Aminated 20 (S)-10, 11-difluoromethylenedioxycamptothecin derivative as well as preparation method and application of aminated 20 (S)-10, 11-difluoromethylenedioxycamptothecin derivative

Abstract

The invention belongs to the technical field of organic synthesis and medicines, and particularly relates to an aminated 20 (S)-10, 11-difluoromethylenedioxy camptothecin derivative as well as a preparation method and application thereof. The derivative is a compound with a structure as shown in a formula (I), and in the formula (I), R is shown in the description; wherein X is an integer from 3 to 10, and n is an integer from 3 to 10; and Z is selected from (amido substitution). The novel aminated 20 (S)-10, 11-difluoromethylenedioxy camptothecin derivative provided by the invention has good in-vivo and in-vitro anti-tumor activity, and can be used for preparing and synthesizing camptothecin drugs for preventing or treating tumors, the preparation method is easy to operate, the post-treatment is simple and convenient, the initial raw materials of the reaction are cheap and easy to obtain, the designability of the reaction substrate is strong, and the method is suitable for industrial production. The reaction efficiency is relatively high, and the practicability is relatively high.

Description

Technical Field

[0001] This invention belongs to the fields of organic synthesis and pharmaceutical technology, specifically relating to the amination of 20(S)-10,11-difluoromethylenedioxycamptothecin derivatives, their preparation methods, and applications. Background Technology

[0002] Camptothecin (CPT) is a naturally occurring quinoline alkaloid from the tree *Camptotheca acuminata*. It exhibits significant antitumor activity by inhibiting DNA topoisomerase I (TOPO I). Its derivatives, topotecan, irinotecan, and beloteccan, have been approved for the clinical treatment of colorectal cancer, small cell lung cancer, and ovarian cancer. Structure-activity relationship studies indicate that modification of camptothecin can significantly affect its antitumor activity, making it an optimal site for further development of novel, highly active antitumor drugs. However, due to its poor water solubility and toxicity under physiological conditions, camptothecin cannot be directly applied clinically.

[0003] Chinese invention patent CN110590796A discloses camptothecin derivatives, their preparation methods, and applications. The compounds provided by this invention are a class of novel camptothecin derivatives with methylenedioxy groups introduced at the 10 and 11 positions of the parent ring and different substituent groups introduced at the 7- position. The preparation method uses readily available raw materials, is simple to synthesize, and is convenient and quick to purify. Furthermore, the compounds exhibit excellent in vitro cytotoxic activity and superior in vivo antitumor effects.

[0004] Invention patents WO2005009347A2 and WO0149291 disclose methods for forming camptothecin compounds that are effective antitumor compounds. These compounds can inhibit the enzyme topoisomerase I and can alkylate the DNA of the associated topoisomerase I-DNA cleavable complex.

[0005] Chinese invention patent application CN117777153A discloses a 20(S)-10,11-difluoromethylenedioxycamptothecin derivative, its preparation method and application. This type of derivative has good solubility, anti-inflammatory activity, in vitro and in vivo antitumor activity and cell transmembrane transport ability.

[0006] The prior art represented by the aforementioned documents has at least the following unresolved technical problems or defects: Currently, camptothecin derivatives approved for anticancer use are susceptible to interactions among several mechanisms that enable cancer cells to tolerate chemotherapy, and drug resistance and toxicity remain bottlenecks in the development of these compounds. There is still a need to develop new camptothecin derivatives, especially those with higher potency, lower toxicity, better permeability, and the ability to overcome tumor drug resistance. Furthermore, the preparation methods for these compounds involve complex reaction procedures, low yields, high costs, and difficulty in large-scale production due to the introduction of substituents in the final reaction step. Because the synthetic route involves introducing substituents into the parent structure in the final step, steric hindrance and the conjugated system make it very difficult to establish negative charges at the introduction site, requiring further improvement. Summary of the Invention

[0007] Drug resistance and toxicity of existing camptothecin derivatives remain key obstacles to their drug development. This invention introduces novel camptothecin derivatives by introducing different types of amino groups at the 7-position of their backbone. This series of derivatives possesses a novel antitumor mechanism, simultaneously targeting Topo I and the RNA helicase DDX5 (p68). While inducing apoptosis by targeting Topo I to induce DNA damage in tumor cells, it simultaneously downregulates DDX5 expression in tumor cells, blocking the DNA damage repair pathway initiated by DDX5, overcoming the risk of tumor drug resistance caused by Topo I inhibitors, and reducing toxicity. Some compounds in this invention have an amino active site at the 7-position, which can be used for toxic loading in ADC drugs. Furthermore, this invention optimizes the synthetic route for these derivatives to address the problems of complex preparation methods and low yields in existing methods. This synthetic route is simple, low-cost, and has a high yield, making it suitable for large-scale preparation.

[0008] To achieve the above objectives, the technical solution adopted by the present invention is as follows: In a first aspect, the present invention provides an aminolated 20(S)-10,11-difluoromethylenedioxycamptothecin derivative, which is a compound having the structure shown in formula (I):

[0009] In equation (Ⅰ), R is ; Where X is n is an integer between 3 and 10; Z is selected from R1 and R2 are each independently selected from H, halogens, substituted or unsubstituted C. 1-30 Alkyl, substituted or unsubstituted C 3-12 alkenyl, substituted or unsubstituted C 1-10 Alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted C 3-20 Ethers, substituted or unsubstituted C 1-10 Ester group, substituted or unsubstituted C 3-10alkoxy, substituted or unsubstituted C 1-10 Alkylsilyl, substituted or unsubstituted C 3-10 alkylthio, substituted or unsubstituted C 3-10 Alkyl, substituted or unsubstituted C 3-10 alkylimine group, substituted or unsubstituted C 7-10 alkenylamine, substituted or unsubstituted C 7-10 alkenyl imine, substituted or unsubstituted C 10-30 adamantanes; The alkyl, alkenyl, alkynyl, alkoxy, ether, alkylthio, alkazyl, alkimino, alkenylamine, and alkenylimino groups are straight-chain, branched, or cyclic structures.

[0010] As some specific embodiments of the present invention, in the compound with the structure shown in formula (I), Z is selected from substituted or unsubstituted C. 3-10 Cycloalkylamines, substituted or unsubstituted C 10-30 adamantane amines, substituted or unsubstituted C 3-12 Cycloalkenylamines, substituted or unsubstituted arylamines, substituted or unsubstituted C 3-20 Ether cyclic amines, or substituted or unsubstituted aromatic heterocyclic amines; Preferably, the aromatic heterocyclic group is selected from amine-substituted pyridine rings, furan rings, thiophene rings, pyrazole rings, indole rings, benzopyrazole rings, piperidine rings, morpholine rings, thiomorpholine rings, naphthalene rings, or triazole rings.

[0011] As some specific embodiments of the present invention, in the compound with the structure shown in formula (I), when R1 and R2 are substituted structures, the substituted groups are each independently selected from halogens, C 1-10 Straight-chain or branched alkyl groups, C 1-10 alkenyl, C 1-10 alkynyl group, C 3-12 cycloalkyl, C 1-10 Alkoxy, C 1-10 Alkylthio, C 1-10 Alkylsilyl, C 1-10 Halogenated alkoxy groups, C 1-10 Ester group, 3-12 membered heterocyclic group, C 6-14 aryl, oxygen C 6-14 aryl, oxygen C 6-14 Aromatic heteroyl, nitrogen C 6-14 Aryl, nitrogen C 5-14 Aromatic heteroyl, 5-14 heteroaryl, -CN, -NO2, -CF2H, -CF2OH, -CF3 or -OCF3.

[0012] As some preferred embodiments of the present invention, in the compound with the structure shown in formula (I), R1 and R2 are each independently selected from H, halogens, and C. 1-10 Straight-chain or branched alkyl, substituted or unsubstituted C 3-12 cycloalkyl, substituted or unsubstituted C 1-10 alkoxy, substituted or unsubstituted C 3-10 Epoxyalkyl, C 1-10 Alkylsilyl, C 1-10 Ester group, substituted or unsubstituted C 6-14 aryl, substituted or unsubstituted C 10-16 adamantane, substituted or unsubstituted C 7-10 alkenylamine, or substituted or unsubstituted C 7-10 Alkenyl imine group.

[0013] As some preferred embodiments of the present invention, in the compound with the structure shown in formula (I), the substituted or unsubstituted C 1-10 The alkoxy group is C 1-10 Halogenated alkoxy groups; and / or the substituted or unsubstituted C groups. 7-10 The alkenyl imine group is either substituted or unsubstituted C. 7-10 Alkyl C 7-10 Alkenyl imine group.

[0014] As some further preferred embodiments of the present invention, in the compound with the structure shown in formula (I), Z is selected from any of the following groups: .

[0015] Secondly, the present invention provides a method for preparing the above-mentioned 20(S)-10,11-difluoromethylenedioxycamptothecin derivative, the synthetic route of which is as follows:

[0016] Includes the following steps: (1) Add the raw material 5-amino-2,2-difluoro-1,3-benzodioxane and brominated C3-10 alkyl nitrile to organic solvent A, add Lewis acid in an inert atmosphere, and react to obtain intermediate I; (2) Intermediate I, tricyclic ketone and catalyst are added to organic solvent B and completely dissolved. The organic solvent is evaporated to dryness to obtain a solid. Vacuum solid-solid reaction is carried out under heating to obtain intermediate II. (3) Add intermediate II and amine compounds to organic solvent C, and add basic compounds to react to obtain the amination of 20(S)-10,11-difluoromethylenedioxycamptothecin derivative.

[0017] As some specific embodiments of the present invention, the 5-amino-2,2-difluoro-1,3-benzodioxane and bromoC 3-10 The molar ratio of alkyl nitrile to Lewis acid is 1:0.1-5:1-50; preferably 1:0.45-0.5:18-22.

[0018] As some specific embodiments of the present invention, the molar ratio of intermediate I, tricyclic ketone and catalyst is 1:0.1-5:0.1-5; preferably 1:0.5-1.2:0.1-0.6.

[0019] As some specific embodiments of the present invention, the molar ratio of intermediate II, amine compound and basic compound is 1:0.1-15:0.1-10; preferably 1:5-10:3-5.

[0020] As some specific embodiments of the present invention, in step (1), the reaction temperature is 80-95℃; the reaction process is monitored by TLC, and after the reaction is completed, an acidic aqueous solution is added to quench the reaction; As some specific embodiments of the present invention, in step (2), the reaction is carried out under vacuum; and / or the temperature of the reaction is 60-150°C and the time is 0.5-20h; preferably, the temperature of the reaction is 115°C and the time is 1h.

[0021] As some specific embodiments of the present invention, in step (3), the reaction temperature is 30-70℃ and the time is 2-30 h; preferably, the reaction temperature is 50℃ and the time is 5 h.

[0022] Preferably, the acidic aqueous solution is an acidic aqueous solution with a concentration of 1-10 mol / ml, and more preferably, a 3 mol / ml HCl aqueous solution.

[0023] As some specific embodiments of the present invention, the Lewis acid is selected from any one of boron trichloride, boron trifluoride, aluminum trichloride, gallium trichloride, trimethylaluminum, ferric chloride, and ferric bromide; preferably boron trichloride.

[0024] As some specific embodiments of the present invention, the catalyst is selected from any one of Lewis acids, iodine, dodecyl sulfate, ferric chloride hexahydrate, aminosulfonic acid, 2,4,6-trichloro-1,3,5-triazine, bismuth trifluoromethanesulfonate, yttrium trifluoromethanesulfonate, lithium bis(trifluoromethanesulfonyl)imide, magnesium bis(trifluoromethanesulfonyl)imide, and p-toluenesulfonic acid or its hydrates; preferably p-toluenesulfonic acid hydrate.

[0025] As some specific embodiments of the present invention, the alkaline compound is selected from one or more of N,N-diisopropylethylamine, sodium bicarbonate, potassium carbonate, sodium carbonate, calcium carbonate, potassium hydroxide, sodium hydroxide, and triethylamine; preferably N,N-diisopropylethylamine, sodium bicarbonate, potassium carbonate, and triethylamine; more preferably N,N-diisopropylethylamine or sodium bicarbonate.

[0026] As some specific embodiments of the present invention, the organic solvent A is selected from one or more of dichloroethane, dichloromethane, methanol and acetonitrile; preferably dichloroethane.

[0027] As some specific embodiments of the present invention, the organic solvent B is selected from methanol, ethanol, n-butanol, ethyl acetate, acetone, dichloromethane, and dichloroethane; preferably dichloromethane or methanol; and more preferably dichloromethane.

[0028] As some specific embodiments of the present invention, the organic solvent C is selected from ethanol, methanol, dichloromethane, and DMF; preferably dichloromethane or DMF; and more preferably DMF.

[0029] Thirdly, the present invention provides the use of the above-mentioned 20(S)-10,11-difluoromethylenedioxycamptothecin derivative in the preparation of medicaments for the prevention and / or treatment of cancer.

[0030] As some preferred embodiments of the present invention, the cancer is any one of bladder cancer, pancreatic cancer, breast cancer, prostate cancer, lung cancer, ovarian cancer, cervical cancer, skin cancer, melanoma, colon cancer, stomach cancer, liver cancer, esophageal cancer, kidney cancer, pharyngeal cancer, thyroid cancer, testicular cancer, brain cancer, bone cancer, and leukemia; preferably bladder cancer.

