A paclitaxel prodrug, its preparation method and application

Abstract

The present invention discloses a paclitaxel prodrug, a preparation method thereof, and an application thereof. The compound of the present invention, its pharmaceutically acceptable salts and esters can block the toxic sites through ROS-responsive linkers, highly respond to the tumor microenvironment, play the role of selective killing and release, have low toxicity to normal cells, while the photosensitive fragment can be used to promote the release of the prodrug, amplify the chemotherapy efficacy, and can also be used as an autoradiographic fragment to play the function of in vivo imaging, and can be used for the precise treatment of breast cancer. Therefore, the paclitaxel prodrug of the present invention can be used to prepare 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 particularly to a paclitaxel prodrug, a preparation method thereof, and an application thereof. Background Art

[0002] Paclitaxel (PTX) is a structurally complex antitumor tricyclic diterpenoid compound initially isolated from the stems and leaves of Taxus. PTX is a white crystalline powder, odorless and tasteless, poorly soluble in water but easily soluble in organic solvents, with the molecular formula C 47 H 51 NO 14 , and the molecular weight is 853.92. Since its discovery, various studies and developments on it have followed one after another. With the in-depth progress of the research on it, the mechanism of action of paclitaxel has been elaborated in detail: PTX can bind to and stabilize β-tubulin in microtubules, resulting in 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 apoptosis in various different ways, involving the activation and regulation of multiple molecular signaling pathways, including the imbalance of Bcl-2 family proteins, the activation of the Caspase cascade reaction, the decrease of mitochondrial membrane potential, and the breakage of DNA, etc.

[0003]

[0004] Paclitaxel is one of the best-selling natural antitumor drugs. Due to its broad-spectrum antitumor activity, paclitaxel has been widely used clinically in the treatment of breast cancer, ovarian cancer, and some head and neck squamous cell carcinomas and lung cancers. However, due to the poor water solubility of paclitaxel, high cardiotoxicity and neurotoxicity, and easy occurrence of myelosuppression and drug resistance, its clinical application has been somewhat limited. Therefore, the research on increasing its effectiveness and druggability has always been a hot spot and a difficult point in the development of paclitaxel.

[0005] Prodrugs generally refer to substances that, through different modification means, convert a compound with drug activity, i.e., the parent drug, into a substance that is inactive in vitro but can release active drug molecules again through specific conversion pathways in vivo.

[0006] During the occurrence and development of tumors, the change of 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 by-products generated during the cell metabolism process due to the change of the redox state. Hydrogen peroxide, singlet oxygen, superoxide anion, and hydroxyl radicals all belong to ROS. In normal cells, it can be rapidly cleared, while due to the higher redox level of tumor cells, the ROS level in tumor cells is much higher than that in normal cells. Many studies have shown that compared with normal cells (<1 mM), tumor cells produce ROS levels (>[10 mM]) that are dozens or even thousands of times higher in concentration.

[0007] Photodynamic therapy is a new method for treating tumors by using photosensitizers and lasers to activate photosensitizers. Irradiating the lesion site with a specific wavelength can activate the photosensitizing drug selectively aggregated in the lesion tissue, triggering a photochemical reaction to destroy the lesion. In photodynamic therapy (PDT), the photosensitizing drug transfers energy to the surrounding oxygen, generating highly reactive singlet oxygen. Singlet oxygen can undergo an oxidation reaction with nearby biological macromolecules, producing cytotoxicity and killing diseased cells. Compared with traditional therapies, the advantage of PDT lies in its ability to perform precise and effective treatment, 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 and chemotherapy. Summary of the Invention

[0009] Object of the Invention: The object of the present invention is to provide a paclitaxel prodrug that combines photodynamic therapy and chemotherapy; another object of the present invention is to provide a preparation method of the paclitaxel prodrug; another object of the present invention is to provide an application of the paclitaxel prodrug.

[0010] Technical Solution: A compound represented by General Formula I or General Formula II according to the present invention, or a pharmaceutically acceptable salt thereof, wherein, R1 is selected from hydrogen, C1-C6 alkyl;

[0011] wherein, R2 is selected from C1-C6 alkyl.

[0012] Further, R1 is selected from hydrogen, C1-C6 n-alkyl.

[0013] Further, R2 is selected from C1-C6 n-alkyl.

[0014] Further, R1 is selected from hydrogen, methyl, ethyl, n-propyl, n-butyl, n-pentyl and n-hexyl.

[0015] Further, R2 is selected from methyl, ethyl, n-propyl, n-butyl, n-pentyl and n-hexyl.

[0016] The embodiments of the present invention provide the following preferred compounds: .

[0017] In some preferred embodiments, the pharmaceutically acceptable salts include, but are not limited to, acid addition salts formed by the compounds represented by General Formula I or General 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 compounds represented by General Formula I or General Formula II and inorganic bases.

[0018] The compounds represented by General Formula I or General Formula II involved in the present invention may also exist in the form of their salts and esters, which are converted into the compounds represented by General Formula I or General Formula II in vivo. For example, within the scope of the present invention, according to the processes known in the art, the compounds of the present invention are converted into the form of pharmaceutically acceptable salts and used in the form of salts.

[0019] On the other hand, the present invention provides a method for preparing the above-mentioned compound or its pharmaceutically acceptable salt,

[0020] wherein, R1 is selected from C1-C6 alkyl.

[0021] On the other hand, the present invention provides a method for preparing the above-mentioned compound or its pharmaceutically acceptable salt,

[0022] wherein, R2 is selected from C1-C6 alkyl.

