Paclitaxel prodrug as well as preparation method and application thereof

Abstract

The invention discloses a paclitaxel prodrug as well as a preparation method and application thereof. The compound and the pharmaceutically acceptable salt and ester thereof can close toxic sites through an ROS responsive connecting arm, are highly responsive to tumor microenvironment, play a selective killing and releasing effect and have low toxicity to normal cells, and a photosensitive fragment can be used for promoting prodrug release and amplifying a chemotherapy curative effect, can also be used as a self-developing fragment, and can be used as a photosensitizer. Therefore, the method can be used for accurate treatment of breast cancer. Therefore, the paclitaxel prodrug can be used for preparing drugs for preventing and / or treating diseases related to breast cancer.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical chemistry, and in particular to a paclitaxel prodrug and a preparation method and application thereof. Background Art

[0002] Paclitaxel (PTX) is a complex anti-tumor tricyclic diterpenoid compound originally isolated from the stems and leaves of Taxus truncatula. PTX is a white crystalline powder, odorless, tasteless, poorly soluble in water, but easily soluble in organic solvents. Its molecular formula is C 47 H 51 NO 14 , with a molecular weight of 853.92. Since its discovery, various research and development have followed. With the in-depth progress of its research, the mechanism of action of paclitaxel has been elaborated in detail: PTX can bind to and stabilize β-tubulin in microtubules, leading to the formation of parallel microtubule bundles and inhibiting cell division in the G2-M phase of the cell cycle; at the same time, paclitaxel can also cause cell apoptosis in a variety of different ways, involving the activation and regulation of multiple molecular signaling pathways, including the imbalance of Bcl-2 family proteins, the activation of Caspase cascade reactions, the decrease of mitochondrial membrane potential, and the breakage of DNA.

[0003]

[0004] Paclitaxel is one of the best-selling natural anti-tumor drugs. Due to its broad-spectrum anti-tumor activity, paclitaxel has been widely used in the treatment of breast cancer, ovarian cancer, some head and neck squamous cell carcinomas, and lung cancer. However, due to its poor water solubility, high cardiac and neurotoxicity, and easy bone marrow suppression and drug resistance, its clinical application is limited to a certain extent. Therefore, research on increasing its effectiveness and drugability has always been a hot spot and difficulty in the development of paclitaxel.

[0005] Prodrug refers to a compound with pharmaceutical activity, i.e., the parent drug, which is converted into a substance that is inactive in vitro through different modification methods, but can re-release active drug molecules in vivo through a specific conversion pathway.

[0006] During the occurrence and development of tumors, changes in the redox state in tumor cells can regulate signal transduction and death regulation. ROS is a general term for a class of free radicals, which are a class of byproducts produced during cell metabolism due to changes in the redox state. Hydrogen peroxide, singlet oxygen, superoxide anions, and hydroxyl radicals are all ROS. In normal cells, they can be quickly cleared, but because tumor cells have higher redox levels, their ROS levels are much higher than normal cells. Many studies have shown that compared with normal cells (<1mM), tumor cells produce ROS levels (>10mM) that are dozens or even thousands of times higher.

[0007] Photodynamic therapy is a new method for treating tumors using photosensitizers and laser-activated photosensitizers. Irradiating the lesion site with a specific wavelength can activate the photosensitive drugs that selectively accumulate in the lesion tissue, triggering a photochemical reaction to destroy the lesion. The photosensitizer in photodynamic therapy (PDT) transfers energy to the surrounding oxygen to generate highly active singlet oxygen. Singlet oxygen can undergo oxidation reactions with nearby biomacromolecules, produce cytotoxicity and kill diseased cells. Compared with traditional therapies, the advantage of PDT is that it can perform precise and effective treatments, and the side effects of this therapy are also very small.

[0008] Therefore, there is an urgent need for a prodrug that combines photodynamic therapy with chemotherapy. Summary of the invention

[0009] Purpose of the invention: The purpose of the present invention is to provide a paclitaxel prodrug that combines photodynamic therapy with chemotherapy; another purpose of the present invention is to provide a method for preparing the paclitaxel prodrug; another purpose of the present invention is to provide an application of the paclitaxel prodrug.

[0010] Technical solution: A compound of the present invention as shown in general formula I or general formula II or a pharmaceutically acceptable salt thereof, Wherein, R1 is selected from hydrogen, C1-C6 alkyl; Wherein, R2 is selected from C1-C6 alkyl.

[0011] Furthermore, the R1 is selected from hydrogen and C1-C6 n-alkyl.

[0012] Furthermore, the R2 is selected from C1-C6 n-alkyl.

[0013] Furthermore, the R1 is selected from hydrogen, methyl, ethyl, n-propyl, n-butyl, n-pentyl and n-hexyl.

[0014] Furthermore, the R2 is selected from methyl, ethyl, n-propyl, n-butyl, n-pentyl and n-hexyl.

[0015] The present invention provides the following preferred compounds: .

[0016] In some preferred embodiments, the pharmaceutically acceptable salts include, but are not limited to, acid addition salts formed by the compound represented by Formula I or Formula II and the following acids: hydrochloric acid, hydrobromic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid or succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid; and also include acid salts formed by the compound represented by Formula I or Formula II and inorganic bases.

[0017] The compounds of the above-mentioned general formula I or general formula II of the present invention may also exist in the form of their salts or esters, which are converted into compounds of the general formula I or general formula II in vivo. For example, within the scope of the present invention, the compounds of the present invention are converted into pharmaceutically acceptable salt forms according to processes known in the art, and they are used in the form of salts.

[0018] In another aspect, the present invention provides a method for preparing the above-mentioned compound or a pharmaceutically acceptable salt thereof.

[0019] Wherein, R1 is selected from C1-C6 alkyl.

[0020] In another aspect, the present invention provides a method for preparing the above-mentioned compound or a pharmaceutically acceptable salt thereof.

[0021] Wherein, R2 is selected from C1-C6 alkyl.

[0022] In another aspect, the present invention provides a pharmaceutical composition comprising the above-mentioned compound or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

[0023] The pharmaceutical composition of the present invention can be administered in various known ways, such as orally, parenterally, by inhalation spray or via an implanted reservoir. The pharmaceutical composition of the present invention can be administered alone or in combination with other drugs. Oral compositions can be in any orally acceptable dosage form, including but not limited to tablets, capsules, emulsions, suspensions, dispersions and solutions. Commonly used pharmaceutically acceptable carriers or excipients include stabilizers, diluents, surfactants, lubricants, antioxidants, adhesives, colorants, fillers, emulsifiers, etc.

[0024] Sterile injectable compositions can be formulated using suitable dispersing agents or wetting agents and suspending agents according to techniques known in the art. Pharmaceutically acceptable carriers and solvents that can be used include water, mannitol, sodium chloride solution, and the like.

[0025] The actual dosage level of the active ingredient in the pharmaceutical composition of the present invention can be varied to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient. The dosage level selected depends on a variety of factors, including the activity of the specific compound of the present invention or its salt used, the route of administration, the time of administration, the excretion rate of the specific composition used, the duration of treatment, other drugs, compounds and / or materials used in combination with the specific composition used, the age, sex, weight, general health and previous medical history of the patient being treated, and similar factors well known in the medical field.

[0026] In another aspect, the present invention provides a use of the above-mentioned compound or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing and / or treating tumor diseases.

[0027] Preferably, the tumor disease is breast cancer.

[0028] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: the toxic sites can be blocked by ROS-responsive linkers, which is highly responsive to the tumor microenvironment, has the effect of selective killing and release, and has low toxicity to normal cells. The photosensitive fragment can be used to promote the release of prodrugs and amplify the efficacy of chemotherapy. It can also be used as a self-imaging fragment to exert in vivo imaging function, and can be used for the precise treatment of breast cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 It simulates the changes of different ROS levels released by compound 2 over time in vitro; Figure 2 This is the in vivo imaging of mice treated with compound 2 for 12 hours, with the left side being the oral group and the right side being the tail vein group; Figure 3 This is the in vivo imaging of mice treated with compound 2 for 24 hours, with the left side showing the oral group and the right side showing the tail vein group; Figure 4 This is the in vivo imaging of mice treated with compound 2 at 36h, with the left side showing the oral group and the right side showing the tail vein group; Figure 5 This is the in vivo imaging of mice treated with compound 2 at 48 hours, with the left side being the oral group and the right side being the tail vein group; Figure 6 These are the in vivo imaging images of mice treated with Compound 2 for 72 hours, with the left side representing the oral group and the right side representing the tail vein group. DETAILED DESCRIPTION

[0030] The preparation methods of the compounds of general formula I or general formula II of the present invention are described below in conjunction with specific examples, but these specific methods do not constitute any limitation to the present invention. The compounds of the present invention can also be conveniently prepared by optionally combining various synthetic methods described in this specification or known in the art, and such combinations can be easily performed by those skilled in the art to which the present invention belongs.

[0031] The starting materials, reaction reagents, etc. used in the specific embodiments of the present invention are all commercially available. The present invention can be prepared in the form of salt by using a salt-forming method commonly used in the art, for example, dissolving the compound in hydrochloric acid ethanol at room temperature to react to generate a hydrochloride; or adding benzenesulfonic acid to react to generate a benzenesulfonate.