[0031] Fourthly, the present invention also provides a pharmaceutical formulation comprising: a therapeutically effective dose of any one of the above-mentioned amino-modified 20(S)-10,11-difluoromethylenedioxycamptothecin derivative, or its active metabolite, prodrug, stereoisomer thereof, pharmaceutically acceptable salt, polymorph, solvate or conjugate, and a pharmaceutically acceptable carrier.

[0032] As some specific embodiments of the present invention, the pharmaceutical formulation is an antibody-drug conjugate using an amino-modified 20(S)-10,11-difluoromethylenedioxycamptothecin derivative, its active metabolite, prodrug, its stereoisomer, pharmaceutically acceptable salt, polymorph, solvate, or conjugate as a cytotoxic agent; the antibody-drug conjugate also includes, but is not limited to, one or more of antibodies, peptides, nucleic acids, and small molecules.

[0033] The pharmaceutical composition can be formulated into liquid preparations such as tablets, capsules, powders, granules, lozenges, suppositories, oral liquids, or sterile parenteral suspensions, as well as injections such as large or small volume injections and lyophilized powders. All of the above dosage forms can be prepared using conventional methods in the pharmaceutical field.

[0034] Compared with the prior art, the present invention has the following beneficial effects: (1) The novel C-introduced at the 7-position of the mother core provided by this invention 3-10 20(S)-10,11-difluoromethylenedioxycamptothecin derivatives with different types of amino groups exhibit good in vitro and in vivo antitumor activity and can be used to prepare and synthesize camptothecin-based drugs for the prevention or treatment of tumors. These compounds have good in vitro and in vivo antitumor activity.

[0035] (2) The synthetic route of the present invention is simple, easy to operate, and easy to post-process. The starting materials are cheap and readily available. Moreover, the synthetic route overcomes the problem of introducing substituents into the parent structure in the last step of the existing synthetic route, which causes the reaction to be difficult due to steric hindrance and the formation of negative electrons at the introduced position caused by the conjugated system. The synthetic route effectively improves the reaction efficiency. At the same time, the reaction substrate of the synthetic route is highly designable and has a wide range of functional groups. Different substituents of 20(S)-10,11-difluoromethylenedioxycamptothecin derivatives can be designed and synthesized according to actual needs, which is highly practical. Attached Figure Description

[0036] Figure 1 This is a synthetic route diagram of the 20(S)-10,11-difluoromethylenedioxycamptothecin derivative of the present invention; Figure 2 Compound 1 prepared in Example 1 of this invention 1 H NMR spectrum; Figure 3 Compound 1 prepared in Example 1 of this invention 13 C NMR spectrum; Figure 4 Compound 7 prepared in Example 7 of this invention 1 H NMR spectrum; Figure 5 Compound 7 prepared in Example 7 of this invention 13 C NMR spectrum; Figure 6 The changes in tumor volume and body weight in mice of different groups were measured in an in vivo experiment to inhibit colon cancer tumor growth. Figure 7 The study investigated changes in tumor volume and body weight in mice across different groups during an in vivo assay to inhibit bladder cancer tumor growth. Detailed Implementation

[0037] Unless otherwise defined, all technical terms in this document have the same meanings as commonly understood by one of ordinary skill in the art to which the subject matter of the claims pertains. Unless otherwise stated, all patents, patent inventions, and publications cited in this document are incorporated herein by reference in their entirety. If multiple definitions exist for terms in this document, the definitions in this chapter shall prevail.

[0038] It should be understood that the above brief description and the following detailed description are exemplary and for illustrative purposes only, and do not limit the subject matter of the invention in any way. In this invention, the singular is used in conjunction with the plural unless otherwise specifically stated. It should also be noted that, unless otherwise stated, the use of “or” or “or” means “and / or”. Furthermore, the use of the term “comprising” and other forms such as “including,” “containing,” and “contains” are not limiting.

[0039] The definition of standard chemical terms can be found in the reference "Basic Organic Chemistry (Volumes 1 & 2)" by Xing Qiyi, Higher Education Press, 3rd Edition, 2005-06.

[0040] Unless otherwise stated, conventional methods within the scope of the art shall be used.

[0041] Unless specifically defined herein, the use of all commercially available products herein employs standard techniques. For example, it may be carried out using the manufacturer's instructions for use with the kit, or in accordance with methods known in the art or the description of this invention. The techniques and methods described herein can generally be implemented according to conventional methods well known in the art, based on the descriptions in the various summary and more specific documents cited and discussed in this specification.

[0042] As used herein, the term "substituted" means that one or more hydrogen atoms have been removed from the chemical group and that it has been substituted by a substituent. As described herein, the term "substituent" has the common meaning consistent in the field and refers to a chemical moiety covalently attached to, or at that time fused to, the parent group. It should be understood that substitution at a given atom is limited by the valence of the atom.

[0043] As used herein, the term “stereoisomer” refers to any of the various stereoisomer configurations (e.g., enantiomers, diastereomers, and racemates) of an asymmetric compound (e.g., a compound having one or more asymmetricly substituted carbon atoms or “asymmetric centers”).

[0044] As used herein, the term "pharmaceutically acceptable salt" includes salts formed by the compounds of this invention with acids or bases.

[0045] As used herein, the term "prodrug" includes compounds that are converted into compounds of the present invention during their residence in the body (e.g., through enzymatic or hydrolytic processes).

[0046] As used herein, the term "solvent" refers to those forms of compounds that, in the solid or liquid state, form complexes with solvent molecules through coordination. Hydrates are a specific form of solvate that coordinates with water. Hydrates are preferred solvates within the scope of this invention.

[0047] As used in this article, the term "polymorph" refers to a compound (or its salt or solvate) that can crystallize in different crystalline stacks, all of which have the same elemental composition.

[0048] As used in this article, the term "complex" refers to a complex formed by two or more compounds bonded together by relatively weak forces, such as hydrogen bonds or van der Waals forces.

[0049] An "intermediate" refers to a compound formed during the process of converting a starting material into a target product through a reaction; it serves as a bridge connecting different reaction steps. It can be a semi-finished product generated from cyclic compounds (such as benzene and naphthalene) or acyclic compounds (such as methane and propylene) through reactions such as sulfonation, nitration, and reduction. Intermediates are relative; they are both the product of one reaction and the starting material for the next.

[0050] The compounds of this invention can be used to treat subjects with a variety of cancers or at risk of developing cancer. Examples of such cancers include pancreatic cancer, 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, testicular cancer, brain cancer, bone cancer, and blood cancers (such as leukemia, chronic lymphocytic leukemia), etc. Other cancers include, but are not limited to, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain and central nervous system (CNS) cancer, cervical cancer, choriocarcinoma, colorectal cancer, connective tissue cancer, digestive system cancer, endometrial cancer, esophageal cancer, eye cancer, head and neck cancer, intraepithelial neoplasia, laryngeal cancer, lung cancer (small cell, large cell), lymphoma (including Hodgkin's lymphoma and non-Hodgkin's lymphoma); melanoma; neuroblastoma; oral cancer (e.g., lip, tongue, mouth and pharynx); retinoblastoma; rhabdomyosarcoma; respiratory system cancer; sarcoma; uterine cancer; urinary system cancer; and other cancers and sarcomas.

[0051] A “therapeutic effective dose” is any amount of a drug, as described below, that, when used alone or in combination with another therapeutic agent, promotes disease regression, manifested as a reduction in the severity of disease symptoms, an increase in the frequency and duration of symptom-free periods, or prevention of impairment or disability resulting from the disease. A therapeutically effective dose or amount of a drug includes a “preventive effective dose,” which is any amount of a drug, as described below, that, when administered alone or in combination with another therapeutic agent to a subject at risk of developing the disease or suffering from a relapse of the disease, inhibits the onset or recurrence of the disease. The ability of a therapeutic agent to promote disease regression or inhibit disease progression or recurrence can be assessed using various methods known to those skilled in the art, such as in animal model systems where efficacy in humans can be predicted or by measuring the activity of the compound reagent in an in vitro assay system.

[0052] As an example of tumor treatment, in the most preferred embodiment, a therapeutically effective amount or dose of the drug can effectively inhibit cell or tumor growth compared to an untreated animal model. The ability of the compound to inhibit tumor growth can be evaluated in animal model systems, which can predict its efficacy in human tumors. Alternatively, this property of the composition can be assessed by testing the compound's ability to inhibit cell growth, such inhibition being measured in vitro using assays known to those skilled in the art. In a preferred embodiment of the invention, tumor regression can be clearly observed.

[0053] The present invention will be further described in detail below with reference to the embodiments.

[0054] This invention synthesizes and tests a series of novel camptothecin derivatives with difluoro-substituted methylenedioxy groups at the 10 and 11 positions of the parent ring and different amino substituent groups at the 7- position. The 20(S)-10,11-difluoromethylenedioxy camptothecin derivatives described in this invention can be listed as some of the compound structures in Table 1, but are not limited to the structures shown in Table 1 (all compound structures are described in the invention description). Table 1

[0055] Figure 1 The synthetic route of the above compounds is shown. The following specific examples illustrate this: Example 1 The preparation of 20(S)-7-[3-(cyclopropylamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 1) is as follows: (1) Preparation of 6-amino-2,2-difluorobenzo[d][1,3]dioxane-4-bromobutanone (intermediate I) The raw material 5-amino-2,2-difluoro-1,3-benzodioxane (1 g, 5.84 mmol) was added to a three-necked flask, dissolved in dichloroethane (5 mL), and stirred in an ice-water bath. Under nitrogen protection, boron trichloride (20 mL, 120 mmol) was added, followed by bromobutyronitrile (569 μL, 2.6 mmol) after 10 min. The mixture was refluxed in an oil bath at 95°C for 3 hours. The reaction was monitored by TLC (using a mixture of petroleum ether and ethyl acetate in a ratio of 800:100, v / v). After the reaction was completed, an appropriate amount of 3 mol / mL HCl aqueous solution was added to quench the reaction, consuming excess bromobutyronitrile. After stirring for ten minutes, the mixture was transferred to a separatory funnel, extracted three times with dichloromethane, and the organic phases were combined. The mixture was dried with anhydrous magnesium sulfate, filtered under reduced pressure, and the filtrate was concentrated to obtain a pale yellow liquid. The liquid was separated by silica gel column chromatography (using a mixture of petroleum ether and dichloromethane in a ratio of 20:1, v / v). After concentration, a yellow oily substance (intermediate I) was obtained, with a yield of 83.2% and a purity of 91.88%.

[0056] (2) Preparation of 20(S)-7-(3-bromopropyl)-10,11-difluoromethylenedioxycamptothecin (intermediate II) The product obtained in step (1) (intermediate I) (110.3 mg, 0.34 mmol), tricyclic ketone (48.5 mg, 0.2 mmol), and p-toluenesulfonic acid hydrate (35.6 mg, 0.2 mmol) were added to a round-bottom flask and dissolved in dichloromethane (3 mL). After reacting at 115 °C under vacuum for 1 h, the reaction was detected by TLC (the developing solvent was a mixed solution of dichloromethane and methanol = 1600:100, v / v). After the reaction was completed, the product was separated by silica gel column chromatography (the eluent was a mixed solution of dichloromethane and methanol = 300:1, v / v, with a small amount of glacial acetic acid added). After concentration, the product was washed with methanol and filtered to obtain a pure white solid product (intermediate II) on the filter cake. The yield was 73.6% and the purity was 97.62%.

[0057] (3) Preparation of 20(S)-7-(3-aminopropylcyclopropyl)-10,11-difluoromethylenedioxycamptothecin (compound 1) 20(S)-7-(3-bromopropyl)-10,11-difluoromethylenedioxycamptothecin (intermediate II) (50 mg, 0.1 mmol) was added to a round-bottom flask, followed by the addition of DMF (2 mL), sodium bicarbonate (3 equivalents, 25 mg), and cyclopropylamine (5 equivalents, 35 μL). The mixed solvent was placed in an oil bath at 50 °C for 5 h. After cooling, the mixture was extracted with DCM and water, and the organic layer was collected. Anhydrous magnesium sulfate was used to remove water, and the solution was then subjected to column chromatography (eluent was a mixed solution of DCM:MeOH = 15:1, v / v) to obtain a white solid powder with a yield of 70% and a purity of 95.34%.

[0058] 1 H NMR (400 MHz, DMSO-d6) δ=8.14 (d, J = 5.6 Hz, 1H), 7.99 – 7.91 (m,1H), 7.25 (d, J = 2.8 Hz, 1H), 6.54 (s, 1H), 5.40 (s, 2H), 5.16 (d, J = 7.2Hz, 2H), 3.16 – 3.10 (m, 2H), 2.65 (t, J = 6.6 Hz, 2H), 2.07 (dq, J = 6.5,3.3 Hz, 1H), 1.84 (ddd, J = 26.4, 13.5, 7.2 Hz, 4H), 0.89 (t, J = 7.3 Hz, 3H), 0.35 (dt, J = 6.0, 2.9 Hz, 2H), 0.22 (p, J = 3.7 Hz, 2H). 13 C NMR (151 MHz, DMSO-d6) δ=172.88, 157.13, 151.71, 150.47, 146.89,146.05, 145.47, 145.17, 143.54, 133.23, 131.55, 129.87, 128.89, 125.03,119.36, 108.87, 103.61, 97.08, 72.81, 65.67, 50.09, 48.89, 30.80, 30.73,29.69, 27.52, 8.20, 6.37. Example 2 The preparation of 20(S)-7-[3-(cyclopropylmethylamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 2) is as follows: Using intermediate II prepared in Example 1, in the third step of the reaction, cyclopropylamine was replaced with methylcyclopropylamine (the molar ratio with the starting material was the same as in Example 1). The types and amounts of each starting material and the operation steps were the same in the remaining steps. Finally, a white solid product (compound 2) was obtained with a yield of 64% and a purity of 95.23%.

