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CN-117304176-B - Enzyme-activated prodrug compound and preparation method and application thereof

CN117304176BCN 117304176 BCN117304176 BCN 117304176BCN-117304176-B

Abstract

The invention discloses an enzyme-activated prodrug compound which has the following structural general formula I: The invention discloses a class of enzyme-sensitive chlorambucil analog prodrugs. The prodrug has high selectivity to hypoxic tumor and can be effectively activated in hypoxic tumor cells. When activated specifically, the prodrug releases the free chemotherapeutic drug chlorambucil and releases the photosensitizer cyanine dye to realize the conversion of the photosensitive property of OFF-ON and generate a large amount of active oxygen species to realize the unification and the synergy of photodynamic therapy and activatable chemotherapy.

Inventors

  • PENG XIAOJUN
  • ZHANG HAN
  • HAN FUPING
  • FAN JIANGLI
  • DU JIANJUN
  • SUN WEN

Assignees

  • 大连理工大学

Dates

Publication Date
20260505
Application Date
20230927

Claims (8)

  1. 1. An enzyme-activated prodrug compound characterized by having the following structural formula I: In the general formula I, the components are shown in the specification, R 1 is-N [ (CH 2 CH 2 ) m X] 2 ) wherein X is selected from halogen chloride and m is 1 An integer of 4; R 2 is nitro; r 3 is-NH; R 4 is O; r 5 is methyl; R 6 is halogen bromide substituted on the 6 membered ring.
  2. 2. The enzyme activated prodrug compound of claim 1 wherein the compound is stimulated release of highly expressed nitroreductase enzyme in anaerobic tumor cells, which compound simultaneously releases free chlorambucil and activated cyanine dye upon nitroreductase hydrolysis in hypoxic tumors.
  3. 3. The enzyme activated prodrug compound of claim 2 wherein the compound releases chlorambucil and activated cyanine dye in a 1:1 relationship.
  4. 4. A process for the preparation of a class of enzyme-activated prodrug compounds according to any of claims 1 to 3, comprising the steps of: (1) 2-hydroxy-5-methyl m-xylylene glycol reacts with a compound with a general formula S-1 according to a molar ratio of 1:1-3 to prepare a compound with a general formula S-2; the reaction time is 4-12 h, and the reaction solvent is at least one of acetone, N-dimethylformamide, dichloromethane, chloroform and ethyl acetate; The catalyst is at least one of potassium carbonate, cesium carbonate, sodium carbonate, triethylamine, 4-dimethylaminopyridine, N' -diisopropylethylamine and pyridine; (2) Reacting the compound prepared in the step (1) with a compound with a general formula S-3 according to a molar ratio of 1:1.2-2 to prepare a compound with a general formula S-4; The reaction time is 12-36 h, the reaction solvent is N, N-dimethylformamide, acetonitrile, dichloromethane, ethanol, ethyl acetate or a mixture thereof, the reaction temperature is 0-40 ℃, and the catalyst is at least one of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, 2- (7-azabenzotriazol) -N, N, N ', N ' -tetramethylurea hexafluorophosphate, O-benzotriazol-tetramethylurea hexafluorophosphate, 4-dimethylaminopyridine, N, N ' -diisopropylethylamine, triethylamine and pyridine; (3) Reacting the compound prepared in the step (2) with benzyl p-nitrochloroformate in a molar ratio of 1:1.5-3 in a nitrogen atmosphere to prepare a compound with a general formula S-5; The reaction time is 2-12 h, the reaction solvent is at least one of dichloromethane, chloroform and N, N-dimethylformamide, the reaction temperature is 0-30 ℃, and the catalyst is at least one of pyridine, piperidine, triethylamine, 4-dimethylaminopyridine, N' -diisopropylethylamine, potassium carbonate and cesium carbonate; (4) Reacting the compound prepared in the step (3) with a compound of the general formula Y-4 according to a molar ratio of 1:1.1-1.5 to prepare an enzyme activated prodrug of the general formula I; The reaction time is 12-48 h, the reaction solvent is at least one of dichloromethane, chloroform, N-dimethylformamide and acetonitrile, the reaction temperature is 0-40 ℃, and the catalyst is at least one of triethylamine, N' -diisopropylethylamine, 4-dimethylaminopyridine, pyridine, potassium carbonate and aniline; 。
  5. 5. the process of claim 4, wherein the compound of formula Y-4 is prepared by: S1, reacting a compound of a general formula Y-1 with an haloalkane compound containing R 5 substitution according to a molar ratio of 1:2-10 to prepare a compound of a general formula Y-2; The reaction time is 12-36 h, the reaction solvent is at least one of acetonitrile, toluene, o-dichlorobenzene, m-dichlorobenzene or DMF, and the reaction temperature is 70-120 ℃; S2, reacting the compound prepared in the step S1 with a compound 2-chloro-3- (hydroxymethylene) -1-cyclohexene-1-formaldehyde according to a molar ratio of 1:0.5-0.75 in a nitrogen atmosphere to prepare a compound with a general formula of Y-3; the reaction time is 12-24 h, the reaction solvent is at least one of ethanol, methanol, n-butanol, toluene, acetonitrile or DMF, the reaction temperature is 70-120 ℃, and the catalyst is at least one of sodium acetate, potassium acetate or potassium carbonate; S3, reacting the compound prepared in the step S2 with m-aminophenol according to a molar ratio of 1:3-4 to prepare a compound with a general formula of Y-4; The reaction time is 4-12 h, the reaction solvent is dichloromethane, chloroform, acetonitrile, DMF or a mixture thereof, the reaction temperature is 25-40 ℃, and the catalyst is one or a mixture of potassium carbonate, sodium carbonate, triethylamine, DIPEA, DMAP and pyridine; 。
  6. 6. use of a compound according to any one of claims 1-3 for the preparation of a diagnostic tumor therapeutic agent that is a biological sample identification marker formulation or a drug responsive to nitroreductase to kill a diagnostic tumor therapeutic; The tumor is hypoxic tumor.
  7. 7. The use according to claim 6, wherein the nitroreductase is a specific reductase highly expressed in hypoxic tumor cells.
  8. 8. The use of claim 6, wherein the nitroreductase-activated prodrug has fluorescence excitation and emission wavelengths greater than 660 nm.