[0032] The design idea of ​​the compounds of general formula I and general formula II of the present invention is as follows: paclitaxel is coupled to the photosensitizer 5-aminolevulinic acid and its derivatives through different redox-responsive linking arms to generate a prodrug combining photodynamic therapy with chemotherapy. In the compounds of the present invention, the linking site of paclitaxel is the C-2' position of paclitaxel, which is connected to one end of the redox-responsive linking arm, and the other end of the redox-responsive linking arm is connected to the amino group of 5-aminolevulinic acid and its derivatives. Among them, the redox-responsive linking arm is a disulfide bond or a thioether bond. The photosensitizer 5-aminolevulinic acid and its derivatives include 5-aminolevulinic acid, 5-aminolevulinic acid methyl ester, 5-aminolevulinic acid ethyl ester, 5-aminolevulinic acid propyl ester, 5-aminolevulinic acid butyl ester, 5-aminolevulinic acid pentyl ester and 5-aminolevulinic acid hexyl ester.

[0033] The thioether prodrug of the present invention is shown in the general formula I:

[0034] Wherein, R1 is selected from hydrogen, and C1-C6 is a normal alkyl group.

[0035] The disulfide bond type prodrug of the present invention is shown in the general formula II:

[0036] Wherein, R2 is selected from C1-C6 n-alkyl.

[0037] The synthetic route of the compound represented by the general formula I of the present invention is as follows:

[0038] Thionyl chloride is added dropwise to the alcohol (R1-OH) solution in an ice bath, and then 5-aminolevulinic acid hydrochloride (5-ALA HCl) is added to react, and the reaction is completed to obtain a 5-aminolevulinic acid ester derivative (5-ALA-R1);

[0039] Paclitaxel (PTX), thiohydroxyacetic anhydride, triethylamine (TEA), and 4-dimethylaminopyridine (DMAP) are added to dichloromethane (DCM) for reaction. After the reaction is completed, the intermediate PTX-S-COOH is obtained; Add the intermediate PTX-S-COOH, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), DIPEA, and 5-aminolevulinic acid ester derivative (5-ALA-R1) into dichloromethane; stir to react, and the target product is obtained after the reaction is completed; Wherein, R1 is selected from C1-C6 n-alkyl.

[0040] When R1 is hydrogen, the synthesis route of the compound represented by general formula I is as follows:

[0041] The intermediate PTX-S-COOH, N-hydroxysuccinimide (NHS) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI) are added to dichloromethane for reaction, and an active ester intermediate is obtained after the reaction is completed; The active ester intermediate, 5-aminolevulinic acid hydrochloride and N,N-diisopropylethylamine (DIPEA) are added to a dichloromethane solution to carry out a reaction. After the reaction is completed, the target product is obtained.

[0042] The synthetic route of the compound shown in the general formula II of the present invention is as follows:

[0043] 2,2'-dithiodiacetic acid, add acetyl chloride, and react under nitrogen protection. The target product 1,4,5-oxydithiophene-2,7-dione (DTDPA) is obtained after the reaction is completed;

[0044] Paclitaxel (PTX), DTDPA, triethylamine and DMAP were added to DCM and reacted overnight. After the reaction, the intermediate PTX-SS-COOH was obtained.

[0045] The intermediate PTX-SS-COOH, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), DIPEA and 5-aminolevulinic acid ester derivative (5-ALA-R2) are added to dichloromethane and stirred for reaction. After the reaction is completed, the target product is obtained; wherein R2 is selected from C1-C6 n-alkyl.

[0046] Synthesis of intermediates The synthesis of the intermediate involved in the embodiment of the present invention is as follows: Synthesis of 1-(((1S,2R)-1-benzamido-3-(((2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS)-6,12b-diacetoxy-12-(benzoyloxy)-4,11-dihydroxy-4a,8,13,13,5-tetramethyl-5-oxo-2a,3,4,4a,5,6,9,11,12,12a,12b-dodecahydro-1H-7,11-methoxycyclodecan[3,4]benzo[1,2-b]oxo-9-yl)oxy)-3-oxo-1-phenylpropane-2-oxoethoxy)-2-oxoethyl)thioacetic acid (PTX-S-COOH)

[0047] Weigh 50 mg of paclitaxel, 15 mg of thiohydroxyacetic anhydride, 10 μl of triethylamine, 1.5 mg of 4-dimethylaminopyridine, and add 2 ml of dichloromethane. React at room temperature overnight. Monitor the reaction by thin layer chromatography. After the reaction is completed, remove the organic solvent under reduced pressure, and extract the residue (dissolve in ethyl acetate, wash with water and saturated sodium chloride in turn). The organic phase is dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to obtain the intermediate PTX-S-COOH, 51 mg, yellow solid. 88% yield.

[0048] 1 H NMR (400 MHz, Chloroform- d ) d 8.03 – 7.97 (m, 2H), 7.85 – 7.79 (m, 2H), 7.63 (dd, J = 8.2, 0.8 Hz, 1H), 7.58 – 7.49 (m, 2H), 7.49 – 7.40 (m, 6H), 7.40 – 7.33 (m, 2H), 7.27 (ddt, J = 8.2, 6.9, 2.1 Hz, 1H), 6.64 (p, J = 1.0 Hz,1H), 6.16 (dddt,J = 7.3, 6.2, 2.6, 1.5 Hz, 1H), 5.69 (d, J = 8.8 Hz, 1H), 5.53(dd, J = 7.6, 0.8 Hz, 1H), 5.53 – 5.46 (m, 1H), 4.82 (ddt, J = 6.4, 3.8, 2.5 Hz,1H), 4.42 – 4.32 (m, 2H), 4.30 (d, J = 12.2 Hz, 1H), 3.84 (d, J = 11.5 Hz, 1H),3.77 (dt, J = 8.8, 2.6 Hz, 1H), 3.54 (d, J = 8.4 Hz, 2H), 3.43 (d, J = 1.4 Hz, 2H), 2.45 (s, 1H), 2.40 – 2.31 (m, 2H), 2.25 – 2.16 (m, 6H), 2.02 (ddd, J = 12.3,7.7, 6.5 Hz, 1H), 1.82 (t, J = 1.2 Hz, 3H), 1.19 (d, J = 1.0 Hz, 5H).

[0049] Electrospray ionization mass spectrometry (ESI-MS) calculated value: C 51 H 55 NO 17 S (MH) - m / z: 984.3, found 984.3.

[0050] 2. Synthesis of 5-aminolevulinic acid ethyl ester

[0051] At 0℃, add 600 microliters of thionyl chloride dropwise to 8 milliliters of ethanol in an ice bath. Stir for 10 minutes. Remove ice and add 1 gram of 5-aminolevulinic acid hydrochloride. React at room temperature for 1 hour. Heat to 78℃ and reflux for 2 hours. Cool and add ether. Solid precipitates and crush and slurry. Filter and wash with ether and acetonitrile. Obtain the target product, 5-aminolevulinic acid ethyl ester, 0.85 grams. White solid. 89% yield. 1 H NMR (400 MHz, Chloroform- d ) d4.13 (q, J = 6.6 Hz, 2H), 3.73 (t, J =6.5 Hz, 2H), 2.90 (q, J = 6.6 Hz, 1H), 2.80 (q, J = 6.5 Hz, 1H), 2.75 – 2.68 (m,2H), 2.63 – 2.56 (m, 2H), 1.24 (t, J = 6.6 Hz, 3H). ESI-MS calculated for: C7H 13 NO3(M+H) + m / z: 160.2, found 160.2.

[0052] 3. Synthesis of 5-aminolevulinic acid propyl ester

[0053] At 0℃, add 600 microliters of thionyl chloride dropwise to 8 milliliters of n-propanol in an ice bath. Stir for 10 minutes. Remove ice and add 1 gram of 5-aminolevulinic acid hydrochloride. React at room temperature for 1 hour. Heat to 98℃ and reflux for 2 hours. Cool and add ether. Solid precipitates and crush and slurry. Filter and wash with ether and acetonitrile. Obtain 0.97 grams of the intermediate 5-aminolevulinic acid propyl ester. White solid. 93.85% yield. 1 H NMR (400 MHz, Chloroform- d ) d 4.07 (t, J = 5.7 Hz, 2H), 3.73 (t, J = 6.5 Hz, 2H), 2.90 (q, J = 6.6 Hz, 1H), 2.80 (q, J = 6.6 Hz, 1H), 2.75 – 2.68(m, 2H), 2.63 – 2.56 (m, 2H), 1.77 – 1.67 (m, 2H), 0.97 (t, J = 8.0 Hz, 3H). ESI-MS calculated: C8H 15 NO3(M+H) + m / z: 174.2, found 174.2.

[0054] 4. Synthesis of butyl 5-aminolevulinate

[0055] At 0℃, add 600 microliters of thionyl chloride dropwise to 8 milliliters of n-butanol in an ice bath. Stir for 10 minutes. Remove ice and add 1 gram of 5-aminolevulinic acid hydrochloride. React at room temperature for 1 hour. Heat to 118℃ and reflux for 2 hours. Cool and add ether. Solid precipitates and crush and slurry. Filter and wash with ether and acetonitrile. Obtain 1.01 grams of the intermediate 5-aminolevulinic acid butyl ester. White solid. 90% yield. 1 H NMR (400 MHz, Chloroform- d ) d 4.07 (t, J = 6.5 Hz, 2H), 3.73 (t, J =6.5 Hz, 2H), 2.90 (q, J = 6.6 Hz, 1H), 2.80 (q, J = 6.6 Hz, 1H), 2.75 – 2.68 (m,2H), 2.63 – 2.56 (m, 2H), 1.61 (p, J = 6.8 Hz, 2H), 1.39 (dt, J = 14.0, 7.0 Hz,2H), 0.97 (t, J = 7.0 Hz, 3H). ESI-MS calculated value: C9H 17 NO3(M+H) + m / z: 188.2, measured value 188.2.