[0059] 1 H NMR (400 MHz, DMSO-d6) δ = 8.32 (s, 1H), 8.05 (d, J = 3.3 Hz, 1H), 7.28 (s, 1H), 6.54 (s, 1H), 5.43 (s, 2H), 5.31 – 5.14 (m, 2H), 3.25 (d, J =9.0 Hz, 2H), 3.09 (d, J = 7.7 Hz, 2H), 2.79 (d, J = 7.1 Hz, 2H), 2.08 – 1.93(m, 2H), 1.88 (tq, J = 14.1, 7.1 Hz, 2H), 1.20 (d, J = 4.2 Hz, 1H), 1.11 –1.03 (m, 1H), 0.89 (t, J = 7.3 Hz, 3H), 0.57 (d, J = 7.4 Hz, 2H), 0.34 (d, J= 4.9 Hz, 2H). 13 C NMR (101 MHz, DMSO-d6) δ = 172.90, 157.19, 152.00, 150.51, 147.02,146.05, 145.55, 143.75, 143.67, 131.58, 129.27, 124.97, 119.49, 109.13,103.75, 97.14, 72.82, 65.69, 51.98, 50.33, 46.61, 30.74, 29.47, 27.05, 26.28,8.23, 7.83, 4.38. Example 3 The preparation of 20(S)-7-[3-(cyclobutamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 3) is as follows: Using intermediate II prepared in Example 1, in the third step of the reaction, cyclopropylamine was replaced with cyclobutylamine (the molar ratio with the starting material was the same as in Example 1). The types and amounts of each starting material and the operation steps were the same in the remaining steps. Finally, a white solid product (compound 3) was obtained with a yield of 72% and a purity of 96.16%.

[0060] 1 H NMR (600 MHz, DMSO-d6) δ = 8.27 (d, J = 2.4 Hz, 1H), 7.97 (dd, J =6.3, 2.6 Hz, 1H), 7.25 (d, J = 2.6 Hz, 1H), 6.52 (s, 1H), 5.40 (s, 2H), 5.23– 5.10 (m, 2H), 3.67 (s, 1H), 3.21 (dd, J = 10.8, 6.4 Hz, 2H), 3.00 (t, J =7.4 Hz, 2H), 2.23 – 2.13 (m, 4H), 1.94 (p, J = 7.6 Hz, 2H), 1.90 – 1.83 (m,2H), 1.82 – 1.72 (m, 2H), 0.88 (t, J = 7.3 Hz, 3H). 13 C NMR (151 MHz, DMSO-d6) δ = 172.87, 157.15, 151.89, 150.48, 146.93,145.96, 145.50, 143.73, 143.60, 133.23, 131.55, 129.86, 129.07, 124.84,119.45, 108.99, 103.61, 97.16, 72.81, 65.69, 55.35, 51.69, 50.23, 44.58,30.82, 27.12, 26.79, 26.40, 15.05, 8.20. Example 4 The preparation of 20(S)-7-[3-(cyclopentanamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 4) is as follows: Using intermediate II prepared in Example 1, in the third step of the reaction, cyclopropylamine was replaced with cyclopentylamine (the molar ratio with the starting material was the same as in Example 1). The types and amounts of each starting material and the operation steps were the same in the remaining steps. Finally, a white solid product (compound 4) was obtained with a yield of 69% and a purity of 95.36%.

[0061] 1H NMR (400 MHz, DMSO-d6) δ = 8.32 (d, J = 3.0 Hz, 1H), 8.07 – 7.96 (m, 1H), 7.28 (d, J = 3.2 Hz, 1H), 6.55 (s, 1H), 5.42 (s, 2H), 5.27 – 5.17(m, 2H), 3.28 – 3.22 (m, 2H), 3.17 – 3.11 (m, 2H), 1.98 (dt, J = 11.9, 7.5Hz, 4H), 1.86 (dq, J = 14.0, 7.0 Hz, 2H), 1.75 – 1.51 (m, 6H), 0.88 (t, J =7.3 Hz, 3H). 13 C NMR (101 MHz, DMSO-d6) δ = 172.91, 157.20, 151.99, 150.51, 146.99,146.03, 145.54, 143.76, 143.52, 134.09, 131.57, 129.21, 124.89, 119.48,109.10, 103.69, 97.19, 72.83, 65.69, 58.93, 50.32, 45.99, 30.75, 29.66,27.08, 26.34, 24.10, 8.22. Example 5 The preparation of 20(S)-7-[3-(N-cyclopentyl-N-methylamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 5) is as follows: Using intermediate II prepared in Example 1, in the third step of the reaction, cyclopropylamine was replaced with cyclopentylmethylamine (the molar ratio with the starting material was the same as in Example 1). The types and amounts of each starting material and the operation steps were the same in the remaining steps. Finally, a white solid product (compound 5) was obtained with a yield of 66% and a purity of 96.16%.

[0062] 1H NMR (400 MHz, DMSO-d6) δ = 8.38 (s, 1H), 8.13 (d, J = 5.8 Hz, 1H), 7.32 (d, J = 2.2 Hz, 1H), 6.56 (s, 1H), 5.45 (s, 2H), 5.36 (s, 2H), 3.23 (s,2H), 3.17 (d, J = 5.0 Hz, 2H), 2.66 (s, 2H), 2.00 (s, 3H), 1.88 (p, J = 7.1Hz, 2H), 1.71 – 1.54 (m, 5H), 1.36 – 1.18 (m, 4H), 0.88 (t, J = 7.3 Hz, 3H). 13 C NMR (101 MHz, DMSO-d6) δ = 172.91, 157.20, 151.99, 150.51, 146.99,146.03, 145.54, 143.76, 143.52, 134.09, 131.57, 129.21, 124.89, 115.28,107.20, 101.89, 92.09, 73.23, 61.64, 56.23, 51.33, 43.24, 31.25, 28.73,26.04, 25.31, 23.95, 14.38, 8.22. Example 6 The preparation of 20(S)-7-[3-(cyclopentylmethylamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 6) is as follows: Using intermediate II prepared in Example 1, in the third step of the reaction, cyclopropylamine was replaced with cyclopentylmethylamine (the molar ratio with the starting material was the same as in Example 1). The types and amounts of each starting material and the operation steps were the same in the remaining steps. Finally, a white solid product (compound 6) was obtained with a yield of 74% and a purity of 97.06%.

[0063] 1H NMR (600 MHz, DMSO-d6) δ = 8.31 (dd, J = 5.9, 2.4 Hz, 1H), 8.05 (dd, J = 14.4, 6.0 Hz, 1H), 7.29 (dd, J = 6.2, 2.6 Hz, 1H), 6.53 (s, 1H), 5.42 (d, J = 2.6 Hz, 2H), 5.25 (dd, J = 12.4, 5.1 Hz, 2H), 3.24 (dt, J = 8.2, 4.0 Hz, 2H), 3.13 – 3.08 (m, 2H), 2.86 (d, J = 7.4 Hz, 2H), 2.13 (dtd, J =10.5, 6.6, 5.4, ​​2.9 Hz, 1H), 2.03 (p, J = 8.0 Hz, 2H), 1.87 (dh, J = 21.5,7.3 Hz, 2H), 1.77 (dq, J = 12.1, 6.9 Hz, 2H), 1.58 (qd, J = 10.4, 9.0, 4.5Hz, 2H), 1.51 (qd, J = 7.9, 7.3, 4.3 Hz, 2H), 1.25 – 1.16 (m, 2H), 0.88 (t, J= 7.3 Hz, 3H). 13 C NMR (151 MHz, DMSO-d6) δ = 172.89, 157.21, 152.00, 150.53, 147.04,146.04, 145.56, 143.76, 143.57, 133.26, 131.58, 129.28, 124.96, 119.50,109.12, 103.73, 97.19, 72.83, 65.70, 52.23, 50.32, 47.36, 40.47, 36.94,30.59, 26.94, 25.93, 25.01, 8.21. Example 7 The preparation of 20(S)-7-[3-(cyclohexylamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 7) is as follows: Using intermediate II prepared in Example 1, in the third step of the reaction, cyclopropylamine was replaced with cyclohexylamine in step 3 (the molar ratio with the starting material was the same as in Example 1). The types and amounts of each starting material and the operation steps were the same in the remaining steps. Finally, a white solid product (compound 7) was obtained with a yield of 74% and a purity of 97.06%.

[0064] 1 H NMR (400 MHz, DMSO-d6) δ = 8.30 (s, 1H), 8.01 (s, 1H), 7.28 (s,1H), 6.53 (s, 1H), 5.42 (s, 2H), 5.21 (s, 2H), 3.24 (dd, J = 10.5, 6.2 Hz,2H), 3.13 (t, J = 7.6 Hz, 2H), 2.96 (dt, J = 10.7, 5.3 Hz, 1H), 2.01 (q, J =9.7, 7.5 Hz, 4H), 1.87 (p, J = 6.9 Hz, 2H), 1.79 – 1.72 (m, 2H), 1.63 – 1.57(m, 1H), 1.34 – 1.08 (m, 6H), 0.89 (t, J = 7.3 Hz, 3H). 13 C NMR (101 MHz, DMSO-d6) δ = 172.90, 157.17, 151.92, 150.51, 146.94,145.99, 145.51, 143.71, 143.56, 134.08, 131.56, 129.16, 124.87, 119.46,109.03, 103.69, 97.17, 72.82, 65.68, 56.50, 50.30, 43.90, 30.77, 29.31,27.10, 26.40, 25.24, 24.40, 8.22. Example 8 The preparation of 20(S)-7-[3-((4-methylcyclohexyl)amino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 8) is as follows: Using intermediate II prepared in Example 1, in the third step of the reaction, cyclopropylamine was replaced with p-methylcyclohexylamine (the molar ratio with the starting material was the same as in Example 1). The types and amounts of each starting material and the operation steps were the same in the remaining steps. Finally, a white solid product (compound 8) was obtained with a yield of 75% and a purity of 96.66%.

[0065] 1 H NMR (400 MHz, DMSO-d6) δ = 8.32 (s, 1H), 8.03 (s, 1H), 7.28 (s,1H), 6.55 (s, 1H), 5.42 (s, 2H), 5.22 (s, 2H), 3.25 (t, J = 8.2 Hz, 2H), 3.15(t, J = 7.5 Hz, 2H), 2.95 (t, J = 11.7 Hz, 1H), 2.02 (q, J = 10.9, 9.0 Hz,3H), 1.88 (h, J = 7.0 Hz, 2H), 1.73 (d, J = 8.6 Hz, 4H), 1.53 – 1.26 (m, 4H),0.89 (dt, J = 22.0, 6.8 Hz, 6H). 13 C NMR (101 MHz, DMSO-d6) δ = 172.90, 157.19, 151.99, 150.52, 146.99,146.03, 145.53, 143.75, 143.50, 134.09, 131.57, 129.24, 124.90, 119.48,109.10, 103.73, 97.18, 72.83, 65.69, 56.37, 55.51, 50.34, 44.26, 43.93,32.89, 31.64, 30.76, 29.07, 28.95, 27.09, 26.26, 24.48, 22.26, 19.15, 8.23. Example 9 The preparation of 20(S)-7-[3-((3-methylcyclohexyl)amino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 9) is as follows: Using intermediate II prepared in Example 1, in the third step of the reaction, cyclopropylamine was replaced with 3-methylcyclohexylamine (the molar ratio with the starting material was the same as in Example 1). The types and amounts of each starting material and the operation steps were the same in the remaining steps. Finally, a white solid product (compound 9) was obtained with a yield of 73% and a purity of 95.03%.

[0066] 1H NMR (400 MHz, DMSO-d6) δ = 8.31 (dd, J = 7.0, 3.4 Hz, 1H), 8.07 –7.95 (m, 1H), 7.31 – 7.22 (m, 1H), 6.54 (s, 1H), 5.41 (s, 2H), 5.20 (dd, J =14.2, 7.3 Hz, 2H), 3.24 (s, 2H), 3.19 – 3.13 (m, 2H), 3.05 – 2.98 (m, 1H), 2.01 (td, J = 16.4, 14.9, 7.2 Hz, 4H), 1.85 (dp, J = 14.4, 7.1 Hz, 2H), 1.79– 1.68 (m, 1H), 1.63 – 1.39 (m, 2H), 1.31 – 1.13 (m, 2H), 1.08 – 0.94 (m,1H), 0.89 (q, J = 7.2 Hz, 6H), 0.79 (dt, J = 12.3, 6.2 Hz, 1H). 13 C NMR (101 MHz, DMSO-d6) δ = 172.89, 157.18, 151.92, 150.51, 146.96,145.99, 145.52, 143.73, 143.50, 131.56, 129.17, 124.87, 119.48, 109.06,103.70, 97.19, 72.82, 65.69, 56.38, 50.31, 43.83, 37.40, 33.89, 31.17, 30.77,28.73, 27.07, 26.29, 24.05, 22.61, 8.22. Example 10 The preparation of 20(S)-7-[3-((2-methylcyclohexyl)amino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 10) is as follows: Using intermediate II prepared in Example 1, in the third step of the reaction, cyclopropylamine was replaced with 2-methylcyclohexylamine (the molar ratio with the starting material was the same as in Example 1). The types and amounts of each starting material and the operation steps were the same in the remaining steps. Finally, a white solid product (compound 10) was obtained with a yield of 65% and a purity of 97.06%.

