (d, J = 8.0 Hz, 1H), 7.48 (s, 1H), 7.38 (s, 1H), 7.28 (dd, J = 8.4, 5.7 Hz, 2H), 7.17 (dd, J = 8.0, 4.7 Hz, 1H), 7.00 (t , J = 8.8 Hz, 2H), 6.95 (s, 1H), 6.26 (d, J = 3.6 Hz, 2H), 5.49 (s, 2H), 5.20 (s, 2H), 4.44 (dd, J = 17.9, 5.9Hz, 1H), 4.30 (dd, J = 17.8, 5.8Hz, 1H), 4.09 (d, J = 14.5Hz, 2H), 2.14–2.07 (m, 2H), 0.85 (t, J = 7.4Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ168.07,167.72,167.40,162.57,160.64,156.95,152.17,151.71,150.09,149.11,147.03,146.90,146.38,145.17,133.80,131.26,131.19,130.81,130.49,128.78,126.08, 123.65,118.43,115.61,115.44,105.05,103.55,103.02,94.49,77.21,66.79,50.63,43.05,37.59,30.74,7.93.

Example 2 Preparation of a 10,11-methylenedioxycamptothecin derivative modified at the 20 position with uracil

The 10,11-methylenedioxycamptothecin derivative modified at the 20th position of the uracil group can be prepared by the following general formula.

Reaction conditions: (a) DMAP, DIC, EDCI, DCM; (b) CF ₃ COOD, CH ₂ Cl ₂ ; (c) 5'-substituted uracil 1 '(N) -acetic acid, DMAP, DIC, EDCI, DCM .

In summary, the preparation steps of the 10,11-methylenedioxycamptothecin derivative modified at the 20 position by uracil group include:

1) 10,11-Methylenedioxycamptothecin undergoes an acylation reaction with Boc-protected glycine under the catalysis of a condensing agent. 10,11-Methylenedioxycamptothecin;

2) The above-mentioned 10,11-methylenedioxycamptothecin modified at the 20-position of glycine undergoes an acylation reaction with uracil acetic acid modified by different substituents under the catalysis of a condensing agent to obtain the 20-positioned uracil Group modified 10,11-methylenedioxycamptothecin derivatives.

The preparation methods of specific compounds are as follows:

1. Preparation of compound w-25

In a 50 mL eggplant-shaped bottle, add compound 3 (60 mg, 0.13 mmol), 2- (2,4-dioxopyrimidin-1-yl) acetic acid (180 mg, 1.1 mmol), and dissolve with a mixed solvent of methylene chloride and methanol until the reaction The solution was clarified, followed by adding DIC, DMAP, EDCI, nitrogen protection, reaction for 24h, the reaction was detected by TLC, the reaction was stopped, the solvent was removed by rotary evaporation, silica gel column chromatography separation and purification (dichloromethane: methanol = 40: 1), to obtain light Yellow powdery solid 60 mg, yield 77%, ie compound w-25. mp> 250 ℃; HRMS (ESI): C ₂₉ H ₂₃ N ₅ O ₁₀ calculated value 602.5280, found value 602.1506.

¹ H NMR (500 MHz, DMSO) δ 11.21 (s, 1H), 8.71 (s, 1H), 8.38 (s, 1H), 7.46 (d, J = 31.5 Hz, 3H), 7.01 (s, 1H), 6.26 (s, 2H), 5.46 (s, 3H), 5.11 (s, 2H), 4.35 (s, 2H), 4.22 (s, 1H), 4.09 (s, 1H), 2.15 (s, 2H), 0.89 (s, 3H).

¹³ C NMR (125MHz, DMSO) δ169.16,167.91,167.52,164.22,156.91,151.84,151.37,150.10,149.12,146.86,145.38,130.58,128.75,127.84,126.08,118.48,114.29,105.07,103.50,103.05,101.07, 94.70,76.84,66.79,50.51,49.67,30.86,7.97.

2. Preparation of compound w-26

Using 2- (2,4-dioxopyrimidin-5-chloro-1-yl) acetic acid instead of 2- (2,4-dioxopyrimidin-1-yl) acetic acid, the preparation and purification methods are the same as compound w-25 To obtain 65 mg of a yellow powdery solid with a yield of 79%, namely compound w-26. mp 242-243 ° C; HRMS (ESI): C ₂₉ H ₂₂ ClN ₅ O ₁₀ calculated value 636.9700, found value 636.1151.

¹ H NMR (500MHz, DMSO) δ 11.79 (s, 1H), 8.74 (s, 1H), 8.38 (s, 1H), 8.03 (s, 1H), 7.42 (s, 2H), 7.01 (s, 1H ), 6.26 (s, 2H), 5.46 (s, 2H), 5.11 (s, 2H), 4.36 (s, 2H), 4.24 (dd, J = 18.3, 5.5Hz, 1H), 4.09 (dd, J = 17.9, 5.3Hz, 1H), 2.19–2.09 (m, 2H), 0.90 (t, J = 7.1Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ169.11,167.61,167.51,159.95,156.91,151.84,150.51,150.09,149.12,146.90,146.86,145.39,144.01,130.55,128.74,126.07,118.45,106.38,105.08,103.48,103.04, 94.70,76.86,66.79,50.50,49.99,40.72,30.85,7.97.

3. Preparation of compound w-27

Using 2- (2,4-dioxopyrimidin-5-fluoro-1-yl) acetic acid instead of 2- (2,4-dioxopyrimidin-1-yl) acetic acid, the preparation and purification methods are the same as compound w-25 , 60 mg of a yellow powdery solid was obtained with a yield of 74%, namely compound w-27. mp 242-243 ° C; HRMS (ESI): calculated value for C ₂₉ H ₂₂ FN ₅ O ₁₀ 620.5184, measured value 620.1409.