[0056] 5. Synthesis of 5-aminolevulinic acid pentyl ester

[0057] At 0℃, add 600 microliters of thionyl chloride dropwise to 8 milliliters of n-pentanol in an ice bath. Stir for 10 minutes. Remove ice and add 1 gram of 5-aminolevulinic acid hydrochloride. React at room temperature for 1 hour. Heat to 139℃ and reflux for 2 hours. Cool and add ether. Solid precipitates and crush and slurry. Filter and wash with ether and acetonitrile. Obtain 1.13 grams of the intermediate 5-aminolevulinic acid pentyl ester. White solid. 94% yield. 1 H NMR (400 MHz, Chloroform- d ) d 4.06 (t, J = 5.9 Hz, 2H), 3.73 (t, J =6.5 Hz, 2H), 2.90 (q, J = 6.6 Hz, 1H), 2.80 (q, J= 6.6 Hz, 1H), 2.75 – 2.68 (m,2H), 2.63 – 2.56 (m, 2H), 1.71 – 1.62 (m, 2H), 1.42 – 1.31 (m, 4H), 0.95 –0.85 (m, 3H). ESI-MS calculated: C 10 H 19 NO3(M+H) + m / z: 202.3, found 202.3.

[0058] 6. Synthesis of 5-aminolevulinic acid hexyl ester

[0059] At 0℃, add 600 microliters of thionyl chloride dropwise to 8 milliliters of n-hexanol in an ice bath. Stir for 10 minutes. Remove ice and add 1 gram of 5-aminolevulinic acid hydrochloride. React at room temperature for 1 hour. Heat to 159℃ and reflux for 2 hours. Cool and add ether. Solid precipitates and crush and slurry. Filter and wash with ether and acetonitrile. Obtain 1.19 grams of the intermediate 5-aminolevulinic acid hexyl ester. White solid. 93% yield. 1 H NMR (400 MHz, Chloroform- d ) d 4.06 (t, J = 6.2 Hz, 2H), 3.73 (t, J =6.5 Hz, 2H), 2.90 (q, J = 6.6 Hz, 1H), 2.80 (q, J = 6.6 Hz, 1H), 2.75 – 2.68 (m,2H), 2.63 – 2.56 (m, 2H), 1.68 – 1.59 (m, 2H), 1.46 – 1.25 (m, 6H), 0.92 –0.86 (m, 3H). ESI-MS calculated: C 11 H21NO3(M+H) + m / z: 216.3, found 216.3.

[0060] 7. Synthesis of 1,4,5-oxodithiophene-2,7-dione (DTDPA)

[0061] 0.5 g of 2,2'-dithiodiacetic acid was added with 2 ml of acetyl chloride. The mixture was reacted at room temperature for 2 hours under nitrogen protection. 20 ml of toluene was added three times, and the mixture was spin-dried to remove the solvent. Ether was added to slurry, and solids were precipitated. The target product 1,4,5-oxodithiophene-2,7-dione (DTDPA) was obtained by suction filtration with a yield of 47% as a white solid.

[0062] 8. Synthesis of 2-(((1S,2R)-1-benzamido-3-(((2aR,4S,4aS,6R,9S,11S,12,12aR,12bS)-6,12b-diacetoxy-12-(benzoyloxy)-4,11-dihydroxy-4a,8,13,13,5-tetramethyl-5-oxo-2a,3,4,4a,5,6,9,11,12,12a,12b-dodecahydro-1H-7,11-methoxycyclodecan[3,4]benzo[1,2-b]oxo-9-yl)oxy)-3-oxo-1-phenylpropane-2-oxoethoxy)-dithio)acetate (PTX-SS-COOH)

[0063] 50 mg of paclitaxel, 15 mg of DTDPA, 10 μl of triethylamine, 1.5 mg of DMAP were added to 2 ml of DCM. The reaction was allowed to proceed overnight. The reaction was monitored by thin layer chromatography. After the reaction was completed, the organic solvent was removed under reduced pressure, and the residue was extracted (dissolved in ethyl acetate, washed with water and saturated sodium chloride in sequence). The organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to obtain the intermediate PTX-SS-COOH, 40 mg. Yellow solid, yield 67%.

[0064] 1 H NMR (400 MHz, Chloroform- d ) d 8.03 – 7.97 (m, 2H), 7.85 – 7.79 (m, 2H), 7.63 (dd, J = 8.3, 0.9 Hz, 1H), 7.58 – 7.49 (m, 2H), 7.49 – 7.40 (m, 2H), 7.44 (s, 2H), 7.40 – 7.33 (m, 2H), 7.27 (ddt, J = 8.2, 6.9, 2.1 Hz, 1H), 6.64(p, J = 1.0 Hz, 1H), 6.16 (dddt, J = 7.3, 6.2, 2.6, 1.5 Hz, 1H), 5.69 (d,J = 8.8Hz, 1H), 5.53 (dd, J = 7.6, 0.7 Hz, 1H), 5.53 – 5.46 (m, 1H), 4.82 (ddt, J = 6.4,3.8, 2.5 Hz, 1H), 4.42 – 4.32 (m, 2H), 4.30 (d, J = 12.2 Hz, 1H), 3.84 (d, J =11.5 Hz, 1H), 3.80 – 3.68 (m, 2H), 3.70 – 3.61 (m, 3H), 2.45 (s, 1H), 2.40 –2.31 (m, 2H), 2.25 – 2.16 (m, 6H), 2.02 (ddd, J = 12.3, 7.7, 6.5 Hz, 1H), 1.82(t, J = 1.2 Hz, 3H), 1.19 (d, J = 1.0 Hz, 5H). ESI-MS calculated value: C 51 H 55 NO 17 S2(MH) - m / z:1016.3, measured value 1016.3.

[0065] Example 1 Synthesis of Compound 1 (3S,4R)-4-(((2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS)-6,12b-diacetoxy-12-(benzoyloxy)-4,11-dihydroxy-4a,8,13,13,13-tetramethyl-5-oxo-2a,3,4,4a,5,6,9,10,11,12,12a,12b-dodecahydro-1H-7,11-methoxycyclodecan[3,4]benzo[1,2-b]oxepin-9-yl)oxycarbonyl)-1,6,10,13-tetraoxo-1,3-diphenyl-5-oxa-8-thia-2,11-diazahexadecane-16-ol

[0066] 60 mg of intermediate PTX-S-COOH, 8 mg of N-hydroxysuccinimide (NHS), 14 mg of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI) were added to 2 ml of dichloromethane in an ice bath, reacted for half an hour, and then returned to room temperature for 24 hours. After the reaction was completed, the organic solvent was removed under reduced pressure, and the residue was extracted (dissolved in ethyl acetate, washed with water and saturated sodium chloride in turn). The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure to obtain an active ester intermediate.

[0067] Then add 12 mg of 5-aminolevulinic acid hydrochloride, 26 μl of N,N-diisopropylethylamine (DIPEA) and active ester in 2 ml of dichloromethane solution, and react overnight. Monitor the reaction by thin layer chromatography. After the reaction is completed, remove the organic solvent under reduced pressure, and extract the residue (dissolve in ethyl acetate, wash with water and saturated sodium chloride in turn). Dry the organic phase with anhydrous sodium sulfate, evaporate to dryness under reduced pressure, and purify by silica gel column chromatography to obtain the target product 1, 21 mg. White solid. 31% yield.

[0068] 1 H NMR (400 MHz, Chloroform- d ) d 8.18 – 8.14 (m, 2H), 7.87 – 7.71 (m,2H), 7.64 – 7.56 (m, 1H), 7.55 – 7.28 (m, 11H), 7.15 (d, J = 6.5 Hz, 1H), 6.33– 6.21 (m, 2H), 6.07 (dd, J = 9.4, 3.3 Hz, 1H), 5.69 (d, J = 7.1 Hz, 1H), 5.44(d, J = 3.3 Hz, 1H), 4.99 (dd, J = 9.8, 2.3 Hz, 1H), 4.44 (dd, J = 10.9, 6.6 Hz,1H), 4.35 – 4.16 (m, 2H), 4.07 (d, J = 5.1 Hz, 2H), 3.81 (d, J = 7.1 Hz, 1H),3.75 – 3.35 (m, 3H), 3.20 (t, J = 14.7 Hz, 2H), 2.57 (s, 4H), 2.51 (s, 3H), 2.44 – 2.14 (m, 7H), 1.94 (d,J = 1.4 Hz, 3H), 1.89 – 1.78 (m, 1H), 1.68 (s, 3H), 1.44 – 1.32 (m, 3H), 1.14 (s, 3H). 13 C NMR (101 MHz, Chloroform- d ) d 203.93, 203.47, 173.43, 171.63,170.35, 168.23, 168.00, 167.09, 162.57, 142.85, 136.93, 133.82, 133.69,132.86, 131.99, 130.40, 129.40, 129.11, 128.89, 128.55, 127.81, 127.03,84.56, 81.28, 79.23, 75.83, 75.68, 75.21, 72.18, 58.56, 52.88, 49.35, 45.79,43.32, 35.95, 34.43, 32.64, 31.27, 29.84, 26.91, 25.91, 22.91, 22.33, 21.02,14.93, 9.80. High resolution mass spectrometry HRMS (ESI) calculated value: C 56 H 62 N2O 19 S (M+Na) + m / z:1121.3559, measured value 1121.3558.