[0067] 1H NMR (400 MHz, DMSO-d6) δ = 8.25 (d, J = 4.1 Hz, 1H), 8.00 (d, J =3.0 Hz, 1H), 7.27 (s, 1H), 6.54 (s, 1H), 5.41 (s, 2H), 5.20 (s, 2H), 3.19(dt, J = 8.3, 4.5 Hz, 2H), 3.02 – 2.79 (m, 3H), 2.37 (d, J = 10.7 Hz, 1H), 1.88 (dq, J = 13.8, 7.0 Hz, 4H), 1.71 – 1.35 (m, 5H), 1.19 (q, J = 12.0, 11.5Hz, 3H), 1.00 (d, J = 6.4 Hz, 3H), 0.92 (d, J = 7.1 Hz, 1H), 0.88 (t, J = 7.3Hz, 3H). 13 C NMR (151 MHz, DMSO-d6) δ = 172.88, 157.19, 151.91, 150.50, 146.97,146.06, 145.52, 144.34, 143.66, 133.25, 131.56, 129.88, 129.15, 129.10,125.01, 119.44, 109.01, 103.70, 97.15, 72.82, 65.69, 62.01, 58.94, 50.26,44.46, 36.10, 34.17, 31.07, 30.80, 29.77, 27.23, 25.37, 24.96, 19.49, 8.20. Example 11 The preparation of 20(S)-7-[3-((4-tert-butylcyclohexyl)amino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 11) is as follows: Using intermediate II prepared in Example 1, in the third step of the reaction, cyclopropylamine was replaced with p-tert-butylcyclohexylamine (the molar ratio with the starting material was the same as in Example 1). The types and amounts of each starting material and the operation steps were the same in the remaining steps. Finally, a white solid product (compound 11) was obtained with a yield of 63% and a purity of 98.26%.

[0068] 1H NMR (400 MHz, DMSO-d6) δ = 8.34 (td, J = 6.1, 2.7 Hz, 1H), 8.11 –8.01 (m, 1H), 7.29 (dd, J = 7.0, 3.3 Hz, 1H), 6.58 – 6.51 (m, 1H), 5.43 (d, J = 3.6 Hz, 2H), 5.25 (dd, J = 14.3, 6.4 Hz, 2H), 3.26 (t, J = 8.8 Hz, 2H), 3.15 (t, J = 7.5 Hz, 2H), 3.04 – 2.84 (m, 1H), 2.15 – 1.95 (m, 4H), 1.88 (dq,J = 14.0, 7.0 Hz, 2H), 1.80 (d, J = 10.6 Hz, 1H), 1.64 – 1.47 (m, 2H), 1.37 –1.20 (m, 2H), 1.00 (dt, J = 16.8, 5.7 Hz, 2H), 0.88 (t, J = 7.3 Hz, 3H), 0.83(d, J = 5.4 Hz, 9H). 13 C NMR (101 MHz, DMSO-d6) δ = 172.92, 157.21, 150.54, 147.00, 146.07,145.55, 143.76, 143.59, 143.52, 131.58, 119.50, 103.76, 97.20, 72.83, 65.69,56.57, 53.33, 50.37, 47.39, 46.78, 45.37, 43.95, 32.77, 32.51, 30.75, 29.34,27.93, 27.79, 25.42, 21.04, 8.23. Example 12 The preparation of 20(S)-7-[3-(cyclohexylmethylamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 12) is as follows: Using intermediate II prepared in Example 1, in the third step of the reaction, cyclopropylamine was replaced with cyclopentylmethylamine (the molar ratio with the starting material was the same as in Example 1). The types and amounts of each starting material and the operation steps were the same in the remaining steps. Finally, a white solid product (compound 12) was obtained with a yield of 65% and a purity of 96.13%.

[0069] 1H NMR (400 MHz, DMSO-d6) δ = 8.32 (t, J = 4.3 Hz, 1H), 8.06 (tdd, J =15.1, 9.3, 4.9 Hz, 1H), 7.29 (q, J = 5.5 Hz, 1H), 6.56 (s, 1H), 5.43 (s, 2H),5.25 (d, J = 8.9 Hz, 2H), 3.24 (d, J = 9.4 Hz, 2H), 3.09 (t, J = 7.7 Hz, 2H),2.75 (d, J = 6.9 Hz, 2H), 2.02 (p, J = 7.8 Hz, 2H), 1.87 (hept, J = 7.1 Hz,2H), 1.77 – 1.59 (m, 6H), 1.24 – 1.10 (m, 3H), 0.94 (td, J = 11.7, 3.0 Hz, 2H), 0.88 (t, J = 7.3 Hz, 3H). 13 C NMR (151 MHz, DMSO-d6) δ = 172.89, 157.22, 152.01, 150.54, 147.05,146.06, 145.57, 143.76, 143.56, 131.58, 129.28, 124.96, 119.51, 109.15,103.74, 97.19, 72.83, 65.70, 53.25, 50.34, 47.48, 34.91, 30.79, 30.43, 26.93,25.99, 25.87, 25.43, 8.21. Example 13 The preparation of 20(S)-7-[3-(cycloheptaneamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 13) is as follows: Using intermediate II prepared in Example 1, in the third step of the reaction, cyclopropylamine was replaced with cycloheptamine. The types and amounts of each raw material and the operation steps were the same in the remaining steps. Finally, a white solid product (compound 13) was obtained with a yield of 92% and a purity of 97.03%.

[0070] 1H NMR (400 MHz, DMSO-d6) δ = 8.31 (d, J = 6.5 Hz, 1H), 8.10 – 7.96(m, 1H), 7.27 (d, J = 6.1 Hz, 1H), 6.54 (d, J = 2.9 Hz, 1H), 5.42 (d, J = 3.5Hz, 2H), 5.30 – 5.12 (m, 2H), 3.23 (t, J = 7.6 Hz, 2H), 3.17 (q, J = 8.1, 7.1Hz, 3H), 2.02 (dp, J = 23.3, 8.0, 6.3 Hz, 4H), 1.87 (hept, J = 7.1 Hz, 2H),1.73 – 1.36 (m, 10H), 0.88 (t, J = 7.3 Hz, 3H). 13 C NMR (101 MHz, DMSO-d6) δ = 172.91, 157.18, 151.94, 150.52, 146.96,145.99, 145.52, 143.74, 143.51, 129.17, 124.88, 119.48, 109.06, 103.70,97.19, 72.83, 65.69, 58.69, 50.32, 44.28, 30.74, 27.85, 27.06, 26.30, 23.81,8.22. Example 14 The preparation of 20(S)-7-[3-(cyclooctylamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 14) is as follows: Using intermediate II prepared in Example 1, in the third step of the reaction, cyclopropylamine was replaced with cyclooctylamine (the molar ratio with the starting material was the same as in Example 1). The types and amounts of each starting material and the operation steps were the same in the remaining steps. Finally, a white solid product (compound 14) was obtained with a yield of 63% and a purity of 95.83%.

[0071] 1H NMR (600 MHz, DMSO-d6) δ = 8.32 (d, J = 2.6 Hz, 1H), 8.06 (dd, J =8.3, 4.8 Hz, 1H), 7.30 (t, J = 3.0 Hz, 1H), 6.53 (s, 1H), 5.43 (s, 2H), 5.30– 5.20 (m, 2H), 3.23 (ddt, J = 18.8, 9.4, 5.7 Hz, 3H), 3.17 (dd, J = 9.3, 6.3Hz, 2H), 1.99 (p, J = 8.2 Hz, 2H), 1.95 – 1.90 (m, 2H), 1.86 (ddt, J = 21.5,14.4, 7.2 Hz, 2H), 1.75 – 1.61 (m, 4H), 1.61 – 1.43 (m, 7H), 1.39 (ddt, J =11.2, 7.9, 3.5 Hz, 1H), 0.88 (t, J = 7.3 Hz, 3H). 13 C NMR (151 MHz, DMSO-d6) δ = 172.90, 157.25, 152.07, 150.55, 147.06,146.08, 145.58, 143.79, 143.52, 133.27, 131.58, 129.89, 129.31, 124.95,119.51, 109.17, 103.71, 97.22, 72.83, 65.70, 58.12, 50.36, 44.32, 30.78,29.13, 29.10, 27.06, 26.40, 26.34, 25.60, 23.74, 8.21. Example 15 The preparation of 20(S)-7-[3-(benzylamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 15) is as follows: Using intermediate II prepared in Example 1, in the third step of the reaction, cyclopropylamine was replaced with benzylamine (the molar ratio with the starting material was the same as in Example 1). The types and amounts of each starting material and the operation steps were the same in the remaining steps. Finally, a white solid product (compound 15) was obtained with a yield of 76% and a purity of 97.03%.

[0072] 1H NMR (400 MHz, DMSO-d6) δ = 8.20 – 8.10 (m, 1H), 8.03 – 7.91 (m,1H), 7.29 – 7.21 (m, 5H), 7.17 (t, J = 6.8 Hz, 1H), 6.57 (s, 1H), 5.40 (d, J= 3.8 Hz, 2H), 5.26 – 5.14 (m, 2H), 3.66 (s, 2H), 3.18 (d, J = 6.5 Hz, 3H), 2.57 (t, J = 6.6 Hz, 2H), 1.85 (dq, J = 13.7, 6.8 Hz, 4H), 0.87 (t, J = 7.3Hz, 3H). 13 C NMR (101 MHz, DMSO-d6) δ = 172.93, 157.15, 151.80, 150.46, 146.94,146.13, 145.48, 145.15, 143.59, 141.12, 131.57, 129.13, 129.03, 128.51,126.99, 125.12, 119.38, 108.94, 103.71, 97.05, 72.82, 65.70, 53.55, 50.19,48.44, 30.75, 29.59, 27.51, 8.23. Example 16 The preparation of 20(S)-7-[3-(propylamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 16) is as follows: Using intermediate II prepared in Example 1, in the third step of the reaction, the compound cyclopropylamine was replaced with n-propylamine (the molar ratio with the starting material was the same as in Example 1), and the remaining steps were the same, finally yielding a white solid product (compound 16) with a yield of 74% and a purity of 96.25%.

[0073] 1H NMR (400 MHz, DMSO-d6) δ = 8.28 (d, J = 6.7 Hz, 1H), 8.15 – 7.90(m, 1H), 7.32 (d, J = 8.0 Hz, 1H), 6.61 (s, 1H), 5.45 – 5.34 (m, 2H), 5.28(dd, J = 14.8, 7.3 Hz, 2H), 3.22 (d, J = 8.1 Hz, 2H), 3.07 (t, J = 7.5 Hz,2H), 2.83 (t, J = 7.7 Hz, 2H), 1.89 (ddd, J = 28.7, 16.1, 7.7 Hz, 4H), 1.58(q, J = 8.2, 7.6 Hz, 2H), 0.89 (dt, J = 13.5, 7.4 Hz, 6H). 13 C NMR (101 MHz, DMSO-d6) δ = 172.88, 157.20, 152.00, 150.52, 147.03,146.04, 145.55, 143.76, 143.60, 131.58, 129.13, 124.25, 119.33, 108.21,102.62, 96.21, 71.95, 67.21, 49.81, 46.08, 28.26, 27.02, 19.80, 8.21. Example 17 The preparation of 20(S)-7-[3-(sec-butylamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 17) is as follows: Using intermediate II prepared in Example 1, in the third step of the reaction, cyclopropylamine was replaced with sec-butylamine (the molar ratio with the starting material was the same as in Example 1). The types and amounts of each starting material and the operation steps were the same in the remaining steps. Finally, a white solid product (compound 17) was obtained with a yield of 79% and a purity of 96.34%.

[0074] 1H NMR (400 MHz, DMSO-d6) δ = 8.37 (s, 1H), 8.16 – 8.11 (m, 1H), 7.32(d, J = 1.7 Hz, 1H), 6.56 (s, 1H), 5.45 (s, 2H), 5.31 (d, J = 3.4 Hz, 2H), 3.29 – 3.24 (m, 2H), 3.17 (d, J = 4.9 Hz, 2H), 2.04 – 1.95 (m, 2H), 1.88 (p,J = 6.9 Hz, 2H), 1.76 (td, J = 7.8, 7.2, 4.0 Hz, 2H), 1.47 (dt, J = 14.1, 7.8Hz, 1H), 1.22 (d, J = 6.5 Hz, 3H), 0.90 (dt, J = 12.1, 7.3 Hz, 6H). 13 C NMR (101 MHz, DMSO-d6) δ = 172.94, 157.28, 152.16, 150.57, 147.11,146.15, 145.61, 143.81, 143.57, 129.47, 125.02, 119.52, 109.27, 97.20, 72.85,67.49, 55.07, 44.09, 30.70, 25.91, 25.59, 15.79, 10.04, 8.23. Example 18 The preparation of 20(S)-7-[3-(isobutylamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 18) is as follows: Using intermediate II prepared in Example 1, in the third step of the reaction, cyclopropylamine was replaced with isobutylamine (the molar ratio with the starting material was the same as in Example 1). The types and amounts of each starting material and the operation steps were the same in the remaining steps. Finally, a white solid product (compound 18) was obtained with a yield of 62% and a purity of 97.04%.

