Azo compound, azo prodrug compound, pharmaceutical composition, lipid molecule responsive to high-energy rays, and liposome responsive to high-energy rays

Abstract

Disclosed in the present invention are an azo compound, an azo prodrug compound, a pharmaceutical composition, a lipid molecule responsive to high-energy rays, and a liposome responsive to high-energy rays. The hydroxyl group end of the azo compound of the present invention can be covalently bonded to a drug, so as to prepare an azo prodrug compound. When the prepared azo prodrug compound is used in combination with radiotherapy, under the irradiation of high-energy rays (e.g. α-rays, β-rays, γ-rays and X-rays), the azo group undergoes a chemical conversion so as to release the drug, thereby achieving a drug chemotherapy while radiotherapy is implemented. Compared with traditional chemotherapeutic drugs, the azo prodrug compound prepared from the azo compound of the present invention has a novel structure and good safety, and can achieve stably-activated radiotherapy responses. In addition, the azo prodrug compound prepared from the azo compound of the present invention has a stable activation efficiency and wide application range of active pharmaceutical ingredients, thereby achieving a good application prospect.

Description

Azo compound, azo prodrug compound, pharmaceutical composition, high-energy ray-responsive lipid molecule, and high-energy ray-responsive liposome Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to an azo compound, an azo prodrug compound, a pharmaceutical composition, a high-energy ray-responsive lipid molecule, and a high-energy ray-responsive liposome. Background Art

[0002] In clinical treatment, combined chemotherapy and radiotherapy are the first-line treatment options for over 50% of cancer patients. While traditional chemotherapy drugs kill tumor cells, they also damage normal cells and tissues, leading to common side effects such as hair loss, cardiotoxicity, and nephrotoxicity. Reducing or eliminating these severe adverse reactions is a pressing issue in the current clinical treatment of cancer.

[0003] Formulating anti-tumor drugs into exogenously responsive prodrugs and then activating them at the tumor site after administration can effectively reduce the drug's systemic toxicity. Compared with other activation methods, X-rays can accurately penetrate deep tissues and have the advantage of high temporal and spatial resolution. X-ray technology currently used in medical radiotherapy is developing rapidly and can accurately locate tumors at the submillimeter level. Therefore, the strategy of X-ray activation is expected to achieve the targeted release of anti-cancer drugs at the tumor site, significantly reducing the killing of normal cells and achieving precise tumor treatment based on combined radiotherapy and chemotherapy.

[0004] According to existing research, the activation of prodrugs by X-rays mainly depends on the up-regulated enzymes (such as caspase-3) expressed in the tumor site after X-ray irradiation and the ionization of water molecules to generate hydroxyl radicals (·OH), hydrogen radicals (·H), hydrated electrons (e - aq Although the above-mentioned prodrug activation system has achieved certain results in in vitro activation and in vivo efficacy experiments, it is still in the early stages of exploration and faces challenges such as unstable activation efficiency and limited application scope of the original drug.

[0005] Therefore, it is very necessary to further develop radiotherapy-responsive prodrug activation systems with novel structures, good safety, and stable activation for a variety of chemotherapy drugs with broad-spectrum anti-tumor activity. Summary of the Invention

[0006] Based on this, it is necessary to provide an azo compound that can solve the above problems.

[0007] In addition, it is also necessary to provide an azo prodrug compound, a pharmaceutical composition, a high-energy ray-responsive lipid molecule, and a high-energy ray-responsive liposome.

[0008] An azo compound having the following general formula I:

[0009] in, It is a single bond, which means that A and B are distributed on both sides or the same side of the azo double bond;

[0010] -A- and -B- are each independently selected from: aryl or heteroaryl;

[0011] The term "aryl" refers to an aromatic hydrocarbon group having 6 to 20 carbon atoms obtained by removing two hydrogen atoms from two carbon atoms in the aromatic nucleus of an aromatic hydrocarbon molecule;

[0012] The term "heteroaryl" refers to aromatic groups that are 5-membered or 6-membered rings and fused ring systems comprising 5 to 20 atoms, wherein at least one ring is aromatic and contains one or more heteroatoms independently selected from nitrogen, oxygen and sulfur, and if the ring contains multiple oxygen atoms, the oxygen atoms are not directly adjacent;

[0013] -C- is -X, -O-, -OC(=O)- or X is Cl, Br or I; when -C- is -X, R2 is an empty bond; when -C- is -O-, R2 is H; when -C- is -OC(=O)-, -R2 is -Cl, When -C- is When -R2 is -Cl;

[0014] -R1 is

[0015] -R3 and -R4 are independently selected from: -CN, -COOR, -CONR'R", -H, halogen, C1-C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group is O or N;

[0016] -R5 and -R6 are independently selected from: -H, C1-C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group is O or N;

[0017] -R7 is H, C1~C 12 Alkyl, C1~C12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group is O or N;

[0018] -R8 is 1 to 4 substituted, and -R8 is selected from -NO2, -F, -CN, -CF3, -COOR, -CONR'R".

[0019] or,

[0020] An azo compound having the following general formula I:

[0021] in, It is a single bond, which means that A and B are distributed on both sides or the same side of the azo double bond;

[0022] -A- and -B- are each independently selected from: aryl or heteroaryl;

[0023] The term "aryl" refers to an aromatic ring group having 6 to 20 carbon atoms obtained by removing two hydrogen atoms from two carbon atoms in the aromatic nucleus of an aromatic ring molecule;

[0024] The term "heteroaryl" refers to aromatic groups that are 5-membered or 6-membered rings and fused ring systems comprising 5 to 20 atoms, wherein at least one ring is aromatic and contains one or more heteroatoms independently selected from nitrogen, oxygen and sulfur, and if the ring contains multiple oxygen atoms, the oxygen atoms are not directly adjacent;

[0025] -C- is -X, -O-, -OC(=O)- or X is Cl, Br or I; when -C- is -X, R2 is an empty bond; when -C- is -O-, R2 is H; when -C- is -OC(=O)-, -R2 is -Cl, When -C- is When -R2 is -Cl;

[0026] -R1 is

[0027] -R3 and -R4 are independently selected from: -CN, -COOR, -CONR'R", -H, halogen, C1-C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20The heteroatom in the aryl group is O or N;

[0028] -R5 is selected from: -H, C1~C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 Aryl, -R6 is selected from C1~C 12 The heteroatom in the heteroalkyl group is O or N;

[0029] -R8 is 1 to 4 substituted, and -R8 is selected from -NO2, -F, -CN, -CF3, -COOR, -CONR'R".

[0030] In one embodiment, the aryl or heteroaryl is substituted with one or more substituents, the substituents including halogen, -R9, -NR9R 10 , -CN, -NO2, -N3, -OR9, -SR9, -NHCOR9, -O-COR9, -CH=CR9R 10 , -C(=O)-R9, -C(=O)-OR9, -C(=O)-Cl, -C(=O)-NH2, -C(=CO)-NH-R9 and -C(=O)-NR9R 10 , among which -R9 and -R 10 Each independently selected from -H, C1-C6 alkyl, C1-C6 haloalkyl, C2-C6 alkenyl, C3-C 10 Cycloalkyl, C6~C 20 The alkyl, alkenyl, cycloalkyl, aryl and heteroaryl groups described for the substituents are optionally replaced by one or more halogen, hydroxyl, mercapto, -NH2, -CN, -NO2, -N3, -NHCOH, -OC(=O)H, -C(=O)H, -C(=O)-OH, -C(=O)-Cl, -C(=O)-NH2, -C(=O)-NH-CH3, -C(=O)-CH3, -C(=O)-OCH3, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C2-C6 alkenyl, C3-C 10 Cycloalkyl, C6~C 20 substituted with an aryl group or a heteroaryl group having 5 to 20 ring atoms;

[0031] -R, -R' and -R" are independently selected from -H, C1-C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group or heteroalkyl group is O or N.

[0032] In one embodiment, the structural formula of the azo compound is:

[0033] In one embodiment, the structural formula of the azo compound is:

[0034] An azo prodrug compound having the following general formula II:

[0035] in, It is a single bond, which means that A and B are distributed on both sides or the same side of the azo double bond;

[0036] -A- and -B- are each independently selected from: aryl or heteroaryl;

[0037] The term "aryl" refers to an aromatic group having 6 to 20 carbon atoms obtained by removing two hydrogen atoms from two carbon atoms in the aromatic nucleus of an aromatic hydrocarbon (aromatic ring) molecule;

[0038] The term "heteroaryl" refers to aromatic groups that are 5-membered or 6-membered rings and fused ring systems comprising 5 to 20 atoms, wherein at least one ring is aromatic and contains one or more heteroatoms independently selected from nitrogen, oxygen and sulfur, and if the ring contains multiple oxygen atoms, the oxygen atoms are not directly adjacent;

[0039] -C- is -O-, -N + (R a R b )-、-OC(=O)- or

[0040] -G1 is a drug group;

[0041] -G2 is a targeting group,

[0042] -R3 and -R4 are independently selected from: -CN, -COOR, -CONR'R", -H, halogen, C1-C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20The heteroatom in the aryl group is O or N;

[0043] -R5 and -R6 are independently selected from: -H, C1-C 12 Alkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 aromatic groups;

[0044] -R7 is -H, C1~C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group or heteroalkyl group is O or N.

[0045] In one embodiment, the aryl or heteroaryl is substituted with one or more substituents, the substituents including halogen, -R9, -NR9R 10 , -CN, -NO2, -N3, -OR9, -SR9, -NHCOR9, -O-COR9, -CH=CR9R 10 , -C(=O)-R9, -C(=O)-OR9, -C(=O)-Cl, -C(=O)-NH2, -C(=CO)-NH-R9 and -C(=O)-NR9R 10 , among which -R9 and -R 10 Each independently selected from -H, C1-C6 alkyl, C1-C6 haloalkyl, C2-C6 alkenyl, C3-C 10 Cycloalkyl, C6~C 20 The alkyl, alkenyl, cycloalkyl, aryl and heteroaryl groups described for the substituents are optionally replaced by one or more halogen, hydroxyl, mercapto, -NH2, -CN, -NO2, -N3, -NHCOH, -OC(=O)H, -C(=O)H, -C(=O)-OH, -C(=O)-Cl, -C(=O)-NH2, -C(=O)-NH-CH3, -C(=O)-CH3, -C(=O)-OCH3, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C2-C6 alkenyl, C3-C 10 Cycloalkyl, C6~C 20 substituted with an aryl group or a heteroaryl group having 5 to 20 ring atoms;

[0046] -R a and -Rb Each independently selected from C1 to C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group is O or N;

[0047] -R, -R' and -R" are independently selected from -H, C1-C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group or heteroalkyl group is O or N.

[0048] In one embodiment, the targeting group is covalently linked to other structures via N, O or S;

[0049] The targeting group is an antibody, a polypeptide, a sugar group, a small molecule ligand or a nucleic acid aptamer;

[0050] The drug group is covalently linked to other structures via N, O or S;

[0051] The drug group is

[0052] A pharmaceutical composition comprises the above-mentioned azo prodrug compound or a pharmaceutically acceptable salt thereof.

[0053] A high-energy ray-responsive lipid molecule has the following general formula III:

[0054] in, It is a single bond, which means that A and B are distributed on both sides or the same side of the azo double bond;

[0055] -A- and -B- are each independently selected from: aryl or heteroaryl;

[0056] The term "aryl" refers to an aromatic group having 6 to 20 carbon atoms obtained by removing one hydrogen atom from each of two carbon atoms in the aromatic nucleus of an aromatic hydrocarbon (aromatic ring) molecule;

[0057] The term "heteroaryl" refers to aromatic groups that are 5-membered or 6-membered rings and fused ring systems comprising 5 to 20 atoms, wherein at least one ring is aromatic and contains one or more heteroatoms independently selected from nitrogen, oxygen and sulfur, and if the ring contains multiple oxygen atoms, the oxygen atoms are not directly adjacent;

[0058] -C- is -O-, -N + (R a R b )-、-OC(=O)- or

[0059] -G3 and -G4 are respectively one of a hydrophilic group and a hydrophobic group, and the high-energy ray-responsive lipid molecule is an amphiphilic molecule;

[0060] -R3 and -R4 are independently selected from: -CN, -COOR, -CONR'R", -H, halogen, C1-C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group or heteroalkyl group is O or N.

[0061] In one embodiment, the lipid molecule is a fatty acid ester or amide derivative, a sterol ester or amide derivative, a glyceride, a glycerophospholipid or a glycolipid containing an azo structural unit;

[0062] The aryl or heteroaryl is substituted by one or more substituents, the substituents including halogen, -R9, -NR9R 10 , -CN, -NO2, -N3, -OR9, -SR9, -NHCOR9, -O-COR9, -CH=CR9R 10 , -C(=O)-R9, -C(=O)-OR9, -C(=O)-Cl, -C(=O)-NH2, -C(=CO)-NH-R9 and -C(=O)-NR9R 10 , among which -R9 and -R 10 Each independently selected from -H, C1-C6 alkyl, C1-C6 haloalkyl, C2-C6 alkenyl, C3-C 10 Cycloalkyl, C6~C 20The alkyl, alkenyl, cycloalkyl, aryl and heteroaryl groups described for the substituents are optionally replaced by one or more halogen, hydroxyl, mercapto, -NH2, -CN, -NO2, -N3, -NHCOH, -OC(=O)H, -C(=O)H, -C(=O)-OH, -C(=O)-Cl, -C(=O)-NH2, -C(=O)-NH-CH3, -C(=O)-CH3, -C(=O)-OCH3, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C2-C6 alkenyl, C3-C 10 Cycloalkyl, C6~C 20 substituted with an aryl group or a heteroaryl group having 5 to 20 ring atoms;

[0063] -R a and -R b Each independently selected from C1 to C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group is O or N;

[0064] -R, -R' and -R" are independently selected from -H, C1-C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group or heteroalkyl group is O or N.

[0065] A high-energy ray-responsive liposome is formed by self-assembly of the above-mentioned high-energy ray-responsive lipid molecules.

[0066] The hydroxyl end of the azo compound of the present invention can be connected to a drug through a substitution reaction to prepare an azo prodrug compound. When the prepared azo prodrug compound is used in combination with radiotherapy, under the irradiation of high-energy rays (for example, α rays, β rays, γ rays, X-rays), the azo group undergoes chemical conversion, thereby releasing the drug, thereby achieving simultaneous radiotherapy and chemotherapy. Beneficial effects:

[0067] Compared with traditional chemotherapy drugs, the azo prodrug compounds prepared using the azo compounds of the present invention have novel structures, excellent safety, and can achieve stable activation and radiotherapy responses. Furthermore, the azo prodrug compounds prepared using the azo compounds of the present invention have stable activation efficiency and a wide range of applicability, showing promising application prospects.

[0068] In addition, the hydroxyl end of the azo compound can be connected to a drug through a substitution reaction to prepare an azo prodrug compound, and the carboxyl end of the azo compound can also be connected to a substance with a targeting function (such as an antibody) through a condensation reaction, thereby making the prepared azo prodrug compound have a targeting function.