¹ H NMR (500 MHz, DMSO) δ 8.73 (s, 1H), 8.37 (s, 1H), 7.97 (d, J = 6.4 Hz, 1H), 7.41 (s, 2H), 7.01 (s, 1H), 6.26 (s, 2H), 5.46 (s, 2H), 5.09 (s, 2H), 4.29 (d, J = 18.1Hz, 2H), 4.26–4.17 (m, 1H), 4.11–4.02 (m, 1H) , 2.19–2.07 (m, 2H), 0.90 (t, J = 6.8Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ169.12,167.68,167.51,156.91,151.84,150.07,150.04,149.11,146.90,146.84,145.40,140.65,131.35,131.07,130.54,128.70,126.05,118.46,105.04,103.47,103.05, 94.71,76.85,66.79,50.48,49.86,40.72,30.86,7.97.

Example 3 Preparation of 10,11-methylenedioxycamptothecin derivative modified by heterocyclic amide at position 20

The 10,11-methylenedioxycamptothecin derivative modified by heterocyclic amide at position 20 of the present invention can be prepared by the following general formula.

Reaction conditions: (a) DMAP, DIC, EDCI, DCM; (b) CF ₃ COOH, CH ₂ Cl ₂ ; (c) CDI, suitable secondary amine (2 ° amine), Et ₃ N, DCM, 0 ° C.

Broadly speaking, the preparation steps of the 10,11-methylenedioxycamptothecin derivative modified with heterocyclic amide at position 20 include:

The preparation methods of specific compounds are as follows:

1. Preparation of compound w-28

Add 3 (50mg, 0.11mmol), 3 drops of triethylamine and dichloromethane (10mL) to a 50mL eggplant-shaped bottle, put them in an ice bath to react. After the reaction temperature drops to 0 ℃, add CDI (23.4mg, 0.14) mmol) for 1 h, morpholine (15 μL, 0.154 mmol) was added, warmed to room temperature, and reacted overnight. After the reaction was detected by TLC, dichloromethane (50 mL) was added for dilution, and the organic layer was washed three times with distilled water and saturated brine in this order, and the organic layer was dried over anhydrous MgSO ₄ . Concentrated by rotary evaporation to obtain a yellow oily liquid, which was purified by silica gel column chromatography (dichloromethane: acetone = 8: 1) to obtain 48 mg of a yellow solid with a yield of 78%, namely compound w-28. mp> 250 ℃; HRMS (ESI): C ₂₈ H ₂₆ N ₄ O ₉ calculated value 563.5350, found value 563.1758.

¹ H NMR (500 MHz, DMSO) δ 8.42 (s, 1H), 7.46 (s, 1H), 7.42 (s, 1H), 7.14 (t, J = 5.7 Hz, 1H), 7.05 (s, 1H), 6.27 (d, J = 4.8 Hz, 2H), 5.45 (s, 2H), 5.15 (s, 2H), 4.02 (dd, J = 17.7, 5.7 Hz, 1H), 3.90 (dd, J = 17.8, 5.8 Hz , 1H), 3.48 (t, J = 4.7 Hz, 4H), 3.25 (dt, J = 13.5, 6.6 Hz, 4H), 2.12 (td, J = 13.8, 7.0 Hz, 2H), 0.89 (t, J = 7.3Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ170.50,167.64,157.76,156.94,151.85,150.20,149.12,146.92,146.68,145.54,130.58,128.76,126.05,118.48,105.01,103.53,103.06,95.08,76.45,66.76,66.35, 50.49,44.21,42.44,30.93,7.93.

2. Preparation of compound w-29

Substituting tetrahydropyrrole for morpholine, the preparation and purification methods are the same as compound w-28, and 46 mg of yellow solid is obtained in 76% yield, ie compound w-29. mp> 250 ℃; HRMS (ESI): C ₂₈ H ₂₆ N ₄ O ₈ calculated value 547.5360, found value 547.1809.

¹ H NMR (500 MHz, DMSO) δ 8.45 (s, 1H), 7.47 (d, J = 22.2 Hz, 2H), 7.03 (s, 1H), 6.64 (s, 1H), 6.27 (s, 2H), 5.45 (s, 2H), 5.20 (s, 2H), 3.98 (s, 1H), 3.90 (d, J = 17.9 Hz, 1H), 3.17 (d, J = 7.5 Hz, 4H), 2.12 (s, 2H ), 1.74 (s, 4H), 0.87 (t, J = 7.1 Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ 170.76,167.64,156.97,156.57,151.86,150.24,149.16,146.97,146.75,145.45,135.04,130.65,128.86,126.13,120.43,118.63,105.09,103.61,103.06,95.00,76.34, 66.82,50.56,45.72,42.16,31.04,25.45,7.89.

3. Preparation of compound w-30

Substituting piperazine for morpholine, the preparation and purification methods were the same as compound w-28, which gave 23 mg of a yellow solid in 36% yield, ie compound w-30. mp 243-244 ℃; HRMS (ESI): C ₂₉ H ₂₉ N ₅ O ₈ calculated value 576.5780, found value 576.2073.

¹ H NMR (500 MHz, DMSO) δ 8.36 (s, 1H), 7.37 (s, 1H), 7.34 (s, 1H), 7.10 (s, 1H), 6.21 (d, J = 2.3 Hz, 2H), 5.42 (q, J = 16.8 Hz, 2H), 5.05 (s, 2H), 4.03 (d, J = 17.9 Hz, 1H), 3.92 (d, J = 17.9 Hz, 1H), 3.36 (s, 4H), 2.59 (s, 4H), 2.36 (s, 3H), 2.08 (dt, J = 24.0, 7.0 Hz, 2H), 0.89 (t, J = 7.3 Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ170.45,167.84,157.51,157.15,151.99,149.78,149.17,146.67,146.48,146.10,130.94,128.51,126.09,118.22,104.60,103.48,103.11,66.58,53.86,53.63,45.17, 44.38,42.33,42.24,30.70,7.86.