[0069] Example 2 Synthesis of Compound 2

[0070] (2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS)-9-((R)-15-(((S)-Benzoylamino(phenyl)methyl)-3,6,9,13-tetraoxo-2,14-dioxa-11-thio-8-azahexadecane-16-yl)oxy)-12-(benzoyloxy)-4,11-dihydroxy-4a,8,13,13-tetramethyl-5-oxo-3,4a,5,6,9,10,12,12-dehydro-1H-7,11-methoxycyclodecan[3,4]benzo[1,2-b]oxa-6,12b(2aH)-diacetic acid diethyl ester

[0071] 51 mg of intermediate PTX-S-COOH, 24 mg of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), 20 μl of DIPEA, and 15 mg of 5-aminolevulinic acid methyl ester were added to 2 ml of dichloromethane. Stir overnight. The reaction was monitored by thin layer chromatography. After the reaction was completed, the organic solvent was removed under reduced pressure, and the residue was extracted (dissolved in ethyl acetate, washed with water and saturated sodium chloride in turn). The organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to obtain the target product 2, 30 mg, yellow solid. The yield was 53%. The purity was 97%. 1 H NMR (400 MHz, Chloroform- d ) d 8.26 (d, J = 9.4 Hz, 1H), 8.22 – 8.10 (m, 2H), 7.90 – 7.82 (m, 2H), 7.65 –7.56 (m, 1H), 7.52 (dd, J = 8.4, 6.9 Hz, 2H), 7.50 – 7.27 (m, 8H), 7.11 (t, J =5.3 Hz, 1H), 6.30 (s, 1H), 6.11 (dd, J = 9.5, 3.2 Hz, 1H), 5.70 (d, J = 7.1 Hz,1H), 5.41 (d, J = 3.2 Hz, 1H), 4.99 (dd, J = 9.7, 2.2 Hz, 1H), 4.33 (d, J = 8.4 Hz,1H), 4.26 – 4.12 (m, 3H), 3.83 (d, J = 7.0 Hz, 1H), 3.78 (d, J = 15.6 Hz, 1H), 3.73 – 3.67 (m, 2H), 3.65 (s, 4H), 3.51 (d, J = 14.0 Hz, 1H), 3.24 – 3.10 (m,4H), 2.93 (s, 1H), 2.80 – 2.54 (m, 6H), 2.53 (s, 4H), 2.42 (dd, J= 15.7, 9.5Hz, 2H), 2.22 (s, 3H), 2.17 (s, 1H), 1.96 (d, J = 1.4 Hz, 3H), 1.69 (s, 3H), 1.24 (s, 3H), 1.14 (s, 3H). 13 C NMR (101 MHz, Chloroform- d ) d 8.40, 127.88,127.00, 84.60, 81.21, 79.28, 76.60, 75.87, 75.76, 75.27, 72.27, 72.18, 73.77, 73.86, 73.18, 73.78, 73.91, 73.76, 73.93, 73.77, 73.97, 73.18, 66.73,58.64, 52.69, 49.34, 45.70, 43.33, 38.76, 35.75, 35.67, 34.85, 34.71, 32.05,27.92, 26.92, 22.94, 22.35, 22.04, 20.96, 14.98, 10.46, 9.76. HRMS (ESI) calculated value: C 57 H 64 N2O 19 S (M+H) + m / z:1113.3897, measured value 1113.3906.

[0072] Example 3 Synthesis of Compound 3

[0073] (2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS)-9-(((R)-2-((S)-Benzamido(phenyl)methyl)-4,8,11,14-tetraoxo-3,15-dioxa-6-thio-9-azaheptadecanoyl)oxy)-12-(benzoyloxy)-4,11-dihydroxy-4a,8,13,13,5-tetramethyl-5-oxo-3,4,4a,5,6,9,11,12,12-dehydro-1H-7,11-methoxycyclodecanoic acid [3,4]benzo[1,2-b]oxa-6,12b(2aH)-diyl diacetate

[0074] 51 mg of intermediate PTX-S-COOH, 24 mg of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), 20 μl of DIPEA, and 16 mg of 5-aminolevulinic acid ethyl ester were added to 2 ml of dichloromethane. Stir overnight. The reaction was monitored by thin layer chromatography. After the reaction was completed, the organic solvent was removed under reduced pressure, and the residue was extracted (dissolved in ethyl acetate, washed with water and saturated sodium chloride in turn). The organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to obtain the target compound 3, 29 mg, yellow solid, with a yield of 50%. The purity was 98%.

[0075] 1 H NMR (400 MHz, Chloroform- d ) d 8.27 (d, J = 9.5 Hz, 1H), 8.21 – 8.14(m, 2H), 7.90 – 7.83 (m, 2H), 7.62 – 7.57 (m, 1H), 7.51 (dd, J = 8.2, 6.8 Hz,2H), 7.45 – 7.31 (m, 8H), 7.08 (t, J = 5.1 Hz, 1H), 6.30 (s, 1H), 6.13 (dd, J =9.5, 3.0 Hz, 1H), 5.70 (d, J = 7.1 Hz, 1H), 5.42 (d, J = 3.0 Hz, 1H), 4.99 (dd, J =9.7, 2.3 Hz, 1H), 4.46 (dd, J = 10.9, 6.6 Hz, 1H), 4.33 (d, J = 8.4 Hz, 1H), 4.23(dd, J = 8.4, 1.1 Hz, 1H), 4.11 (q, J = 7.1 Hz, 3H), 3.86 – 3.76 (m, 2H), 3.51(d, J = 13.9 Hz, 1H), 3.15 (dd, J= 14.7, 4.2 Hz, 2H), 2.83 (s, 2H), 2.81 – 2.69(m, 1H), 2.71 – 2.66 (m, 1H), 2.64 (dq, J = 3.1, 2.0, 1.5 Hz, 2H), 2.54 (s,3H), 2.44 (dd, J = 15.5, 9.4 Hz, 1H), 2.23 (s, 5H), 1.97 (d, J = 1.5 Hz, 3H), 1.69 (s, 3H), 1.27 – 1.21 (m, 9H), 1.14 (s, 3H). 13 C NMR (101 MHz, Chloroform- d ) d 203.72, 203.56, 172.20, 171.09,169.74, 168.79, 168.40, 167.86, 167.53, 166.84, 142.68, 136.78, 133.46,132.55, 131.52, 130.11, 129.09, 128.72, 128.57, 128.14, 128.09, 127.56,126.68, 84.28, 80.89, 78.98, 75.56, 75.44, 74.96, 71.95, 71.87, 60.80, 58.33,52.37, 49.02, 45.39, 43.02, 38.51, 35.44, 35.36, 34.55, 34.38, 31.76, 29.52,27.65, 26.60, 22.64, 22.05, 20.65, 14.67, 13.96, 9.45. HRMS (ESI) calculated value: C 58 H 66 N2O 19 S (M+H) + m / z:1127.4053, measured value 1127.4050.

[0076] Example 4 Synthesis of Compound 4

[0077] (2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS)-9-(((R)-2-((S)-Benzamido(phenyl)methyl)-4,8,11,14-tetraoxo-3,15-dioxa-6-thioxo-9-azaoctadecanoyl)oxy)-12-(benzoyloxy)-4,11-dihydroxy-4a,8,13,13,5-tetramethyl-5-oxo-3,4,4a,5,6,9,11,12,12-dehydro-1H-7,11-methoxycyclodeca[3,4]benzo[1,2-b]oxa-6,12b(2aH)-diacetic acid diethyl ester

[0078] 51 mg of intermediate PTX-S-COOH, 24 mg of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), 20 μL of DIPEA, and 18 mg of 5-aminolevulinic acid propyl ester were added to 2 ml of dichloromethane. Stir overnight. The reaction was monitored by thin layer chromatography. After the reaction was completed, the organic solvent was removed under reduced pressure, and the residue was extracted (dissolved in ethyl acetate, washed with water and saturated sodium chloride in turn). The organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to obtain the target compound 4, 30 mg, yellow solid, yield 50%. Purity 95%.

[0079] 1 H NMR (400 MHz, Chloroform- d ) d 8.28 (d, J = 9.5 Hz, 1H), 8.21 – 8.15(m, 2H), 7.89 – 7.83 (m, 2H), 7.63 – 7.56 (m, 1H), 7.51 (dd, J = 8.3, 6.8 Hz,2H), 7.45 – 7.31 (m, 8H), 7.08 (t, J = 5.2 Hz, 1H), 6.30 (s, 1H), 6.13 (dd, J =9.5, 3.0 Hz, 1H), 5.70 (d, J = 7.1 Hz, 1H), 5.42 (d, J = 3.0 Hz, 1H), 4.99 (dd, J =9.7, 2.3 Hz, 1H), 4.46 (dd, J = 10.9, 6.6 Hz, 1H), 4.33 (d,J = 8.4 Hz, 1H), 4.25– 4.12 (m, 3H), 4.01 (t, J = 6.7 Hz, 2H), 3.87 – 3.74 (m, 2H), 3.51 (d, J = 13.9Hz, 1H), 3.15 (dd, J = 14.7, 4.8 Hz, 2H), 2.81 (s, 7H), 2.54 (s, 4H), 2.44 (dd, J = 15.5, 9.4 Hz, 1H), 2.22 (s, 4H), 1.96 (d, J = 1.4 Hz, 3H), 1.69 (s, 3H),1.63 (q, J = 7.1 Hz, 2H), 1.42 (s, 1H), 1.24 (s, 3H), 1.14 (s, 3H), 0.92 (t, J =7.4 Hz, 3H). 13 C NMR (101 MHz, Chloroform- d ) d 204.04, 203.89, 172.64, 171.40,170.09, 169.15, 168.77, 168.24, 167.91, 167.12, 142.97, 137.06, 133.77,132.85, 131.84, 130.40, 129.41, 129.04, 128.87, 128.45, 128.42, 127.85,127.03, 84.58, 81.18, 79.23, 76.59, 75.87, 75.76, 75.26, 72.25, 72.18, 65.31,58.62, 55.86, 52.78, 49.33, 45.70, 43.80, 43.33, 38.75, 35.71, 34.85, 34.68,32.08, 28.35, 28.11, 27.92, 26.90, 22.92, 22.41, 22.34, 20.96, 18.72, 17.33,14.96, 14.07, 12.65, 9.76. HRMS (ESI) Calculated: C 59 H 68 N2O19 S (M+H) + m / z:1141.4210, measured value 1141.4211.