[0069] In addition, the hydroxyl end and carboxyl end of the azo compound of the present invention can be connected to a hydrophilic group and a hydrophobic group through a substitution reaction and a condensation reaction, respectively, to form an amphiphilic lipid molecule. The lipid molecule can achieve high-energy ray response through the structural transformation of the azo group before and after irradiation. The amphiphilic property of the lipid molecule enables it to self-assemble to form high-energy ray-responsive liposomes. The interior of the high-energy ray-responsive liposomes can be coated with drugs. Under the irradiation of high-energy rays (for example, α rays, β rays, γ rays, and X-rays), the azo group undergoes structural transformation, the hydrophilic group and the hydrophobic group separate, and the liposome structure decomposes, thereby releasing the internally coated drug, thereby achieving radiotherapy and drug chemotherapy at the same time. BRIEF DESCRIPTION OF THE DRAWINGS

[0070] FIG1 to FIG4 are comparative test results of toxicity of various azo prodrug compounds and the original drug in various cells in Test Example 2.

[0071] FIG5 is a diagram showing the hydrophobic interaction chromatography (HIC-HPLC) test results of the antibody-drug conjugate XPADC-106-011 prepared in Example 24.

[0072] FIG6 is a graph showing the LCMS chromatographic detection results of the 107-008 liposomes prepared in Example 25 before irradiation with 60 Gy and after the nanoparticles were demulsified with ethanol.

[0073] FIG7 is a graph showing the LCMS chromatographic detection results of the 107-008 liposomes prepared in Example 25 after irradiation with 60 Gy and demulsification of the nanoparticles with ethanol.

[0074] FIG8 is a graph showing the particle size distribution of 115-001 liposomes prepared in Example 26 before irradiation with 60 Gy.

[0075] FIG9 is a graph showing the particle size distribution of the 115-001 liposomes prepared in Example 26 after irradiation with 60 Gy. DETAILED DESCRIPTION

[0076] The following will be combined with specific embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts shall fall within the scope of protection of the present invention.

[0077] It should be noted that all directional indications in the embodiments of the present invention (such as up, down, left, right, front, back, etc.) are only used to explain the relative position relationship and movement status of each component in a certain specific posture. If the specific posture changes, the directional indication will also change accordingly.

[0078] In addition, the descriptions of "first", "second", etc. in the present invention are for descriptive purposes only and should not be understood as indicating or implying their relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined as "first" or "second" may explicitly or implicitly include at least one of the said features. In addition, the technical solutions between the various embodiments can be combined with each other, but this must be based on the fact that they can be implemented by ordinary technicians in this field. When the combination of technical solutions is contradictory or cannot be implemented, it should be deemed that such combination of technical solutions does not exist and is not within the scope of protection required by the present invention.

[0079] The present invention discloses an azo compound according to one embodiment, which has the following general formula I:

[0080] in, It is a single bond, which means that A and B are distributed on both sides or the same side of the azo double bond;

[0081] -A- and -B- are each independently selected from: aryl or heteroaryl;

[0082] The term "aryl" refers to an aromatic group having 6 to 20 carbon atoms obtained by removing two hydrogen atoms from two carbon atoms in the aromatic nucleus of an aromatic ring molecule;

[0083] The term "heteroaryl" refers to aromatic groups that are 5-membered or 6-membered rings and fused ring systems comprising 5 to 20 atoms, wherein at least one ring is aromatic and contains one or more heteroatoms independently selected from nitrogen, oxygen and sulfur, and if the ring contains multiple oxygen atoms, the oxygen atoms are not directly adjacent;

[0084] -C- is -X, -O-, -OC(=O)- or X is Cl, Br or I; when -C- is -X, R2 is an empty bond; when -C- is -O-, R2 is H; when -C- is -OC(=O)-, -R2 is -Cl, When -C- is When -R2 is -Cl;

[0085] -R1 is

[0086] -R3 and -R4 are independently selected from: -CN, -COOR, -CONR'R", -H, halogen, C1-C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group is O or N;

[0087] -R5 and -R6 are independently selected from: -H, C1-C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group is O or N;

[0088] -R7 is H, C1~C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group is O or N;

[0089] -R8 is 1 to 4 substituted, and -R8 is selected from -NO2, -F, -CN, -CF3, -COOR, -CONR'R".

[0090] in, etc. all represent the connection site between the drug group and other structures, that is, half of a single bond “-”.

[0091] The hydroxyl end of the azo compound of the present invention can be connected to a drug through a substitution reaction to prepare an azo prodrug compound. When the prepared azo prodrug compound is used in combination with radiotherapy, under the irradiation of high-energy rays (for example, α rays, β rays, γ rays, X-rays), the azo group undergoes chemical conversion, thereby releasing the drug, thereby achieving simultaneous radiotherapy and chemotherapy.

[0092] Compared with traditional chemotherapy drugs, the azo prodrug compounds prepared using the azo compounds of the present invention have novel structures, excellent safety, and can achieve stable activation and radiotherapy responses. Furthermore, the azo prodrug compounds prepared using the azo compounds of the present invention have stable activation efficiency and a wide range of applicability, showing promising application prospects.

[0093] In addition, the hydroxyl end of the azo compound can be connected to a drug through a substitution reaction to prepare an azo prodrug compound, and the carboxyl end of the azo compound can also be connected to a substance with a targeting function (such as an antibody) through a condensation reaction, thereby making the prepared azo prodrug compound have a targeting function.

[0094] In addition, the hydroxyl end and carboxyl end of the azo compound of the present invention can be connected to a hydrophilic group and a hydrophobic group through a substitution reaction and a condensation reaction, respectively, to form an amphiphilic lipid molecule. The lipid molecule can achieve high-energy ray response through the structural transformation of the azo group before and after irradiation. The amphiphilic property of the lipid molecule enables it to self-assemble to form high-energy ray-responsive liposomes. The interior of the high-energy ray-responsive liposomes can be coated with drugs. Under the irradiation of high-energy rays (for example, α rays, β rays, γ rays, and X-rays), the azo group undergoes structural transformation, the hydrophilic group and the hydrophobic group separate, and the liposome structure decomposes, thereby releasing the internally coated drug, thereby achieving radiotherapy and drug chemotherapy at the same time.

[0095] The heteroaryl group can be a monocyclic heteroaryl group, a bicyclic heteroaryl group, and a tricyclic heteroaryl group.

[0096] Exemplary monocyclic heteroaryl groups include pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, tetrazinyl, furanyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, oxatriazolyl, furazanyl, thiazolyl, isothiazolyl, and the like.

[0097] Exemplary bicyclic heteroaryl groups include indolyl, benzothiazolyl, benzodioxolyl, benzoxazolyl, benzothiophenyl, quinolinyl, tetrahydroisoquinolinyl, isoquinolinyl, benzimidazolyl, benzopyranyl, indolizinyl, benzofuranyl, chromonyl, coumarinyl, benzopyranyl, cinnolinyl, quinoxalinyl, indazolyl, pyrrolopyridinyl, furopyridinyl, dihydroisoindolyl, tetrahydroquinolinyl, and the like.

[0098] Exemplary tricyclic heteroaryl groups include carbazolyl, dibenzofuranyl, dibenzothiophenyl, benzindolyl, phenanthrolinyl, acridinyl, phenanthridinyl, xanthene, and the like.

[0099] It should be noted that when -C- is -X and R2 is a null bond, -C-R2 is -X.

[0100] In one embodiment, the aryl or heteroaryl group has no substituents other than the linking azo and A / B groups.

[0101] Preferably, in this embodiment, the aryl or heteroaryl group is substituted by one or more substituents, and the substituents include halogen, -R9, -NR9R 10 , -CN, -NO2, -N3, -OR9, -SR9, -NHCOR9, -O-COR9, -CH=CR9R 10 , -C(=O)-R9, -C(=O)-OR9, -C(=O)-Cl, -C(=O)-NH2, -C(=CO)-NH-R9 and -C(=O)-NR9R 10 , among which -R9 and -R 10 Each independently selected from -H, C1-C6 alkyl, C1-C6 haloalkyl, C2-C6 alkenyl, C3-C 10 Cycloalkyl, C6~C 20 The alkyl, alkenyl, cycloalkyl, aryl and heteroaryl groups described for the above substituents are optionally substituted by one or more halogen, hydroxyl, mercapto, -NH2, -CN, -NO2, -N3, -NHCOH, -OC(=O)H, -C(=O)H, -C(=O)-OH, -C(=O)-Cl, -C(=O)-NH2, -C(=O)-NH-CH3, -C(=O)-CH3, -C(=O)-OCH3, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C2-C6 alkenyl, C3-C 10 Cycloalkyl, C6~C 20 or a heteroaryl group having 5 to 20 ring atoms.

[0102] Preferably, in this embodiment, -R, -R' and -R" are independently selected from -H, C1-C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group or heteroalkyl group is O or N.

[0103] More preferably, in one embodiment, the aryl or heteroaryl group is substituted with one or more substituents, which include alkoxy (eg, methoxy, ethoxy, propoxy, butoxy).

[0104] More preferably, in another embodiment, the aryl or heteroaryl group is substituted with one or more substituents, which include halogen, such as fluorine, chlorine, bromine, iodine, and more preferably fluorine.

[0105] In a particular embodiment, -A- is -B- is

[0106] The azo compound with the general formula I disclosed in the present invention can be prepared and synthesized through existing processes.

[0107] Specifically, the azo compound ON-3(tert-butyl(E)-N-(4-((4-(hydroxymethyl)phenyl)diazenyl)benzoyl)-N-methylglycinate), the azo compound ON-4(tert-butyl(E)-N-methyl-N-(4-((4-((((4-nitrophenoxy)carbonyl)oxy)methyl)phenyl)diazenyl)benzoyl)glycinate) and the azo compound ON-5((E)-N-methyl-N-(4-((4-((((4-nitrophenoxy)carbonyl)oxy)methyl)phenyl)diazenyl)benzoyl)glycine) can be prepared by the following synthetic route:

[0108] Specifically, the azo compound YJY-2-67((E)-4-((4-((2-(dimethylamino)ethyl)carbamoyl)phenyl)diazenyl)benzyl(4-nitrophenyl)carbonate) can be prepared by the following synthetic route:

[0109] Specifically, the azo compound 2-MZL-Cl-2(tert-butyl(E)-N-(3-methoxy-4-((4-((((4-nitrophenoxy)carbonyl)oxy)methyl)phenyl)diazenyl)benzoyl)-N-methylglycinate) can be prepared by the following synthetic route:

[0110] Specifically, the azo compound ON-2F-4(tert-butyl(E)-N-(3,5-difluoro-4-((4-((((4-nitrophenoxy)carbonyl)oxy)methyl)phenyl)diazenyl)benzoyl)-N-methylglycinate) can be prepared by the following synthetic route:

[0111] Specifically, the azo compound HJA-ON-4(tert-butyl(E)-N-(4-((2-methoxy-4-((((4-nitrophenoxy)carbonyl)oxy)methyl)phenyl)diazenyl)benzoyl)-N-methylglycinate) can be prepared by the following synthetic route:

[0112] Specifically, the azo compound ON-3-Cl (tert-butyl (E) -N- (4- ((4- (((chlorocarbonyl)oxy)methyl)phenyl)diazenyl)benzoyl) -N-methylglycinate) can be prepared by the following synthetic route:

[0113] Specifically, the azo compound ON-3-IMZ((E)-4-((4-((2-(tert-butoxy)-2-oxoethyl)(methyl)carbamoyl)phenyl)diazenyl)benzyl 1H-imidazole-1-carboxylate) can be prepared by the following synthetic route:

[0114] Specifically, the azo compound 12 (tert-butyl(E)-N-methyl-N-(5-((6-((((4-nitrophenoxy)carbonyl)oxy)methyl)pyridin-3-yl)diazenyl)picolinoyl)glycinate) can be prepared by the following synthetic route:

[0115] Specifically, the azo compound 17 (tert-butyl(E)-N-methyl-N-(4-((2-((((4-nitrophenoxy)carbonyl)oxy)methyl)furan-3-yl)diazenyl)furan-2-carbonyl)glycinate) can be prepared by the following synthetic route:

[0116] Specifically, the azo compound 19 (tert-butyl(E)-N-(4-((4-(((((4-nitrophenoxy)carbonyl)oxy)methyl)phenyl)diazenyl)benzoyl)-N-(tetrahydrofuran-2-yl)glycinate) can be prepared by the following synthetic route:

[0117] Specifically, the azo compound 21 (1-(2-(tert-butoxy)-2-oxoethyl)piperidin-4-yl(E)-4-((4-((((4-nitrophenoxy)carbonyl)oxy)methyl)phenyl)diazenyl)benzoate) can be prepared by the following synthetic route:

[0118] The present invention also discloses an azo prodrug compound according to one embodiment, which has the following general formula II:

[0119] in, It is a single bond, which means that A and B are distributed on both sides or the same side of the azo double bond;

[0120] -A- and -B- are each independently selected from: aryl or heteroaryl;

[0121] The term "aryl" refers to an aromatic group having 6 to 20 carbon atoms obtained by removing two hydrogen atoms from two carbon atoms in the aromatic nucleus of an aromatic ring molecule;

[0122] The term "heteroaryl" refers to aromatic groups that are 5-membered or 6-membered rings and fused ring systems comprising 5 to 20 atoms, wherein at least one ring is aromatic and contains one or more heteroatoms independently selected from nitrogen, oxygen and sulfur, and if the ring contains multiple oxygen atoms, the oxygen atoms are not directly adjacent;

[0123] -C- is -O-, -N + (R a R b)-、-OC(=O)- or

[0124] -G1 is a drug group;

[0125] -G2 is a targeting group,

[0126] -R3 and -R4 are independently selected from: -CN, -COOR, -CONR'R", -H, halogen, C1-C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group is O or N;

[0127] -R5 and -R6 are independently selected from: -H, C1-C 12 Alkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 aromatic groups;

[0128] -R7 is -H, C1~C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group or heteroalkyl group is O or N.

[0129] in, etc. all represent the connection site between the drug group and other structures, that is, half of a single bond “-”.

[0130] When the azo prodrug compound of the present invention is used in combination with radiotherapy, under the irradiation of high-energy rays (such as α rays, β rays, γ rays, and X rays), the azo group undergoes chemical conversion, thereby releasing the drug, thereby achieving simultaneous radiotherapy and chemotherapy.

[0131] The azo prodrug compound of the present invention can be prepared by connecting a drug through a substitution reaction at the hydroxyl end of the above-mentioned azo compound.

[0132] It should be noted that the specific structure of the aryl or heteroaryl group in the azo prodrug compound is as described above and will not be repeated here.

[0133] Preferably, in this embodiment, -Ra and -R b Each independently selected from C1 to C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group or heteroalkyl group is O or N.