[0023] On the other hand, the present invention provides a pharmaceutical composition, which comprises the above-mentioned compound or its pharmaceutically acceptable salt, and one or more pharmaceutically acceptable carriers.

[0024] 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. The oral composition can be any orally acceptable dosage form, including but not limited to tablets, capsules, emulsions, and suspensions, dispersions and solutions. Commonly used pharmaceutically acceptable carriers or excipients include stabilizers, diluents, surfactants, lubricants, antioxidants, binders, colorants, fillers, emulsifiers, etc.

[0025] The sterile injectable composition can be formulated according to the techniques known in the art using suitable dispersing or wetting agents and suspending agents. Pharmaceutically acceptable carriers and solvents that can be used include water, mannitol, sodium chloride solution, etc.

[0026] 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 in achieving the desired therapeutic response for a particular patient, composition, and mode of administration and that is non-toxic to the patient. The selected dosage level depends on a variety of factors, including the activity of the specific compound or its salt of the present invention employed, the route of administration, the time of administration, the rate of excretion of the specific composition employed, the duration of the treatment, other drugs, compounds, and / or materials used in combination with the specific composition employed, the age, sex, weight, general health, and medical history of the patient being treated, and like factors well known in the medical arts.

[0027] On the other hand, the present invention provides the use of the above-mentioned compound or its pharmaceutically acceptable salt in the preparation of a drug for preventing and / or treating tumor diseases.

[0028] Preferably, the tumor disease is breast cancer.

[0029] Beneficial effects: Compared with the prior art, the present invention has the following remarkable advantages: The toxicity site can be blocked by a ROS-responsive linker, highly responsive to the tumor microenvironment, achieving the effect of selective killing and release, having low toxicity to normal cells, while the photosensitive fragment can be used to promote the release of the prodrug, amplify the chemotherapy efficacy, and can also be used as an autoradiographic fragment to play the function of in vivo imaging, and can be used for the precise treatment of breast cancer. Description of the Drawings

[0030] Figure 1 It shows the release of Compound 2 in vitro simulating different ROS levels over time;

[0031] Figure 2 It is the in vivo imaging diagram of Compound 2 in mice at 12 h, with the left side being the oral group and the right side being the tail vein group;

[0032] Figure 3 It is the in vivo imaging diagram of Compound 2 in mice at 24 h, with the left side being the oral group and the right side being the tail vein group;

[0033] Figure 4 It is the in vivo imaging diagram of Compound 2 in mice at 36 h, with the left side being the oral group and the right side being the tail vein group;

[0034] Figure 5 It is the in vivo imaging diagram of Compound 2 in mice at 48 h, with the left side being the oral group and the right side being the tail vein group;

[0035] Figure 6 It is the in vivo imaging diagram of Compound 2 in mice at 72 h, with the left side being the oral group and the right side being the tail vein group. Detailed Description of the Invention

[0036] The preparation methods of the compounds of general formula I or general formula II of the present invention will be described below in combination with specific embodiments, 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 synthesis methods described in this specification or known in the art, and such combinations can be easily carried out by those skilled in the art to which the present invention pertains.

[0037] 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 a salt by using the common salt-forming methods in the art. For example, at room temperature, the compound is dissolved in hydrochloric acid ethanol for reaction to form a hydrochloride salt; or benzenesulfonic acid is added thereto for reaction to form a benzenesulfonate salt.

[0038] The design concept of the compounds of general formula I and general formula II of the present invention is as follows: Paclitaxel is coupled with the photosensitizer 5-aminolevulinic acid and its derivatives through different redox-responsive linkers to generate a prodrug combining photodynamic therapy and 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 linker, and the other end of the redox-responsive linker is connected to the amino group of 5-aminolevulinic acid and its derivatives. Among them, the redox-responsive linker is a disulfide bond or a thioether bond. The photosensitizer 5-aminolevulinic acid and its derivatives include 5-aminolevulinic acid, methyl 5-aminolevulinate, ethyl 5-aminolevulinate, propyl 5-aminolevulinate, butyl 5-aminolevulinate, pentyl 5-aminolevulinate, and hexyl 5-aminolevulinate.

[0039] The thioether-type prodrugs of the present invention are as shown in general formula I:

[0040]

[0041] Among them, R1 is selected from hydrogen, and C1-C6 is a normal alkyl group.

[0042] The disulfide bond-type prodrugs of the present invention are as shown in general formula II:

[0043]

[0044] Among them, R2 is selected from C1-C6 normal alkyl groups.

[0045] The synthetic route of the compound shown in general formula I of the present invention is as follows:

[0046]

[0047] Thionyl chloride is dropped into a solution of alcohol (R1-OH) in an ice bath, and then 5-aminolevulinic acid hydrochloride (5-ALA HCl) is added for reaction. After the reaction is completed, a 5-aminolevulinate derivative (5-ALA-R1) is obtained;

[0048] Paclitaxel (PTX), thiohydroxyacetic anhydride, triethylamine (TEA), 4-dimethylaminopyridine (DMAP) are added to dichloromethane (DCM) for reaction. After the reaction is completed, the intermediate PTX-S-COOH is obtained;

[0049] The intermediate PTX-S-COOH, 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), DIPEA, and 5-aminolevulinic acid ester derivative (5-ALA-R1) are added to dichloromethane; stirring reaction is carried out, and the target product is obtained after the reaction is completed;

[0050] Among them, R1 is selected from C1-C6 normal alkyl groups.