[0075] 1H NMR (400 MHz, DMSO-d6) δ = 8.43 (s, 1H), 8.32 (d, J = 3.9 Hz, 1H), 8.08 (dd, J = 17.0, 9.5 Hz, 1H), 7.31 (d, J = 3.2 Hz, 1H), 6.55 (s, 1H), 5.43(d, J = 3.9 Hz, 2H), 5.29 (d, J = 5.7 Hz, 2H), 3.25 (d, J = 8.0 Hz, 2H), 3.10(dt, J = 10.6, 5.5 Hz, 2H), 2.75 (q, J = 6.5 Hz, 2H), 2.03 (p, J = 8.4 Hz,2H), 1.91 (dp, J = 28.1, 6.6 Hz, 3H), 0.93 (d, J = 6.7 Hz, 6H), 0.88 (t, J =7.3 Hz, 3H). 13 C NMR (101 MHz, DMSO-d6) δ = 172.93, 157.25, 150.55, 146.10, 145.59,143.79, 143.53, 125.00, 119.50, 103.72, 97.20, 72.83, 65.69, 54.40, 50.35,47.37, 30.73, 26.87, 25.91, 25.84, 20.43, 8.21. Example 19 The preparation of 20(S)-7-[3-(Butamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 19) is as follows: Using intermediate II prepared in Example 1, in the third step of the reaction, cyclopropylamine was replaced with n-butylamine (the molar ratio with the starting material was the same as in Example 1). The types and amounts of each starting material and the operation steps were the same in the remaining steps. Finally, a white solid product (compound 19) was obtained with a yield of 73% and a purity of 96.56%.

[0076] 1H NMR (400 MHz, DMSO-d6) δ = 8.31 (d, J = 3.9 Hz, 1H), 8.07 – 8.01(m, 1H), 7.28 (q, J = 2.3 Hz, 1H), 6.55 (s, 1H), 5.43 (s, 2H), 5.28 – 5.16(m, 2H), 3.23 (d, J = 8.5 Hz, 2H), 3.09 (t, J = 7.6 Hz, 2H), 2.88 (t, J = 7.7Hz, 2H), 1.99 (p, J = 8.8, 8.0 Hz, 2H), 1.86 (dq, J = 14.1, 6.9 Hz, 2H), 1.58(p, J = 7.8 Hz, 2H), 1.33 (q, J = 7.5 Hz, 2H), 1.20 (s, 1H), 0.89 (td, J =7.4, 3.6 Hz, 6H). 13 C NMR (101 MHz, DMSO-d6) δ = 172.88, 157.20, 152.00, 150.52, 147.03,146.04, 145.55, 143.76, 143.60, 131.58, 129.23, 124.95, 119.49, 109.10,103.72, 97.16, 72.83, 65.71, 50.31, 47.19, 46.97, 30.80, 28.26, 27.02, 26.21,19.80, 13.96, 8.21. Example 20 The preparation of 20(S)-7-[3-(pentanamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 20) is as follows: Using intermediate II prepared in Example 1, in the third step of the reaction, cyclopropylamine was replaced with n-pentylamine (the molar ratio with the starting material was the same as in Example 1). The types and amounts of each starting material and the operation steps were the same in the remaining steps. Finally, a white solid product (compound 20) was obtained with a yield of 62% and a purity of 96.33%.

[0077] 1H NMR (400 MHz, DMSO-d6) δ = 8.34 (s, 1H), 8.11 (s, 1H), 7.31 (s,1H), 6.56 (s, 1H), 5.44 (s, 2H), 5.29 (s, 2H), 3.26 (d, J = 3.2 Hz, 2H), 3.06(t, J = 7.3 Hz, 2H), 2.88 – 2.82 (m, 2H), 1.98 (p, J = 7.5 Hz, 2H), 1.87 (dt,J = 14.3, 6.6 Hz, 2H), 1.58 (p, J = 7.3 Hz, 2H), 1.29 (h, J = 3.9, 3.5 Hz, 4H), 1.22 (s, 1H), 0.88 (td, J = 7.2, 3.8 Hz, 6H). 13 C NMR (101 MHz, DMSO-d6) δ = 172.91, 162.81, 157.26, 152.10, 150.56,147.10, 146.12, 145.60, 143.80, 143.63, 131.60, 129.40, 125.02, 119.52,109.22, 103.74, 97.19, 72.84, 65.71, 50.39, 47.44, 46.96, 30.76, 28.59,26.99, 26.28, 25.91, 22.10, 14.17, 8.22. Example 21 The preparation of 20(S)-7-[3-(((3R,5R,7R)-adamantane-1-yl)amino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 21) is as follows: Using intermediate II prepared in Example 1, in the third step of the reaction, cyclopropylamine was replaced with adamantaneamine, wherein adamantaneamine was in a 3-equivalent weight, DIPEA was in a 2-equivalent weight, and sodium bicarbonate was in a 6-equivalent weight. The types and amounts of each raw material and the operation steps were the same in the remaining steps, and a white solid product (compound 21) was finally obtained with a yield of 82% and a purity of 96.33%.

[0078] 1H NMR (400 MHz, DMSO-d6) δ = 8.37 (s, 1H), 8.03 (dd, J = 7.3, 3.4 Hz,1H), 7.28 (s, 1H), 6.53 (s, 1H), 5.43 (s, 2H), 5.22 (d, J = 7.3 Hz, 2H), 3.26(dd, J = 10.7, 6.1 Hz, 2H), 3.18 (t, J = 7.8 Hz, 2H), 2.15 (s, 3H), 2.03 –1.81 (m, 10H), 1.65 (q, J = 12.5 Hz, 6H), 0.89 (t, J = 7.3 Hz, 3H). 13 C NMR (101 MHz, DMSO-d6) δ = 143.66 (d, J = 18.9 Hz), 109.12,103.75, 97.16, 72.83, 65.70, 56.83, 50.34, 39.80, 39.23, 38.32, 35.67, 30.77,28.92, 27.28, 26.75, 8.23. Example 22 The preparation of 20(S)-7-[3-(((1R,3R,5S,7S)-adamantane-2-yl)amino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 22) is as follows: Using intermediate II prepared in Example 1, in the third step of the reaction, cyclopropylamine was replaced with 2-adamantaneamine, wherein 2-adamantaneamine was in 7 equivalents, DIPEA was in 6 equivalents, and sodium bicarbonate was in 4 equivalents; the types and amounts of each raw material and the operation steps were the same in the remaining steps, and finally a white solid product (compound 22) was obtained with a yield of 85% and a purity of 98.13%.

[0079] 1H NMR (400 MHz, DMSO-d6) δ = 8.33 (d, J = 3.7 Hz, 1H), 8.06 – 7.97(m, 1H), 7.28 (d, J = 3.2 Hz, 1H), 6.53 (s, 1H), 5.42 (s, 2H), 5.21 (d, J =5.4 Hz, 2H), 3.23 (d, J = 11.5 Hz, 3H), 3.16 (t, J = 8.3 Hz, 2H), 2.21 – 2.02(m, 6H), 1.93 – 1.79 (m, 6H), 1.71 (d, J = 14.7 Hz, 4H), 1.56 (d, J = 13.0Hz, 2H), 0.89 (t, J = 7.3 Hz, 3H). 13 C NMR (101 MHz, DMSO-d6) δ = 172.88, 157.19, 152.02, 150.52, 147.03,146.05, 145.53, 143.72, 143.68, 134.11, 131.58, 129.27, 124.96, 119.51,109.13, 103.78, 97.15, 72.83, 65.70, 62.02, 55.40, 50.34, 45.04, 37.14,36.64, 30.77, 30.16, 29.03, 27.09, 26.93, 26.68, 26.15, 8.23. Example 23 The preparation of 20(S)-7-[3-(((1R,3R,5S,7S)-3,5-dimethyladamantane-1-yl)amino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 23) is as follows: Using intermediate II prepared in Example 1, in the third step of the reaction, cyclopropylamine was replaced with dimethyladamantaneamine, wherein dimethyladamantaneamine was 3 equivalents, DIPEA was 4 equivalents, and sodium bicarbonate was 5 equivalents; the amount of each raw material and the operation steps were the same in the remaining steps, and finally a white solid product (compound 23) was obtained with a yield of 92% and a purity of 98.65%.

[0080] 1H NMR (400 MHz, DMSO-d6) δ = 8.37 (s, 1H), 8.06 (s, 1H), 7.29 (s,1H), 6.53 (s, 1H), 5.43 (s, 2H), 5.23 (s, 2H), 3.26 (dd, J = 10.5, 6.3 Hz,2H), 3.18 (t, J = 7.6 Hz, 2H), 2.25 – 2.18 (m, 1H), 1.98 (p, J = 10.2, 9.0Hz, 2H), 1.87 (h, J = 6.9 Hz, 2H), 1.75 (d, J = 3.2 Hz, 2H), 1.57 (t, J = 8.2Hz, 4H), 1.33 (s, 3H), 1.17 (q, J = 12.7 Hz, 2H), 0.90 (d, J = 8.8 Hz, 9H). 13 C NMR (101 MHz, DMSO-d6) δ = 172.89, 157.21, 152.07, 150.52, 147.02,146.07, 145.54, 143.77, 143.61, 134.10, 131.58, 129.21, 124.92, 119.51,109.16, 103.71, 97.17, 72.83, 65.70, 58.26, 50.33, 49.90, 44.26, 41.97,39.48, 37.06, 32.60, 30.74, 30.12, 29.64, 27.22, 26.83, 8.24. Example 24 The preparation of 20(S)-7-[4-(propylamino)butyl]-10,11-difluoromethylenedioxycamptothecin (compound 24) is as follows: (1) Preparation of 6-amino-2,2-difluorobenzo[d][1,3]dioxacyclopentene-5-bromobutanone (intermediate I) Add 5-amino-2,2-difluoro-1,3-benzodioxane (1 g, 5.84 mmol) to a three-necked flask, dissolve in dichloroethane (5 mL), and stir in an ice-water bath. Under nitrogen protection, add boron trichloride (20 mL, 120 mmol) and aluminum trichloride (3 mL). After 10 min, add bromopentonitrile (673 μL, 3... The mixture was refluxed in an oil bath at 85°C for 3 hours. The reaction was monitored by TLC (using a mixture of petroleum ether and ethyl acetate in a ratio of 1000:100, v / v). After the reaction was completed, an appropriate amount of 3 mol / mL HCl aqueous solution was added to quench the reaction, consuming excess bromopentonitrile. After stirring for ten minutes, the mixture was transferred to a separatory funnel and extracted three times with dichloromethane (DCM). The organic phases were combined, dried over anhydrous magnesium sulfate, and filtered under reduced pressure. The filtrate was concentrated to obtain a pale yellow liquid, which was separated by silica gel column chromatography (using a mixture of petroleum ether and ethyl acetate in a ratio of 10:1, v / v). After concentration, a yellow oily substance (intermediate I) was obtained, with a yield of 93.2% and a purity of 96.88%.

[0081] (2) Preparation of 20(S)-7-(4-bromobutyl)-10,11-difluoromethylenedioxycamptothecin (intermediate II) The product obtained in step (1) (intermediate I) (329 mg, 1 mmol), tricyclic ketone (133.6 mg, 0.5 mmol), and p-toluenesulfonic acid hydrate (32.3 mg, 0.17 mmol) were added to a round-bottom flask, dissolved in dichloromethane, and reacted under vacuum at 115 °C for 1 h. The reaction was then detected by TLC (using a mixed solution of dichloromethane and methanol = 1600:100 as the eluent, v / v). After the reaction was completed, the product was separated by silica gel column chromatography (using a mixed solution of dichloromethane and methanol = 300:1 as the eluent, v / v, with a small amount of glacial acetic acid added). The product was concentrated, washed with methanol, and filtered to obtain a pure white solid product (intermediate II) on the filter cake. The yield was 86.6%, and the purity was 98.82%.

[0082] (3) Preparation of 20(S)-7-[4-(propylamino)butyl]-10,11-difluoromethylenedioxycamptothecin (compound 24) 20(S)-7-(4-bromobutyl)-10,11-difluoromethylenedioxycamptothecin (intermediate II) (50 mg, 0.1 mmol) was added to a round-bottom flask, followed by the addition of DMF (2 mL), sodium bicarbonate (3 equivalents, 25 mg), and n-propylamine (5 equivalents, 41 μL). The mixed solvent was placed in an oil bath at 70 °C for 3 h. After cooling, the mixture was extracted with DCM and water, and the organic layer was collected. Anhydrous magnesium sulfate was used to remove water, and the solution was then subjected to column chromatography (eluent was a mixed solution of DCM:MeOH = 15:1, v / v) to obtain a white solid powder with a yield of 70% and a purity of 95.34%.