[0134] Preferably, in this embodiment, -R, -R' and -R" are independently selected from -H, C1-C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group or heteroalkyl group is O or N.

[0135] It should be noted that when -G2 is a targeting group, the azo prodrug compound has a targeting function. In this case, a substance having a targeting function (e.g., an antibody) can be connected to the carboxyl end of the azo compound through a condensation reaction to prepare an azo prodrug compound having a targeting function.

[0136] Preferably, the targeting group is covalently linked to other structures via N, O or S.

[0137] Generally speaking, the targeting group can be any group having a targeting effect.

[0138] Specifically, the targeting group is an antibody, a peptide, a sugar group, a small molecule ligand or a nucleic acid aptamer.

[0139] When -G2 is not a targeting group, the azo prodrug compound does not have a targeting function.

[0140] In general, the drug group can be any therapeutic agent that can be coupled to the azo structure, as long as it can be detached from the azo structure and release a therapeutically active molecule under the action of radiation.

[0141] In the present invention, the released therapeutically active molecule generally has the same structure as the therapeutic agent G1 in Formula II. In some embodiments, the released therapeutically active molecule has a slightly different structure than the therapeutic agent G1 in Formula II, i.e., the therapeutic agent molecule undergoes a chemical reaction under the action of radiation that does not significantly affect its therapeutic activity.

[0142] Active molecules that can be used as therapeutic agents include, but are not limited to, polypeptides, oligopeptides, peptidomimetics, amino acids, enzyme inhibitors, hormones, toxins, antibiotics, anti-inflammatory substances, and the like.

[0143] Preferably, the active molecule is a therapeutic agent for treating cancer.

[0144] When the active molecule is a therapeutic agent for treating cancer, this radiotherapy-activated prodrug approach can be used to achieve combined radiotherapy and chemotherapy, that is, while radiation is treating cancer, irradiation also activates the release of chemical anti-tumor drugs to achieve chemotherapy.

[0145] More preferably, the present invention uses a drug containing a primary amine or a secondary amine as a therapeutic agent. Specifically, the present invention uses an anticancer drug containing a primary amine or a secondary amine as a therapeutic agent. Drugs containing primary amines or secondary amines include, for example, ibrutinib, acalabrutinib, zanubrutinib, doxorubicin, mitomycin-C, mitomycin-A, daunorubicin, aminopterin, actinomycin, bleomycin, 9-aminocamptothecin, N8-acetylspermidine, bis(2-chloroethyl)amine, yunnanmycin, gemcitabine, cytarabine, dolastatin, dacarbazine, 5-fluorouracil, and derivatives thereof.

[0146] Drugs containing primary or secondary amines also include amino derivatives of drugs that do not naturally contain amino groups. In other words, drugs that do not naturally contain amino groups can be chemically modified to have amino groups, and then coupled to radiation-responsive groups or to radiation-responsive groups and linkers using the primary or secondary amine coupling methods described herein.

[0147] In particular, the therapeutic agent is monomethyl auristatin E.

[0148] In another embodiment, the present invention uses a hydroxyl-containing anticancer drug as a therapeutic agent. Examples of hydroxyl-containing therapeutic agents include paclitaxel, docetaxel, gemcitabine, cytarabine, and the like. Hydroxyl-containing therapeutic agents also include hydroxyl derivatives of drugs that do not naturally contain hydroxyl groups. In other words, drugs that do not originally contain hydroxyl groups can be chemically modified to have hydroxyl groups, and then coupled to a radiation-responsive group or to a radiation-responsive group and a linker using the hydroxyl coupling method described in this disclosure.

[0149] In another embodiment, the present invention utilizes a thiol-containing drug as a therapeutic agent. Preferably, the present disclosure utilizes a thiol-containing anticancer drug as a therapeutic agent. Examples of thiol-containing therapeutic agents include 6-mercaptopurine. Thiol-containing therapeutic agents also include thiol derivatives of drugs that do not naturally contain thiol groups. In other words, drugs that do not naturally contain thiol groups can be chemically modified to have thiol groups, and then coupled to a radiation-responsive group or to a radiation-responsive group and a linker using the thiol coupling methods described herein.

[0150] Preferably, the drug group is covalently linked to other structures via N or O.

[0151] Specifically, in this embodiment, the drug group is

[0152] in, Represents the connection site between the drug group and other structures, that is, half of a single bond "-".

[0153] The present invention also discloses a pharmaceutical composition according to one embodiment, comprising the above-mentioned azo prodrug compound or a pharmaceutically acceptable salt thereof.

[0154] Specifically, pharmaceutically acceptable salts of azo prodrug compounds include inorganic salts or organic salts.

[0155] Among them, inorganic salts include hydrochloride, hydrobromide, hydroiodide, perchlorate, sulfate, bisulfate, nitrate, phosphate, acid phosphate, sodium salt, calcium salt, potassium salt, magnesium salt, silver salt, and lithium salt; organic salts include formate, acetate, trifluoroacetate, propionate, pyruvate, glycolate, oxalate, malonate, succinate, glutarate, fumarate, maleate, lactate, malate, citrate, tartrate, methanesulfonate, ethanesulfonate, benzenesulfonate, salicylate, p-toluenesulfonate, ascorbate, meglumine salt, tromethamine salt, diethylamine salt, lysine salt, choline salt, arginine salt, terbutylamine salt, and N,N-dibenzylethylenediamine salt.

[0156] Preferably, the above-mentioned pharmaceutical composition further comprises pharmaceutical excipients.

[0157] The pharmaceutical excipients include: at least one of: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integrators, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoaming agents, thickeners, inclusion agents, humectants, flocculants and deflocculating agents, filter aids and release retardants.

[0158] The present invention also discloses a high-energy ray-responsive lipid molecule according to an embodiment of the present invention, which has the following general formula III:

[0159] in, It is a single bond, which means that A and B are distributed on both sides or the same side of the azo double bond;

[0160] -A- and -B- are each independently selected from: aryl or heteroaryl;

[0161] The term "aryl" refers to an aromatic group having 6 to 20 carbon atoms obtained by removing one hydrogen atom from each of two carbon atoms in the aromatic nucleus of an aromatic hydrocarbon (aromatic ring) molecule;

[0162] The term "heteroaryl" refers to aromatic groups that are 5-membered or 6-membered rings and fused ring systems comprising 5 to 20 atoms, wherein at least one ring is aromatic and contains one or more heteroatoms independently selected from nitrogen, oxygen and sulfur, and if the ring contains multiple oxygen atoms, the oxygen atoms are not directly adjacent;

[0163] -C- is -O-, -N + (R a R b )-、-OC(=O)- or

[0164] -G3 and -G4 are respectively one of a hydrophilic group and a hydrophobic group, and the high-energy ray-responsive lipid molecule is an amphiphilic molecule;

[0165] -R3 and -R4 are independently selected from: -CN, -COOR, -CONR'R", -H, halogen, C1-C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group or heteroalkyl group is O or N.

[0166] in, etc. all represent the connection site between the drug group and other structures, that is, half of a single bond “-”.

[0167] The high-energy ray-responsive lipid molecules of the present invention have lipid functions, so that the high-energy ray-responsive lipid molecules can self-assemble to form high-energy ray-responsive liposomes. The interior of the high-energy ray-responsive liposomes can be coated with drugs. Under the irradiation of high-energy rays (for example, α rays, β rays, γ rays, and X-rays), the azo groups undergo structural transformation, the hydrophilic groups and the hydrophobic groups separate, and the high-energy ray-responsive liposome structure decomposes, thereby releasing the internally coated drugs, thereby achieving radiotherapy and drug chemotherapy simultaneously.

[0168] The high-energy ray-responsive lipid molecule of the present invention can be prepared by connecting a hydrophilic group and a hydrophobic group at the hydroxyl end and the carboxyl end of an azo compound through a substitution reaction and a condensation reaction, respectively.

[0169] It should be pointed out that the specific structure of the aromatic or heteroaromatic group in the high-energy ray-responsive lipid molecule is as described above and will not be repeated here.

[0170] Preferably, in this embodiment, -R a and -R b Each independently selected from C1 to C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group or heteroalkyl group is O or N.

[0171] Preferably, in this embodiment, -R, -R' and -R" are independently selected from C1 to C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group or heteroalkyl group is O or N.

[0172] Preferably, the hydrophilic groups include cationic groups, such as tertiary amino groups, quaternary amino groups, etc.; anionic groups, such as carboxylic acid groups, sulfonic acid groups, phosphoric acid groups, sulfate groups, etc.; polar nonionic groups, such as hydroxyl groups, ether groups, amino groups, amide groups, etc.

[0173] Preferably, the hydrophobic group comprises C 10 ~C 20 hydrocarbon groups; hydrocarbon groups containing aromatic, ester, ether, amine, amide and other groups; hydrocarbon groups containing double bonds; it can also be a lipophilic polymer, such as polyoxypropylene, long-chain perfluoroalkyl, polysiloxane, etc.; it can also be a chemotherapeutic agent with hydrophobic effect, such as bufalin, camptothecin, etc.

[0174] Specifically, the lipid molecule is a fatty acid ester or amide derivative, a sterol ester or amide derivative, a glyceride, a glycerophospholipid or a glycolipid containing an azo structural unit.

[0175] The present invention also discloses a high-energy ray-responsive liposome according to an embodiment of the present invention, which is formed by self-assembly of the high-energy ray-responsive lipid molecules.

[0176] Specifically, the high-energy ray-responsive liposomes can be prepared by mixing the aqueous phase with the organic phase containing high-energy ray-responsive lipid molecules through the LNP intelligent synthesizer to prepare liposome nanoparticles (LNP).

[0177] The interior of the high-energy ray-responsive liposome of the present invention can be coated with drugs. Under the irradiation of high-energy rays (for example, α rays, β rays, γ rays, and X rays), the azo group undergoes structural transformation, the hydrophilic group and the hydrophobic group separate, and the high-energy ray-responsive liposome structure decomposes, thereby releasing the internally coated drugs, thereby achieving radiotherapy and drug chemotherapy at the same time.

[0178] The following are specific examples.

[0179] In a specific embodiment, the chromatographic conditions involved in the HPLC test process are shown in Table 1 below.

[0180] Table 1

[0181] The irradiation parameters used in the specific examples are as follows: the instrument used is a small animal X-ray irradiator, model RS2000-PRO-225, and the selected irradiation parameters are 225 kV, 17.7 mA, position 5, 60 Gy, and duration of 9-10 minutes. The sample is first dissolved in DMSO, with an appropriate amount of the surfactant Triton X-100 added, and then diluted with pure water to a concentration of 5-10 μM. The prepared solution is placed in a glass bottle and sonicated for 5-10 minutes. A 1 mL sample is then removed and the remaining liquid is purged with argon for 20 minutes, then sealed and irradiated at room temperature.

[0182] Example 1

[0183] Preparation of azo prodrug XP-106-MMAE-010:

[0184] 1) Preparation of azo compound ON-2, azo compound ON-3 and azo compound ON-4:

[0185] 1.1)

[0186] Procedure: p-Aminobenzoic acid (2.0 g) was dissolved in dry DCM (40 mL), and aqueous potassium persulfate solution (8.64 g, 1.57 eq) was added thereto and stirred at room temperature for 2 hours.

[0187] Detection: The reaction changes from white suspension to yellow suspension.

[0188] Post-treatment: The reaction mixture was directly filtered, the filter cake was rinsed with water 3-5 times, and the filter cake was dried to obtain the product ON-1 (2 g, 90% yield) as a yellow solid. (Used directly in the next step without purification)

[0189] 1.2)

[0190] Procedure: Dissolve ON-1 (2 g, 1.0 eq) in dry DMF (200 mL), add p-aminobenzyl alcohol (2 g, 1.1 eq) and glacial acetic acid (50 mL) to the stirred reaction solution, and stir at room temperature for 24 hours.

[0191] Detection: LCMS detection.

[0192] Post-treatment: The reaction solution was spin-dried to give the product ON-2 (4.6 g, crude) as a brown solid. (Used directly in the next step without purification)

[0193] 1.3)

[0194] Procedure: Dissolve ON-2 (2.3 g, 1.0 eq) in dry DCM (50 mL) and add sarcosine tert-butyl ester hydrochloride (1.95 g, 1.2 eq), DCC (2.32 g, 1.2 eq), HOBt (1.52 g, 1.2 eq) and triethylamine (2.6 mL, 2.0 eq) to the reaction. Stir the reaction at room temperature for 24 h.

[0195] Detection: LCMS showed that the starting material was completely reacted.

[0196] Post-treatment: the reaction was quenched with water, extracted with DCM (80 mL x 3), the combined organic phases were washed once with saturated brine, and spin-dried.

[0197] Purification: The product ON-3 (2.3 g, 66% yield) was isolated by column purification (EA:PE = 0:1, 1:10, 1:5, 1:2 to produce the product, 1:1) as an orange-yellow solid.

[0198] 1.4)

[0199] Procedure: ON-3 (1.3 g, 1.0 eq) was dissolved in dry DCM (26 mL), p-nitrophenylcarbonyl chloride (820 mg, 1.2 eq), DMAP (41 mg, 0.1 eq) and triethylamine (565 μL, 1.2 eq) were added to the reaction, and the reaction was stirred at room temperature for 3 h.

[0200] Detection: LCMS showed that the starting material was completely reacted.

[0201] Post-treatment: the reaction was quenched with water, extracted with DCM (80 mL x 3), the combined organic phases were washed once with saturated brine, and spin-dried.

[0202] Purification: The product ON-4 (1.3 g, 69% yield) was isolated by column purification (E:P = 0:1, 1:10, 1:5, 1:3, 1:2 to start the product, 1:1) as an orange-yellow semi-oil and semi-solid state.

[0203] 2) Preparation of azo prodrug XP-106-MMAE-010

[0204] Synthesis method:

[0205] Procedure: MMAE (350 mg, 1.0 eq) was dissolved in dry DMF (10 mL), ON-4 (580 mg, 1.8 eq), HOBt (188 mg, 2 eq) and triethylamine (100 μL, 2 eq) were added to the reaction, and the reaction was stirred at room temperature for 24 h.

[0206] Detection: LCMS showed the reaction was complete.

[0207] Post-treatment and purification: DMF was spin-dried, dissolved in DCM and purified by column (E:P = 0:1, 1:10, 1:5, 1:3, 1:1, 1:0) to give the product XP-106-MMAE-010 (350 mg, 63% yield) as a yellow solid.