4. Preparation of compound w-31

Substituting 2-methylpiperidine for morpholine, the preparation and purification methods were the same as compound w-28, and 49 mg of yellow solid was obtained in 78% yield, ie compound w-31. mp 240-242 ° C; HRMS (ESI): calculated value of C ₃₀ H ₃₀ N ₄ O ₈ 575.5900, actual value 575.2121.

¹ H NMR (500 MHz, DMSO) δ 8.28 (d, J = 3.2 Hz, 1H), 7.32–7.28 (m, 1H), 7.25 (d, J = 10.6 Hz, 1H), 7.13 (d, J = 8.0 Hz, 1H), 6.20 (d, J = 5.0 Hz, 2H), 5.41 (dd, J = 30.3, 16.6 Hz, 2H), 4.94 (dd, J = 22.9, 12.8 Hz, 2H), 4.07–3.97 (m , 1H), 3.97–3.85 (m, 1H), 3.71 (d, J = 14.6Hz, 1H), 2.69 (dt, J = 26.3, 12.7Hz, 1H), 2.14–2.01 (m, 2H), 1.87– 1.79 (m, 1H), 1.53–1.30 (m, 5H), 0.98 (dd, J = 21.3, 6.8 Hz, 3H), 0.93–0.83 (m, 3H).

¹³ C NMR (125MHz, DMSO) δ172.97,170.74,157.28,156.98,151.79,150.51,150.40,149.09,147.02,146.36,130.61,128.86,126.05,118.53,105.27,103.56,103.03,96.34,76.40,72.86,65.69, 55.37, 50.62, 45.60, 38.35, 30.68, 18.75, 15.74, 8.24.

5. Preparation of compound w-32

Substituting thiomorpholine for morpholine, the preparation and purification methods were the same as compound w-28, and 45 mg of yellow solid was obtained in 70% yield, ie compound w-32. mp> 250 ℃; HRMS (ESI): C ₂₈ H ₂₆ N ₄ O ₈ S calculated value 579.5860, found value 579.1544.

¹ H NMR (500 MHz, DMSO) δ 8.43 (s, 1H), 7.47 (s, 1H), 7.42 (s, 1H), 7.17 (t, J = 5.8 Hz, 1H), 7.09 (d, J = 5.9 Hz, 1H), 6.27 (d, J = 4.9 Hz, 2H), 5.46 (s, 2H), 5.17 (d, J = 9.6 Hz, 2H), 4.01 (dd, J = 17.7, 5.7 Hz, 1H), 3.89 (dd, J = 17.7, 5.7 Hz, 1H), 3.59 (ddd, J = 12.9, 9.1, 4.3 Hz, 3H), 3.29-3.25 (m, 1H), 2.84-2.80 (m, 1H), 2.46 ( t, J = 4.9 Hz, 3H), 2.13 (tt, J = 14.2, 7.2 Hz, 2H), 0.91 (t, J = 7.4 Hz, 3H).

¹³ C NMR (125MHz, DMSO) δ 170.56,167.66,157.18,156.97,151.85,150.23,149.13,146.94,146.69,145.65,130.58,128.78,126.05,118.42,105.03,103.55,103.06,95.20,76.49,66.74,50.52, 46.54,45.21,42.70,30.91,26.55,24.08,7.97.

6. Preparation of compound w-33

Substituting piperidine for morpholine, the preparation and purification methods are the same as compound w-28, to obtain a yellow solid 51mg, yield 74%, that is compound w-33. mp 240-242 ° C; HRMS (ESI): calculated value for C ₂₉ H ₂₈ N ₄ O ₈ 560.5580, found value 560.2821.

Example 4 The 10,11-methylenedioxycamptothecin derivative modified with carboxylic acid at position 20

1. Preparation of compound z-1-9

The compound z-1-9 of the present invention can be prepared by the following reaction.

Reaction: (a) DMAP, DIC, EDCI, DCM; (b) CF ₃ COOH, CH ₂ Cl ₂ .

Specifically, add 20 (S) -10,11-methylenedioxycamptothecin (200mg, 0.5mmol) to a 100mL eggplant-shaped bottle, dissolve with anhydrous dichloromethane (45mL), and then add EDCI ( 0.84g, 4.4mmol), DMAP (124mg, 1.02mmol), 2S- (4'-butyric acid amide) -3R-tert-butyldimethylsilyl nonadecanol (1.81g, 4.4mmol), nitrogen protection Reaction at room temperature for 12h. After the reaction was detected by TLC, it was cooled to room temperature, diluted with dichloromethane (100 mL), washed with 0.1 M dilute hydrochloric acid, distilled water, and saturated saline three times, and concentrated by rotary evaporation to obtain a yellow oily liquid. Silica gel column chromatography (dichloromethane : Acetone = 20: 1) Separately pure to obtain 0.27g of yellow solid, yield 60%, that is compound 4. mp 223-224 ° C; HRMS (ESI): calculated value for C ₅₁ H ₇₅ N ₃ O ₈ Si 885.5341, found value 885.2656.

1HNMR (500MHz, dmso) δ 8.45 (s, 1H), 7.55 (d, J = 6.2Hz, 1H), 7.50 (s, 1H), 7.45 (s, 1H), 7.00 (s, 1H), 6.26 (s, 2H), 5.44 (s, 2H), 5.19 (s, 2H), 3.01 (s, 2H), 2.99–2.93 (m, 1H), 2.74 (s, 2H), 2.64 (d, J = 21.2 Hz, 2H), 2.11 (s, 3H), 1.99 (s, 2H), 1.19 (t, J = 14.5Hz, 34H), 0.88 (t, J = 7.3Hz, 3H), 0.84-0.80 (m, 4H ), 0.75 (d, J = 10.1 Hz, 9H), -0.04 (t, J = 13.3 Hz, 6H).