[0051] When R1 is hydrogen, the synthetic route of the compound shown in general formula I is as follows:

[0052] 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;

[0053] The above-mentioned active ester intermediate, 5-aminolevulinic acid hydrochloride, and N,N-diisopropylethylamine (DIPEA) are added to a dichloromethane solution for reaction, and the target product is obtained after the reaction is completed.

[0054] The synthetic route of the compound shown in general formula II of the present invention is as follows:

[0055] 2,2'-Dithiobis(acetic acid) is added with acetyl chloride, and the reaction is carried out under nitrogen protection. After the reaction is completed, the target product 1,4,5-oxodithiophene-2,7-dione (DTDPA) is obtained;

[0056] Paclitaxel (PTX), DTDPA, triethylamine, and DMAP are added to DCM, and the reaction is carried out overnight. After the reaction is completed, the intermediate PTX-S-S-COOH is obtained;

[0057] The intermediate PTX-S-S-COOH, 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), DIPEA, and 5-aminolevulinic acid ester derivative (5-ALA-R2) are added to dichloromethane, and stirring reaction is carried out. After the reaction is completed, the target product is obtained; among them, R2 is selected from C1-C6 normal alkyl groups.

[0058] Synthesis of Intermediate

[0059] The synthesis of the intermediate involved in the embodiments of the present invention is as follows:

[0060] 1. Synthesis of 2 -(((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-methanocyclodeca[3,4]benzo[1,2-b]oxocin-9-yl)oxy)-3-oxo-1-phenylpropan-2-oxoethoxy)-2-oxoethyl)thioacetic acid (PTX-S-COOH)

[0061]

[0062] Weigh 50 mg of paclitaxel, 15 mg of thiohydroxyacetic anhydride, 10 μL of triethylamine, and 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 and wash successively with water and saturated sodium chloride). Dry the organic phase with anhydrous sodium sulfate, evaporate to dryness under reduced pressure, and purify by silica gel column chromatography to obtain the intermediate PTX-S-COOH, 51 mg, yellow solid. 88% yield.

[0063] 1 H NMR (400 MHz, Chloroform- d ) δ 8.03 – 7.97 (m, 2H), 7.85 – 7.79 (m,2H), 7.63 (dd, J 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 J = 8.2, 6.9, 2.1 Hz, 1H), 6.64 (p, J J = 1.0 Hz,1H), 6.16 (dddt, J J = 7.3, 6.2, 2.6, 1.5 Hz, 1H), 5.69 (d, J 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).

[0064] Calculated value of electrospray mass spectrometry (ESI-MS): C 51 H 55 NO 17 S (M-H) - m / z: 984.3, measured value 984.3.

[0065] 2. Synthesis of ethyl 5-aminolevulinate

[0066]

[0067] At 0 °C, 600 μL of thionyl chloride was added dropwise to 8 mL of ethanol in an ice bath. Stir for 10 min. Remove the ice and add 1 g of 5-aminolevulinic acid hydrochloride. React at room temperature for 1 h. Heat to 78 °C and reflux for 2 h. Cool, add ether, and a solid precipitates. Grind and slurry. Filter and wash with ether and acetonitrile. 0.85 g of the target product, ethyl 5-aminolevulinate, was obtained. White solid. 89% yield. 1 H NMR (400 MHz, Chloroform- d ) δ 4.13 (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 value: C7H 13 NO3(M+H) + m / z: 160.2, found 160.2.

[0068] 3. Synthesis of propyl 5-aminolevulinate

[0069]

[0070] At 0 °C, 600 μL of thionyl chloride was added dropwise to 8 mL of n-propanol in an ice bath. Stir for 10 min. Remove the ice and add 1 g of 5-aminolevulinic acid hydrochloride. React at room temperature for 1 h. Heat to 98 °C and reflux for 2 h. Cool, add ether, and a solid precipitates. Grind and slurry. Filter and wash with ether and acetonitrile. 0.97 g of the intermediate propyl 5-aminolevulinate was obtained. White solid. Yield 93.85%. 1 1H NMR (400 MHz, Chloroform- d ) δ 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 value: C8H 15 NO3(M+H) + m / z: 174.2, found 174.2.

[0071] 4. Synthesis of butyl 5-aminolevulinate

[0072]

[0073] At 0 °C, 600 μL of thionyl chloride was added dropwise to 8 mL of n-butanol in an ice bath. Stir for 10 min. Remove the ice and add 1 g of 5-aminolevulinic acid hydrochloride. React at room temperature for 1 hour. Heat to 118 °C and reflux for 2 hours. Cool, add ether, and a solid precipitates. Grind it into a slurry. Filter and wash with ether and acetonitrile. 1.01 g of the intermediate 5-aminolevulinic acid butyl ester was obtained. White solid. 90% yield. 1 H NMR (400 MHz, Chloroform- d ) δ 4.07 (t, J J = 6.5 Hz, 2H), 3.73 (t, J J=6.5 Hz, 2H), 2.90 (q, J J = 6.6 Hz, 1H), 2.80 (q, J J = 6.6 Hz, 1H), 2.75 – 2.68 (m,2H), 2.63 – 2.56 (m, 2H), 1.61 (p, J J = 6.8 Hz, 2H), 1.39 (dt, J J = 14.0, 7.0 Hz,2H), 0.97 (t, J J = 7.0 Hz, 3H). ESI-MS calculated value: C9H 17 NO3(M+H) + m / z: 188.2, found 188.2.