[0083] 1 H NMR (400 MHz, DMSO-d6) δ=8.31 ​​(d, J = 6.7 Hz, 1H), 8.17 – 8.05 (m,1H), 7.36 (d, J = 8.0 Hz, 1H), 6.65 (s, 1H), 5.47 – 5.38 (m, 2H), 5.31 (dd, J= 14.8, 7.5 Hz, 2H), 3.26 (d, J = 8.0 Hz, 2H), 3.17 (t, J = 7.2 Hz, 2H), 3.05(t, J = 7.4 Hz, 2H), 2.85 (t, J = 7.7 Hz, 2H), 1.93 (ddd, J = 28.7, 16.1,8.1Hz, 4H), 1.62 (q, J = 8.2, 7.9 Hz, 2H), 0.94 (dt, J = 13.5, 7.7Hz, 6H). 13 C NMR (101 MHz, DMSO-d6) δ=172.88, 157.20, 152.00, 150.52, 147.03,146.04, 145.55, 143.76, 143.60, 131.58, 129.13, 124.25, 119.33, 108.21,102.62, 96.21, 71.95, 67.21, 49.81, 46.08, 39.13, 28.26, 27.02, 19.80,8.21. Example 25 The preparation of 20(S)-7-[1-(cyclopropylamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 26) is as follows: (1) Preparation of 6-amino-2,2-difluorobenzo[d][1,3]dioxane-4-bromoethyl ketone (intermediate I) Add 1 g (5.84 mmol) of 5-amino-2,2-difluoro-1,3-benzodioxane to a three-necked flask, dissolve in 5 mL of dichloroethane, stir in an ice-water bath, and add boron trichloride (20 mL) under nitrogen protection. After 10 min, bromoacetonitrile (367 μL, 1.8 mmol) was added and the mixture was refluxed in an oil bath at 95 °C for 3 h. The reaction was monitored by TLC (using a mixture of petroleum ether and ethyl acetate in a ratio of 800:100, v / v). After the reaction was completed, an appropriate amount of 3 mol / mL HCl aqueous solution was added to quench the reaction, consuming excess bromoacetonitrile. After stirring for 10 min, the mixture was transferred to a separatory funnel and extracted three times with dichloromethane (DCM). The organic phases were combined, dried with anhydrous magnesium sulfate, and filtered under reduced pressure. The filtrate was concentrated to obtain a pale yellow liquid, which was separated by silica gel column chromatography (using a mixture of petroleum ether and dichloromethane in a ratio of 20:1, v / v). After concentration, a yellow oily substance (intermediate I) was obtained with a yield of 87.2% and a purity of 97.18%.

[0084] (2) Preparation of 20(S)-7-(2-bromomethyl)-10,11-difluoromethylenedioxycamptothecin (intermediate II) The product obtained in step (1) (intermediate I) (104.2 mg, 0.33 mmol), tricyclic ketone (48.5 mg, 0.18 mmol), and p-toluenesulfonic acid hydrate (35.6 mg, 0.19 mmol) were added to a round-bottom flask and dissolved in dichloromethane (3 mL). After reacting at 115 °C under vacuum for 1 h, the reaction was detected by TLC (the developing solvent was a mixed solution of dichloromethane and methanol = 1600:100, v / v). After the reaction was completed, the product was separated by silica gel column chromatography (the eluent was a mixed solution of dichloromethane and methanol = 300:1, v / v, with a small amount of glacial acetic acid added). After concentration, the product was washed with methanol and filtered to obtain a pure white solid product (intermediate II) on the filter cake. The yield was 69.3% and the purity was 96.12%.

[0085] (3) Preparation of 20(S)-7-[1-(cyclopropylamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 26) 20(S)-7-(2-bromomethyl)-10,11-difluoromethylenedioxycamptothecin (intermediate II) (50 mg, 0.1 mmol) was added to a round-bottom flask, followed by the addition of DMF (2 mL), sodium bicarbonate (3 equivalents, 25 mg), and cyclopropylamine (5 equivalents, 35 μL). The mixed solvent was placed in an oil bath at 50 °C for 5 h. After cooling, the mixture was extracted with DCM and water, and the organic layer was collected. Anhydrous magnesium sulfate was used to remove water, and the solution was then subjected to column chromatography (eluent was a mixed solution of DCM:MeOH = 15:1, v / v) to obtain a white solid powder with a yield of 70% and a purity of 95.34%.

[0086] 1 H NMR (400 MHz, DMSO-d6) δ=8.23 (d, J = 5.6 Hz, 1H), 7.68 – 7.62 (m,1H), 7.23 (d, J = 2.8 Hz, 1H), 6.43 (s, 1H), 5.40 (s, 2H), 5.03 (d, J = 7.2Hz, 2H), 2.05 (dq, J = 6.5, 3.3 Hz, 1H), 1.74 (ddd, J = 25.8, 12.7, 7.2 Hz, 4H), 0.77(t, J = 7.1 Hz, 3H), 0.21(dt, J = 6.0, 2.6 Hz, 2H), 0.17 (p, J = 3.2Hz, 2H). 13 C NMR (151 MHz, DMSO-d6) δ=172.09, 156.77, 150.71, 150.47, 146.89,146.05, 145.47, 145.17, 143.54, 133.23, 131.55, 129.87, 128.89, 125.03,118.23, 107.61, 102.51, 96.98, 70.21, 65.67, 50.09, 29.56, 28.57, 26.12,8.16, 6.21. Comparative Example 1 The preparation of 20(S)-7-[3-(cyclopropylamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 1) is as follows: (1) Preparation of 6-amino-2,2-difluorobenzo[d][1,3]dioxane-4-bromobutanone (intermediate I) The raw material 5-amino-2,2-difluoro-1,3-benzodioxane (1 g, 5.84 mmol) was added to a three-necked flask, dissolved in dichloroethane (5 mL), and stirred in an ice-water bath. Under nitrogen protection, aluminum trichloride (18 mg, 0.13 mmol) was added, followed by bromobutyronitrile (569 μL, 2.6 mmol) after 10 min. The mixture was refluxed in an oil bath at 95°C for 3 hours. The reaction was monitored by TLC (using a mixture of petroleum ether and ethyl acetate in a ratio of 800:100, v / v). After the reaction was completed, an appropriate amount of 3 mol / mL HCl aqueous solution was added to quench the reaction, consuming excess bromobutyronitrile. After stirring for ten minutes, the mixture was transferred to a separatory funnel and extracted three times with dichloromethane (DCM). The organic phases were combined, dried with anhydrous magnesium sulfate, and filtered under reduced pressure. The filtrate was concentrated to obtain a pale yellow liquid, which was separated by silica gel column chromatography (using a mixture of petroleum ether and dichloromethane in a ratio of 20:1, v / v). After concentration, a yellow oily substance (intermediate I) was obtained, with a yield of 70.2% and a purity of 90.43%.

[0087] Comparative Example 2 The preparation of 20(S)-7-[3-(cyclopropylamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 1) is as follows: (1) Preparation of 6-amino-2,2-difluorobenzo[d][1,3]dioxane-4-bromobutanone (intermediate I) The raw material 5-amino-2,2-difluoro-1,3-benzodioxane (A) (1 g, 5.84 mmol) was added to a three-necked flask, dissolved in dichloroethane, and stirred in an ice-water bath. Gallium trichloride (21 mg, 0.12 mmol) was added under nitrogen protection. After 10 min, bromobutyronitrile (569 μL, 2.6 mmol) was added. The mixture was refluxed in an oil bath at 95°C for 3 hours. The reaction was monitored by TLC (using a mixture of petroleum ether and ethyl acetate in a ratio of 800:100, v / v). After the reaction was completed, an appropriate amount of 3 mol / mL HCl aqueous solution was added to quench the reaction, consuming excess bromobutyronitrile. After stirring for ten minutes, the mixture was transferred to a separatory funnel and extracted three times with dichloromethane (DCM). The organic phases were combined, dried with anhydrous magnesium sulfate, and filtered under reduced pressure. The filtrate was concentrated to obtain a pale yellow liquid, which was separated by silica gel column chromatography (using a mixture of petroleum ether and dichloromethane in a ratio of 20:1, v / v). After concentration, a yellow oily substance (intermediate I) was obtained, with a yield of 75.1% and a purity of 92.03%.

[0088] Comparative Example 3 The preparation of 20(S)-7-[3-(cyclopropylamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 1) is as follows: (1) Preparation of 6-amino-2,2-difluorobenzo[d][1,3]dioxane-4-bromobutanone (intermediate I) Same as Example 1; (2) Preparation of 20(S)-7-(3-bromopropyl)-10,11-difluoromethylenedioxycamptothecin (intermediate II) The product obtained in step (1) (intermediate I) (110.3 mg, 0.34 mmol), tricyclic ketone (48.5 mg, 0.2 mmol), and p-toluenesulfonic acid hydrate (35.6 mg, 0.2 mmol) were added to a round-bottom flask, dissolved in glacial acetic acid (3 mL), and reacted under vacuum at 115 °C for 1 h. The reaction was then detected by TLC (using a mixture of dichloromethane and methanol as the eluent, v / v). After the reaction was completed, the product was separated by silica gel column chromatography (using a mixture of dichloromethane and methanol as the eluent, v / v, with a small amount of glacial acetic acid added). The product was concentrated, washed with methanol, and filtered to obtain a pure white solid product (intermediate II) on the filter cake. The yield was 60.3% and the purity was 95.32%.

[0089] Comparative Example 4 The preparation of 20(S)-7-[3-(cyclopropylamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 1) is as follows: (1) Preparation of 6-amino-2,2-difluorobenzo[d][1,3]dioxane-4-bromobutanone (intermediate I) Same as Example 1.

[0090] (2) Preparation of 20(S)-7-(3-bromopropyl)-10,11-difluoromethylenedioxycamptothecin (intermediate II) Same as Example 1.

[0091] (3) Preparation of 20(S)-7-[3-(cyclopropylamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 1) 20(S)-7-(3-bromopropyl)-10,11-difluoromethylenedioxycamptothecin (intermediate II) (50 mg, 0.1 mmol) was added to a round-bottom flask, followed by the addition of DMF (2 mL), sodium bicarbonate (6 equivalents, 50 mg), and cyclopropylamine (5 equivalents, 35 μL). The mixed solvent was placed in an oil bath at 50 °C for 5 h. After cooling, the mixture was extracted with DCM and water, and the organic layer was collected. Anhydrous magnesium sulfate was used to remove water, and the solution was then subjected to column chromatography (eluent was a mixed solution of DCM:MeOH = 15:1, v / v) to obtain a white solid powder with a yield of 62% and a purity of 94.04%.

[0092] Comparative Example 5 The preparation of 20(S)-7-[3-(cyclopropylamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 1) is as follows: (1) Preparation of 6-amino-2,2-difluorobenzo[d][1,3]dioxane-4-bromobutanone (intermediate I) Same as Example 1.

[0093] (2) Preparation of 20(S)-7-(3-bromopropyl)-10,11-difluoromethylenedioxycamptothecin (intermediate II) Same as Example 1.

[0094] (3) Preparation of 20(S)-7-[3-(cyclopropylamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 1) 20(S)-7-(3-bromopropyl)-10,11-difluoromethylenedioxycamptothecin (intermediate II) (50 mg, 0.1 mmol) was added to a round-bottom flask, followed by the addition of DMF (2 mL), sodium hydroxide (3 equivalents, 12 mg), and cyclopropylamine (5 equivalents, 35 μL). The mixed solvent was placed in an oil bath at 50 °C for 5 h. After cooling, the mixture was extracted with DCM and water, and the organic layer was collected. Anhydrous magnesium sulfate was used to remove water, and the solution was then subjected to column chromatography (eluent was a mixed solution of DCM:MeOH = 15:1, v / v) to obtain a white solid powder with a yield of 41% and a purity of 93.02%.

[0095] Comparative Example 6 The preparation of 20(S)-7-[3-(cyclopropylamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 1) is as follows: (1) Preparation of 6-amino-2,2-difluorobenzo[d][1,3]dioxane-4-bromobutanone (intermediate I) Same as Example 1.

[0096] (2) Preparation of 20(S)-7-(3-bromopropyl)-10,11-difluoromethylenedioxycamptothecin (intermediate II) Same as Example 1.

[0097] (3) Preparation of 20(S)-7-[3-(cyclopropylamino)propyl]-10,11-difluoromethylenedioxycamptothecin (compound 1) 20(S)-7-(3-bromopropyl)-10,11-difluoromethylenedioxycamptothecin (intermediate II) (50 mg, 0.1 mmol) was added to a round-bottom flask, followed by the addition of DMF (2 mL), DIPEA (3 equivalents, 52 μL), and cyclopropylamine (5 equivalents, 35 μL). The mixed solvent was placed in an oil bath at 50 °C for 5 h. After cooling, the mixture was extracted with DCM and water, and the organic layer was collected. Anhydrous magnesium sulfate was used to remove water, and the solution was then subjected to column chromatography (eluent was a mixed solution of DCM:MeOH = 15:1, v / v) to obtain a white solid powder with a yield of 47% and a purity of 95.13%.

[0098] Experimental Example 1: In vitro antitumor activity test of the compound of the present invention Tumor cells are characterized by rapid growth and unlimited reproduction. Normal cells are invaded by tumor cells, which can metastasize and spread within the body, causing fatal damage. Therefore, the primary condition for screening anti-tumor drugs is that they have a significant lethal effect on tumor cells.