[0080] Example 5 Synthesis of Compound 5

[0081] (2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS)-9-(((R)-2-((S)-Benzamido(phenyl)methyl)-4,8,11,14-tetraoxo-3,15-dioxa-6-thio-9-azanonadecanoyl)oxy)-12-(benzoyloxy)-4,11-dihydroxy-4a,8,13,13,5-tetramethyl-5-oxo-3,4,4a,5,6,10,11,12,12-dehydro-1H-7,11-methoxycyclodecanoyl[3,4]benzo[1,2-b]oxa-6,12b(2aH)-diyl diacetate

[0082] 51 mg of intermediate PTX-S-COOH, 24 mg of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), 20 μL of DIPEA, and 19 mg of 5-aminolevulinic acid butyl ester were added to 2 ml of dichloromethane. Stir overnight. The reaction was monitored by thin layer chromatography. After the reaction was completed, the organic solvent was removed under reduced pressure, and the residue was extracted (dissolved in ethyl acetate, washed with water and saturated sodium chloride in turn). The organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to obtain the target compound 5, 32 mg, yellow solid, with a yield of 54%. The purity was 96%.

[0083] 1 H NMR (400 MHz, Chloroform- d ) d 8.27 (d, J = 9.5 Hz, 1H), 8.18 (d, J = 7.6Hz, 2H), 7.86 (d, J = 7.6 Hz, 2H), 7.59 (d, J = 7.3 Hz, 1H), 7.51 (t, J = 7.6 Hz,2H), 7.46 – 7.31 (m, 8H), 7.07 (t, J = 5.1 Hz, 1H), 6.30 (s, 1H), 6.13 (dd, J =9.1, 2.8 Hz, 1H), 5.70 (d,J = 7.1 Hz, 1H), 5.42 (d, J = 2.8 Hz, 1H), 5.03 – 4.95(m, 1H), 4.46 (dd, J = 11.0, 6.6 Hz, 1H), 4.33 (d, J = 8.5 Hz, 1H), 4.27 – 4.13(m, 3H), 4.06 (t, J = 6.7 Hz, 2H), 3.87 – 3.75 (m, 2H), 3.51 (d, J = 13.8 Hz,1H), 3.15 (dd, J = 14.7, 4.9 Hz, 2H), 2.72 – 2.61 (m, 4H), 2.54 (s, 4H), 2.45(dd, J = 15.5, 9.4 Hz, 1H), 2.23 (s, 4H), 1.97 (s, 3H), 1.93 – 1.85 (m, 1H),1.69 (s, 3H), 1.60 (q, J = 7.2 Hz, 3H), 1.36 (h, J = 7.5 Hz, 3H), 1.24 (s, 4H),1.14 (s, 3H), 0.92 (t, J = 7.4 Hz, 3H). 13 C NMR (101 MHz, Chloroform- d ) d84.58, 81.18, 79.23, 76.59, 75.87, 75.76, 75.26, 72.25, 72.18, 73.70, 73.97, 73.87, 73.96, 73.77, 73.84, 73.96, 73.86, 73.94, 73.78, 73.96, 73.87, 73.86, 73.97, 73.87, 73.84, 73.96, 73.86, 73.87, 73.84, 73.84, 73.84, 65.31,58.62, 55.86, 52.78, 49.33, 45.70, 43.80, 43.33, 38.75, 35.71, 34.85, 34.68,32.08, 28.35, 28.11, 27.92, 26.90, 22.92, 22.41, 22.34, 20.96, 18.72, 17.33,14.96, 14.07, 12.65, 9.76. HRMS (ESI) calculated value: C 60 H 70 N2O 19 S (M+H) + m / z:1155.4366, measured value 1155.4368.

[0084] Example 6 Synthesis of Compound 6

[0085] (2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS)-9-(((R)-2-((S)-Benzamido(phenyl)methyl)-4,8,11,14-tetraoxo-3,15-dioxa-6-thio-9-azaeicosyl)oxy)-12-(benzoyloxy)-4,11-dihydroxy-4a,8,13,13,5-tetramethyl-5-oxo-3,4,4a,5,6,10,11,12,12-dehydro-1H-7,11-methoxycyclodecan[3,4]benzo[1,2-b]oxa-6,12b(2aH)-diacetic acid diethyl ester

[0086] 51 mg of intermediate PTX-S-COOH, 24 mg of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), 20 μl of DIPEA, and 21 mg of 5-aminolevulinic acid pentyl ester were added to 2 ml of dichloromethane. Stir overnight. The reaction was monitored by thin layer chromatography. After the reaction was completed, the organic solvent was removed under reduced pressure, and the residue was extracted (dissolved in ethyl acetate, washed with water and saturated sodium chloride in turn). The organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to obtain the target compound 6, 34 mg, yellow solid, yield 56%. Purity 96%.

[0087] 1 H NMR (400 MHz, Chloroform- d ) d 8.27 (d, J = 9.4 Hz, 1H), 8.20 – 8.16(m, 2H), 7.89 – 7.84 (m, 2H), 7.60 (t, J = 7.3 Hz, 1H), 7.51 (dd, J = 8.4, 6.8Hz, 2H), 7.45 – 7.31 (m, 8H), 7.09 (t, J = 5.2 Hz, 1H), 6.30 (s, 1H), 6.12 (dd, J = 9.5, 3.1 Hz, 1H), 5.70 (d, J = 7.1 Hz, 1H), 5.41 (d, J = 3.1 Hz, 1H), 4.99(dd, J = 9.7, 2.3 Hz, 1H), 4.46 (dd, J = 11.0, 6.6 Hz, 1H), 4.33 (d, J = 8.5 Hz,1H), 4.25 – 4.13 (m, 3H), 4.04 (t, J = 6.8 Hz, 2H), 3.87 – 3.76 (m, 2H), 3.51(d, J = 13.9 Hz, 1H), 3.17 (t, J = 5.1 Hz, 2H), 2.84 (s, 4H), 2.64 (p, J = 2.4 Hz,2H), 2.54 (s, 3H), 2.43 (dd, J= 15.5, 9.3 Hz, 1H), 2.23 (s, 4H), 1.96 (d, J =1.4 Hz, 3H), 1.69 (s, 3H), 1.60 (q, J = 7.0 Hz, 3H), 1.45 (dd, J = 17.5, 6.6 Hz,6H), 1.24 (s, 3H), 1.14 (s, 3H), 0.90 (d, J = 6.7 Hz, 3H). 13 C NMR (101 MHz, Chloroform- d ) d 84.58, 81.18, 79.23, 76.59, 75.87, 75.76, 75.26, 72.25, 72.18, 73.70, 73.97, 73.87, 73.96, 73.77, 73.84, 73.96, 73.86, 73.94, 73.78, 73.96, 73.87, 73.86, 73.97, 73.87, 73.84, 73.96, 73.86, 73.87, 73.84, 73.84, 73.84, 65.31,58.62, 55.86, 52.78, 49.33, 45.70, 43.80, 43.33, 38.75, 35.71, 34.85, 34.68,32.08, 28.35, 28.11, 27.92, 26.90, 22.92, 22.41, 22.34, 20.96, 18.72, 17.33,14.96, 14.07, 12.65, 9.76. HRMS (ESI) calculated value: C 61 H 72 N2O 19 S (M+H) + m / z:1169.4523, measured value 1169.4519.

[0088] Example 7 Synthesis of Compound 7

[0089] (2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS)-9-(((R)-2-((S)-Benzamido(phenyl)methyl)-4,8,11,14-tetraoxo-3,15-dioxa-6-thio-9-azaeicosanoyl)oxy)-12-(benzoyloxy)-4,11-dihydroxy-4a,8,13,13,5-tetramethyl-5-oxo-3,4,4a,5,6,10,11,12,12-dehydro-1H-7,11-methoxycyclodecan[3,4]benzo[1,2-b]oxo-6,12b(2aH)-diyl diacetate

[0090] 51 mg of intermediate PTX-S-COOH, 24 mg of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), 20 μl of DIPEA, and 23 mg of 5-aminolevulinic acid hexyl ester were added to 2 ml of dichloromethane. Stir overnight. The reaction was monitored by thin layer chromatography. After the reaction was completed, the organic solvent was removed under reduced pressure, and the residue was extracted (dissolved in ethyl acetate, washed with water and saturated sodium chloride in turn). The organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to obtain the target compound 7, 31 mg, yellow solid, with a yield of 51%. The purity was 97%.