[0208] 1 H NMR (400MHz, CDCl3) δ7.95(dd,J=25.6,13.2Hz,3H),7.62(d,J=8.0Hz,1H),7.52(dd,J=19.6,7.6Hz,2H),7.43-7.29(m,4H),7.24(t ,J=7.2Hz,1H),6.85(t,J=10.8Hz,1H),6.58(dd,J=24.0,8.4Hz,1H),5.38-5.06(m,2H),4.94(d,J=2.4Hz,1H),4.87-4.60(m,2H),4. 31-4.00(m,5H),3.86(dd,J=8.8,7.2Hz,1H),3.48(d,J=8.8Hz,3H),3.39(t,J=4.8Hz,4H),3.32(d,J=10.0Hz,3H),3.19-2.84(m,8H) ,2.52–2.15(m,4H),2.13-1.89(m,7H),1.82(dd,J=10.4,6.8Hz,2H),1.58-1.41(m,9H),1.26(t,J=10.4Hz,5H),1.08-0.65(m,21H).

[0209] LCMS(ESI):m / z Calcd for C 61 H 91 N8O 12 ,[M+H] + :1127.7,found:1128.1.

[0210] HPLC analysis: The HPLC peak of the original drug molecule MMAE appeared around 17.0 minutes. The prodrug peak area (HPLC 315 nm) decreased by 96.4% before and after irradiation (60 Gy, 10 μM). Irradiation produced the original drug MMAE (t = 17.02 minutes) and the detached linker 1_a (t = 20.491 minutes). The HPLC analysis results are shown in Table 2 below.

[0211] Table 2 HPLC test results of XP-106-MMAE-010 before and after irradiation

[0212] Conversion rate involved = (peak area of ​​azo prodrug before irradiation - peak area of ​​azo prodrug after irradiation) / peak area of ​​azo prodrug before irradiation.

[0213] Example 2

[0214] Preparation of azo prodrug XP-106-MMAE-011:

[0215] Procedure: MMAE-010 (330 mg) was dissolved in dry DCM (8 mL), 0.5 mL of TMSOTf was added to the reaction and stirred at room temperature for 30 min.

[0216] Detection: TLC detection (PE:EA=1:1).

[0217] Post-treatment: the reaction was quenched with NaHCO 3 , extracted with EA, washed with water, washed with salt, filtered, and dried to give a crude product (242 mg, 84% yield).

[0218] Purification: 20 mg was purified by Pre-HPLC to obtain XP-106-MMAE-011 (13 mg, 98.7% purity) as a yellow solid.

[0219] LCMS(ESI):m / z Calcd for C 57 H 83 O 12 N8,[M+H] + :1071.6,found:1072.0.

[0220] HPLC analysis: The HPLC peak time of the original drug molecule MMAE is approximately 17.0 minutes. The peak area (HPLC 315nm) of the prodrug decreased by 66.4% before and after irradiation (60Gy, 10uM), resulting in the original drug MMAE and linker peak 2_a (HPLC 254nm peak time of 15.793 minutes). The HPLC analysis results are shown in Table 3 below.

[0221] Table 3 HPLC test results of XP-106-MMAE-011 before and after irradiation

[0222] Example 3

[0223] Preparation of azo prodrug XP-107-BUF-009:

[0224] 1) Preparation of azo compound ON-5:

[0225] Procedure: 200 mg of compound ON-4 was dissolved in 4 mL of DCM / TFA (1 / 1) under ice bath, and then reacted at room temperature for two hours. TLC (DCM:MeOH=10:1) showed that the reaction of the raw material was complete, and a highly polar spot was generated.

[0226] Detection: TLC detection (DCM:MeOH=10:1).

[0227] Post-treatment: The reaction solution was rotary evaporated at 40°C to remove dichloromethane, and then pumped dry with an oil pump to obtain 140 mg of yellow solid ON-5 (used directly in the next step without purification).

[0228] 2) Preparation of azo compound ON-6:

[0229] Procedure: Dissolve 120 mg of compound ON-5 in 5 mL of anhydrous DCM, add 51 mg of piperazine and 72 uL of TEA, and react at room temperature for 2 h.

[0230] Detection: TLC detection (PE:EA=1:1)

[0231] Post-treatment and purification: The product was spin-dried at 37°C and re-separated by wet column chromatography using dichloromethane (PE:EA = 1:0, 0:1), (EA:MeOH = 20:1, 10:1, 5:1) and 1 / 1000 ammonia solution to obtain product 6 (92.2 mg) as a yellow solid, ON-6.

[0232] 3) Preparation of compound 7:

[0233] Procedure: Dissolve 100 mg of bufalin in 1 mL of anhydrous DCM, add 120 mg of p-nitrophenyl chloroformate, add 38 mg of DMAP and 60 μL of TEA at 0°C, and react at room temperature for 24 h.

[0234] Detection: TLC detection (PE:EA=1:1)

[0235] Post-treatment and purification: The product was dried by rotary evaporation at 37°C, re-dissolved in dichloromethane, and separated by wet column chromatography (PE:EA = 1:0, 5:1, 3:1, 1:1). The product, compound 7 (110 mg, 77.1% yield), was obtained as a white solid.

[0236] 2) Preparation of azo prodrug XP-107-BUF-009:

[0237] Procedure: 100 mg of compound 7 was dissolved in 3 mL of DCM at room temperature, and then 92 mg of compound 2 was added. The reaction was allowed to proceed overnight at room temperature.

[0238] Detection: LCMS showed that the reaction of compound 2 was complete and product MS was generated.

[0239] Post-treatment and purification: After the reaction is complete, add 10 mL of water and 20 mL of dichloromethane to precipitate a solid. Filter and collect the filter cake. The filtrate is extracted with dichloromethane. The organic phase is dried over anhydrous sodium sulfate and the dichloromethane is removed by rotary evaporation at 40°C to yield a yellow solid. The filter cake and solid are combined. The product is analyzed by TLC (DCM / MeOH = 10:1, 1 / 1000 formic acid) and pumped dry using an oil pump. 4.0 mg (99% purity) of XP-107-BUF-0096 was obtained as a yellow solid.

[0240] LCMS(ESI):m / z Calcd for C 47 H 58 O 10 N5,[M+H] + :852.4,found:852.2.

[0241] HPLC analysis: The HPLC peak time of the original drug molecule 3-2-3 was approximately 17.2 minutes. The prodrug peak area (HPLC 315 nm) decreased by 88.4% before and after irradiation (60 Gy, 10 μM), resulting in the original drug 3-2-3 (315 nm) and the linker peak 2_a (HPLC 254 nm peak time of 15.796 minutes). The HPLC analysis results are shown in Table 4 below.

[0242] Table 4 HPLC test results of XP-107-BUF-009 before and after irradiation

[0243] Example 4

[0244] Preparation of azo prodrug XP-101-DAU-002:

[0245] Procedure: Dissolve 50 mg of DAU in 2 mL of anhydrous DMF, add 58 mg of ON-4, 14.3 mg of HOBt and 37 uL of TEA, and react at room temperature for 24 h.

[0246] Detection: TLC detection (DCM:MeOH=10:1)

[0247] Post-treatment and purification: The product was spin-dried at 37°C, redissolved in dichloromethane, and separated on a silica gel plate (DCM:MeOH=10:1). The product XP-101-DAU-002 (22.7 mg) was obtained as a red solid.

[0248] LCMS(ESI):m / z Calcd for C 49 H 53 O4N 15 ,[M+H] + :937.3,found:937.1.

[0249] HPLC analysis: The HPLC peak time of the original drug molecule DAU was 17.2 min. The prodrug peak area (HPLC 315 nm) decreased by 50.1% before and after irradiation (60 Gy, 10 μM), indicating the production of the original drug DAU and linker 1_a. The HPLC analysis results are shown in Table 5 below.

[0250] Table 5 HPLC test results of XP-101-DAU-002 before and after irradiation

[0251] Example 5

[0252] Preparation of azo prodrug XP-104-CPT-010:

[0253] Procedure: 200 mg of CPT was dissolved in 5 mL of DCM, and 200 mg of compound 3, 20 mg of DMAP and 40 μL of TEA were added, and the mixture was reacted at room temperature for 24 h.

[0254] Detection: TLC detection (PE:EA=1:1)

[0255] Post-treatment and purification: The product was dried by spin drying at 37°C, redissolved in dichloromethane, and separated on a silica gel plate (DCM:MeOH=30:1). The product CPT_1 (55.6 mg) was obtained as a yellow solid.

[0256] Synthesis method:

[0257] Procedure: Dissolve 55.6 mg of CPT_1 in 2 mL of DCM, add 50 mg of ON-3 and 30 μL of TEA, and react at room temperature for 24 h.

[0258] Detection: TLC detection (EA)

[0259] Post-treatment and purification: spin-dry at 37°C, redissolve in MeOH and separate by preparative HPLC to obtain the product XP-104-CPT-010 (10 mg) as a white solid.

[0260] LCMS(ESI):m / z Calcd for C 42 H 40 O9N5,[M+H] + :758.3,found:758.1.

[0261] HPLC analysis: The HPLC peak time of the original drug molecule was 18.5 min. The prodrug peak area (HPLC 315 nm) decreased by 61.1% before and after irradiation (60 Gy, 10 μM), yielding the original drug camptothecin and linker compound 1_a. The HPLC analysis results are shown in Table 6 below.

[0262] Table 6 HPLC test results of XP-104-CPT-010 before and after irradiation

[0263] Example 6

[0264] Azo prodrug XP-109-EXA-002:

[0265] Procedure: Dissolve 15 mg of isotecan in 0.5 mL of DMF and 0.5 mL of DCM, add ON-4 (23 mg) and 5 mg of DMAP, and triethylamine (9 uL) to the reaction mixture. React at room temperature for 48 hours.

[0266] Detection: LCMS detection showed that the reaction of the starting material was incomplete (about 40% remained).

[0267] Post-treatment and purification: Preparative HPLC separation gave 2 mg of yellow solid XP-109-EXA-002.

[0268] 1 H NMR(400MHz,d6-DMSO)δ8.48(s,1H),8.22(d,J=9.2Hz,1H),8.16-7.92(m,4H),7.87-7.47(m,4H),7 .35(d,J=20.0Hz,1H),6.53(s,1H),5.45(s,1H),5.37-5.19(m,4H),4.20(d,J=26.4Hz,1H),3.00(d, J=16.7Hz,2H),2.39(s,2H),2.31-2.15(m,1H),2.00(dd,J=15.2,7.2Hz,2H),1.86(tt,J=14.0,7.2 Hz,1H),1.75(s,1H),1.47(s,3H),1.37(d,J=12.8Hz,3H),1.24(s,9H),0.85(dd,J=7.2,3.2Hz,3H).

[0269] LCMS(ESI):m / z Calcd for C 46 H 46 O9N6F1,[M+H] + :845.3,found:845.1.

[0270] HPLC analysis: The HPLC peak time of the original drug molecule was approximately 15.2 minutes. The peak area (HPLC 254nm) of the prodrug decreased by -% before and after irradiation (60 Gy, 10 μM), but no new peaks were observed. The HPLC analysis results are shown in Table 7 below.

[0271] Table 7 HPLC test results of XP-109-EXA-002 before and after irradiation

[0272] Example 7

[0273] Preparation of azo prodrug XP-111-RES-004:

[0274] Procedure: RES (50 mg) was dissolved in dry DMF (40 mL) and stirred. To the stirred solution were added ON-4 (96 mg), HOBt (30 mg) and triethylamine (66 μL), and the mixture was stirred at room temperature for 24 hours.

[0275] Detection: LCMS detection, the raw material has not reacted completely.

[0276] Workup and purification: The product (4 mg, 3% yield) was directly obtained as a yellow solid.

[0277] 1 H NMR (400MHz, CDCl3) δ8.46(s,1H),8.23(s,1H),8.17(d,J=8.4Hz,1H),7.98(d,J=8.4H z,3H),7.70-7.63(m,3H),7.58(d,J=7.6Hz,1H),7.50(d,J=8.0Hz,1H),7.43-7.38(m,1 H),5.43(s,2H),4.93(s,2H),4.81(s,2H),4.22(s,1H),3.90(s,1H),3.68(q,J=6.8Hz ,2H),3.12(d,J=36.8Hz3H),1.55(s,5H),1.49(s,4H),1.35(s,5H),1.30-1.24(m,4H).

[0278] LCMS(ESI):m / z Calcd for C 39 H 46 O7N7,[M+H] + :724.3,found:724.2.

[0279] HPLC analysis: The peak time of the original drug molecule (HPLC (254 nm) was approximately 15.1 minutes. The peak area of ​​the prodrug (HPLC 315 nm) decreased by 27.1% before and after irradiation (60 Gy, 10 μM), yielding the original resiquimod drug and linker 1_a. The HPLC analysis results are shown in Table 8 below.

[0280] Table 8 HPLC test results of XP-111-RES-004 before and after irradiation

[0281] Example 8

[0282] Preparation of azo prodrug XP-113-PTX-002:

[0283] Procedure: Dissolve 25 mg of paclitaxel in 2 mL of DCM, add 20 mg of ON-4, then add 3 mg of DMAP and 7 uL of TEA, and react at room temperature for 24 h.

[0284] Detection: LCMS detection, the raw material has not reacted completely.

[0285] Post-treatment and purification: After removing a portion of the solvent at 37°C, the product was separated on a large silica gel plate (DCM:MeOH=20:1). The obtained solid contained the raw material, which was then separated by HPLC to obtain the product XP-113-PTX-002 as a yellow solid (7 mg).

[0286] 1 H NMR(400MHz, CDCl3) δ8.15(d,J=7.6Hz,2H),7.94(dd,J=12.4,8.4Hz,4H),7.73( d,J=7.6Hz,2H),7.67-7.46(m,7H),7.46-7.32(m,6H),6.88(dd,J=33.2,8.4Hz,2 H),6.38-6.23(m,2H),6.01(dd,J=9.2,2.4Hz,1H),5.70(d,J=7.2Hz,1H),5.48(d ,J=2.4Hz,1H),5.36-5.18(m,2H),4.98(d,J=8.4Hz,1H),4.45(s,1H),4.32(d,J= 8.4Hz,1H),4.21(d,J=8.4Hz,2H),3.88(s,1H),3.83(d,J=7.2Hz,1H),3.72(dd, J=14.0,6.8Hz,1H),3.15(s,1H),3.05(s,1H),2.63-2.52(m,1H),2.51-2.43(m,3 H),2.43-2.35(m,1H),2.26-2.18(m,3H),2.06-1.98(m,1H),1.96-1.87(m,3H),1 .87-1.79(m,1H),1.69(s,3H),1.59(s,9H),1.50(d,J=22.0Hz,6H),1.14(s,3H).

[0287] LCMS(ESI):m / z Calcd for C 69 H 75 N4O 19 ,[M+H] + :1263.5,found:1263.1.

[0288] HPLC analysis: The peak time of the original drug molecule at HPLC (254 nm) was approximately 24.006 min. The peak area of ​​the prodrug (HPLC (315 nm)) decreased by 39.2% before and after irradiation (60 Gy, 10 μM), yielding the paclitaxel original drug and linker 1_a. The HPLC analysis results are shown in Table 9 below.