Compound 4 (100 mg, 0.11 mmol) was added to a 100 mL eggplant-shaped bottle, dissolved with CF ₃ COOH, and reacted at room temperature for 12 h. After the reaction was detected by TLC, the solution was concentrated by rotary evaporation to obtain a yellow oily liquid, which was purified by silica gel column chromatography (dichloromethane: acetone = 5: 1) to obtain a light yellow solid 0.07g, yield 90%, that is compound z-1- 9. mp 240-241 ℃; HRMS (ESI): C ₄₄ H ₅₉ N ₃ O ₉ calculated value 773.4351, found value 773.2159.

1H NMR (500MHz, cdcl3) δ 8.23 (s, 1H), 7.62 (s, 1H), 7.17 (s, 1H), 6.21 (s, 2H), 5.67 (d, J = 16.9Hz, 1H), 5.38 (d, J = 17.1Hz, 1H), 5.21 (s, 2H), 3.25–3.14 (m, 2H), 2.96–2.84 (m, 2H), 2.63–2.53 (m, 2H), 2.30–2.07 (m , 4H), 1.25 (s, 47H), 0.98 (t, J = 7.0Hz, 3H), 0.88 (t, J = 6.7Hz, 4H).

2. Preparation of compound z-2-5

The compound z-2-5 of the present invention can be prepared by the following reaction.

Reaction conditions: (a) DMAP, DIC, EDCI, DCM.

Specifically, 20 (S) -10,11-methylenedioxycamptothecin (200 mg, 0.5 mmol) was added to a 100 mL eggplant-shaped bottle, dissolved with anhydrous dichloromethane (45 mL), and then EDCI ( 0.84g, 4.4mmol), DMAP (124mg, 1.02mmol), (4E, 8E, 12E, 16E) -4,8,12,16,2021-pentamethyl-4,8,12,16,20-penta En-docosapentaenoic acid (1.76g, 4.4mmol), reacted at room temperature under nitrogen for 12h. After the reaction was detected by TLC, it was cooled to room temperature, diluted with dichloromethane (100 mL), washed with 0.1 M dilute hydrochloric acid, distilled water, and saturated saline three times, and concentrated by rotary evaporation to obtain a yellow oily liquid. Silica gel column chromatography (dichloromethane : Acetone = 15: 1) Separately pure, to obtain 0.25g of brown solid, yield 65%, namely compound z-2-5. mp 223-224 ° C; HRMS (ESI): C ₄₈ H ₅₈ N ₂ O ₇ calculated value 774.4251, found value 774.3124.

1H NMR (500MHz, cdcl3) δ 8.17 (s, 1H), 7.48 (s, 1H), 7.16 (s, 1H), 7.12 (s, 1H), 6.19 (s, 2H), 5.67 (d, J = 17.1Hz, 1H), 5.40 (d, J = 17.1Hz, 1H), 5.22–5.05 (m, 6H), 3.50 (s, 2H), 2.65–2.54 (m, 2H), 2.37–2.23 (m, 3H ), 2.21–2.11 (m, 2H), 2.11–1.88 (m, 14H), 1.71–1.55 (m, 15H), 1.53 (s, 3H), 0.97 (t, J = 7.5Hz, 3H).

Example 5 Preparation of carbonate modified 10,11-methylenedioxycamptothecin derivative at position 20

1. Preparation of compound z-3-3

The compound z-3-3 of the present invention can be prepared by the following reaction.

Reaction conditions: (a) Triphosgene, CH ₂ Cl ₂ , DMAP, ₂ , 2'-dithiobis-Ethanol; (b) DMAP, DIC, EDCI, DCM; (c) CF ₃ COOH, CH ₂ Cl ₂ .

Specifically, under the protection of N ₂ , triphosgene (0.45 g, 1.53 mmol) dissolved in anhydrous DCM was added dropwise to 20 (S) -10,11-methylenedioxyl dissolved in anhydrous DCM at 0 ° C. To the alkaloid (1.5g, 3.82mmol) and DMAP (1.4g, 4.59mmol), after stirring at room temperature for 1h, bis (2-hydroxyethyl) disulfide (11.7g, 76.3mmol) dissolved in anhydrous THF was added, The reaction was carried out at room temperature for 12h under the protection of nitrogen. After the reaction was completed, the reaction was stopped, and the temperature was lowered to room temperature. The solvent was removed by rotary evaporation and concentration. It was separated and purified by silica gel column chromatography (dichloromethane: acetone = 10: 1) to obtain a yellow-brown powdery solid 1.3g, with a yield of 67%. 5. mp 211-212 ° C; HRMS (ESI): C ₂₆ H ₂₄ N ₂ O ₉ S ₂ calculated value 572.0952, found value 572.0142.

1H NMR (500MHz, dmso) δ 8.43 (s, 1H), 7.48 (d, J = 4.2Hz, 1H), 6.95 (s, 1H), 6.27 (s, 2H), 5.47 (d, J = 17.2Hz , 2H), 5.19 (d, J = 4.7 Hz, 2H), 4.32 (t, J = 6.1 Hz, 2H), 3.55 (t, J = 6.4 Hz, 2H), 2.98 (dd, J = 13.7, 7.5 Hz , 2H), 2.80-2.72 (m, 2H), 2.15 (tq, J = 14.2, 7.0 Hz, 2H), 0.90 (dd, J = 16.4, 9.0 Hz, 3H).