[0099] This study used the MTT assay to detect the lethality of the compound of this invention and the control drug SN-38 against three types of tumor cells: 5637 cells (human bladder cancer cells), T24 cells (human bladder transitional cell carcinoma cells), and UC3 cells (human bladder transitional cell carcinoma cells), as well as normal human cells SV cells (human ureteral epithelial immortalized cells).

[0100] Test method: (1) Preparation before the experiment: 10% complete culture medium: 10% serum + 1% penicillin and streptomycin mixture + DMEM culture medium; 0.5% MTT solution: weigh 250 mg MTT and dissolve it in 50 mL PBS buffer, filter it through a 0.22 μm sterile filter membrane for sterilization, and store the drug storage solution in a sealed container at 4℃ away from light.

[0101] The experimental compounds (compounds 1-26 and SN-38 of this invention (a topoisomerase I inhibitor, the active metabolite of irinotecan)) were dissolved in dimethyl sulfoxide (DMSO) to prepare a stock solution with a concentration of 1 mM. Before administration, the solutions were diluted with complete DMEM medium containing 1% FBS buffer to prepare drug solutions with concentration gradients of 6.25, 12.5, 25, 50, 100, and 200 nM. (2) Cell culture: When the cells reach 80-90% confluence, digest the cells (5637 cells, T24 cells, UC3 cells and normal human SV cells) with trypsin-EDTA (0.02%), count the cells with a hemocytometer, and adjust the cell density to 5×10⁻⁶ cells based on the counting results. 4The fractions were seeded at a rate of 100 μL / mL into each well of a 96-well plate. 200 μL of PBS buffer was added to each well around the outermost edge of the 96-well plate, and the plates were incubated for 24 h.

[0102] (3) Drug addition: When the cells reach 80% confluence, discard the supernatant and add 200 μL of different drug solutions to each well. The drugs are the compound of this invention and SN-38 (a topoisomerase I inhibitor, the active metabolite of irinotecan). Three parallel wells are set for each sample concentration. Add 200 μL of complete DMEM medium containing 1% FBS buffer as a blank control. Place the 96-well plate in an incubator at 37°C and 5% CO2 and continue to incubate for 72 h. Add 20 μL of 0.5% MTT solution to each well and continue to incubate for 4 h. Remove the plate, carefully discard the supernatant, and add 150 μL of DMSO solution to each well to dissolve the blue-purple crystal product. Place the 96-well plate on a microplate shaker and shake for 20 min to dissolve. Set the reference wavelength to 630 nm using a microplate reader and detect the absorbance OD value at 570 nm.

[0103] Calculate the inhibition rate of cell proliferation by the drug using the following formula: Proliferation inhibition rate % = (OD blank group - OD experimental group) / OD blank group × 100%.

[0104] In the formula, OD experimental group is the absorbance value of the drug-treated group; OD blank group is the absorbance value of the cell group cultured normally in drug-free medium.

[0105] Table 2 below shows the IC50 values ​​of compounds 1-26 and SN-38 of the present invention against 5637 cell lines cultured for 48 hours, 5637 cell lines cultured for 72 hours, T24 cell lines cultured for 72 hours, and UC3 cell lines cultured for 72 hours. 50 value.

[0106] Table 2

[0107] Analysis of the data in Table 2 shows that: ①Most of the compounds of this invention have better inhibitory activity against the above three tumor cell lines than the positive control SN-38, and most of the compounds have lower inhibitory activity against normal human cell lines than the positive control SN-38, with lower toxic side effects.

[0108] ② Compounds 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, 15, 18, 19, 20, and 22 of this invention exhibit excellent inhibitory activity against 5637 cells cultured for 72 hours, among which compounds 1 and 7 have the highest IC50 values. 50The value is less than 5.0 nM, which is far superior to similar compounds, and it has higher inhibitory activity than SN-38.

[0109] ③ Compounds 1, 2, 4, 5, 6, 7 and 15 of this invention exhibit superior inhibitory activity against T24 cells compared to SN-38.

[0110] ④ Compounds 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20 and 22 of the present invention exhibit superior inhibitory activity against UC3 cells compared to SN-38.

[0111] ⑤ Compounds 2, 5, and 6 in this invention have lower inhibitory activity against SV cells and lower toxicity than SN-38. Other compounds are similar to SN-38 and have comparable toxicity.

[0112] ⑥ Compound 25 (n=1) in Example 1 showed lower inhibitory activity against 5637, T24 and UC3 cells compared to Compound 1 (n=3) in this invention.

[0113] The above in vitro experiments show that the compounds of the present invention have selective and good antitumor activity, possess antitumor advantages, and can be used to prepare drugs for treating cancer.

[0114] Experimental Example 2: Experimental data on dual-target selection and transmembrane uptake capacity of the compounds of this invention The cell membrane serves as a barrier, facilitating transmembrane transport, transmembrane information transmission, and energy conversion, functions determined by its chemical composition and molecular structure. The transmembrane transport capacity of drugs to tumor cells significantly impacts drug efficacy and bioavailability. Therefore, this study evaluates the transmembrane transport capacity of the compound by observing its absorption and transport from the simulated intestinal lumen (AP side) to the plasma (BL side) and its secretion and efflux from the BL side to the AP side.

[0115] (1) Preparation of sample solution: Based on the results of in vitro antitumor activity test, compounds 1, 4 and 7 with better in vitro antitumor activity were selected for cell transmembrane transport ability test. The positive control drug SN-38 was used for this test. Each drug was accurately weighed to 1 μM and placed in a 1 mL volumetric flask. It was dissolved and diluted with DMSO to obtain the drug stock solution (1 mM). It was placed in a refrigerator at 4℃ for later use. Before use, each drug stock solution was diluted to 1 μM with HBSS buffer.

[0116] (2) Caco 2 monolayer cell model: Caco 2 cell monolayer membrane model was established according to the literature method (CHEMICAL & PHARMACEUTICAL BULLETIN, 69(11), 1054-1060) to simulate intestinal absorption and to conduct bidirectional transmembrane transport experiments of the compound in this experiment.

[0117] (3) Assay method for AP to BL side absorption and transport: Discard the original culture medium and wash with an appropriate amount of balanced salt solution (HBSS). Add 0.5 mL of 1 μM drug solution to the AP side and 1.5 mL of HBSS buffer to the BL side. Incubate the Transwell cell culture plate in an incubator. After 3 h of incubation, aspirate 0.5 mL of the transported drug solution from the BL side and add the same volume of pre-warmed HBSS buffer to the BL side. Add an equal volume of acetonitrile to the transported drug solution, vortex, and centrifuge (10000 rpm / min, 10 min). Filter through a 0.22 μm microporous membrane, collect the supernatant, analyze by HPLC, and calculate the drug content.

[0118] (4) Method for determining the secretion and efflux from BL to AP side: Discard the original culture medium and wash with an appropriate amount of balanced salt solution (HBSS). Add 1.5 mL of 1 μM drug solution to the BL side and 0.5 mL of HBSS buffer to the AP side. Incubate the Transwell cell culture plate in an incubator. After 3 h of incubation, aspirate 0.25 mL of the transported drug solution from the AP side and add the same volume of pre-warmed HBSS buffer to the AP side. Add an equal volume of acetonitrile to the transported drug solution, vortex, and centrifuge (10000 rpm, 10 min). Filter through a 0.22 μm microporous membrane, collect the supernatant, analyze by HPLC, and calculate the drug content.

[0119] (5) Calculation of Papp value and ER value: The apparent permeability coefficient (Papp) is calculated to assess the ability of compounds to permeate biofilms.

[0120] Calculation formula: Papp=(V / (C0×A))×(ΔQ / Δt); In the formula, V represents the volume of the solution on the receiving side, in mL; A represents the area of ​​the polycarbonate membrane, in 1.12 cm². 2 C0 represents the initial concentration of the analyte, in μg / mL; ΔQ / Δt represents the mass concentration of the analyte transported per unit time at the receiving end, in μg / mL·s. -1 .

[0121] Efflux rate (ER) is calculated to reflect the efflux and secretion of the tested compound.

[0122] Calculation formula: ER = Papp( BL→AP) / Papp （AP→BL） ×100%; In the formula, Papp （BL→AP） Papp represents the apparent permeability coefficient from the BL side to the AP side, dimensionless; （AP→BL） The apparent permeability coefficient from the AP side to the BL side is dimensionless.

[0123] For the efficacy assay of the compounds against the target DDX5, cells were co-incubated with DMSO or compounds 1, 4, 7, and SN-38 (100 nM) for 48 hours. Cells were then washed three times with pre-chilled PBS and lysed with RIPA buffer containing protease inhibitors. Lysates were sonicated on ice and then centrifuged at 12000×g for 20 min at 4°C. Total protein concentration was quantified using a BCA kit. Protein samples (30 μg each) were loaded with 5×SDS-PAGE loading buffer at 100°C for 10 min and then analyzed by SDS-PAGE. The isolated proteins were transferred to PVDF membranes, blocked with 5% skim milk powder for 2 h at room temperature, then incubated overnight with primary antibody at 4°C, and incubated for 2 h with HRP-labeled secondary antibody at room temperature. The membranes were washed three times with TBST between incubations. Protein bands were detected using enhanced chemiluminescence (ECL) substrate (Bio-Rad) and captured using a ChemiDoc MP imaging system (Bio-Rad). Signal intensity was measured using ImageJ software (V 1.42) and normalized using β-actin as a reference.

[0124] The efficacy assay (DNA relaxation assay) of compounds against target topoisomerase I (Topo I) determined their inhibitory effects on topoisomerase I by evaluating the ability of compounds 1, 4, 7, and SN-38 to loosen supercoiled (SC) pBR322 plasmid DNA. Each 20 μL reaction system contained 35 mM Tris-HCl (pH 8.0), 72 mM KCl, 5 mM MgCl2, 5 mM dithiothreitol, 5 mM spermine, 0.01% bovine serum albumin, 0.5 μg pBR322 plasmid DNA, and 1 unit of topoisomerase I (purchased from Takara Bio Engineering (Dalian) Co., Ltd.), along with either compound 1, 4, 7, or SN-38 at a concentration of 5 μM. The mixture was incubated at 37°C for 30 min, and then the reaction was terminated by adding 5% sodium dodecyl sulfate (SDS) at a concentration of 5 mg / mL proteinase K. DNA loading buffer (6:1 volume ratio) was then added, and the sample was electrophoresed on a 1% agarose gel at 75 V for 2 h in TAE buffer. The gel was stained with 1 μg / mL ethidium bromide (EB), rinsed with deionized water, and then observed using a gel imaging system. The inhibition rate was calculated by comparing the fluorescence intensity of the bands using ImageJ software.

[0125] The experimental results are shown in Table 3.

[0126] Table 3. Inhibition rates of the compounds of this invention and SN-38 against the two targets of topoisomerase 1 and DDX5. and cross-biomembrane Papp and ER data

[0127] It can be seen that compounds 1, 4, and 7 exhibited stronger DDX5 target activity than the control drug SN-38. The inhibition rates of compounds 1, 4, and 7 in inhibiting topoisomerase 1-induced DNA damage leading to cell death in tumor cells were 18.9%, 21.4%, and 19.8%, respectively, while the inhibition rate of SN-38 was 73.1%. The activation of the DNA damage repair pathway can lead to drug resistance in tumor cells. However, the compounds simultaneously inhibited DDX5, which can block the activation of the DNA damage repair pathway. The inhibition rates of compounds 1, 4, and 7 were 68.8%, 68.3%, and 70.7%, respectively, which were 14.5% higher than the inhibition rate of the control drug SN-38, indicating a lower risk of drug resistance in tumor cells and greater sensitivity to drug-resistant tumor cells.

[0128] Meanwhile, based on the comparative experiments of compounds 1, 4, 7, and SN-38 in the Caco-2 monolayer cell model simulating the mesentery, the apparent permeability coefficients (Papp) of compounds 1, 4, and 7 were significantly better than those of the control drug SN-38, indicating good transmembrane absorption. Moreover, the efflux rates (ER) of compounds 1, 4, and 7 were all less than 1, indicating that they are not efflux pump substrates that cause tumor drug resistance and are more sensitive to drug-resistant tumor cells. In contrast, the ER of the control drug SN-38 was 5.89, showing obvious efflux pump substrate characteristics. This characteristic is closely related to the resistance to topotecan, the prodrug of SN-38, in clinical practice.

[0129] The above results show that the structural features of the compound of the present invention determine its dual-target characteristics, excellent transmembrane absorption capacity and sensitivity to drug-resistant tumors, which are significantly superior to the current clinical drug SN-38.

[0130] Experimental Example 3: Comparison of the inhibitory activity of the compound of the present invention and SN-38 on drug-resistant tumor cells The inhibitory activity of compounds 1, 4, 7, and SN-38 on the proliferation of drug-resistant tumor cells was evaluated using the MTT cell proliferation inhibition method described in Experiment 1. The administration time was 72 h, and the cell culture method was the same as in Experiment 1. The difference between Experiment 1 and Experiment 1 was the cell lines used. In this experiment, HCT116 (oxaliplatin-resistant human colon cancer), T24 (cisplatin-resistant human bladder cancer), and 5637 (gemcitabine-resistant human bladder cancer) were used. The results are shown in Table 4.