[0289] Table 9 HPLC test results of XP-113-PTX-002 before and after irradiation

[0290] Example 9

[0291] Preparation of azo prodrug XP-102-DOX-011:

[0292] Procedure: Dissolve 50 mg of DOX in 2 mL of anhydrous DMF, add 55 mg of ON-4, 15 mg of HOBt and 37 μL of TEA, and react at room temperature for 24 h.

[0293] Detection: TLC detection (DCM:MeOH=10:1)

[0294] Post-treatment and purification: The product was spin-dried at 37°C, redissolved in dichloromethane, and separated on a silica gel plate (DCM:MeOH=10:1). The product XP-102-DOX-011 (11 mg) was obtained as a red solid.

[0295] LCMS(ESI):m / z Calcd for C 49 H 53 O 16 N4,[M+H] + :953.3,found:952.9.

[0296] 1 H NMR (400MHz, CDCl3) δ13.99(d,J=9.1Hz,1H),13.24(s,1H),8.03(d,J=7.6Hz,1H),7.96–7.69(m,5H),7.58(dd,J=30.0,8.0Hz,2H ),7.42(dd,J=26.1,8.2Hz,3H),5.51(d,J=3.5Hz,1H),5.36–5.26(m,1H),4.77(d,J=11.8Hz,2H),4.16(dd,J=17.8,11.3Hz,2H),4 .07(s,3H),3.87(s,2H),3.68(s,1H),3.28(d,J=18.2Hz,1H),3.14(s,1H),3.08–2.92(m,3H),2.34(d,J=14.8Hz,1H),2.23–2.10( m,1H),2.06–1.98(m,1H),1.96–1.86(m,1H),1.84–1.74(m,2H),1.68(s,3H),1.47(dd,J=24.4,14.2Hz,9H),1.29(d,J=6.5Hz,3H)

[0297] HPLC analysis: The HPLC peak time of the original drug molecule DOX was approximately 16.2 minutes. The prodrug peak area (HPLC 315nm) decreased by 95.2% before and after irradiation (60 Gy, 5 μM), indicating the production of the original drug DOX and linker 1_a. The HPLC analysis results are shown in Table 10 below.

[0298] Table 10 HPLC test results of XP-102-DOX-011 before and after irradiation

[0299] Example 10

[0300] Preparation of azo prodrug XP-107-BUF-008

[0301] 1) Preparation of azo compound YJY-2-67:

[0302] 1.1)

[0303] Procedure: ON-2 (200 mg) was dissolved in DCM (5 mL), 200 mg DCC, 120 mg HOBt and 200 uL TEA were added, and then 80 mg of compound 2 was added, and the mixture was stirred at room temperature for 24 hours.

[0304] Detection: TLC detection (DCM:MeOH=10:1).

[0305] Post-treatment: spin-dry, dissolve with DCM:MeOH=5:1 solution, apply to a large plate, and then climb with DCM:MeOH=10:1. Scrape the yellow band in the middle to obtain the product YJY-2-61 (180 mg) as a yellow solid.

[0306] 1.2)

[0307] Procedure: 120 mg of YJY-2-61 was dissolved in DCM (2 mL), 10 mg of DMAP and 130 uL of TEA were added, and then 60 mg of compound 3 was added, and the mixture was stirred at room temperature for 24 hours.

[0308] Detection: TLC detection (DCM:MeOH=10:1).

[0309] Post-treatment: The product mixture was used directly in the next step without purification.

[0310] 2) Preparation of azo prodrug XP-107-BUF-008:

[0311] Synthesis method:

[0312] Procedure: Add 32 mg of 3-2-3 to the reaction mixture YJY-2-67 from the previous step and react at room temperature for 24 h.

[0313] Detection: TLC detection (DCM:MeOH=10:1)

[0314] Post-treatment and purification: Concentrate at 37°C and separate using a large silica gel plate (DCM:MeOH=10:1) to obtain the product YJY-2-69 (40 mg) as a yellow solid.

[0315] LCMS(ESI):m / z Calcd for C 48 H 63 O8N6,[M+H] + :851.5,found:851.6

[0316] Procedure: 40 mg of YJY-2-69 was dissolved in 2 mL of DCM, 0.5 mL of iodomethane was added, and the mixture was reacted at room temperature for 16 h.

[0317] Detection: LCMS detection.

[0318] Post-treatment and purification: A yellow solid was produced during the reaction, and the precipitate was collected by centrifugation. The precipitate was washed with DCM and PE in sequence to obtain the product XP-107-BUF-008 (40 mg) as a yellow solid.

[0319] LCMS(ESI):m / z Calcd for C 49 H 65 O8N6 + ,[M] + :865.5,found:865.1.

[0320] 1H NMR(400MHz,d6-DMSO)δ8.98(t,J=5.6Hz,1H),8.13–7.79(m,6H),7.66–7.47(m,3H),6.29(d,J=9.8Hz,1H),5.75(s, 1H),5.22(s,2H),4.90(s,1H),4.15(s,1H),3.73(d,J=6.0Hz,1H),3.53(t,J=6.5Hz,1H),3.39(d,J=14.6Hz,8H),3.1 5(d,J=7.6Hz,9H),2.86(d,J=8.3Hz,2H),2.67(s,1H),2.33(s,1H),2.13–1.89(m,4H),1.77(d,J=10.2Hz,2H),1.58 (t,J=21.8Hz,9H),1.34(t,J=12.0Hz,4H),1.19(ddd,J=36.9,18.4,11.7Hz,5H),0.91(d,J=4.2Hz,3H),0.60(s,3H).

[0321] HPLC analysis revealed that the HPLC peak of the prodrug molecule 3-2-3 appeared at approximately 17.142 min. The prodrug peak area (HPLC 315 nm) decreased by 88.4% before and after irradiation (60 Gy, 10 μM), indicating the production of the prodrug molecule 3-2-3. The HPLC analysis results are shown in Table 11.

[0322] Table 11 HPLC test results of XP-107-BUF-008 before and after irradiation

[0323] Example 11

[0324] Preparation of azo prodrug XP-102-DOX-012:

[0325] Procedure: Dissolve 50 mg of DOX in 2 mL of anhydrous DMF, add 76 mg of ON-5, 30 mg of HOBt and 30 μL of TEA, and react at room temperature for 24 h.

[0326] Detection: TLC detection (DCM:MeOH=10:1)

[0327] Post-treatment and purification: The product was dried by spin drying at 37°C, redissolved in dichloromethane, and separated on a large silica gel plate (DCM:MeOH=10:1). The product XP-102-DOX-012 (73 mg) was obtained as a red solid.

[0328] LCMS(ESI):m / z Calcd for C 45H 45 O 16 N4,[M+H] + :897.3,found:896.9.

[0329] 1 H NMR(400MHz,d6-DMSO)δ13.97(d,J=38.1Hz,1H),13.26(s,1H),8.07–7.76(m,5H),7.76–7.34(m,5H),7.01(t,J= 19.3Hz,1H),5.47(s,1H),5.28(d,J=34.4Hz,1H),5.08(q,J=13.7Hz,2H),4.98–4.69(m,3H),4.58(s,2H),4.18( s,2H),3.92(d,J=33.3Hz,3H),3.78(d,J=24.8Hz,1H),3.45(d,J=21.8Hz,2H),3.10–2.83(m,5H),2.73(s,1H),2 .28–2.06(m,2H),1.88(dd,J=12.6,9.4Hz,1H),1.50(d,J=8.1Hz,1H),1.22(d,J=10.0Hz,1H),1.21–1.06(m,3H).

[0330] Example 12

[0331] Preparation of azo prodrug XP-102-DOX-013:

[0332] Procedure: Dissolve 73 mg of XP-102-DOX-012 in 2 mL of anhydrous DMF, add 40 mg of compound 5 ((2R,3R,4R,5R,6S)-2-(2-aminoethoxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol), 20 mg of HATU and 20 uL of DIPEA, and react at room temperature for 2 h.

[0333] Detection: LCMS detection

[0334] Post-treatment and purification: Filter through a 0.22 μM filter membrane and then prepare and purify the product XP-102-DOX-013 (26 mg) as a red solid.

[0335] LCMS(ESI):m / z Calcd for C 53 H 59 O 21 N5Na,[M+Na] +:1124.4,found:1123.8.

[0336] 1 H NMR (400MHz, d6-DMSO) δ14.20–13.92(m,1H),13.32–13.07(m,1H),8.05(s,1H),7.91(dd,J=16.4,5.3Hz,4H),7.58(dd,J=31.7,21. 4Hz,5H),7.03(d,J=8.1Hz,1H),5.76(s,1H),5.47(s,1H),5.24(s,1H),5.08(s,2H),4.88(dd,J=28.4,22.3Hz,3H),4.80–4.46(m,6H ),4.24–4.07(m,3H),4.01(d,J=20.5Hz,3H),3.84(s,1H),3.74(s,1H),3.60(s,3H),3.17(d,J=5.2Hz,2H),2.97(d,J=6.3Hz,5H),2 .33(s,1H),2.26–2.07(m,2H),1.99(s,1H),1.92–1.82(m,1H),1.76(s,1H),1.47(s,2H),1.23(s,2H),1.16(dd,J=20.6,6.7Hz,3H).

[0337] HPLC analysis: The HPLC peak time of the original drug molecule DOX was approximately 16.2 minutes. The prodrug peak area (HPLC 254nm) decreased by 22.1% before and after irradiation (24 Gy, 5 μM), indicating the production of the original drug DOX. The HPLC analysis results are shown in Table 12 below.

[0338] Table 12 HPLC test results of XP-102-DOX-013 before and after irradiation

[0339] Example 13

[0340] Preparation of azo prodrug XP-106-MMAE-019

[0341] 1) Preparation of azo compounds:

[0342] 1.1)

[0343] Procedure: 100 mg of 4-amino-3-methoxybenzoic acid was dissolved in dry DCM (2 mL), and aqueous potassium persulfate solution (307.38 mg, 1.6 eq) was added thereto and stirred at room temperature for 2 hours.

[0344] Detection: LC-MS detection.

[0345] Post-treatment: water was added to the reaction mixture and the product was filtered to obtain 74.5 mg of a yellow solid with a yield of 68.8%.

[0346] 1.2)

[0347] Procedure: 2-MZL-4 (74.5 mg, 1.0 eq) was dissolved in dry DMF (3 mL). p-Aminobenzyl alcohol (55.4 mg, 1.1 eq) and formic acid (3 mL) were added to the stirred reaction solution and stirred at room temperature for 24 hours.

[0348] Detection: LC-MS detection.

[0349] Work-up: extraction with EA and H2O three times to remove DMF to give the product 2-MZL-5 (114.8 mg, crude) as a brown solid.

[0350] 1.3)

[0351] Procedure: 2-MZL-5 (114.8 mg, 1.0 eq) was dissolved in dry DCM (5 mL). Sarcosine tert-butyl ester hydrochloride (87.2 mg, 1.2 eq), DCC (99.04 mg, 1.2 eq), HOBt (64.9 mg, 1.2 eq), and triethylamine (167 μL, 3.0 eq) were added to the reaction. The reaction was stirred at room temperature for 24 h.

[0352] Detection: LC-MS detection.

[0353] Post-treatment: The reaction solution was purified by silica gel chromatography to obtain 37.7 mg of an orange-yellow solid with an eluent ratio of (EA:PE=1:5) and a yield of 22.8%.

[0354] 1.4)

[0355] Procedure: 2-MZL-O-1 (37.7 mg, 1.0 eq) was dissolved in dry DCM (2 mL). p-Nitrophenylcarbonyl chloride (27.2 mg, 1.5 eq), DMAP (22 mg, 2 eq) and triethylamine (38 μL, 3 eq) were added to the reaction. The reaction was stirred at room temperature for 16 h.

[0356] Detection: LC-MS detection.

[0357] Post-treatment: The reaction solution was purified by silica gel chromatography to obtain 41.3 mg of an orange-yellow solid with an eluent ratio of (EA:PE=1:5) and a yield of 80.5%.

[0358] 2) Preparation of azo prodrug XP-106-MMAE-019:

[0359] Procedure: MMAE (57 mg, 1.1 eq) was dissolved in dry DCM (2 mL), 2-MZL-Cl-2 (41.3 mg, 1 eq), HOBt (18.9 mg, 2 eq) and triethylamine (20 μL, 2 eq) were added to the reaction, and the reaction was stirred at room temperature for 24 h.

[0360] Detection: LC-MS detection.

[0361] Post-treatment: DCM was spin-dried with a water pump, dissolved in DMF and MeOH, and purified by semi-preparative purification to obtain 21 mg of an orange-yellow solid with a yield of 24.7%.

[0362] LCMS(ESI):m / z Calcd for C 62 H 93 N8O 13 ,[M+H] + :1157.7,found:1157.1.

[0363] HPLC analysis: The HPLC peak of the original drug molecule MMAE appeared around 17.0 minutes. The prodrug peak area (HPLC 315 nm) decreased by 45.2% before and after irradiation (18 Gy, 5 μM). Irradiation produced the original drug MMAE (t = 16.957 minutes) and the dropped linker 4_a (t = 21.125 minutes). HPLC analysis results are shown in Table 13 below.

[0364] Table 13 HPLC test results of XP-106-MMAE-019 before and after irradiation

[0365] Example 14

[0366] Preparation of azo prodrug XP-106-MMAE-020:

[0367] 1) Preparation of azo compound ON-2F-2, azo compound ON-2F-3 and azo compound ON-2F-4:

[0368] 1.1)

[0369] Procedure: The starting material 6 (500 mg) was dissolved in dry DCM (10 mL), and aqueous potassium persulfate solution (600 mg, 1.5 eq) was added thereto and stirred at room temperature for 2 hours.

[0370] Detection: LC-MS detection.

[0371] Post-treatment: Filtration gave the solid product ON-2F-1 (520 mg, 96% yield) as a white solid.

[0372] 1.2)

[0373] ON-2F-1 (260 mg, 1.0 eq) was dissolved in dry DMF (10 mL). p-Aminobenzyl alcohol (256 mg, 1.5 eq) and glacial ethyl (15 mL) were added to the stirred reaction solution, and the mixture was stirred at room temperature for 24 hours.

[0374] Detection: LC-MS detection.

[0375] Post-treatment: the reaction was quenched with water, extracted with EA, and dried in spun-drying to give the product ON-2F-2 (380 mg, crude) as a brown oil.

[0376] 1.3)

[0377] Procedure: Dissolve ON-2F-2 (200 mg, 1.0 eq) in dry DCM (4 mL), add sarcosine tert-butyl ester hydrochloride (149 mg, 1.2 eq), HATU (312 mg, 1.2 eq) and DIPEA (177 mg, 2 eq) to the reaction, and stir at room temperature for 4 h.

[0378] Detection: LC-MS detection.

[0379] Post-treatment: The reaction solution was separated by prep-TLC (E:P=1:2.5) to obtain the product ON-2F-3 (47 mg, 17% yield) as an orange oily solid.