Add compound 5 (100 mg, 0.17 mmol) in a 100 mL eggplant-shaped bottle, dissolve with anhydrous dichloromethane (45 mL), and then add EDCI (0.84 g, 4.4 mmol), DMAP (124 mg, 1.02 mmol), 2S- ( 4'-butyric acid amide) -3R-tert-butyldimethylsilyl nonadecanol (1.81g, 4.4mmol), reacted at room temperature under nitrogen for 12h. After the reaction was detected by TLC, it was cooled to room temperature, diluted with dichloromethane (100 mL), washed with 0.1 M dilute hydrochloric acid, distilled water, and saturated saline three times, and concentrated by rotary evaporation to obtain a yellow oily liquid. Silica gel column chromatography (dichloromethane : Acetone = 20: 1) Separately pure, to obtain 0.17g of yellow solid, yield 65%, that is compound 6. mp 254-255 ℃; HRMS (ESI): C ₅₅ H ₈₁ N ₃ O ₁₂ S ₂ Si calculated value 1067.5124, found value 1067.1245.

1H NMR (500MHz, dmso) δ 8.42 (s, 1H), 7.55 (s, 1H), 7.46 (s, 1H), 7.44 (s, 1H), 6.93 (d, J = 4.9Hz, 1H), 6.24 (d, J = 20.3 Hz, 2H), 5.46 (d, J = 19.1 Hz, 2H), 5.17 (s, 2H), 4.30 (dd, J = 16.7, 10.8 Hz, 2H), 4.15 (s, 2H) , 3.16 (d, J = 13.7 Hz, 1H), 3.07-3.02 (m, 2H), 3.01-2.96 (m, 2H), 2.91 (t, J = 6.2 Hz, 2H), 2.45-2.41 (m, 2H ), 2.34 (d, J = 6.3 Hz, 2H), 2.14 (dt, J = 22.2, 7.3 Hz, 2H), 1.33–1.11 (m, 31H), 1.06 (s, 3H), 0.90 (t, J = 7.3Hz, 3H), 0.82-0.78 (m, 9H), -0.00 (dd, J = 25.7, 6.1Hz, 6H).

Compound 6 (100 mg, 0.09 mmol) was added to a 100 mL eggplant-shaped bottle, dissolved with CF ₃ COOH, and stirred at room temperature for 12 h. After the reaction was detected by TLC, the solution was concentrated by vacuum rotary evaporation to obtain a yellow oily liquid, which was purified by silica gel column chromatography (dichloromethane: acetone = 5: 1) to obtain a pale yellow solid 0.08g, yield 90%, that is, compound z-3 -3. mp 251-252 ° C; HRMS (ESI): C ₄₉ H ₆₇ N ₃ O ₁₂ S ₂ calculated value 953.4215, found value 953.3215.

1H NMR (500MHz, dmso) δ 8.43 (s, 1H), 7.62 (s, 1H), 7.46 (d, J = 14.0 Hz, 2H), 6.94 (s, 1H), 6.26 (s, 2H), 5.48 (s, 2H), 5.18 (s, 2H), 4.31 (s, 2H), 4.15 (s, 2H), 2.97 (s, 4H), 2.90 (t, J = 6.1Hz, 2H), 2.42 (d, J = 6.6 Hz, 2H), 2.35 (d, J = 6.5 Hz, 2H), 2.14 (dd, J = 14.5, 7.0 Hz, 2H), 1.21 (d, J = 23.7 Hz, 30H), 0.90 (dd, J = 15.8, 8.6 Hz, 6H), 0.82 (t, J = 6.5 Hz, 3H).

2. Preparation of compound z-4-2

The compound z-4-2 of the present invention can be prepared by the following reaction.

Reaction conditions: (a) Triphosgene, CH ₂ Cl ₂ , DMAP, 2,2'-dithiobis-Ethanol; (b) DMAP, DIC, EDCI, DCM.

Add 5 (100mg, 0.17mmol) to a 100mL eggplant-shaped bottle, dissolve with anhydrous dichloromethane (45mL), then add EDCI (0.84g, 4.4mmol), DMAP (124mg, 1.02mmol), (4E, 8E , 12E, 16E) -4,8,12,16,21-pentamethyl-4,8,12,16,20-pentaene-docosanoic acid (1.76g, 4.4mmol), stirring at room temperature under nitrogen 12h. After the reaction was detected by TLC, it was cooled to room temperature, diluted with dichloromethane (100 mL), washed with 0.1 M diluted hydrochloric acid, distilled water, and saturated saline three times, and concentrated by vacuum rotary evaporation to obtain a yellow oily liquid. Methane: acetone = 20: 1) was pure, to obtain 0.17g of yellow solid, yield 67%, that is compound z-4-2. mp 254-255 ℃; HRMS (ESI): C ₅₃ H ₆₆ N ₂ O ₁₀ S ₂ calculated value 954.4219, found value 954.3258.

1H NMR (500MHz, dmso) δ 8.44 (s, 1H), 7.48 (s, 1H), 7.44 (s, 1H), 6.94 (s, 1H), 6.26 (d, J = 4.6Hz, 2H), 5.47 (t, J = 9.4Hz, 2H), 5.19 (s, 2H), 5.04 (d, J = 7.5Hz, 4H), 4.30 (dd, J = 9.8, 4.1Hz, 2H), 4.15 (t, J = 6.1 Hz, 2H), 2.99 (t, J = 6.0 Hz, 2H), 2.91 (t, J = 6.2 Hz, 2H), 2.29 (t, J = 7.6 Hz, 2H), 2.17–2.08 (m, 4H) , 1.94 (ddd, J = 37.3, 14.3, 6.9 Hz, 13H), 1.59 (d, J = 6.1 Hz, 3H), 1.53-1.48 (m, 12H), 0.89 (t, J = 7.4 Hz, 3H).

3. Preparation of compound z-5-1

The compound z-5-1 of the present invention can be prepared by the following reaction.