[0091] 1 H NMR (400 MHz, Chloroform- d ) d 8.34 (d, J = 9.5 Hz, 1H), 8.23 ​​– 8.18(m, 2H), 7.91 – 7.86 (m, 2H), 7.64 – 7.58 (m, 1H), 7.54 (dd, J = 8.3, 6.8 Hz,2H), 7.47 – 7.34 (m, 8H), 7.14 – 7.10 (m, 1H), 6.32 (d, J = 4.7 Hz, 2H), 6.15(dd, J = 9.5, 2.9 Hz, 1H), 5.72 (d, J = 7.1 Hz, 1H), 5.43 (d, J = 3.0 Hz, 1H), 5.02(dd, J = 9.8, 2.3 Hz, 1H), 4.48 (dd, J= 10.9, 6.6 Hz, 1H), 4.34 (s, 1H), 4.26(s, 3H), 4.07 (t, J = 6.8 Hz, 2H), 3.86 (d, J = 7.2 Hz, 1H), 3.81 (s, 1H), 3.54(d, J = 13.9 Hz, 1H), 3.18 (s, 2H), 2.66 (d, J = 2.3 Hz, 2H), 2.57 (s, 3H), 2.52– 2.39 (m, 2H), 2.25 (s, 4H), 1.99 (d, J = 1.5 Hz, 3H), 1.92 (ddd, J = 13.9,11.0, 2.4 Hz, 2H), 1.72 (s, 3H), 1.66 – 1.56 (m, 3H), 1.45 (d, J = 1.6 Hz, 1H),1.39 (s, 2H), 1.28 (s, 5H), 1.27 (s, 3H), 1.16 (s, 3H), 0.91 – 0.89 (m, 3H). 13 C NMR (101 MHz, Chloroform- d ) d84.60, 81.19, 79.30, 76.60, 75.90, 75.76, 75.26, 72.28, 72.17, 73.03, 73.77, 73.83, 73.76, 73.83, 73.91, 73.17, 73.04, 73.76, 73.83, 73.83, 73.43, 73.05, 73.87, 73.83, 73.16, 73.03, 73.76, 73.83, 73.83, 73.43, 29.83, 28.63, 27.93, 26.91, 25.65,22.96, 22.83, 22.66, 22.36, 20.97, 15.00, 14.14, 9.76. HRMS (ESI) calculated value: C 61 H 72 N2O 19 S (M+H) + m / z:1183.4679, measured value 1183.4677.

[0092] Example 8 Synthesis of Compound 8

[0093] (2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS)-9-(((R)-16-((S)-Benzoylamino(phenyl)methyl)-3,6,9,14-tetraoxo-2,15-dioxa-11,12-dithio-8-azaheptadecan-17-yl)oxy)-12-(benzoyloxy)-4,11-dihydroxy-4a,8,13,13-tetramethyl-5,4a,5,6,9,11,12,12a-dehydro-1H-7,11-methoxycyclodecan[3,4]benzo[1,2-b]oxa-6,12b(2aH)-diacetic acid diethyl ester

[0094] 51 mg of intermediate PTX-SS-COOH, 24 mg of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), 20 μL of DIPEA, and 15 mg of 5-aminolevulinic acid methyl ester were added to 2 ml of dichloromethane. Stir overnight. The reaction was monitored by thin layer chromatography. After the reaction was completed, the organic solvent was removed under reduced pressure, and the residue was extracted (dissolved in ethyl acetate, washed with water and saturated sodium chloride in turn). The organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to obtain the target compound 8, 24 mg, yellow solid, with a yield of 43%. The purity was 96%.

[0095] 1 H NMR (400 MHz, Chloroform- d ) d 8.16 – 8.09 (m, 2H), 7.87 – 7.81 (m,2H), 7.78 (d, J = 8.6 Hz, 1H), 7.67 – 7.59 (m, 1H), 7.58 – 7.35 (m, 9H), 7.28(d, J = 3.3 Hz, 1H), 6.99 (t, J = 4.7 Hz, 1H), 6.30 (s, 1H), 6.21 – 6.13 (m, 1H), 5.91 (dd, J = 8.5, 5.6 Hz, 1H), 5.65 (d, J = 7.1 Hz, 1H), 5.48 (d, J = 5.6 Hz, 1H),4.96 (dd, J = 9.7, 2.3 Hz, 1H), 4.44 (dd, J = 10.9, 6.6 Hz, 1H), 4.33 – 3.99 (m,4H), 3.76 (d, J = 7.1 Hz, 1H), 3.61 – 3.50 (m, 2H), 3.42 – 3.20 (m, 2H), 2.85 (s, 3H), 2.75 – 2.56 (m, 4H), 2.51 (ddd, J = 14.6, 9.7, 6.5 Hz, 1H), 2.44 (s, 3H), 2.22 (s, 4H), 2.14 (dd, J= 15.6, 9.3 Hz, 2H), 2.09 – 1.99 (m, 1H), 1.95 –1.84 (m, 5H), 1.68 (s, 3H), 1.46 (dd, J = 22.6, 6.5 Hz, 2H), 1.13 (s, 3H). 13 C NMR (101 MHz, Chloroform- d ) d 8.66, 128.60,128.25, 84.57, 81.17, 79.13, 76.54, 75.74, 75.68, 75.11, 72.14, 71.96, 58.48,53.73, 52.19, 49.65, 45.88, 43.27, 42.36, 40.82, 38.84, 35.80, 35.34, 34.72,27.58, 26.86, 22.99, 22.14, 20.98, 18.78, 17.44, 15.01, 9.75. HRMS (ESI) calculated value: C 57 H 64 N2O 19 S2(M+H) + m / z:1145.3617, measured value 1145.3615.

[0096] Example 9 Synthesis of Compound 9

[0097] (2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS)-9-(((R)-2-((S)-benzamido(phenyl)methyl)-4,9,12,15-tetraoxo-3,16-dioxa-6,7-dithia-10-octadecanoyl)oxy)-12-(benzoyloxy)-4,11-dihydroxy-4a,8,13,13-tetramethyl-5,4a,5,6,9,10,11,12,12-dehydro-1H-7,11-methoxycyclodecanoic acid [3,4]benzo[1,2-b]oxa-6,12b(2aH)-diyl diacetate

[0098] 51 mg of intermediate PTX-SS-COOH, 24 mg of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), 20 μL of DIPEA, and 15 mg of ethyl 5-aminolevulinate were added to 2 ml of dichloromethane. Stir overnight. The reaction was monitored by thin layer chromatography. After the reaction was completed, the organic solvent was removed under reduced pressure, and the residue was extracted (dissolved in ethyl acetate, washed with water and saturated sodium chloride in turn). The organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to obtain the target compound 9, 24 mg, yellow solid, with a yield of 42%. The purity was 97%.

[0099] 1 H NMR (400 MHz, Chloroform- d ) d 8.17 – 8.09 (m, 2H), 7.87 – 7.80 (m, 2H), 7.78 (d, J = 8.5 Hz, 1H), 7.68 – 7.58 (m, 1H), 7.57 – 7.33 (m, 10H), 6.97(t, J = 5.7 Hz, 1H), 6.30 (s, 1H), 6.22 – 6.08 (m, 1H), 5.92 (dd, J = 8.5, 5.5Hz, 1H), 5.66 (d, J = 7.1 Hz, 1H), 5.48 (d, J = 5.5 Hz, 1H), 4.97 (dd, J = 9.7, 2.3Hz, 1H), 4.44 (dd, J = 10.9, 6.6 Hz, 1H), 4.32 – 4.16 (m, 3H), 4.12 – 4.00 (m,3H), 3.77 (d, J = 7.0 Hz, 1H), 3.64 – 3.46 (m, 2H), 3.42 – 3.20 (m, 2H), 2.73 –2.66 (m, 2H), 2.60 (ddd, J= 7.4, 5.8, 3.4 Hz, 2H), 2.44 (s, 3H), 2.34 – 2.10(m, 5H), 1.97 – 1.86 (m, 6H), 1.68 (s, 4H), 1.42 (d, J = 5.5 Hz, 2H), 1.37 (s,1H), 1.33 (s, 3H), 1.13 (s, 3H). 13 C NMR (101 MHz, Chloroform- d ) d 8.51, 79.17, 75.74, 75.13, 72.16, 71.99, 61.18, 58.51, 62.77, 63.00, 64.73, 65.71, 66.99, 61.18, 67.24, 68.83, 68.38, 67.63, 67.08, 142.63, 136.95, 134.27,133.86, 133.00, 132.00, 130.37, 129.43, 129.16, 128.81, 128.68, 127.61,127.24, 84.60, 81.20, 79.17, 75.74, 75.13, 72.16, 71.99, 61.18, 58.51, 53.70,49.70, 45.89, 43.29, 42.40, 40.80, 35.81, 35.37, 34.74, 32.07, 31.58, 30.46,30.34, 29.84, 29.51, 27.88, 26.88, 23.01, 22.84, 22.16, 20.99, 15.03, 14.27,14.24, 9.76. HRMS (ESI) calculated value: C 58 H 66 N2O 19 S2(M+H) + m / z:1159.3774, measured value 1159.3773.

[0100] Example 10 Synthesis of Compound 10 (2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS)-9-(((R)-2-((S)-benzamido(phenyl)methyl)-4,9,12,15-tetraoxo-3,16-dioxa-6,7-dithia-10-azanonanoyl)oxy)-12-(benzoyloxy)-4,11-dihydroxy-4a,8,13,13,13-tetramethyl-5-oxo-3,4,4a,5,6,9,10,12,12-12a-decahydro-1H-7,11-methoxycyclodeca[3,4]benzo[1,2b]oxa-6,12b(2aH)-diacetic acid diethyl ester

[0101] 51 mg of intermediate PTX-SS-COOH, 24 mg of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), 20 μl of DIPEA, and 15 mg of 5-aminolevulinic acid propyl ester were added to 2 ml of dichloromethane. Stir overnight. The reaction was monitored by thin layer chromatography. After the reaction was completed, the organic solvent was removed under reduced pressure, and the residue was extracted (dissolved in ethyl acetate, washed with water and saturated sodium chloride in turn). The organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to obtain the target compound 10, 27 mg, yellow solid, with a yield of 47%. The purity was 95%.