[0074] 5. Synthesis of 5-aminolevulinic acid pentyl ester

[0075]

[0076] At 0 °C, 600 μL of thionyl chloride was added dropwise to 8 mL of n-pentanol in an ice bath. Stir for 10 min. Remove the ice and add 1 g of 5-aminolevulinic acid hydrochloride. React at room temperature for 1 hour. Heat to 139 °C and reflux for 2 hours. Cool, add ether, and a solid precipitates. Grind it into a slurry. Filter and wash with ether and acetonitrile. 1.13 g of the intermediate 5-aminolevulinic acid pentyl ester was obtained. White solid. 94% yield. 1 H NMR (400 MHz, Chloroform- d ) δ 4.06 (t, J J = 5.9 Hz, 2H), 3.73 (t, J J=6.5 Hz, 2H), 2.90 (q, J 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 value: C 10 H 19 NO3(M + H) + m / z: 202.3, found 202.3.

[0077] 6. Synthesis of hexyl 5 - aminolevulinate

[0078]

[0079] At 0 °C, 600 μL of thionyl chloride was added dropwise to 8 mL of n - hexanol in an ice bath. Stir for 10 min. Remove the ice and add 1 g of 5 - aminolevulinic acid hydrochloride. React at room temperature for 1 h. Heat to 159 °C and reflux for 2 h. Cool, add ether, and a solid precipitates. Grind and slurry. Filter and wash with ether and acetonitrile. 1.19 g of the intermediate hexyl 5 - aminolevulinate was obtained. White solid. Yield 93%. 1 H NMR (400 MHz, Chloroform - d ) δ 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 value: C 11 H21NO3(M + H) + m / z: 216.3, found 216.3.

[0080] 7. Synthesis of 1,4,5 - oxodithiophene - 2,7 - dione (DTDPA)

[0081]

[0082] 0.5 g of 2,2'-dithiobis(acetic acid) was added with 2 mL of acetyl chloride. Under nitrogen protection, the reaction was carried out at room temperature for 2 hours. Toluene (20 mL) was added three times, and the mixture was rotary evaporated to remove the solvent. Ether was added for pulping, and the solid precipitated. The target product 1,4,5-oxadithiophene-2,7-dione (DTDPA), 270 mg, was obtained by suction filtration. It was a white solid with a yield of 47%.

[0083] 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-methano-cyclodeca[3,4]benzo[1,2-b]oxocin-9-yl)oxy)-3-oxo-1-phenylpropan-2-yl)oxy)-disulfanyl)acetic acid ester (PTX-S-S-COOH)

[0084]

[0085] 50 mg of paclitaxel, 15 mg of DTDPA, 10 μL of triethylamine, and 1.5 mg of DMAP were added to 2 mL of DCM. The reaction was carried out 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 and washed successively with water and saturated sodium chloride). The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to obtain the intermediate PTX-S-S-COOH, 40 mg. It was a yellow solid with a yield of 67%.

[0086] 1 H NMR (400 MHz, Chloroform- d ) δ 8.03 – 7.97 (m, 2H), 7.85 – 7.79 (m,2H), 7.63 (dd, J 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 J = 8.2, 6.9, 2.1 Hz, 1H), 6.64(p, J 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).

[0087] ESI-MS calculated value: C 51 H 55 NO 17 S2(M-H) - m / z:1016.3, measured value 1016.3.

[0088] Example 1 Synthesis of Compound 1

[0089] (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-methano-cyclodeca[3,4]benzo[1,2-b]oxocin-9-yl)oxycarbonyl)-1,6,10,13-tetraoxo-1,3-diphenyl-5-oxa-8-thia-2,11-diazahexadec-16-oic acid

[0090]

[0091] 60 mg of intermediate PTX-S-COOH, 8 mg of N-hydroxysuccinimide (NHS), and 14 mg of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI) were added to 2 mL of dichloromethane in an ice bath. After reacting for half an hour, the reaction was allowed to resume at room temperature for 24 h. After the reaction was completed, the organic solvent was removed under reduced pressure, and the residue was extracted (dissolved in ethyl acetate and washed successively with water and saturated sodium chloride). The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure to obtain the active ester intermediate.

[0092] Then, 12 mg of 5-aminolevulinic acid hydrochloride and 26 μL of N,N-diisopropylethylamine (DIPEA) were added to the active ester in 2 mL of dichloromethane solution, and the reaction was carried out 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 and washed successively with water and saturated sodium chloride). The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure, and the target product 1, 21 mg, was obtained by silica gel column chromatography purification. White solid. 31% yield.

[0093] 1 H NMR (400 MHz, Chloroform- d ) δ 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).

[0094] 13 C NMR (101 MHz, Chloroform- d ) δ 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.

[0095] High resolution mass spectrometry HRMS (ESI) calculated value: C 56 H 62 N2O 19 S (M+Na) + m / z: 1121.3559, found 1121.3558.

[0096] Example 2 Synthesis of Compound 2

[0097] (2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS)-9-((R)-15-(((S)-benzamido(phenyl)methyl)-3,6,9,13-tetraoxo-2,14-dioxa-11-thia-8-azapentadec-16-yl)oxy)-12-(benzoyloxy)-4,11-dihydroxy-4a,8,13,13-tetramethyl-5-oxo-3,4a,5,6,9,10,12,12a-dehydro-1H-7,11-methanocyclodeca[3,4]benzo[1,2-b]oxepine-6,12b(2aH)-diacetic acid diethyl ester

[0098] 51 mg of intermediate PTX-S-COOH, 24 mg of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), 20 μL of DIPEA, and 15 mg of methyl 5-aminolevulinate were added to 2 mL of dichloromethane. The mixture was stirred 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 and washed successively with water and saturated sodium chloride). The organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to obtain 30 mg of the target product 2 as a yellow solid. The yield was 53% and the purity was 97%. 1 H NMR (400 MHz, Chloroform- d ) δ 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).