[0131] The experimental results (Table 4) show that compounds 1, 4, and 7 protected by this invention exhibit strong inhibitory activity against oxaliplatin-resistant human colon cancer HCT116, cisplatin-resistant human bladder cancer T24, and gemcitabine-resistant human bladder cancer 5637. In contrast, the positive control drug SN-38 is insensitive to these resistant cell lines. In summary, the results in Table 4 show that compounds 1, 4, and 7 protected by this invention are effective against resistant cells at low doses, and their toxicity-to-efficacy relationship is far superior to that of the clinical drug SN38.

[0132] Table 4. Inhibitory activity of compounds 1, 4, 7, and SN-38 against the proliferation of drug-resistant tumor cells at 48 h.

[0133] Test Example 4: In vivo test of the inhibitory activity of the compound of the present invention on colon cancer tumor growth The in vivo antitumor activity of the compounds of the present invention was tested using a nude mouse HT-29 (colon cancer cell) xenograft model. Based on the results of the in vitro antitumor activity test, compound 7 of the present invention was selected for in vivo antitumor activity testing, and the positive control group was irinotecan, a clinical drug for colon cancer.

[0134] Test method: Five-week-old Balb / c nude mice weighing 18±2 g were used in the experiment. Colon cancer cells (HT-29) were inoculated into the axilla of the nude mice, and the tumors grew to 50-100 mm. 3 Drug administration began around 2:00 PM. The experiment was divided into a saline group, an irinotecan control group, and an experimental drug group (including compound 7 of this invention), with 8 nude mice in each group. Mice were fasted for 2 hours before the experiment. The experimental drug group received oral administration at doses of 4, 8, and 16 mg / kg. The irinotecan control group received intraperitoneal injection at a dose of 80 mg / kg. Administration was once weekly on days 0, 7, 14, and 21, for a total of four administrations. The feeding period was 28 days. Tumor volume and body weight were measured every other day. Results are as follows: Figure 6 As shown.

[0135] It can be seen that compound 7 of the present invention has good inhibitory activity against HT-29 tumors, and its activity is better than that of irinotecan. The mice in the compound 7 group of the present invention showed no significant change in body weight, and all physical signs of the mice were normal.

[0136] Experimental Example 5: In vivo test of the inhibitory activity of the compound of the present invention on bladder cancer tumor growth The in vivo antitumor activity of the compounds of the present invention was tested using a nude mouse T24 (bladder cancer cell) xenograft model. Based on the results of the in vitro antitumor activity test, compound 1 of the present invention was selected for in vivo antitumor activity testing. The positive control group consisted of irinotecan and cisplatin, clinical drugs for bladder cancer.

[0137] Test method: Five-week-old Balb / c nude mice weighing 18±2 g were selected for the experiment. T24 (bladder cancer cells) were inoculated into the axilla of the nude mice, and the tumors grew to 50-100 mm. 3 Drug administration began around 10:00 AM. The experiment was divided into a saline group, an irinotecan control group, a cisplatin control group, and an experimental drug group (including compound 1 of this invention), with 8 nude mice in each group. Mice were fasted for 2 hours before the experiment. The experimental drug group received oral administration at doses of 0.5 mg / kg and 1 mg / kg, respectively, once a week on days 0, 7, 14, and 21. The irinotecan control group received intraperitoneal injection once a week on days 0, 7, 14, and 21, at a dose of 40 mg / kg. The cisplatin group received intraperitoneal injection once a week on days 0, 7, 14, and 21, at a dose of 5 mg / kg. The feeding period was 30 days, and tumor volume and body weight were measured every other day. The results are as follows: Figure 7 As shown in Table 5, the tumor inhibition rate against bladder cancer is also shown.

[0138] Table 5. Inhibitory activity of the compounds of this invention, irinotecan, and cisplatin against bladder cancer tumor growth.

[0139] according to Figure 7 As can be seen from Table 5, compound 1 of the present invention has excellent inhibitory activity against T24 tumors. In the experimental group, the tumors of 3 mice disappeared completely. The activity was significantly better than that of irinotecan and cisplatin. The body weight of the mice in the compound 1 group of the present invention did not change significantly, and all the signs of the mice were normal.

[0140] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. An amination-modified 20(S)-10,11-difluoromethylenedioxycamptothecin derivative, characterized in that, Compounds having the structure shown in formula (Ⅰ): In equation (Ⅰ), R is ; Where X is n is an integer between 3 and 10; Z is selected from R1 and R2 are each independently selected from H, halogens, substituted or unsubstituted C. 1-30 Alkyl, substituted or unsubstituted C 3-12 alkenyl, substituted or unsubstituted C 1-10 Alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted C 3-20 Ethers, substituted or unsubstituted C 1-10 Ester group, substituted or unsubstituted C 3-10 alkoxy, substituted or unsubstituted C 1-10 Alkylsilyl, substituted or unsubstituted C 3-10 alkylthio, substituted or unsubstituted C 3-10 Alkyl, substituted or unsubstituted C 3-10 alkylimine group, substituted or unsubstituted C 7-10 alkenylamine, substituted or unsubstituted C 7-10 alkenyl imine, substituted or unsubstituted C 10-30 adamantanes; The alkyl, alkenyl, alkynyl, alkoxy, ether, alkylthio, alkazyl, alkimino, alkenylamine, and alkenylimino groups are straight-chain, branched, or cyclic structures.

2. The amination-modified 20(S)-10,11-difluoromethylenedioxycamptothecin derivative according to claim 1, characterized in that, Z is selected from either substituted or unsubstituted C. 3-10 Cycloalkylamines, substituted or unsubstituted C 10-30 adamantane amines, substituted or unsubstituted C 3-12 Cycloalkenylamines, substituted or unsubstituted arylamines, substituted or unsubstituted C 3-20 Ether cyclic amines, or substituted or unsubstituted aromatic heterocyclic amines; Preferably, the aromatic heterocyclic group is selected from amine-substituted pyridine rings, furan rings, thiophene rings, pyrazole rings, indole rings, benzopyrazole rings, piperidine rings, morpholine rings, thiomorpholine rings, naphthalene rings, or triazole rings.

3. The amination-modified 20(S)-10,11-difluoromethylenedioxycamptothecin derivative according to claim 1 or 2, characterized in that, When R1 and R2 are substituted structures, the substituted groups are each independently selected from halogens, C 1-10 Straight-chain or branched alkyl groups, C 1-10 alkenyl, C 1-10 alkynyl group, C 3-12 cycloalkyl, C 1-10 Alkoxy, C 1-10 Alkylthio, C 1-10 Alkylsilyl, C 1-10 Halogenated alkoxy groups, C 1-10 Ester group, 3-12 membered heterocyclic group, C 6-14 aryl, oxygen C 6-14 aryl, oxygen C 6-14 Aromatic heteroyl, nitrogen C 6-14 Aryl, nitrogen C 5-14 Aromatic heteroyl, 5-14 heteroaryl, -CN, -NO2, -CF2H, -CF2OH, -CF3 or -OCF3.

4. The amination-modified 20(S)-10,11-difluoromethylenedioxycamptothecin derivative according to claim 1, characterized in that, R1 and R2 are each independently selected from H, halogens, and C. 1-10 Straight-chain or branched alkyl, substituted or unsubstituted C 3-12 cycloalkyl, substituted or unsubstituted C 1-10 alkoxy, substituted or unsubstituted C 3-10 Epoxyalkyl, C 1-10 Alkylsilyl, C 1-10 Ester group, substituted or unsubstituted C 6-14 aryl, substituted or unsubstituted C 10-16 adamantane, substituted or unsubstituted C 7-10 alkenylamine, or substituted or unsubstituted C 7-10 alkenylimino group; Preferably, the substituted or unsubstituted C 1-10 The alkoxy group is C 1-10 Halogenated alkoxy groups; and / or the substituted or unsubstituted C groups. 7-10 The alkenyl imine group is either substituted or unsubstituted C. 7-10 Alkyl C 7-10 alkenylimino group; More preferably, the Z is selected from any of the following groups: 。 5. A method for preparing the amino-modified 20(S)-10,11-difluoromethylenedioxycamptothecin derivative as described in any one of claims 1-4, characterized in that, The synthesis route is as follows: Includes the following steps: (1) The raw material 5-amino-2,2-difluoro-1,3-benzodioxane was reacted with bromoC 3-10 Alkyl nitrile is added to organic solvent A, and a Lewis acid is added under an inert atmosphere to carry out the reaction, yielding intermediate I; (2) Intermediate I, tricyclic ketone and catalyst are added to organic solvent B and completely dissolved. The organic solvent is evaporated to dryness to obtain a solid. Vacuum solid-solid reaction is carried out under heating to obtain intermediate II. (3) Add intermediate II and amine compounds to organic solvent C, and add basic compounds to react to obtain the amination of 20(S)-10,11-difluoromethylenedioxycamptothecin derivative.

6. The preparation method according to claim 5, characterized in that, The 5-amino-2,2-difluoro-1,3-benzodioxane, brominated C 3-10 The molar ratio of alkyl nitrile to Lewis acid is 1:0.1-5:1-50; preferably 1:0.45-0.5:18-22. And / or the molar ratio of intermediate I, tricyclic ketone and catalyst is 1:0.1-5:0.1-5; preferably 1:0.5-1.2:0.1-0.6; And / or the molar ratio of intermediate II, amine compound and basic compound is 1:0.1-15:0.1-10; preferably 1:5-10:3-5.

7. The preparation method according to claim 5, characterized in that, In step (1), the reaction temperature is 80-95℃; the reaction process is monitored by TLC, and after the reaction is completed, an acidic aqueous solution is added to quench the reaction. And / or in step (2), the reaction is carried out under vacuum; and / or the temperature of the reaction is 60-150°C and the time is 0.5-20h; preferably, the temperature of the reaction is 115°C and the time is 1h. In step (3), the reaction temperature is 30-70°C and the reaction time is 2-30 h; preferably, the reaction temperature is 50°C and the reaction time is 5 h. Preferably, the acidic aqueous solution is an acidic aqueous solution with a concentration of 1-10 mol / mL, and more preferably, a 3 mol / mL HCl aqueous solution.

8. The preparation method according to claim 5, characterized in that, The Lewis acid is selected from any one of boron trichloride, boron trifluoride, aluminum trichloride, gallium trichloride, trimethylaluminum, ferric chloride, and ferric bromide; preferably boron trichloride; And / or the catalyst is selected from any one of Lewis acids, iodine, dodecyl sulfate, ferric chloride hexahydrate, aminosulfonic acid, 2,4,6-trichloro-1,3,5-triazine, bismuth trifluoromethanesulfonate, yttrium trifluoromethanesulfonate, lithium bis(trifluoromethanesulfonyl)imide, magnesium bis(trifluoromethanesulfonyl)imide, and p-toluenesulfonic acid or its hydrates; preferably p-toluenesulfonic acid hydrate; The alkaline compound is selected from one or more of N,N-diisopropylethylamine, sodium bicarbonate, potassium carbonate, sodium carbonate, calcium carbonate, potassium hydroxide, sodium hydroxide, and triethylamine; preferably N,N-diisopropylethylamine, sodium bicarbonate, potassium carbonate, and triethylamine; more preferably N,N-diisopropylethylamine or sodium bicarbonate.

9. The preparation method according to claim 5, characterized in that, The organic solvent A is selected from one or more of dichloroethane, dichloromethane, methanol, and acetonitrile; preferably dichloroethane. And / or the organic solvent B is selected from methanol, ethanol, n-butanol, ethyl acetate, acetone, dichloromethane, and dichloroethane; preferably dichloromethane or methanol; more preferably dichloromethane; And / or the organic solvent C is selected from ethanol, methanol, dichloromethane, DMF; preferably dichloromethane or DMF; more preferably DMF.

10. The use of an amino-modified 20(S)-10,11-difluoromethylenedioxycamptothecin derivative as described in any one of claims 1-4 in the preparation of a medicament for the prevention and / or treatment of cancer.

11. The application according to claim 10, characterized in that, The cancer is any one of bladder cancer, pancreatic cancer, breast cancer, prostate cancer, lung cancer, ovarian cancer, cervical cancer, skin cancer, melanoma, colon cancer, stomach cancer, liver cancer, esophageal cancer, kidney cancer, pharyngeal cancer, thyroid cancer, testicular cancer, brain cancer, bone cancer, and leukemia; preferably bladder cancer.

12. A pharmaceutical preparation, characterized in that, The pharmaceutical preparation comprises: any one of the following: an amino-modified 20(S)-10,11-difluoromethylenedioxycamptothecin derivative, or its active metabolite, prodrug, stereoisomer, pharmaceutically acceptable salt, polymorph, solvate, or conjugate, as described in any one of claims 1-4, in a therapeutically effective dose, and a pharmaceutically acceptable carrier.

13. The pharmaceutical preparation according to claim 12, characterized in that, The pharmaceutical formulation is a conjugate drug that uses an amino-modified 20(S)-10,11-difluoromethylenedioxycamptothecin derivative, its active metabolite, prodrug, its stereoisomer, pharmaceutically acceptable salt, polymorph, solvate, or conjugate as a cytotoxic agent. The conjugated drug also includes one or more of the following: antibodies, peptides, nucleic acids, and small molecules.

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