[0102] 1 H NMR (300 MHz, Chloroform- d ) d 8.17 – 8.07 (m, 2H), 7.86 – 7.82 (m,2H), 7.74 – 7.30 (m, 11H), 7.28 (t, J = 1.7 Hz, 1H), 6.99 (t, J = 4.6 Hz, 1H), 6.30 (s, 1H), 6.16 (t, J = 8.8 Hz, 1H), 5.91 (dd, J = 8.5, 5.6 Hz, 1H), 5.65 (d, J = 7.0 Hz, 1H), 5.47 (d, J = 5.6 Hz, 1H), 5.02 – 4.90 (m, 1H), 4.44 (dd, J = 10.9,6.6 Hz, 1H), 4.33 – 4.06 (m, 4H), 3.98 (t, J= 6.7 Hz, 2H), 3.76 (d, J = 7.1 Hz,1H), 3.66 – 3.47 (m, 2H), 3.43 – 3.17 (m, 2H), 2.74 – 2.48 (m, 5H), 2.44 (s,3H), 2.21 (s, 3H), 1.91 (d, J = 9.9 Hz, 4H), 1.67 (s, 3H), 1.59 (p, J = 7.1 Hz,2H), 1.25 (s, 3H), 1.21 (s, 3H), 1.12 (s, 3H), 0.90 (t, J = 7.4 Hz, 3H). 13 C NMR (75 MHz, Chloroform- d ) d 203.88, 203.57, 172.77, 171.17, 170.09,169.11, 168.76, 168.40, 167.64, 167.05, 142.62, 136.93, 134.26, 133.87,132.96, 131.98, 130.36, 129.39, 129.14, 128.80, 128.65, 127.61, 127.25,84.57, 81.16, 79.12, 75.68, 75.09, 72.14, 66.76, 58.46, 53.72, 52.73, 49.68,45.88, 43.25, 42.35, 40.74, 36.94, 35.33, 34.73, 30.51, 29.85, 29.82, 29.77,27.81, 26.86, 22.99, 21.99, 20.98, 17.83, 15.02, 10.46, 9.74. HRMS (ESI) calculated value: C 59 H 68 N2O 19 S2(M+H) + m / z:1173.3931, measured value 1173.3934.

[0103] Example 11 Synthesis of Compound 11 (2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS)-9-(((R)-2-((S)-benzamido(phenyl)methyl)-4,9,12,15-tetraoxo-3,16-dioxa-6,7-dithia-10-azacycloalkyl)oxy)-12-(benzoyloxy)-4,11-dihydroxy-4a,8,13,13,5-tetramethyl-5-oxo-3,4,4a,5,6,10,11,12,12-dehydro-1H-7,11-methoxycyclodeca[3,4]benzo[1,2-b]oxa-6,12b(2aH)-diyl diacetate

[0104] 51 mg of intermediate PTX-SS-COOH, 24 mg of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), 20 μl of DIPEA, and 15 mg of 5-aminolevulinic acid butyl ester were added to 2 ml of dichloromethane. Stir overnight. The reaction was monitored by thin layer chromatography. After the reaction was completed, the organic solvent was removed under reduced pressure, and the residue was extracted (dissolved in ethyl acetate, washed with water and saturated sodium chloride in turn). The organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to obtain the target compound 11, 26 mg, yellow solid, with a yield of 45%. The purity was 96%.

[0105] 1 H NMR (300 MHz, Chloroform- d ) d 8.18 – 8.06 (m, 2H), 7.87 – 7.28 (m,14H), 6.98 (t, J = 4.6 Hz, 1H), 6.30 (s, 1H), 6.22 – 6.08 (m, 1H), 5.91 (dd, J =8.5, 5.5 Hz, 1H), 5.65 (d, J = 7.1 Hz, 1H), 5.47 (d, J = 5.6 Hz, 1H), 4.96 (dd, J =9.6, 2.2 Hz, 1H), 4.44 (t, J = 8.8 Hz, 1H), 4.33 – 4.06 (m, 4H), 4.03 (t, J = 6.7Hz, 2H), 3.76 (d, J= 7.1 Hz, 1H), 3.64 – 3.49 (m, 2H), 3.45 – 3.20 (m, 2H), 2.75 – 2.48 (m, 6H), 2.44 (s, 3H), 2.22 (s, 3H), 2.14 (dd, J = 15.6, 9.3 Hz,1H), 1.90 (d, J = 11.1 Hz, 4H), 1.67 (s, 4H), 1.61 – 1.50 (m, 2H), 1.23 (d, J =12.5 Hz, 5H), 1.13 (s, 3H), 0.91 (t, J = 7.3 Hz, 4H). 13 C NMR (75 MHz, Chloroform- d ) d 8.47, 79.13, 75.73, 74.77, 71.80, 71.42, 65.08, 58.47, 79.13, 75.74, 74.77, 71.81, 71.42, 65.08, 58.47, 79.13, 75.73, 74.77, 71.80, 71.42, 65.08, 58.47, 54.98, 49.69,45.88, 43.26, 42.29, 40.98, 35.91, 34.74, 30.65, 29.96, 29.75, 29.38, 27.84,26.87, 22.99, 21.93, 20.99, 19.20, 15.03, 13.83, 9.76. HRMS (ESI) calculated value: C 60 H 70 N2O 19 S2(M+H) + m / z:1187.4087, measured value 1187.4087.

[0106] Example 12 Synthesis of Compound 12 (2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS)-9-(((R)-2-((S)-benzamido(phenyl)methyl)-4,9,12,15-tetraoxo-3,16-dioxa-6,7-dithia-10-azaeicosanoyl)oxy)-12-(benzoyloxy)-4,11-dihydroxy-4a,8,13,13,13-tetramethyl-5-oxo-3,4,4a,5,6,9,10,12,12a-decahydro-1H-7,11-methoxycyclodeca[3,4]benzo[1,2b]oxa-6,12b(2aH)-diacetic acid diethyl ester

[0107] 51 mg of intermediate PTX-SS-COOH, 24 mg of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), 20 μL of DIPEA, and 15 mg of 5-aminolevulinic acid pentyl ester were added to 2 ml of dichloromethane. Stir overnight. The reaction was monitored by thin layer chromatography. After the reaction was completed, the organic solvent was removed under reduced pressure, and the residue was extracted (dissolved in ethyl acetate, washed with water and saturated sodium chloride in turn). The organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to obtain the target compound 12, 28 mg, yellow solid, with a yield of 48%. The purity was 96%.

[0108] 1 H NMR (300 MHz, Chloroform- d ) d 8.17 – 8.10 (m, 2H), 7.75 – 7.68 (m,2H), 7.65 – 7.30 (m, 12H), 7.10 (d, J = 9.4 Hz, 1H), 6.27 (s, 1H), 6.01 (dd, J =9.2, 3.4 Hz, 1H), 5.67 (d, J = 7.1 Hz, 1H), 5.55 (d, J = 3.5 Hz, 1H), 5.01 – 4.93(m, 1H), 4.43 (dd, J = 10.8, 6.6 Hz, 1H), 4.34 – 4.15 (m, 2H), 3.80 (d, J = 7.0Hz, 1H), 3.50 (s, 2H), 2.55 (ddd, J= 15.4, 9.6, 6.4 Hz, 1H), 2.44 (s, 3H), 2.34 (dd, J = 15.6, 9.0 Hz, 2H), 2.22 (s, 3H), 2.18 – 1.99 (m, 3H), 1.89 (d, J =1.3 Hz, 4H), 1.68 (s, 3H), 1.42 (d, J = 4.4 Hz, 2H), 1.33 (s, 3H), 1.27 (d, J =4.8 Hz, 6H), 1.21 (s, 4H), 1.13 (s, 3H), 0.86 (dt, J = 8.6, 6.2 Hz, 4H). 13 C NMR (75 MHz, Chloroform- d ) d 8.33, 167.75, 167.21, 142.61, 137.70, 136.71, 134.09,133.61, 133.06, 132.26, 130.37, 129.30, 128.85, 128.35, 127.30, 126.86,83.76, 81.21, 79.25, 75.68, 74.98, 72.03, 69.94, 68.01, 58.63, 52.85, 51.22,43.31, 40.43, 39.25, 39.17, 35.78, 34.98, 31.94, 31.55, 30.80, 29.84, 26.94,23.30, 22.44, 20.47, 20.06, 14.95, 13.88, 9.80. HRMS (ESI) calculated value: C 61 H 72 N2O 19 S2(M+Na) + m / z:1223.4063, measured value 1223.4056.

[0109] Example 13 Synthesis of Compound 13 (2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS)-9-(((R)-2-((S)-benzamido(phenyl)methyl)-4,9,12,15-tetraoxo-3,16-dioxa-6,7-dithia-10-azadocosabutanoyl)oxy)-12-(benzoyloxy)-4,11-dihydroxy-4a,8,13,13,13-tetramethyl-5-oxo-3,4,4a,5,6,10,11,12,12-dehydro-1H-7,11-methoxycyclodecan[3,4]benzo[1,2-b]oxa-6,12b(2aH)-diacetic acid diethyl ester

[0110] 51 mg of intermediate PTX-SS-COOH, 24 mg of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), 20 μl of DIPEA, and 15 mg of 5-aminolevulinic acid hexyl ester were added to 2 ml of dichloromethane. Stir overnight. The reaction was monitored by thin layer chromatography. After the reaction was completed, the organic solvent was removed under reduced pressure, and the residue was extracted (dissolved in ethyl acetate, washed with water and saturated sodium chloride in turn). The organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to obtain the target compound 13, 28 mg, yellow solid, with a yield of 47%. The purity was 98%.