Add 5 (100mg, 0.17mmol) to a 100mL eggplant-shaped bottle, dissolve with anhydrous dichloromethane (45mL), then add EDCI (0.84g, 4.4mmol), DMAP (124mg, 1.02mmol), 9-eighteen The enoic acid (1.23g, 4.4mmol) was reacted at room temperature under nitrogen for 12h. After the reaction was detected by TLC, it was cooled to room temperature, diluted with dichloromethane (100 mL), washed with 0.1 M dilute hydrochloric acid, distilled water, and saturated saline three times, and concentrated by rotary evaporation to obtain a yellow oily liquid. : Acetone = 20: 1) Separately pure to obtain 0.23g of yellow solid, yield 70%, that is compound z-5-1. mp 223-225 ° C; HRMS (ESI): C ₄₄ H ₅₆ N ₂ O ₁₀ S ₂ calculated value 836.5119, found value 836.3258.

1H NMR (500MHz, dmso) δ 8.44 (s, 1H), 7.48 (s, 1H), 7.44 (s, 1H), 6.94 (s, 1H), 6.26 (d, J = 4.6Hz, 2H), 5.47 (t, J = 9.4Hz, 2H), 5.19 (s, 2H), 5.04 (d, J = 7.5Hz, 4H), 4.30 (dd, J = 9.8, 4.1Hz, 2H), 4.15 (t, J = 6.1 Hz, 2H), 2.99 (t, J = 6.0 Hz, 2H), 2.91 (t, J = 6.2 Hz, 2H), 2.29 (t, J = 7.6 Hz, 2H), 2.17–2.08 (m, 4H) , 1.59 (d, J = 6.1 Hz, 3H), 1.53-1.48 (m, 12H), 0.89 (t, J = 7.4 Hz, 3H).

4. Preparation of compound z-6-1

Substituting 9,12-dieneoctadecanoic acid for 9-octadecenoic acid, the preparation and purification methods are the same as compound z-5-1, to obtain a yellow solid 200mg, yield 71%, that is compound z-6-1. mp 238-240 ° C; HRMS (ESI): C ₄₄ H ₅₆ N ₂ O ₁₀ S ₂ calculated value 836.1200, actual value 836.2121.

1H NMR (500MHz, dmso) δ 8.44 (s, 1H), 7.48 (s, 1H), 7.44 (s, 1H), 6.94 (s, 1H), 6.26 (d, J = 4.6Hz, 2H), 5.47 (t, J = 9.4Hz, 2H), 5.19 (s, 2H), 5.04 (d, J = 7.5Hz, 4H), 4.30 (dd, J = 9.8, 4.1Hz, 2H), 4.15 (t, J = 6.1Hz, 2H), 2.99 (t, J = 6.0Hz, 2H), 2.91 (t, J = 6.2Hz, 2H), 2.29 (t, J = 7.6Hz, 1H), 2.17–2.08 (m, 4H) , 1.59 (d, J = 6.1 Hz, 3H), 1.53-1.48 (m, 12H), 0.89 (t, J = 7.4 Hz, 3H).

5. Preparation of compound z-7-1

The compound z-7-1 of the present invention can be prepared by the following reaction.

Specifically, under N ₂ protection, solid triphosgene (0.45 g, 1.53 mmol) dissolved in anhydrous DCM was added dropwise to 20 (S) -10,11-methylenedioxy dissolved in anhydrous DCM at 0 ° C. After camptothecin (1.5g, 3.82mmol) and DMAP (1.4g, 4.59mmol) were stirred at room temperature for 1h, bis (2-hydroxyethyl) disulfide (11.7g, 76.3mmol) dissolved in anhydrous THF was added At room temperature under nitrogen protection for 12h. After the reaction was completed, the reaction was stopped, and the temperature was lowered to room temperature. The solvent was removed by rotary evaporation and concentration. It was separated and purified by silica gel column chromatography (dichloromethane: acetone = 10: 1) to obtain a yellow-brown powdery solid 1.3g, with a yield of 67%. 5. mp 211-212 ° C; HRMS (ESI): C ₂₆ H ₂₄ N ₂ O ₉ S ₂ calculated value 572.0952, found value 572.0142.

Add 5 (100mg, 0.17mmol) in a 100mL eggplant-shaped bottle, dissolve with anhydrous dichloromethane (45mL), then add EDCI (0.84g, 4.4mmol), DMAP (124mg, 1.02mmol), 6 (1.82g , 4.4mmol), under the protection of nitrogen at room temperature for 12h. After the reaction was detected by TLC, it was cooled to room temperature, diluted with dichloromethane (100 mL), washed with 0.1 M dilute hydrochloric acid, distilled water, and saturated saline three times, and concentrated by rotary evaporation to obtain a yellow oily liquid. Silica gel column chromatography (dichloromethane : Acetone = 20: 1) Separately pure to obtain 0.31g of yellow solid, yield 70%, namely compound z-7-1. mp 250-252 ° C; HRMS (ESI): C ₄₂ H ₄₃ N ₅ O ₁₄ S ₄ calculated value 969.1519, found value 969.3258.

6. Preparation of compound z-8-1

The compound 6 was taken up with the bromo compound at the 3-position of the benzene ring of the compound 6, and the preparation and purification methods were the same as that of compound z-7-1 to obtain 320 mg of a yellow solid in a yield of 69%, that is, compound z-8-1. mp 252-254 ° C; HRMS (ESI): C ₄₂ H ₄₂ BrN ₂ O ₁₄ S ₄ calculated value 1047.1430, found value 1047.2131.

Example 6 In vitro anti-tumor activity test of the 10,11-methylenedioxycamptothecin derivative of the present invention

To preliminarily evaluate the cytotoxic activity of the 20-position modified camptothecin derivative, we determined the inhibition rate of compound 2, 3 and w-1-w-33 on human lung cancer A549 cells at a concentration of 50 nM according to conventional test methods in the art . The experimental results are shown in Table 1. At a concentration of 50 nM, the 20-position sulfonyl amidine-modified compound w-1-w-24 with glycine as the connecting arm, and the 20-position uracil-modified compound w- with glycine as the connecting arm The 25-w-27 and 20-position heterocyclic amide-modified compounds w-28-w-33 with glycine as the connecting arm have an inhibition rate of more than 50% on A549 cells, showing good cytotoxic activity and significant Better than irinotecan.