[0099] 13 C NMR (101 MHz, Chloroform- d ) δ 204.04, 203.87, 172.61, 171.40,170.06, 169.13, 168.71, 168.18, 167.86, 167.15, 143.00, 137.10, 133.77,132.86, 131.83, 130.43, 129.41, 129.04, 128.88, 128.45, 128.40, 127.88,127.00, 84.60, 81.21, 79.28, 76.60, 75.87, 75.76, 75.27, 72.27, 72.18, 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.

[0100] HRMS (ESI) Calcd for: C 57 H 64 N2O 19 S (M+H) + m / z: 1113.3897, found 1113.3906.

[0101] Example 3 Synthesis of Compound 3

[0102] (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-thia-9-azapentadecanoyl)oxy)-12-(benzoyloxy)-4,11-dihydroxy-4a,8,13,13,5-pentamethyl-5-oxo-3,4,4a,5,6,9,11,12,12a-dehydro-1H-7,11-methanoazacyclododecano[3,4]benzo[1,2-b]oxepin-6,12b(2aH)-diyl diacetate

[0103] 51 mg of intermediate PTX-S-COOH, 24 mg of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), 20 μL of DIPEA, and 16 mg of ethyl 5-aminolevulinate were added to 2 mL of dichloromethane. The mixture was stirred 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 and washed successively with water and saturated sodium chloride). The organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to obtain 29 mg of the target compound 3 as a yellow solid, with a yield of 50% and a purity of 98%.

[0104] 1 H NMR (400 MHz, Chloroform- d ) δ 8.27 (d, J 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 J = 8.2, 6.8 Hz,2H), 7.45 – 7.31 (m, 8H), 7.08 (t, J J = 5.1 Hz, 1H), 6.30 (s, 1H), 6.13 (dd, J J =9.5, 3.0 Hz, 1H), 5.70 (d, J J = 7.1 Hz, 1H), 5.42 (d, J J = 3.0 Hz, 1H), 4.99 (dd, J J =9.7, 2.3 Hz, 1H), 4.46 (dd, J 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).

[0105] 13 C NMR (101 MHz, Chloroform- d ) δ 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.

[0106] HRMS (ESI) Calcd: C 58 H 66 N2O19 S (M+H) + m / z: 1127.4053, measured value 1127.4050

[0107] Example 4 Synthesis of Compound 4

[0108] (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-thia-9-azaoctadecanoyl)oxy)-12-(benzoyloxy)-4,11-dihydroxy-4a,8,13,13,5-tetramethyl-5-oxo-3,4,4a,5,6,9,11,12,12a-dehydro-1H-7,11-methoxycyclodeca[3,4]benzo[1,2-b]oxa-6,12b(2aH)-diacetic acid diethyl ester

[0109] 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. Stirred 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 and washed successively with water and saturated sodium chloride). The organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to obtain 30 mg of the target compound 4, as a yellow solid, with a yield of 50% and a purity of 95%.

[0110] 1 H NMR (400 MHz, Chloroform- d ) δ 8.28 (d, J 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 J = 8.3, 6.8 Hz,2H), 7.45 – 7.31 (m, 8H), 7.08 (t, J J = 5.2 Hz, 1H), 6.30 (s, 1H), 6.13 (dd, J J =9.5, 3.0 Hz, 1H), 5.70 (d, J 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).

[0111] 13 C NMR (101 MHz, Chloroform- d ) δ204.04, 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.

[0112] HRMS (ESI) Calculated for: C 59 H 68 N2O 19 S (M+H) + m / z: 1141.4210, Found 1141.4211.

[0113] Example 5 Synthesis of Compound 5

[0114] (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-thia-9-azanonadecanoyl)oxy)-12-(benzoyloxy)-4,11-dihydroxy-4a,8,13,13,5-pentamethyl-5-oxo-3,4,4a,5,6,10,11,12,12a-decahydro-1H-7,11-methano-cyclodeca[3,4]benzo[1,2-b]oxepin-6,12b(2aH)-diyl diacetate

[0115] 51 mg of intermediate PTX-S-COOH, 24 mg of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), 20 μL of DIPEA, and 19 mg of butyl 5-aminolevulinate were added to 2 mL of dichloromethane. The mixture was stirred 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 and washed successively with water and saturated sodium chloride). The organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to obtain 32 mg of the target compound 5 as a yellow solid, with a yield of 54% and a purity of 96%.

[0116] 1 H NMR (400 MHz, Chloroform- d ) δ 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).

[0117] 13 C NMR (101 MHz, Chloroform- d ) δ 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.

[0118] HRMS (ESI) Calcd: C 60 H 70 N2O 19 S (M+H) + m / z:1155.4366, Found 1155.4368.