[0111] 1 H NMR (400 MHz, Chloroform- d ) d 8.13 (dt, J = 7.0, 1.3 Hz, 2H), 7.88 –7.82 (m, 2H), 7.79 (d, J = 8.5 Hz, 1H), 7.68 – 7.60 (m, 1H), 7.59 – 7.29 (m,10H), 6.96 (t, J = 4.5 Hz, 1H), 6.30 (s, 1H), 6.17 (t, J = 9.0 Hz, 1H), 5.92 (dd, J = 8.5, 5.6 Hz, 1H), 5.65 (d, J = 7.1 Hz, 1H), 5.48 (d, J = 5.6 Hz, 1H), 4.96(dd, J = 9.7, 2.3 Hz, 1H), 4.44 (dd, J= 10.9, 6.6 Hz, 1H), 4.32 – 4.15 (m, 4H), 4.11 – 3.98 (m, 4H), 3.77 (d, J = 7.0 Hz, 1H), 3.64 – 3.51 (m, 3H), 3.42 3.21(m, 2H), 2.77 (dd, J = 7.3, 5.5 Hz, 1H), 2.72 – 2.58 (m, 5H), 2.56 – 2.48 (m, 1H), 2.44 (s, 3H), 2.22 (s, 3H), 2.14 (dd, J = 15.5, 9.0 Hz, 1H), 1.94 – 1.89(m, 4H), 1.68 (s, 3H), 1.62 – 1.55 (m, 3H), 1.21 (s, 3H), 1.13 (s, 3H), 0.93– 0.85 (m, 6H). 13 C NMR (101 MHz, Chloroform- d ) d 204.19, 203.88, 173.07, 171.48, 170.41, 169.41, 169.15, 168.70, 167.94, 167.39, 142.96, 137.27, 134.59, 134.18, 133.30, 132.29, 130.69, 129.74, 129.47, 129.12, 128.98, 127.93, 127.57, 84.91, 81.50, 79.48, 76.06, 76.01, 75.44, 72.47, 72.28, 65.71, 65.55, 58.81, 54.04, 50.02, 46.21, 43.59, 43.14, 42.70, 41.09, 36.14, 35.66, 35.07, 31.86, 31.85, 30.15, 28.97, 28.92, 28.24, 28.17, 27.20, 26.00, 25.96, 23.31, 22.97, 22.46, 21.29, 15.34, 14.58, 14.45, 10.07. HRMS (ESI) Calculated: C 62 H 74 N2O 19S2(M+H) + m / z:1215.4400, measured value 1215.4404.

[0112] Example 14 Compound Prodrug Release Experiment Compound 2 was prepared into a stock solution of 10 millimolar per liter (mM) in dimethyl sulfoxide (DMSO) as a solvent and stored in a refrigerator at -20 °C. An appropriate amount of the compound stock solution was diluted to a final concentration of 100 micromol per liter with phosphate buffered saline (PBS), and then H2O2 (0, 1, 2, 10 mM) was added and incubated at 37 °C for 3 times. Samples were collected at appropriate time intervals (0, 1, 2, 4, 6, 8, 10, 12, 24 h) and directly analyzed by high performance liquid chromatography (HPLC) and mass spectrometry (MS). The following conditions were used for the Agilent 1100 HPLC and UV detector: Agilent C18 column (4.6 × 150 mm, 3.5 μm); mobile phase: 85% methanol: 0.1% trifluoroacetic acid; flow rate: 1.0 mL / min.

[0113] like Figure 1 As shown, after 24 hours of incubation, the prodrug was basically not decomposed in the case of 0mmol / LH2O2, showing good stability. With the increase of ROS concentration, the rate and degree of prodrug release increased significantly, showing a positive correlation. At low concentration (1mmol / L), the single sulfide bond prodrug released 50.07%±0.02, and the disulfide bond prodrug released 53.06%±0.06. At high concentration (10mmol / L), the single sulfide bond prodrug released 68.83%±0.02, and the disulfide bond prodrug released 69.23%±0.02.

[0114] The difference between high and low concentrations showed that the single-sulfide bond prodrug released 18.76% ± 0.04 more, and the disulfide bond prodrug released 16.17% ± 0.03 more. These data indicate that the release of this type of prodrug is a ROS-responsive prodrug.

[0115] Example 15 Anti-cell proliferation assay (MTT assay) Take the MDA MB-231 cells in the logarithmic growth phase (cells were purchased from the cell bank of the Wuhan Institute of Virology, Chinese Academy of Sciences), digest them with 0.25% trypsin in a clean bench, discard the trypsin, and terminate the digestion. Use a pipette to suck up the culture medium, blow off the adherent cells, and transfer them to a 10 ml centrifuge tube. Centrifuge at 1000 rpm for five minutes and discard the supernatant. Then add an appropriate amount of culture medium and blow evenly to make a single cell suspension. Pipette 20 microliters and add it to the cell counting plate. After counting, dilute with culture medium to adjust the cell concentration to 3000 / well. Inoculate 200 microliters per well of the 96-well plate. After inoculation, culture for 12 hours. After the cells are attached, aspirate the culture medium and add 200 microliters of culture medium containing 1μM drug. Set up 3 parallel wells for each compound concentration, and add an equal amount of blank culture medium to the control group. 24 hours after administration, 20 μl of MTT was added to each well, and the cells were incubated for 3 hours before being taken out and shaken on an oscillator for 10 minutes. The absorbance of each well was detected using an ELISA reader at 490 / 570 nm as the detection wavelength, and the cell survival rate after the action of the test compound at different concentrations was calculated according to the corresponding formula: Cell survival rate (%) = (1- A 实验组 / A 对照组 )×100%; where A 实验组 Indicates the absorbance value of the experimental group, A 对照组 represents the absorbance value of the control group. The experimental results are shown in Table 1. The data show that the prodrugs exhibited similar or even stronger killing effects on cancer cells. The preferred compound 13 exhibited an inhibition rate 9% higher than that of the positive drug.

[0116] Table 1 Antiproliferative activity of prodrugs against breast cancer cells

[0117] The inhibition rate (1 μM) data results are the average of three parallel experiments, and ± represents the standard deviation (SD); Example 16 Acute toxicity test of compounds in mice ICR mice of about five weeks old were raised normally for nearly a week, and then randomly assigned to 6 dose groups of compound 2 and paclitaxel, corresponding to different doses (10 mg / kg, 20 mg / kg, 50 mg / kg, 100 mg / kg, 150 mg / kg, 200 mg / kg), with 6 mice (half male and half female) in each group. The compound was injected through the tail vein. Abnormal behavior and death were recorded within two weeks after administration. The maximum tolerated dose (MTD value) was used as an indicator for acute toxicity judgment, that is, the maximum dose that keeps the mice in the entire dose group alive after a single dose.

[0118] The MTD value of the positive drug paclitaxel group is 20 mg / kg. It can be observed that the weight of mice at 20 mg / kg decreased significantly two weeks after administration; while the paclitaxel prodrug group did not die even at the highest dose of 200 mg / kg, and the body weight of the low-dose group did not change significantly. This result shows that the MTD value of paclitaxel prodrug in ICR mice is at least 10 times higher than that of the positive drug paclitaxel, and its in vivo safety is greatly improved.

[0119] Example 17 In vivo imaging experiment of compound 2 in mice

[0120] The purpose of this example is to explore the fluorescence imaging ability of the compound in tumor-bearing mice. After the mice with 4T1 tumor cells were subcutaneously implanted with tumors, compound 2 was administered intravenously and orally, and then scanned with the Tanon ABL imaging system at different time points (0, 1, 2, 3, 4, 6, 8, 12, 24, 36, 48, 72h). After 12h, 24h, 36h, 48h and 72h of administration, significant near-infrared fluorescence signals were observed at the tumor site, such as Figure 2-6 The quantitative values ​​of the fluorescence signals of in vivo imaging in mice are shown in Table 2. It is worth noting that at 12h after injection, the near-infrared fluorescence signals in the tumors of the tail vein injection group have been clearly identified. In contrast, certain fluorescence signals can be identified in the tumor-bearing mice of the oral administration group at 24h. This indicates that compound 2 will have an accumulation effect in the tumors of mice and has a certain oral activity.

[0121] Table 2 Quantitative response values ​​of compound 2 in vivo imaging in mice

[0122] As described above, although the present invention has been shown and described with reference to specific preferred embodiments, it should not be construed as limiting the present invention itself. Various changes in form and details may be made without departing from the spirit and scope of the present invention as defined in the appended claims.

Claims

1. A compound as shown in general formula I or general formula II or a pharmaceutically acceptable salt thereof, characterized in that: Wherein, R1 is selected from hydrogen, C1-C6 alkyl; Wherein, R2 is selected from C1-C6 alkyl.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The R1 is selected from hydrogen and C1-C6 n-alkyl.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The R2 is selected from C1-C6 n-alkyl.

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The R1 is selected from hydrogen, methyl, ethyl, n-propyl, n-butyl, n-pentyl and n-hexyl.

5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The R2 is selected from methyl, ethyl, n-propyl, n-butyl, n-pentyl and n-hexyl.

6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: Select any of the following compounds: 。 7. A method for preparing the compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: Wherein, R1 is selected from C1-C6 alkyl.

8. A method for preparing the compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: Wherein, R2 is selected from C1-C6 alkyl.

9. A pharmaceutical composition comprising the compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

10. Use of the compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing and / or treating tumor diseases.

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