Table 1 A549 cell screening results 1

Table 2 IC ₅₀ values of some compounds on in vitro cell lines A549, HCT-116 and NCI-H1975

In addition, we have tested the compound w-25-w-32 on human lung adenocarcinoma cell line A549, human colon cancer cell line HCT-116 and human non-small cell lung adenocarcinoma cell line NCI-H1975 in accordance with conventional test methods in the art Cytotoxic activity (Table 2), the results show that the 20-modified derivatives have strong anti-tumor activity, A549, HCT-116 and NCI-H1975 cells on compounds w-25, w-27, w-30 and w- All of 31 are very sensitive, in which the compound w-31 modified by 2-methylpiperidine amide at position 20 has an inhibitory activity on A549 cells (IC ₅₀ = 9.76 nM) 1000 times better than that of irinotecan (IC ₅₀ = 9.14 uM).

Table 3 in vitro cell lines of some compounds SW620, HCT-8, IC ₅₀ values of NCI-H446

Note: ^a table shows the inhibition rate of irinotecan at 10uM on the corresponding cell lines.

In addition, we have tested compounds w-31, z-1-9, z-2-5, z-3-3, and z-4-2 on human colon cancer cells SW620, human colon cancer in accordance with conventional test methods in the art. Cell HCT-8 and human small cell lung cancer cell NCI-H446 cytotoxic activity (Table 3), the data shows that SW620, HCT-8, NCI-H446 cells are very sensitive to compound w-31, IC ₅₀ can reach 0.11nM, which The activity is higher than irinotecan, and the other four compounds are also higher than irinotecan.

Example 7 In vivo anti-tumor activity test of compound w-31

Using a similar test method, we tested the growth inhibitory effect of compound w-31 on mouse A549 xenografts.

Human lung cancer A549 cells were subcutaneously inoculated into the right armpit of mice, and when they grew to about 100mm ³ , they were divided into groups. The w-31 high, medium and low dose groups were given 20mg / kg, 10mg / kg and 5mg / kg compound w-31, twice a week, the solvent control group was gavaged with an equal volume of solvent. The body weight of mice was measured three times a week. 19 days after the administration, the control mice were sacrificed. The mice in the remaining groups stopped the administration and continued to measure until 28 days, and the mice were sacrificed.

Table 5. Effect of compound W-31 on the growth of A549 xenografts in mice

Note: ^* p <0.01 vs solvent control group

^aCalculated based on data 19 days after administration

The results show that in Table 5, Figure 1 and Figure 2, compound w-31 has a significant inhibitory effect on the growth of A549 solid tumors in mice, and it is dose-dependent; 5, 10 mg / kg dose administration has no significant effect on mouse body weight, The 20mg / kg dose has a certain effect on the body weight of mice, and the toxic and side effects are improved compared with similar compounds.

Example 8 In vivo anti-tumor activity test of compound z-1-9

Using techniques well known in the art, the present inventors tested the in vivo antitumor activity of the compound z-1-9 of the present invention through a mouse xenograft model.

Test methods include:

Human lung cancer A549 cells were cultured to the required number in vitro, collected cells were centrifuged, the supernatant was washed, washed once with PBS, resuspended the cells with PBS, and subcutaneously inoculated into the armpit of the right forelimb of BALB / c-nu mice, each 8 × 10 ⁶ cells. When the volume of the axillary tumor of the mouse grew to about 100 mm ³ , the mice were randomly divided into 3 groups according to the tumor volume, the solvent control group, the z-1-9 high-dose group and the low-dose group. The z-1-9 high-dose group and the low-dose group were given intragastrically 24 mg / kg and 12 mg / kg z-1-9, once every 2 days. The changes in body weight and tumor volume were recorded three times a week.

Observation of results:

After BALB / c-nu mice were subcutaneously inoculated with A549 cells, the tumor volume reached 100 mm ^{3 in} about 10 days, and were randomly administered in groups. On the 19th day, the tumor tissue of the solvent control group grew to 1157.21 ± 370.81 mm ³ , the mice were sacrificed, the tumor tissue was stripped, and frozen at -80 ° C. The rest of the groups stopped the drug administration, continued observation, and recorded the tumor volume. All mice were sacrificed on the 28th day after the drug administration, the tumor tissue was stripped, and frozen at -80 ° C.

Using the days of administration as the abscissa, draw the tumor volume growth curve, as shown in Figure 3. Compared with the solvent control group, the high-dose and low-dose administration of z-1-9 compound can significantly inhibit the growth of A549 transplanted tumors in mice, and it has a certain dose dependence. The tumor inhibition rates after administration for 19 days were 73.79, respectively. % And 54.66%.

Table 4. Effect of compound z-1-9 on the growth of A549 xenografts in mice

Note: ^* p <0.01 vs solvent control group

^aCalculated based on data 19 days after administration

Taking the number of days of administration as the abscissa, the mouse body weight change curve is drawn. The results are shown in Figure 4. The body weight of the mice in the solvent control group gradually increased. After z-1-9 administration, the body weight of mice in the high and low dose groups decreased significantly. It gradually recovered after 5 days, but was still lower than the body weight of the control mice, especially in the high-dose group, which indicated that it had certain toxic and side effects. Slight toxic and side effects are common problems of camptothecin compounds.

Example 9 In vivo anti-tumor activity test of compound z-3-3

Using a similar test method, we tested the growth inhibitory effect of compound z-3-3 on mouse A549 xenografts.