[0380] 1.4)

[0381] Procedure: ON-2F-3 (47 mg, 1.0 eq) was dissolved in dry DCM (2 mL), p-nitrophenylcarbonyl chloride (26 mg, 1.2 eq), DMAP (4 mg, 0.1 eq) and triethylamine (30 μL, 2 eq) were added to the reaction, and the reaction was stirred at room temperature for 24 h.

[0382] Detection: LC-MS detection.

[0383] Post-treatment: The reaction solution was separated by prep-TLC (EA:PE=1:2) to obtain the product ON-2F-4 (60 mg, 90% yield) as a yellow solid.

[0384] 2) Preparation of azo prodrug XP-106-MMAE-020:

[0385] Procedure: MMAE (50 mg, 1.0 eq) was dissolved in dry DMF (2 mL), ON-4 (60 mg, 1.5 eq), HOBt (19 mg, 2 eq) and triethylamine (19 μL, 2 eq) were added to the reaction, and the reaction was stirred at room temperature for 24 h.

[0386] Detection: LC-MS detection.

[0387] Post-treatment: DMF was spin-dried, dissolved in MeOH and separated by prep-TLC (M:D=1:10) to obtain the product MMAE-020 (17 mg, 20% yield) as a yellow solid.

[0388] LCMS(ESI):m / z Calcd for C 61 H 89 N8O 12 F2,[M+H] + :1163.6,found:1163.0.

[0389] 1 H NMR (400MHz, CDCl3) δ7.94(d,J=8.1Hz,1H),7.54(t,J=11.2Hz,1H),7.44–7.30(m,4H),7.27–7.09(m,2H),6.96(dd,J=15.9,7.7H z,1H),6.57(s,1H),5.24(ddt,J=44.5,30.9,8.7Hz,2H),4.97(s,1H),4.80–4.62(m,1H),4.27(d,J=6.7Hz,1H),4.13(dd,J=27.5 ,16.5Hz,4H),3.94–3.70(m,2H),3.50(dd,J=12.5,5.9Hz,1H),3.46–3.30(m,6H),3.16–2.88(m,9H),2.54–2.17(m,4H),2.03(dd ,J=48.6,16.9Hz,4H),1.91–1.60(m,7H),1.49(dd,J=23.5,12.8Hz,8H),1.33(d,J=13.4Hz,2H),1.27(s,6H),1.07–0.71(m,20H).

[0390] HPLC analysis: The HPLC peak time of the original drug molecule MMAE is approximately 17.0 minutes. The prodrug peak area (HPLC 315nm) decreased by 47.5% before and after irradiation (18 Gy, 5 μM). After irradiation, the original drug MMAE was generated (t = 16.942 minutes). The HPLC analysis results are shown in Table 14 below.

[0391] Table 14 HPLC test results of XP-106-MMAE-020 before and after irradiation

[0392] Example 15

[0393] Preparation of azo prodrug XP-106-MMAE-023:

[0394] 1) Preparation of azo compound HJA-ON-2, azo compound HJA-ON-3 and azo compound HJA-ON-4:

[0395] 1.1)

[0396] Procedure: ON-1 (39.0 mg, 2.0 eq) was dissolved in dry DMF (1 mL). 4-amino-3-methoxybenzyl alcohol (20 mg, 1.0 eq) and glacial acetic acid (1 mL) were added to the stirred reaction solution and stirred at room temperature for 24 hours.

[0397] Detection: LC-MS detection.

[0398] Post-treatment: The reaction solution was spin-dried to give the product HJA-ON-2 (37.3 mg, crude) as a yellow solid.

[0399] 1.2)

[0400] Procedure: HJA-ON-2 (37.3 mg, 1.0 eq) was dissolved in dry DCM (2 mL). Sarcosine tert-butyl ester hydrochloride (22.0 mg, 1.2 eq), DCC (25.0 mg, 1.2 eq), HOBT (16.2 mg, 1.2 eq), and triethylamine (17 uL, 2.0 eq) were added to the reaction. The reaction was stirred at room temperature for 24 h.

[0401] Detection: LC-MS detection.

[0402] Post-treatment: spin-drying, p-TLC (PE:EA = 1:1) purification and isolation of the product HJA-ON-3 (13.8 mg) as a yellow solid.

[0403] 1.3)

[0404] Procedure: HJA-ON-3 (13.8 mg, 1.0 eq) was dissolved in dry DCM (2 mL), p-nitrophenylcarbonyl chloride (8.04 mg, 1.2 eq), DMAP (1.00 mg, 0.1 eq) and triethylamine (5.6 μL, 1.2 eq) were added to the reaction, and the reaction was stirred at room temperature for 3 h.

[0405] Detection: LC-MS detection.

[0406] Post-treatment: spin-drying, and purification by prep-TLC (PE:EA=1:1) to isolate the product ON-4 (10.0 mg, 50% yield) as a yellow solid.

[0407] 2) Preparation of azo prodrug XP-106-MMAE-023

[0408] Procedure: MMAE (9.00 mg, 1.0 eq) was dissolved in dry DMF (1 mL), HJA-ON-4 (10.0 mg, 2.0 eq), HOBt (3.39 mg, 2 eq) and triethylamine (3.5 μL, 2 eq) were added to the reaction, and the reaction was stirred at room temperature for 24 h.

[0409] Detection: LC-MS detection.

[0410] Post-treatment: DMF was spin-dried, dissolved in DCM, and purified by prep-TLC (EA=100%) to isolate the product XP-106-MMAE-023 (4.20 mg, 28% yield, 95% purity) as a yellow solid.

[0411] LCMS(ESI):m / z Calcd for C 62 H 93 N8O 13 ,[M+H] + :1157.7,found:1157.1.

[0412] HPLC analysis: The HPLC peak of the original drug molecule, MMAE, was observed at approximately 17.0 min. The prodrug peak area (HPLC 315 nm) decreased by 36.6% before and after irradiation (18 Gy, 5 μM). After irradiation, the original drug, MMAE, was generated (t = 16.954 min). The HPLC analysis results are shown in Table 15 below.

[0413] Table 15 HPLC test results of XP-106-MMAE-023 before and after irradiation

[0414] Example 16

[0415] 1) Preparation of azo compound ON-3-Cl:

[0416] Procedure: To a solution of trichloromethyl chloroformate (158 mg, 0.8 mmol) in dry tetrahydrofuran (15 mL) at 0°C was added approximately ON-3 (383 mg) dissolved in dry THF (40 mL) and TEA (102 mg), and stirred at room temperature for 2 hours.

[0417] Detection: TLC (PE:EA=1:1).

[0418] Work-up: the resulting suspension was filtered, the filter cake was washed with a little tetrahydrofuran, and the combined organic phases were concentrated in vacuo to give ON-3-Cl (200 mg) as a crude yellow oil.

[0419] 2) Preparation of azo compound XP-106-MMAE-010:

[0420] Procedure: MMAE (50 mg, 1.0 eq) was dissolved in DCM (1 mL), ON-3-Cl (50 mg) was added to the reaction, and the reaction was stirred at room temperature for 3 h.

[0421] Detection: LCMS detection.

[0422] Post-treatment and purification: DCM was partially concentrated and then purified by prep-TLC (EA=100%) to isolate the product XP-106-MMAE-010 (10 mg) as a yellow solid.

[0423] LCMS(ESI):m / z Calcd for C 61 H 91 N8O 12 ,[M+H] + :1127.7,found:1128.1.

[0424] Example 17

[0425] 1) Preparation of azo compound ON-3-IMZ:

[0426] Workup: ON-3 (383 mg) was dissolved in DCM (40 mL) at room temperature, CDI (162 mg) was added and stirred at room temperature for 2 hours.

[0427] Detection: TLC (PE:EA=1:1).

[0428] Post-treatment: spin-drying and purification by prep-TLC gave ON-3-IMZ (300 mg) as a yellow solid.

[0429] 2) Preparation of azo compound XP-106-MMAE-010:

[0430] Procedure: MMAE (50 mg, 1.0 eq) was dissolved in DCM (1 mL), ON-3-IMZ (50 mg) was added to the reaction, and the reaction was stirred at room temperature for 3 h.

[0431] Detection: LCMS detection.

[0432] Post-treatment and purification: DCM was partially concentrated and then purified by prep-TLC (EA=100%) to isolate the product XP-106-MMAE-010 (10 mg) as a yellow solid.

[0433] LCMS(ESI):m / z Calcd for C 61 H 91 N8O 12 ,[M+H] + :1127.7,found:1128.1.

[0434] Example 18

[0435] 1) Preparation of azo compound 10, azo compound 11 and azo compound 12:

[0436] 1.1)

[0437] Procedure: 5-Amino-2-pyridinecarboxylic acid (100 mg) was dissolved in dry DCM (2 mL), and aqueous potassium hydrogen persulfate solution (450 mg) was added thereto and stirred at room temperature for 2 hours.

[0438] Detection: The reaction changes from white suspension to yellow suspension.

[0439] Post-treatment: Filter and wash with water 3-5 times to obtain product 9 (80 mg) as a yellow solid. (Used directly in the next step without purification)

[0440] 1.2)

[0441] Procedure: 9 (80 mg, 1.0 eq) was dissolved in dry DMF (2 mL). 3-Amino-6-pyridinemethanol (70 mg) and glacial ethyl acetate (2 mL) were added to the stirred reaction solution and stirred at room temperature for 24 hours.

[0442] Detection: LCMS detection.

[0443] Post-treatment: The reaction solution was spun down to dryness and pumped dry with an oil pump to obtain product 10 (160 mg, crude) as a brown solid. (Used directly in the next step without purification)

[0444] 1.3)

[0445] Procedure: 10 (130 mg, 1.0 eq) was dissolved in dry DCM (5 mL), and sarcosine tert-butyl ester hydrochloride (80 mg), DCC (85 mg), HOBt (65 mg) and triethylamine (40 uL) were added to the reaction. The reaction was stirred at room temperature for 24 h.

[0446] Detection: LCMS showed that the reaction of the starting material was almost complete.

[0447] Post-treatment: the reaction was quenched with water, extracted with DCM (8 mL x 3), the combined organic phases were washed once with saturated brine, and spin-dried.

[0448] Purification: pre-TLC (EA:PE=2:1) ​​separated the product 11 (60 mg) as a yellow solid.

[0449] 1.4)

[0450] Procedure: 11 (60 mg, 1.0 eq) was dissolved in dry DCM (2 mL), p-nitrophenylcarbonyl chloride (38 mg, 1.2 eq), DMAP (6 mg) and triethylamine (30 μL) were added to the reaction, and the reaction was stirred at room temperature for 3 h.

[0451] Detection: TLC (EA:PE=2:1) ​​detection.

[0452] Post-treatment: the reaction was quenched with water, extracted with DCM (8 mL x 3), the combined organic phases were washed once with saturated brine, and spin-dried.

[0453] Purification: prep-TLC (EA:PE=1:1) separated the product 12 (30 mg) as an orange-yellow oily solid.

[0454] 2) Preparation of azo compound XP-106-MMAE-024:

[0455] Procedure: MMAE (50 mg, 1.0 eq) was dissolved in DMF (1 mL), 12 (50 mg), HOBt (33 mg) and triethylamine (30 μL) were added to the reaction, and the reaction was stirred at room temperature for 3 h.

[0456] Detection: LCMS detection.

[0457] Post-treatment and purification: DMF was concentrated and then purified by prep-TLC (EA=100%) to isolate the product XP-106-MMAE-024 (5 mg) as a yellow solid.

[0458] LCMS(ESI):m / z Calcd for C 59 H 89 N 10 O 12 ,[M+H] + :1129.7,found:1129.2.

[0459] Example 19

[0460] 1) Preparation of azo compound 15, azo compound 16 and azo compound 17:

[0461] 1.1)

[0462] Procedure: 4-Amino-2-furancarboxylic acid (100 mg) was dissolved in dry DCM (2 mL), and aqueous potassium persulfate solution (400 mg) was added thereto and stirred at room temperature for 2 hours.

[0463] Detection: The reaction changes from white suspension to brown suspension.

[0464] Post-treatment: Filter and wash with water 3-5 times to obtain product 14 (60 mg) as a yellow solid. (Used directly in the next step without purification)

[0465] 1.2)

[0466] Procedure: 14 (60 mg, 1.0 eq) was dissolved in dry DMF (2 mL). 3-Aminofuran-2-methanol (50 mg) and glacial acetic acid (2 mL) were added to the stirred reaction solution and stirred at room temperature for 24 hours.

[0467] Detection: LCMS detection.

[0468] Post-treatment: The reaction solution was spun down to dryness and pumped dry with an oil pump to obtain product 15 (120 mg, crude) as a brown solid. (Used directly in the next step without purification)

[0469] 1.3)

[0470] Procedure: 15 (100 mg, 1.0 eq) was dissolved in dry DCM (5 mL), and sarcosine tert-butyl ester hydrochloride (70 mg), DCC (75 mg), HOBt (60 mg) and triethylamine (30 uL) were added to the reaction. The reaction was stirred at room temperature for 24 h.

[0471] Detection: LCMS showed that the reaction of the starting material was almost complete.

[0472] Post-treatment: the reaction was quenched with water, extracted with DCM (8 mL x 3), the combined organic phases were washed once with saturated brine, and spin-dried.

[0473] Purification: pre-TLC (EA:PE=2:1) ​​separated the product 16 (40 mg) as a yellow solid.

[0474] 1.4)

[0475] Procedure: 16 (40 mg, 1.0 eq) was dissolved in dry DCM (2 mL), p-nitrophenylcarbonyl chloride (28 mg, 1.2 eq), DMAP (4 mg) and triethylamine (15 μL) were added to the reaction, and the reaction was stirred at room temperature for 3 h.

[0476] Detection: TLC (EA:PE=2:1) ​​detection.

[0477] Post-treatment: the reaction was quenched with water, extracted with DCM (8 mL x 3), the combined organic phases were washed once with saturated brine, and spin-dried.

[0478] Purification: prep-TLC (EA:PE=1:1) separated the product 17 (15 mg) as an orange-yellow oily solid.

[0479] 2) Preparation of azo compound XP-106-MMAE-025:

[0480] Procedure: MMAE (50 mg, 1.0 eq) was dissolved in DMF (1 mL), 12 (50 mg), HOBt (30 mg) and triethylamine (25 μL) were added to the reaction, and the reaction was stirred at room temperature for 3 h.

[0481] Detection: LCMS detection.

[0482] Post-treatment and purification: DMF was concentrated and purified by prep-TLC (EA=100%), and then separated by prep-HPLC to obtain the product XP-106-MMAE-025 (6 mg) as a yellow solid.

[0483] LCMS(ESI):m / z Calcd for C57 H 87 N8O 14 ,[M+H] + :1107.7,found:1107.1.

[0484] Example 20

[0485] 1) Preparation of azo compound 18 and azo compound 19:

[0486] 1.1)

[0487] Procedure: ON-2 (50 mg) was dissolved in dry DCM (2 mL), and tert-butyl (tetrahydrofuran-2-yl) glycinate (40 mg), DCC (35 mg), HOBt (30 mg) and triethylamine (10 uL) were added thereto, and the mixture was stirred at room temperature for 24 hours.