[0119] Example 6 Synthesis of Compound 6

[0120] (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-thia-9-azaeicosyl)oxy)-12-(benzoyloxy)-4,11-dihydroxy-4a,8,13,13,5-tetramethyl-5-oxo-3,4,4a,5,6,10,11,12,12a-dehydro-1H-7,11-methano-cyclodeca[3,4]benzo[1,2-b]oxa-6,12b(2aH)-diacetic acid diethyl ester

[0121] 51 mg of intermediate PTX-S-COOH, 24 mg of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), 20 μL of DIPEA, and 21 mg of pentyl 5-aminolevulinate were added to 2 mL of dichloromethane. The mixture was stirred 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 and washed successively with water and saturated sodium chloride). The organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to obtain 34 mg of the target compound 6 as a yellow solid, with a yield of 56% and a purity of 96%.

[0122] 1 H NMR (400 MHz, Chloroform- d ) δ 8.27 (d, J J = 9.4 Hz, 1H), 8.20 – 8.16(m, 2H), 7.89 – 7.84 (m, 2H), 7.60 (t, J J = 7.3 Hz, 1H), 7.51 (dd, J J = 8.4, 6.8Hz, 2H), 7.45 – 7.31 (m, 8H), 7.09 (t, J J = 5.2 Hz, 1H), 6.30 (s, 1H), 6.12 (dd, J J = 9.5, 3.1 Hz, 1H), 5.70 (d, J J = 7.1 Hz, 1H), 5.41 (d, J J = 3.1 Hz, 1H), 4.99(dd, J J = 9.7, 2.3 Hz, 1H), 4.46 (dd, J 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).

[0123] 13 C NMR (101 MHz, Chloroform - d ) δ204.04, 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.

[0124] HRMS (ESI) Calculated for: C 61 H 72 N2O 19 S (M+H) + m / z: 1169.4523, found 1169.4519.

[0125] Example 7 Synthesis of Compound 7

[0126] (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-thia-9-azadocosanoyl)oxy)-12-(benzoyloxy)-4,11-dihydroxy-4a,8,13,13,5-tetramethyl-5-oxo-3,4,4a,5,6,10,11,12,12a-decahydro-1H-7,11-methano-cyclodeca[3,4]benzo[1,2-b]oxocin-6,12b(2aH)-diyl diacetate

[0127] 51 mg of intermediate PTX-S-COOH, 24 mg of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), 20 μL of DIPEA, and 23 mg of hexyl 5-aminolevulinate were added to 2 mL of dichloromethane. The mixture was stirred 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 and washed successively with water and saturated sodium chloride). The organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to obtain 31 mg of the target compound 7 as a yellow solid, with a yield of 51% and a purity of 97%.

[0128] 1 H NMR (400 MHz, Chloroform- d ) δ 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).

[0129] 13 C NMR (101 MHz, Chloroform - d ) δ 204.05, 203.87, 172.63, 171.42, 170.05, 169.12, 168.70, 168.17, 167.83, 167.16, 143.03, 137.10, 133.76, 132.83, 131.83, 130.43, 129.38, 129.03, 128.88, 128.44, 128.39, 127.89, 127.00, 84.60, 81.19, 79.30, 76.60, 75.90, 75.76, 75.26, 72.28, 72.17, 65.35, 58.64, 52.66, 49.34, 45.69, 43.32, 35.74, 35.66, 34.82, 34.71, 32.06, 32.02, 31.64, 31.57, 31.52, 30.44, 30.32, 30.26, 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.

[0130] HRMS (ESI) Calcd for C 61 H 72 N2O 19 S (M + H) + m / z: 1183.4679, Found 1183.4677.

[0131] Example 8 Synthesis of Compound 8

[0132] (2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS)-9-(((R)-16-((S)-benzamido(phenyl)methyl)-3,6,9,14-tetraoxo-2,15-dioxa-11,12-dithia-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-methanocyclodeca[3,4]benzo[1,2-b]oxa-6,12b(2aH)-diacetic acid diethyl ester

[0133] 51 mg of intermediate PTX-S-S-COOH, 24 mg of 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), 20 μL of DIPEA, and 15 mg of methyl 5-aminolevulinate were added to 2 mL of dichloromethane. The mixture was stirred 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 and washed successively with water and saturated sodium chloride). The organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to obtain 24 mg of the target compound 8 as a yellow solid, with a yield of 43% and a purity of 96%.

[0134] 1 H NMR (400 MHz, Chloroform- d ) δ 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).

[0135] 13 C NMR (101 MHz, Chloroform- d ) δ 203.86, 203.51, 173.11, 171.18, 170.11, 169.12, 168.89, 168.40, 167.61, 167.05, 142.60, 136.93, 134.24, 133.86, 132.98, 131.98, 130.35, 129.42, 129.15, 128.79, 128.66, 127.60, 127.25, 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.

[0136] HRMS (ESI) Calcd for C 57 H 64 N2O 19 S2(M+H) + m / z: 1145.3617, found 1145.3615.

[0137] Example 9 Synthesis of Compound 9

[0138] (2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS)-9-(((R)-2-((S)-benzamidophenyl)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-methano-cyclodeca[3,4]benzo[1,2-b]oxa-6,12b(2aH)-diyl diacetate

[0139] 51 mg of intermediate PTX-S-S-COOH, 24 mg of 2-(7-azabenzotriazol)-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. The mixture was stirred 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 and washed successively with water and saturated sodium chloride). The organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to obtain 24 mg of the target compound 9 as a yellow solid, with a yield of 42% and a purity of 97%.

[0140] 1 H NMR (400 MHz, Chloroform- d ) δ 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).

[0141] 13 C NMR (101 MHz, Chloroform- d ) δ 203.88, 203.56, 172.67, 171.19, 170.10, 169.10, 168.83, 168.38, 167.63, 167.08, 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.