Description

Enzyme-activated prodrug compound and preparation method and application thereof Technical Field The invention relates to release of targeted antitumor drugs, in particular to an enzyme-activated prodrug compound, a preparation method and application thereof. Background For a long time, the conventional chemotherapy drugs have poor prognosis effect on cancer patients due to the limitation of factors such as poor enrichment power in tumors, serious toxic and side effects, frequent drug resistance and the like. In contrast, selective activation of prodrugs changes the state of the art of traditional chemotherapy. Prodrugs are nontoxic in normal cells, but can be specifically activated as highly toxic factors in tumor cells. This not only improves the selectivity of the tumor, reduces cell killing to non-target spots, but also minimizes the risk of drug resistance. Existing prodrugs can be activated by a variety of triggers in the tumor microenvironment, such as specific pH values, reactive oxygen species, reactive thiols, and the like. Among them, prodrugs activated by enzymes endogenous to cells are most attractive because of their advantages of high affinity, high specificity, and rapid responsiveness. The enzyme activated prodrug not only greatly improves the targeting force on tumors, but also overcomes the complexity of manually introducing exogenous animals. However, the range of applications of existing enzyme-activated prodrugs is generally limited by tumor heterogeneity. Furthermore, most enzyme-activated prodrugs involve only a single chemotherapy. Therefore, research and development of enzyme-activated prodrugs that can accurately identify tumors and can be combined with other therapeutic modalities is of great significance. Chlorambucil (Chlorambucil) is used clinically in a variety of malignancies, including chronic lymphocytic leukemia, ovarian cancer, non-hodgkin's lymphoma, and the like. It induces apoptosis by causing inter-strand cross-linking in DNA. However, the drawbacks associated with chemotherapy have limited their widespread use, including relatively poor selectivity for tumor cells, irreversible damage to normal cells and tissues, and the like. Therefore, the development of a chlorambucil-based multimode enzyme-activated prodrug is of great significance. The cyanine dye photosensitizer has a plurality of special physicochemical properties, such as larger molar extinction coefficient and fluorescence quantum yield, good thermal stability and photostability, and better phototoxicity, can control the photosensitive property of OFF-ON through chemical modification, and can efficiently carry out photodynamic therapy (PDT) and realize tumor inhibition. Disclosure of Invention Based on the current situation of lack of enzyme-activated prodrugs capable of realizing accurate chemotherapy and considering excellent tumor selectivity and inhibition effect of photodynamic therapy, the invention constructs a class of enzyme-sensitive chlorambucil analogue prodrugs. The prodrug has high selectivity to hypoxic tumor and can be effectively activated in hypoxic tumor cells. When activated specifically, the prodrug releases the free chemotherapeutic drug chlorambucil and releases the photosensitizer cyanine dye to realize the conversion of the photosensitive property of OFF-ON and generate a large amount of active oxygen species to realize the unification and the synergy of photodynamic therapy and activatable chemotherapy. In order to achieve the aim, the technical scheme of the invention is that a class of enzyme-activated prodrug compounds has the following structural general formula I: In the general formula I, the components are shown in the specification, R 1 is-N [ (CH 2CH2)mX]2), wherein X is selected from halogen, hydroxy, mercapto or nitro; m is an integer of 1-4; r 2 is selected from nitro or any of the groups of formulas i-iii; r 3 is selected from-NH or O; r 4 is selected from O or S; R 5 is selected from hydrogen, alkyl having 1-6 carbons, carboxyalkyl having 1-6 carbons, hydroxyalkyl having 1-6 carbons, or alkylsulfonate having 1-6 carbons; r 6 is optionally substituted on the 6-membered ring hydrogen, halogen, hydroxy, mercapto, cyano, nitro, alkyl having 1-6 carbons, carboxyalkyl having 1-6 carbons, hydroxyalkyl having 1-6 carbons or alkylsulfonate having 1-6 carbons. Further, the stimulus release factor of the compound is the highly expressed nitroreductase in the anaerobic tumor cells, and the compound simultaneously releases free chlorambucil and activated cyanine dye under the hydrolysis of nitroreductase in the anaerobic tumor. Further, the compounds release chlorambucil and activated cyanine dye in a 1:1 relationship. A method for preparing an enzyme-activated prodrug compound comprising the steps of: (1) 2-hydroxy-5-methyl m-xylylene glycol reacts with a compound with a general formula S-1 according to a molar ratio of 1:1-3 to prepare a compound with a general formula S-2; the