[0488] Detection: LCMS detection.

[0489] Post-treatment: prep-TLC (EA:PE=1:1) separated the product 18 (20 mg) as a yellow solid.

[0490] 1.2)

[0491] Procedure: 18 (20 mg, 1.0 eq) was dissolved in dry DCM (2 mL), p-nitrophenylcarbonyl chloride (15 mg, 1.2 eq), DMAP (2 mg) and triethylamine (8 μL) were added to the reaction, and the reaction was stirred at room temperature for 3 h.

[0492] Detection: TLC (EA:PE=1:1) detection.

[0493] Post-treatment: the reaction was quenched with water, extracted with DCM (8 mL x 3), the combined organic phases were washed once with saturated brine, and spin-dried.

[0494] Purification: prep-TLC (EA:PE=1:1) separated the product 19 (10 mg) as an orange-yellow oily solid.

[0495] 2) Preparation of azo compound XP-106-MMAE-026:

[0496] Procedure: MMAE (15 mg, 1.0 eq) was dissolved in DMF (1 mL), and 19 (10 mg), HOBt (8 mg) and triethylamine (6 μL) were added to the reaction, and the reaction was stirred at room temperature for 3 h.

[0497] Detection: LCMS detection.

[0498] Post-treatment and purification: The product XP-106-MMAE-026 (2 mg) was isolated by prep-HPLC as a yellow solid.

[0499] LCMS(ESI):m / z Calcd for C 64 H 95 N8O 13 ,[M+H] + :1183.7,found:1183.2.

[0500] Example 21

[0501] 1) Preparation of azo compound 20 and azo compound 21:

[0502] 1.1)

[0503] Procedure: ON-2 (50 mg) was dissolved in dry DCM (2 mL), and tert-butyl peroxy-2-(4-hydroxypiperidin-1-yl)acetate (40 mg), DCC (35 mg), and DMAP (5 mg) were added thereto, and the mixture was stirred at room temperature for 24 hours.

[0504] Detection: LCMS detection.

[0505] Post-treatment: prep-TLC (EA:PE=1:1) separated the product 17 (15 mg) as a yellow solid.

[0506] 1.2)

[0507] Procedure: 20 (15 mg, 1.0 eq) was dissolved in dry DCM (2 mL), p-nitrophenylcarbonyl chloride (10 mg, 1.2 eq), DMAP (2 mg) and triethylamine (5 μL) were added to the reaction, and the reaction was stirred at room temperature for 3 h.

[0508] Detection: TLC (EA:PE=1:1) detection.

[0509] Post-treatment: the reaction was quenched with water, extracted with DCM (5 mL x 2), the combined organic phases were washed once with saturated brine, and spin-dried.

[0510] Purification: prep-TLC (EA:PE=1:1) separated the product 17 (15 mg) as an orange-yellow oily solid.

[0511] 2) Preparation of azo compound XP-106-MMAE-027:

[0512] Procedure: MMAE (20 mg, 1.0 eq) was dissolved in DMF (1 mL), 21 (15 mg), HOBt (8 mg) and triethylamine (6 μL) were added to the reaction, and the reaction was stirred at room temperature for 3 h.

[0513] Detection: LCMS detection.

[0514] Post-treatment and purification: DMF was concentrated and purified by prep-TLC (EA=100%), and then separated by prep-HPLC to obtain the product XP-106-MMAE-027 (3 mg) as a yellow solid.

[0515] LCMS(ESI):m / z Calcd for C 65 H 97 N8O 13 ,[M+H] + :1197.7,found:1197.1.

[0516] Example 22

[0517] 1) Preparation of azo compound 22:

[0518] Procedure: Dissolve ON-2 (50 mg) in dry DCM (2 mL), add tert-butyl 2-(4-hydroxypiperidin-1-yl)acetate (40 mg), DCC (35 mg), and DMAP (5 mg). Stir at room temperature for 24 hours. Dissolve ON-3 (1040 mg, 1.0 eq) in dry DCM (10 mL), add pyridine (110 μL, 0.5 eq), and stir in a 45°C oil bath. Slowly add phosphorus tribromide (50 μL x 3) dropwise and reflux for 3 hours. TLC confirms complete reaction. Cool the reaction to room temperature, add isopropanol, and stir for 10 minutes. Quench the reaction with saturated NaHCO₃ and extract with EA (10 mL x 3). Combine the organic phases and spin dry to obtain the product (1126 mg, crude).

[0519] 2) Preparation of azo compound XP-103-SN38-028:

[0520] Procedure: SN-38 (50 mg, 1.0 eq) was dissolved in dry DCM (2 mL) and DMF (2 mL), and 73 mg of compound 22 was added. K2CO3 (18 mg) was added to the stirred reaction solution and stirred at room temperature for 24 hours.

[0521] Detection: TLC detection (DCM:MeOH=10:1).

[0522] Post-treatment and purification: DMSO was added to the reaction solution to completely dissolve it, and the filtrate was filtered to obtain the product (4 mg, 5% Yield) as a yellow solid.

[0523] LCMS(ESI):m / z Calcd for C 43 H 44 N5O8,[M+H] + :758.3,found:758.2.

[0524] Example 23

[0525] 1) Preparation of azo compound XP-114-NM-029:

[0526] Procedure: NM (119 mg, 1.0 eq) was dissolved in dry THF (2 mL), 490 mg of compound 22 (1.1 eq) was added, and the mixture was stirred under reflux for 24 hours.

[0527] Detection: TLC detection (DCM:MeOH=10:1).

[0528] Post-treatment and purification: Silica gel column purification gave 411 mg of NM-1 (yield 85%) as a yellow solid.

[0529] Procedure: NM-1 (242 mg, 0.5 mmol) was added dropwise to a dichloromethane solution of thionyl chloride (10 eq.) and stirred at room temperature for 12 hours.

[0530] Post-treatment and purification: The reaction mixture was filtered to obtain a yellow solid as the target product XP-114-NM-029 (247 mg, yield 95%).

[0531] LCMS(ESI):m / z Calcd for C 26 H 36 C l2 N4O3,[M] + :521.2,found:521.1.

[0532] Test Example 1

[0533] The azo prodrug compounds prepared in Examples 1 to 9 (irradiation dose of 60 Gy), 12 (irradiation dose of 24 Gy) and 13 to 15 (irradiation dose of 18 Gy) were subjected to irradiation release. The test results are shown in Table 16 below:

[0534] Table 16

[0535] As can be seen from Table 12, after irradiation, the prodrug peak areas of the azo prodrug compounds prepared in Examples 1 to 15 were significantly reduced, and obvious prodrugs were detected.

[0536] Test Example 2

[0537] The cytotoxicity of the azo prodrug compounds prepared above was compared with that of the original drug. The corresponding cells (e.g., HeLa, HT29, LOVO, HepG2, NPC / HK-1, SCC-090, 143B, purchased from ATCC) were seeded at a density of 4000 per well in a 96-well cell culture plate and incubated at 37°C, 4% carbon dioxide for 18 hours to ensure cell adhesion. The cells were then treated with the corresponding drugs at different concentrations and incubated for another 72 hours. After 72 hours, the original culture medium was aspirated and the cells were washed three times with PBS. 100 μL of diluted CCK-8 solution (1:10 ratio to culture medium) was added to each well. After incubation for 2 hours, the ultraviolet absorbance of each well at 450 nm was measured using a microplate reader. The absorbance of the treated cells was compared with that of the untreated cells to obtain the cell viability value.

[0538] The test results are shown in Tables 17 to 23 and Figures 1 to 4. The higher the prodrug / original drug IC50 ratio, the lower the cytotoxicity of the prodrug.

[0539] Table 17

[0540] Table 18

[0541] Table 19

[0542] Table 20

[0543] Note: “-” means that the XP-104-CPT-010 drug did not reach the half-lethal concentration of HT29 cells at a concentration of 10000 nM.

[0544] Table 21

[0545] Table 22

[0546] Table 23

[0547] Example 24

[0548] Preparation and analysis of antibody-drug conjugate XPADC-106-011:

[0549] The synthetic route of XPADC-106-011 is as follows:

[0550] Procedure: 106-011 (100 mg) was dissolved in dry DCM (4 mL), and DCC (23.4 mg), TEA (16.7 ul) and HOBt (16.7 mg) were added thereto. After stirring evenly, piperazine (16.7 mg) was added and stirred at room temperature for 7 hours.

[0551] Detection: LCMS detection.

[0552] Post-treatment: DCM was dried and purified by PLC (DCM:MeOH = 10:1) to give the product 106-011-piperazine (90.1 mg, 86% yield) as a yellow solid. (Used directly in the next step without purification)

[0553] Step 2

[0554] Procedure: 106-011-piperazine (45 mg) was dissolved in dry DCM (10 mL), and TEA (11 ul, 2 eq.) and HOBt (6.4 mg, 1.2 eq.) were added. SMCC (39.7 mg, 3 eq.) dissolved in 1.5 ml of DCM was then added under an ice-water bath. After stirring at room temperature for 24 hours, LC-MS showed that approximately 40% of the 106-011-piperazine had not reacted. 1.5 ml of SMCC (39.7 mg, 3 eq.) dissolved in DCM was then added and the reaction continued for 2 hours.

[0555] Detection: LCMS detection.

[0556] Post-treatment: the reaction solution was spin-dried and purified by Pre-HPLC to obtain 106-011-SMCC (12.3 mg) as a yellow solid.

[0557] Step 3:

[0558] TCEP (20 mM His) solution was added to the antibody solution and reacted at room temperature for 3 hours. Then, 10% v / v DMSO-dissolved 106-011-SMCC solution was added and reacted for 1 hour. Finally, samples were taken for characterization. The characterization results are shown in Table 24 below.

[0559] Antibody name: Cetuximab injection, antibody concentration: 5 mg / mL, antibody purchase source: Merck Pharmaceuticals (Jiangsu) Co., Ltd.

[0560] UV-Vis test sample pretreatment: The sample needs to remove unreacted linker-drug and DMSO. This can be removed by filtration followed by ultrafiltration or desalting. HIC test sample pretreatment: The sample can be injected after filtration.

[0561] Table 24

[0562] The results of hydrophobic interaction chromatography (HIC-HPLC) are shown in FIG5 .

[0563] The DAR value was calculated based on the above test results, and the calculation results are shown in Table 25 below.

[0564] Table 25

[0565] The antibody-conjugated azo prodrug prepared above was subjected to a comparative cytotoxicity test with the original drug. The corresponding cells (such as LOVO, HepG2, NPC / HK-1, Panc-1, purchased from ATCC) were seeded in a 96-well cell culture plate at a density of 4000 per well and incubated at 37°C, 4% carbon dioxide for 18 hours to ensure cell adhesion; the cells were then treated with different concentrations of the corresponding drugs and incubated at a constant temperature for 72 hours; after 72 hours, the original culture medium was aspirated and the cells were washed 3 times with PBS, 100 μL of diluted CCK-8 solution (1:10 ratio with culture medium) was added to each well, and after incubation at a constant temperature for 2 hours, the ultraviolet absorption at 450 nm in each well was detected with an enzyme marker, and the absorbance of the treated cells was compared with that of the untreated cells to obtain the cell activity value. The test results are shown in Table 26 below:

[0566] Table 26: Tumor cell inhibitory activity

[0567] Example 25

[0568] Synthesis and analysis of lipid component XP-107-BUF-008:

[0569] Synthesis method:

[0570] Procedure: Add 32 mg of 3-2-3 to the reaction mixture YJY-2-67 from the previous step and react at room temperature for 24 h.

[0571] Detection: TLC detection (DCM:MeOH=10:1)

[0572] Post-treatment and purification: Concentrate at 37°C and separate using a large silica gel plate (DCM:MeOH=10:1) to obtain the product YJY-2-69 (40 mg) as a yellow solid.

[0573] LCMS(ESI):m / z Calcd for C 48 H 63 O8N6,[M+H] + :851.5,found:851.6

[0574] Procedure: 40 mg of YJY-2-69 was dissolved in 2 mL of DCM, 0.5 mL of iodomethane was added, and the mixture was reacted at room temperature for 16 h.

[0575] Detection: LCMS detection.

[0576] Post-treatment and purification: A yellow solid was produced during the reaction, and the precipitate was collected by centrifugation. The precipitate was washed with DCM and PE in sequence to obtain the product XP-107-BUF-008 (40 mg) as a yellow solid.

[0577] LCMS(ESI):m / z Calcd for C49H65O8N6 + ,[M] + :865.5,found:865.1.

[0578] 1 H NMR(400MHz,d6-DMSO)δ8.98(t,J=5.6Hz,1H),8.13–7.79(m,6H),7.66–7.47(m,3H),6.29(d,J=9.8Hz,1H),5.75(s, 1H),5.22(s,2H),4.90(s,1H),4.15(s,1H),3.73(d,J=6.0Hz,1H),3.53(t,J=6.5Hz,1H),3.39(d,J=14.6Hz,8H),3.1 5(d,J=7.6Hz,9H),2.86(d,J=8.3Hz,2H),2.67(s,1H),2.33(s,1H),2.13–1.89(m,4H),1.77(d,J=10.2Hz,2H),1.58 (t,J=21.8Hz,9H),1.34(t,J=12.0Hz,4H),1.19(ddd,J=36.9,18.4,11.7Hz,5H),0.91(d,J=4.2Hz,3H),0.60(s,3H).

[0579] Preparation of XP-107-BUF-008 liposomes (107-008-LNPs):

[0580] Methods: 69.3% cholesterol, 18% DSPC, 2.7% PEG-2000-DMG, and 10% XP-107-BUF-008 (hereinafter referred to as 107-008) were prepared in ethanol to a 12 mM solution according to the molar ratio. The organic phase was filtered through a 0.22 μm filter. The aqueous phase was PBS.

[0581] Liposome nanoparticles (LNPs) were prepared using an LNP intelligent synthesizer at a ratio of organic phase to aqueous phase of 1:3. The prepared LNPs were passed through a 30 kD ultrafiltration tube for solution exchange. The ethanol concentration was diluted to below 0.5% with PBS and concentrated to the target concentration before storage at 4°C.

[0582] Radiological testing:

[0583] The particle size distribution of the XP-107-BUF-008 liposomes prepared above was tested before and after irradiation with 60 Gy. The test results are shown in Table 27 below.

[0584] Table 27: Particle size intensity distribution before and after irradiation

[0585] Combined with Table 27, it can be seen that the particle size distribution of the XP-107-BUF-008 liposome mixture with a content of 10% changes significantly after irradiation with 60 Gy. This is because the XP-107-BUF-008 liposomes rupture after 60 Gy irradiation, and the ruptured mixture reorganizes to form new aggregates.