Human lung cancer A549 cells were subcutaneously inoculated into the right armpit of mice, and when they grew to about 100mm ³ , they were administered in groups. The two dose groups of z-3-3 high and low were given 5mg / kg and 2.5mg / kg in the tail vein. Compound z-3-3 was administered three times a week for a total of six times for two weeks. The solvent control group was gavaged with an equal volume of solvent. The positive control was irinotecan (3 mg / kg). The body weight of mice was measured three times a week. 28 days after the administration, the mice were sacrificed.

Table 6. Effect of compound z-3-3 on the growth of A549 xenografts in mice

Note: ^* p <0.05vs solvent control group

^** p <0.01 vs solvent control group

^a calculated based on the data 28 days after administration

The results show that in Table 6, Figure 5 and Figure 6, compound z-3-3 has a significant inhibitory effect on the growth of mouse A549 solid tumors in a dose-dependent manner; two doses do not have a significant effect on the body weight of mice, and are toxic Compared with similar compounds (for example: compound z-1-9), the side effects were improved to a certain extent; at a dose of 5 mg / kg, the tumor suppression rate could reach 76.2%, and the growth status of mice was good.

Although cell tests show that the in vitro tumor suppressive activity of the two compounds is significantly lower than that of w-31 and other compounds; but animal experimental data shows that z-1-9 and z-3-3 also have good antitumor activity, which may be Their prodrug properties are related; z-3-3 has both good safety and good prospects for drug development.

The above are only the preferred embodiments of the present invention, and are not intended to limit the present invention in other forms. Any person skilled in the art may use the technical content disclosed above to change or modify the equivalent of equivalent changes. Examples. However, any simple modifications, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention without departing from the content of the technical solution of the present invention still fall within the protection scope of the technical solution of the present invention.

Claims

A compound having the structure represented by formula (I), its isomers and pharmaceutically acceptable salts:

Where X is-(CH 2 ) n -,-(CH 2 ) n O-,-(CH 2 ) n NH-,-(CH 2 ) n CONH-,-(CH 2 ) n NHCO (CH 2 ) m- , (CH 2 ) n- (CH = CH) p- (CH 2 ) m -,-(CH 2 ) n- (CH = CH) p- (CH 2 ) m O-, -O (CH 2 ) n SS (CH 2 ) m O- or -O (CH 2 ) n SS (CH 2 ) m OCOY-;

Y is - (CH 2) n - or - (CH 2) n CONH-;

n and m are independently 0 or positive integer;

p is a positive integer;

R is selected from any one of a hydrogen atom, a substituted or unsubstituted sulfonamide group, a substituted or unsubstituted base, a substituted or unsubstituted saturated nitrogen-containing heterocyclic ring, and a substituted or unsubstituted alkyl group;

When the group is a substituted group, the substituent is selected from hydrogen atom, halogen atom, hydroxyl group, nitro group, amino group, alkyl group, substituted alkyl group, alkoxy group, substituted alkoxy group, aromatic Any one of a group, a substituted aryl group, a heteroaryl group and a substituted heteroaryl group.
The compound according to claim 1, wherein X is-(CH 2 ) n NH-, n is 1, R is a substituted or unsubstituted sulfonamide group, and R has a structure represented by formula (II):

among them,

R 1 is alkyl, aryl, substituted aryl, heteroaryl or substituted heteroaryl;

R 2 is aryl or substituted aryl.
The compound according to claim 2, characterized in that R 1 is C1-C4 alkyl, preferably methyl, ethyl, n-propyl or n-butyl.
The compound according to claim 2, wherein R 1 is phenyl, substituted phenyl, naphthyl, thiophene or pyridine, preferably para-fluorine substitution, chlorine substitution, nitro substitution, methyl substitution or Methoxy substituted phenyl.
The compound according to claim 2, wherein R 2 is a phenyl group or a substituted phenyl group, preferably a para-fluorine-substituted or methoxy-substituted phenyl group.
The compound according to claim 2, wherein the compound is w-1, w-2, w-3, w-4, w-5, w-6, w-7, w-8, w -9, w-10, w-11, w-12, w-13, w-14, w-15, w-16, w-17, w-18, w-19, w-20, w-21 , W-22, w-23 or w-24.
The compound according to claim 1, wherein X is-(CH 2 ) n NHCO (CH 2 ) m- , n and m are both 1, R is a substituted or unsubstituted uracil, and R has the formula (III) Structure shown:

Wherein R 3 is H, F or Cl.
The compound according to claim 7, wherein the compound is w-25, w-26 or w-27.
The compound according to claim 1, wherein X is-(CH 2 ) n NHCO (CH 2 ) m- , n is 1, m is 0, and R is a substituted or unsubstituted saturated nitrogen-containing heterocyclic ring.
The compound according to claim 9, wherein the compound is w-28, w-29, w-30, w-31, w-32 or w-33.
The compound according to claim 1, wherein, X is - (CH 2) n - or - (CH 2) n CONH-, n is 2, R is a substituted or unsubstituted alkyl group.
The compound according to claim 11, wherein the compound is z-1-9 or z-2-5.
The compound according to claim 1, characterized in that

X is -O (CH 2 ) n SS (CH 2 ) m OCOY-, Y is-(CH 2 ) n -or-(CH 2 ) n CONH-;

Both n and m are 2, and R is substituted or unsubstituted alkyl.
The compound according to claim 13, wherein the compound is z-3-3, z-4-2, z-5-1, z-6-1, z-7-1 or z-8 -1.
The compound according to claim 1, wherein X is -CH 2 NH- or -OCH 2 CH 2 SSCH 2 CH 2 O-, and R is H.
A pharmaceutical composition comprising a therapeutically effective dose of the compound of any one of claims 1-15, its isomer or pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier.
Use of a compound according to any one of claims 1-15, an isomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 16 in the preparation of a medicament for preventing and / or treating cancer.
The use according to claim 17, wherein the cancer is lung cancer or colon cancer.