[0142] HRMS (ESI) Calcd for C 58 H 66 N2O 19 S2(M+H) + m / z: 1159.3774, Found 1159.3773.

[0143] Example 10 Synthesis of Compound 10

[0144] (2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS)-9-(((R)-2-((S)-benzamidophenyl)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-methano-cyclodeca[3,4]benzo[1,2b]oxa-6,12b(2aH)-diacetic acid diethyl ester

[0145]

[0146] 51 mg of intermediate PTX-S-S-COOH, 24 mg of 2-(7-azabenzotriazol)-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. Stirred 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 and washed successively with water and saturated sodium chloride). 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, yield 47%. Purity 95%.

[0147] 1 H NMR (300 MHz, Chloroform- d ) δ 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).

[0148] 13 C NMR (75 MHz, Chloroform - d ) δ 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.

[0149] HRMS (ESI) Calcd: C 59 H 68 N2O 19 S2(M + H) + m / z: 1173.3931, Found 1173.3934.

[0150] Example 11 Synthesis of Compound 11

[0151] (2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS)-9-(((R)-2-((S)-benzamidophenyl)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

[0152]

[0153] 51 mg of intermediate PTX-S-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 butyl ester were added to 2 mL of dichloromethane. The mixture was stirred 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 and washed successively with water and saturated sodium chloride). The organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to obtain 26 mg of the target compound 11 as a yellow solid, with a yield of 45% and a purity of 96%.

[0154] 1 H NMR (300 MHz, Chloroform- d ) δ 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 = 6.7 Hz, 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).

[0155] 13 C NMR (75 MHz, Chloroform- d ) δ 203.88, 203.58, 172.77, 171.18, 170.09, 169.11, 168.85, 168.40, 167.66, 167.06, 142.63, 136.93, 134.26, 133.57, 132.97, 132.07, 130.36, 129.40, 129.14, 128.79, 128.66, 127.61, 127.24, 84.33, 81.17, 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.

[0156] HRMS (ESI) Calcd for C 60 H 70 N2O 19 S2 (M+H) + m / z: 1187.4087, found 1187.4087.

[0157] Example 12 Synthesis of Compound 12

[0158] (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-azadocosanoyl)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-methano-cyclodeca[3,4]benzo[1,2b]oxa-6,12b(2aH)-diacetic acid diethyl ester

[0159]

[0160] 51 mg of intermediate PTX-S-S-COOH, 24 mg of 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), 20 μL of DIPEA, and 15 mg of pentyl 5-aminolevulinate were added to 2 mL of dichloromethane. The mixture was stirred 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 and washed successively with water and saturated sodium chloride). The organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to obtain 28 mg of the target compound 12 as a yellow solid, with a yield of 48% and a purity of 96%.

[0161] 1 H NMR (300 MHz, Chloroform- d ) δ 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).

[0162] 13 C NMR (75 MHz, Chloroform- d ) δ 203.94, 203.82, 171.19, 169.89, 169.28, 168.60, 168.50, 168.33, 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.

[0163] HRMS (ESI) Calcd for C 61 H 72 N2O 19 S2(M+Na) + m / z: 1223.4063, Found 1223.4056.

[0164] Example 13 Synthesis of Compound 13

[0165] (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-azadocosanoyl)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-methano-cyclodeca[3,4]benzo[1,2-b]oxa-6,12b(2aH)-diacetic acid diethyl ester

[0166]

[0167] 51 mg of intermediate PTX-S-S-COOH, 24 mg of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), 20 μL of DIPEA, and 15 mg of hexyl 5-aminolevulinate were added to 2 mL of dichloromethane. The mixture was stirred 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 and washed successively with water and saturated sodium chloride). The organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to obtain 28 mg of the target compound as a yellow solid, with a yield of 47% and a purity of 98%.

[0168] 1 H NMR (400 MHz, Chloroform- d ) δ 8.13 (dt, J J = 7.0, 1.3 Hz, 2H), 7.88 –7.82 (m, 2H), 7.79 (d, J J = 8.5 Hz, 1H), 7.68 – 7.60 (m, 1H), 7.59 – 7.29 (m,10H), 6.96 (t, J J = 4.5 Hz, 1H), 6.30 (s, 1H), 6.17 (t, J J = 9.0 Hz, 1H), 5.92 (dd, J J = 8.5, 5.6 Hz, 1H), 5.65 (d, J J = 7.1 Hz, 1H), 5.48 (d, J J = 5.6 Hz, 1H), 4.96(dd, J 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).

[0169] 13 C NMR (101 MHz, Chloroform- d ) δ 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.

[0170] HRMS (ESI) Calcd: C 62 H74 N2O 19 S2(M+H) + m / z: 1215.4400, measured value 1215.4404.

[0171] Example 14 Compound Prodrug Release Experiment

[0172] Using dimethyl sulfoxide (DMSO) as the solvent, compound 2 was prepared into a stock solution of 10 millimoles per liter (mM) and stored in a refrigerator at -20 °C. An appropriate amount of the compound stock solution was diluted with phosphate buffer (PBS) to a final concentration of 100 micromoles per liter, and then H2O2 (0, 1, 2, 10 mM) was added, and incubation was carried out at 37 °C, repeated 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). Agilent 1100 HPLC and ultraviolet detector were used under the following conditions: Agilent C18 chromatographic column (4.6×150 mm, 3.5 μm); mobile phase: 85% methanol: 0.1% trifluoroacetic acid; flow rate: 1.0 ml / min.