[0586] The XP-107-BUF-008 liposomes prepared above were subjected to 60 Gy irradiation and irradiation, and the nanoparticles were demulsified with ethanol and then detected by LCMS. The detection results are shown in Figures 6 and 7.

[0587] 6 and 7 , it can be seen that after 60 Gy irradiation, the prodrug essentially disappears.

[0588] Example 26

[0589] Synthesis of lipid component 115-001:

[0590] The specific steps include:

[0591] first step

[0592] Procedure: DSPE (50 mg) was dissolved in dry chloroform (3 mL), and ON-4 (50 mg) and TEA (40 uL) were added thereto, followed by stirring at 50° C. for 3 hours.

[0593] Detection: TLC (DCM:MeOH=10:1).

[0594] Post-treatment: The residue was concentrated to 1 mL and directly scraped on a plate (DCM:MeOH=10:1) to obtain 25 mg of YJY-2-87 as a yellow solid (yield 32.3%).

[0595] Step 2

[0596] Procedure: YJY-2-87 (25 mg, 1.0 eq) was dissolved in DCM (2 mL), TFA (0.5 mL) was added to the stirred reaction solution at 0°C, and the mixture was stirred at room temperature for 1 hour.

[0597] Detection: TLC (DCM:MeOH=10:1).

[0598] Post-treatment: The reaction solution was concentrated to obtain the product YJY-2-98 (22 mg, crude) as a yellow-brown solid. (Used directly in the next step without purification)

[0599] Step 3

[0600] Procedure: YJY-2-98 (30 mg, 1.0 eq) was dissolved in DCM (2 mL), and compound 4 (10 mg), HATU (12 mg), and DIPEA (60 uL) were added to the reaction mixture. The reaction was stirred at room temperature for 24 h.

[0601] Detection: TLC (DCM:MeOH=10:1).

[0602] Post-treatment: The residue was concentrated to 1 mL and directly scraped on a plate (DCM:MeOH=10:1) to obtain 26 mg of the product as a yellow solid (yield 81.5%).

[0603] HRMS(ESI):m / z Calcd for C63H108O12N6P,[M+H]+:1171.7685,found:1171.7790.

[0604] Step 4

[0605] Procedure: YJY-2-98 (38 mg, 1.0 eq) was dissolved in DCM (5 mL), 1 mL of iodomethane was added to the reaction, and the reaction was stirred at 50 °C overnight.

[0606] Detection: HRMS showed that the reaction of the raw material was complete.

[0607] Post-treatment: spin-drying, washing the solid with ethyl acetate and centrifuging three times, dissolving it with a small amount of DCM, and adding a large amount of petroleum ether to precipitate a yellow solid, which was centrifuged to obtain 30 mg of product 115-001.

[0608] HRMS(ESI):m / z Calcd for C 64 H 111 O 12 N6P,[M+H] + :1185.7914,found:1185.7917.

[0609] 1 H NMR (400MHz, CDCl3) δ8.32 (s, 1H), 7.83 (d, J = 12.5Hz, 3H), 7.71–7.35 (m, 4H), 6. 45(s,1H),5.19(d,J=41.1Hz,2H),4.37(s,1H),4.14(dd,J=12.9,5.7Hz,2H),4.0 6(s,4H),3.87(s,3H),3.69(s,4H),3.42(s,9H),3.37(s,6H),3.07(s,13H),2.96 (s,1H),2.31(d,J=7.6Hz,3H),1.59(s,4H),1.26(s,46H),0.89(t,J=6.6Hz,6H).

[0610] 115-001 Liposome Preparation:

[0611] Methods: 40% cholesterol, 50% Dlin-MC3-DMA, 1.5% PEG-2000-DMG, and 10% 115-001 were prepared in ethanol to a 12 mM solution, which was then filtered through a 0.22 μm filter to obtain the organic phase. The aqueous phase was PBS. Liposome nanoparticles (LNPs) were prepared using an LNP intelligent synthesizer at a ratio of 1:3 organic phase:aqueous phase. The prepared LNPs were then passed through a 30 kD ultrafiltration tube for solvent exchange. The ethanol concentration was diluted with PBS to below 0.5%, concentrated to the target concentration, and stored at 4°C.

[0612] The particle size distribution of the 115-001 liposomes prepared above was tested before and after irradiation with 60 Gy. The test results are shown in Figures 8 and 9.

[0613] 8 and 9 , it can be seen that the particle size distribution of the 115-001 liposomes prepared above changed significantly after irradiation with 60 Gy. This is because the 115-001 liposomes ruptured after 60 Gy irradiation, and the ruptured mixture reorganized to form new aggregates.

Claims

1. An azo compound, characterized in that It has the following general formula I: in, It is a single bond, which means that A and B are distributed on both sides or the same side of the azo double bond; -A- and -B- are each independently selected from: aryl or heteroaryl; The term "aryl" refers to an aromatic ring group having 6 to 20 carbon atoms obtained by removing two hydrogen atoms from two carbon atoms in the aromatic nucleus of an aromatic ring molecule; The term "heteroaryl" refers to aromatic groups that are 5-membered or 6-membered rings and fused ring systems comprising 5 to 20 atoms, wherein at least one ring is aromatic and contains one or more heteroatoms independently selected from nitrogen, oxygen and sulfur, and if the ring contains multiple oxygen atoms, the oxygen atoms are not directly adjacent; -C- is -X, -O-, -OC(=O)- or X is Cl, Br or I; when -C- is -X, R2 is an empty bond; when -C- is -O-, R2 is H; when -C- is -OC(=O)-, -R2 is -Cl, When -C- is When -R2 is -Cl; -R1 is -R3 and -R4 are independently selected from: -CN, -COOR, -CONR'R", -H, halogen, C1-C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group is O or N; -R5 is selected from: -H, C1~C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 Aryl, -R6 is selected from C1~C 12 The heteroatom in the heteroalkyl group is O or N; -R8 is 1 to 4 substituted, and -R8 is selected from -NO2, -F, -CN, -CF3, -COOR, -CONR'R".

2. An azo compound, characterized in that It has the following general formula I: in, It is a single bond, which means that A and B are distributed on both sides or the same side of the azo double bond; -A- and -B- are each independently selected from: aryl or heteroaryl; The term "aryl" refers to an aromatic hydrocarbon group having 6 to 20 carbon atoms obtained by removing two hydrogen atoms from two carbon atoms in the aromatic nucleus of an aromatic hydrocarbon molecule; The term "heteroaryl" refers to aromatic groups that are 5-membered or 6-membered rings and fused ring systems comprising 5 to 20 atoms, wherein at least one ring is aromatic and contains one or more heteroatoms independently selected from nitrogen, oxygen and sulfur, and if the ring contains multiple oxygen atoms, the oxygen atoms are not directly adjacent; -C- is -X, -O-, -OC(=O)- or X is Cl, Br or I; when -C- is -X, R2 is an empty bond; when -C- is -O-, R2 is H; when -C- is -OC(=O)-, -R2 is -Cl, When -C- is When -R2 is -Cl; -R1 is -R3 and -R4 are independently selected from: -CN, -COOR, -CONR'R", -H, halogen, C1-C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group is O or N; -R5 is selected from: -H, C1~C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The aryl group, -R6 is selected from C1~C 12 The heteroalkyl group of the heteroalkyl group is O or N; -R8 is 1 to 4 substituted, and -R8 is selected from -NO2, -F, -CN, -CF3, -COOR, -CONR'R"; The aryl or heteroaryl is substituted by one or more substituents, the substituents including halogen, -R9, -NR9R 10 , -CN, -NO2, -N3, -OR9, -SR9, -NHCOR9, -O-COR9, -CH=CR9R 10 , -C(=O)-R9, -C(=O)-OR9, -C(=O)-Cl, -C(=O)-NH2, -C(=CO)-NH-R9 and -C(=O)-NR9R 10 , among which -R9 and -R 10 Each independently selected from -H, C1-C6 alkyl, C1-C6 haloalkyl, C2-C6 alkenyl, C3-C 10 Cycloalkyl, C6~C 20 The alkyl, alkenyl, cycloalkyl, aryl and heteroaryl groups described for the substituents are optionally replaced by one or more halogen, hydroxyl, mercapto, -NH2, -CN, -NO2, -N3, -NHCOH, -OC(=O)H, -C(=O)H, -C(=O)-OH, -C(=O)-Cl, -C(=O)-NH2, -C(=O)-NH-CH3, -C(=O)-CH3, -C(=O)-OCH3, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C2-C6 alkenyl, C3-C 10 Cycloalkyl, C6~C 20 substituted with an aryl group or a heteroaryl group having 5 to 20 ring atoms; -R, -R' and -R" are independently selected from -H, C1-C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group or heteroalkyl group is O or N.

3. The azo compound according to claim 1, characterized in that The structural formula of the azo compound is:

4. An azo compound, characterized in that The structural formula of the azo compound is:

5. An azo prodrug compound, characterized in that The azo prodrug compound has the following general formula II: in, It is a single bond, which means that A and B are distributed on both sides or the same side of the azo double bond; -A- and -B- are each independently selected from: aryl or heteroaryl; The term "aryl" refers to an aromatic group having 6 to 20 carbon atoms obtained by removing two hydrogen atoms from two carbon atoms in the aromatic nucleus of an aromatic hydrocarbon (aromatic ring) molecule; The term "heteroaryl" refers to aromatic groups that are 5-membered or 6-membered rings and fused ring systems comprising 5 to 20 atoms, wherein at least one ring is aromatic and contains one or more heteroatoms independently selected from nitrogen, oxygen and sulfur, and if the ring contains multiple oxygen atoms, the oxygen atoms are not directly adjacent; -C- is -O-, -N + (R a R b )-、-OC(=O)- or -G1 is a drug group; -G2 is a targeting group, -R3 and -R4 are independently selected from: -CN, -COOR, -CONR'R", -H, halogen, C1-C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group is O or N; -R5 and -R6 are independently selected from: -H, C1-C 12 Alkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 aryl groups; -R7 is -H, C1~C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group or heteroalkyl group is O or N.

6. The azo prodrug compound according to claim 5, characterized in that The aryl or heteroaryl is substituted by one or more substituents, the substituents including halogen, -R9, -NR9R 10 , -CN, -NO2, -N3, -OR9, -SR9, -NHCOR9, -O-COR9, -CH=CR9R 10 , -C(=O)-R9, -C(=O)-OR9, -C(=O)-Cl, -C(=O)-NH2, -C(=CO)-NH-R9 and -C(=O)-NR9R 10 , among which -R9 and -R 10 Each independently selected from -H, C1-C6 alkyl, C1-C6 haloalkyl, C2-C6 alkenyl, C3-C 10 Cycloalkyl, C6~C 20 The alkyl, alkenyl, cycloalkyl, aryl and heteroaryl groups described for the substituents are optionally replaced by one or more halogen, hydroxyl, mercapto, -NH2, -CN, -NO2, -N3, -NHCOH, -OC(=O)H, -C(=O)H, -C(=O)-OH, -C(=O)-Cl, -C(=O)-NH2, -C(=O)-NH-CH3, -C(=O)-CH3, -C(=O)-OCH3, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C2-C6 alkenyl, C3-C 10 Cycloalkyl, C6~C 20 substituted with an aryl group or a heteroaryl group having 5 to 20 ring atoms; -R a and -R b Each independently selected from C1 to C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group is O or N; -R, -R' and -R" are independently selected from -H, C1-C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group or heteroalkyl group is O or N.

7. The azo prodrug compound according to claim 5, characterized in that The targeting group is covalently linked to other structures via N, O or S; The targeting group is an antibody, a polypeptide, a sugar group, a small molecule ligand or a nucleic acid aptamer; The drug group is covalently linked to other structures via N, O or S; The drug group is 8. A pharmaceutical composition, characterized in that The invention comprises the azo prodrug compound or a pharmaceutically acceptable salt thereof according to any one of claims 5 to 7.

9. A high-energy ray-responsive lipid molecule, characterized in that: It has the following general formula III: in, It is a single bond, which means that A and B are distributed on both sides or the same side of the azo double bond; -A- and -B- are each independently selected from: aryl or heteroaryl; The term "aryl" refers to an aromatic group having 6 to 20 carbon atoms obtained by removing one hydrogen atom from each of two carbon atoms in the aromatic nucleus of an aromatic hydrocarbon (aromatic ring) molecule; The term "heteroaryl" refers to aromatic groups that are 5-membered or 6-membered rings and fused ring systems comprising 5 to 20 atoms, wherein at least one ring is aromatic and contains one or more heteroatoms independently selected from nitrogen, oxygen and sulfur, and if the ring contains multiple oxygen atoms, the oxygen atoms are not directly adjacent; -C- is -O-, -N + (R a R b )-、-OC(=O)- or -G3 and -G4 are respectively one of a hydrophilic group and a hydrophobic group, and the high-energy ray-responsive lipid molecule is an amphiphilic molecule; -R3 and -R4 are independently selected from: -CN, -COOR, -CONR'R", -H, halogen, C1-C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group or heteroalkyl group is O or N.

10. The high-energy ray-responsive lipid molecule according to claim 9, characterized in that The lipid molecule is a fatty acid ester or amide derivative, a sterol ester or amide derivative, a glyceride, a glycerophospholipid or a glycolipid containing an azo structural unit; The aryl or heteroaryl is substituted by one or more substituents, the substituents including halogen, -R9, -NR9R 10 , -CN, -NO2, -N3, -OR9, -SR9, -NHCOR9, -O-COR9, -CH=CR9R 10 , -C(=O)-R9, -C(=O)-OR9, -C(=O)-Cl, -C(=O)-NH2, -C(=CO)-NH-R9 and -C(=O)-NR9R 10 , among which -R9 and -R 10 Each independently selected from -H, C1-C6 alkyl, C1-C6 haloalkyl, C2-C6 alkenyl, C3-C 10 Cycloalkyl, C6~C 20 The alkyl, alkenyl, cycloalkyl, aryl and heteroaryl groups described for the substituents are optionally replaced by one or more halogen, hydroxyl, mercapto, -NH2, -CN, -NO2, -N3, -NHCOH, -OC(=O)H, -C(=O)H, -C(=O)-OH, -C(=O)-Cl, -C(=O)-NH2, -C(=O)-NH-CH3, -C(=O)-CH3, -C(=O)-OCH3, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C2-C6 alkenyl, C3-C 10 Cycloalkyl, C6~C 20 substituted with an aryl group or a heteroaryl group having 5 to 20 ring atoms; -R a and -R b Each independently selected from C1 to C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group is O or N; -R, -R' and -R" are independently selected from -H, C1-C 12 Alkyl, C1~C 12 Heteroalkyl, C3~C 12 Cycloalkyl, C2~C 12 Alkenyl, C3~C 12 Cycloalkyl or C6~C 20 The heteroatom in the aryl group or heteroalkyl group is O or N.

11. A high-energy radiation-responsive liposome, characterized in that: It is formed by self-assembly of the high-energy ray-responsive lipid molecules according to claim 9 or 10.

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