[0173] As Figure 1 shown, after 24-hour incubation, the prodrug hardly decomposed in the case of 0 mmol / L H2O2, showing good stability. As the concentration of ROS increased, the rate and extent of prodrug release increased significantly, showing a positive correlation. In the case of low concentration (1 mmol / L), the monothiol prodrug released 50.07% ± 0.02, and the disulfide prodrug released 53.06% ± 0.06. In the case of high concentration (10 mmol / L), the monothiol prodrug released 68.83% ± 0.02, and the disulfide prodrug released 69.23% ± 0.02.

[0174] The monothiol prodrug released 18.76% ± 0.04 more and the disulfide prodrug released 16.17% ± 0.03 more at high concentration than at low concentration. These data indicate that the release of such prodrugs is ROS-responsive prodrugs.

[0175] Example 15 Anti-cell Proliferation Assay (MTT assay)

[0176] Take MDA MB-231 cells in the logarithmic growth phase (the cells were purchased from the cell bank of Wuhan Institute of Virology, Chinese Academy of Sciences). In the laminar flow hood, after digestion with 0.25% trypsin, discard the trypsin and terminate the digestion. Use a pipette to aspirate the culture medium to blow down the adherent cells and transfer them into a 10-ml centrifuge tube. Centrifuge at 1000 revolutions per minute (rpm) for five minutes, then discard the supernatant. Add an appropriate amount of culture medium and pipette gently to make a single-cell suspension. Aspirate 20 μl and add it to the cell counting chamber. After counting, dilute with the culture medium to adjust the cell concentration to 3000 cells per well. Inoculate 200 μl per well in a 96-well plate. After inoculation, culture routinely for 12 h. After the cells adhere to the wall, aspirate the culture medium and re-add 200 μl of the culture medium containing 1 μM of the drug. Set 3 parallel wells for each compound concentration, and add an equal amount of blank culture medium to the control group. After 24 h of drug administration, add 20 μl of MTT to each well, continue to incubate for 3 h, then take out and shake on a shaker for 10 min. Using 490 / 570 nm as the detection wavelength, use an enzyme-linked immunosorbent assay (ELISA) reader to detect the absorbance of each well. Calculate the cell viability after the action of the test compound at different concentrations according to the corresponding formula: Cell viability (%) = (1 - A 实验组 / A 对照组 )× 100%; where A 实验组 represents the absorbance of the experimental group, and A 对照组 represents the absorbance of the control group. The experimental results are shown in Table 1. The data show that the prodrug exhibits a similar or even stronger killing effect on cancer cells. The preferred compound 13 exhibits an inhibition rate 9% higher than that of the positive drug.

[0177] Table 1 Anti-proliferative activity of prodrugs against breast cancer cells

[0178]

[0179] The inhibition rate (1 μM) data results are the average of three parallel experiments, and ± represents the standard deviation (SD);

[0180] Example 16 Acute toxicity detection of compounds in mice

[0181] Raise ICR mice about five weeks old normally for nearly one week, and then randomly assign the mice so that there are 6 dose groups for compound 2 and the paclitaxel group, corresponding to different administration doses (10 mg / kg, 20 mg / kg, 50 mg / kg, 100 mg / kg, 150 mg / kg, 200 mg / kg), and there are 6 mice in each group (half male and half female). Inject the compound through the tail vein. Record abnormal behaviors and deaths within two weeks after drug administration. Use the maximum tolerated dose (MTD value) as an indicator for acute toxicity judgment, that is, the maximum administration dose that keeps the mice in the whole dose group alive after a single administration.

[0182] The MTD value of the positive drug paclitaxel group was 20 mg / kg. It was observed that the body weight of mice at 20 mg / kg decreased significantly two weeks after drug administration; even at the highest dose of 200 mg / kg, no death occurred in the paclitaxel prodrug group, and there was no significant change in the body weight of the low-dose group. The results showed that the MTD value of the paclitaxel prodrug in ICR mice was at least 10 times higher than that of the positive drug paclitaxel, and the in vivo safety was greatly improved.

[0183] Example 17 In vivo imaging experiment of Compound 2 in mice

[0184] The purpose of this example was to explore the fluorescence imaging ability of the compound in tumor-bearing mice. After subcutaneous implantation of 4T1 tumor cells in mice to form tumors, Compound 2 was administered by intravenous injection and orally, and then scanned with a Tanon ABL imaging system at different time points (0, 1, 2, 3, 4, 6, 8, 12, 24, 36, 48, 72 h). After 12 h, 24 h, 36 h, 48 h, and 72 h of drug administration, significant near-infrared fluorescence signals were observed at the tumor site, as Figures 2 - 6 shown. The quantitative values of the fluorescence signals in the in vivo imaging of mice are shown in Table 2. It should be noted that 12 h after injection, the near-infrared fluorescence signal in the tumor of the tail vein injection group had been clearly identified. In contrast, a certain fluorescence signal could be identified in the tumor-bearing mice of the oral administration group at 24 h. This indicated that Compound 2 had an accumulation effect in the tumors of mice and had certain oral activity.

[0185] Table 2 Quantitative response values of in vivo imaging of Compound 2 in mice

[0186]

[0187] As described above, although the present invention has been shown and described with reference to specific preferred embodiments, it should not be construed as a limitation of the present invention itself. Various changes in form and detail may be made without departing from the spirit and scope of the present invention defined by 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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