Photosensitizer conjugate compound as well as pharmaceutical composition and application thereof

Abstract

The invention belongs to the field of precise diagnosis and treatment, and relates to a photosensitizer conjugate compound, in particular to a ligand photosensitizer conjugate (especially a double-ligand photosensitizer conjugate shown as a formula I), a pharmaceutical composition based on the photosensitizer conjugate compound and application of the pharmaceutical composition in the field of medicine. The photosensitizer conjugate compound combines the advantages of a ligand drug conjugate and a photodynamic technology, the photosensitizer is coupled to a ligand with a targeting effect, the targeting property of the photosensitizer is improved while the advantages of small side effect and low drug resistance of a photosensitizer drug are utilized, and the drug forming possibility of the photosensitizer drug is provided. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of precision diagnosis and treatment, and relates to a photosensitizer conjugate compound, specifically to a ligand-photosensitizer conjugate (especially a dual-ligand photosensitizer conjugate), a pharmaceutical composition based on the photosensitizer conjugate compound (or a ligand-photosensitizer conjugate, a dual-ligand photosensitizer conjugate), and its application in the medical field. Background Art

[0002] Photodynamic therapy (PDT) is a new technology that uses the photodynamic effect to diagnose and treat diseases. Photodynamic therapy consists of three major elements: light, oxygen, and photosensitizer (a compound with photosensitivity), which work together to achieve its effects (for example, inducing tumor cell death). Its action process is roughly as follows: first, the photosensitizer selectively accumulates in the tumor tissue, and then is exposed to light of an appropriate wavelength, allowing the photosensitizer to capture light energy. Finally, in the presence of molecular oxygen, the light energy is converted into a chemical reaction to produce singlet oxygen ( 1 O2) and other highly reactive oxygen species (ROS) and induce cell damage through direct or indirect cytotoxicity.

[0003] Compared with traditional tumor treatment methods, PDT is a non-invasive therapy with a series of significant advantages: small side effects, low drug resistance, good selectivity; it can be used alone or in combination with other treatment methods. PDT's selectivity for target tissues and degree of damage reduces damage to normal tissues. It has the advantages of small trauma, strong applicability, auxiliary treatment, repeatable treatment, no drug resistance, elimination of tiny lesions, and preservation of important organ functions. In particular, it provides a new treatment method for cancer patients in the middle and late stages, especially those who cannot (or refuse) to use traditional treatment methods. Compared with traditional therapies, PDT can avoid the impact of multiple surgeries on patients, while also being able to exert good therapeutic effects.

[0004] Currently, PDT has been widely used in the treatment of malignant tumors and precancerous lesions such as skin cancer, esophageal cancer, colorectal cancer, lung cancer, cervical cancer, and bladder cancer. It is mainly used clinically for tumors in cavities, cavities, and body surfaces, as well as some abnormal proliferations. However, there is only one APC (antibody photosensitizer conjugate) drug (i.e., cetuximab sarotalocan / developed by Rakuten Medical) that can effectively treat malignant tumors and precancerous lesions. ) has been approved for marketing, so there is an urgent need to develop more photosensitizer conjugates to meet the growing demand in the field of precision diagnosis and treatment. Summary of the invention

[0005] Problem that the invention aims to solve

[0006] In view of the problems that the current photosensitizer conjugates have few types, lack of targeting, and are difficult to develop into drugs, the present invention combines the advantages of ligand drug conjugates (PDC) and photodynamic therapy (PDT) to discover a series of new ligand photosensitizer conjugates (ligand photosensitizer conjugates, LPC), especially bi-ligand photosensitizer conjugates (bi-ligand photosensitizer conjugates, BLPC) constructed based on dual targeting technology. By coupling the photosensitizer to a ligand with a targeting effect, while utilizing the advantages of the photosensitizer drug itself with few side effects and low drug resistance, the targeting and drugability of the photosensitizer are improved, and the types of photosensitizer conjugates are further enriched.

[0007] Solutions for solving problems

[0008] <First aspect>

[0009] The present invention provides a coupled compound, which may comprise a photosensitizer and a targeting ligand, preferably 1 to 6 targeting ligands, more preferably 2 or 4 targeting ligands.

[0010] In one embodiment of the present invention, the photosensitizer in the above-mentioned coupled compound can be formed by porphyrins, phthalocyanines, cyanines or Bodipy compounds, preferably formed by phthalocyanine compounds, more preferably formed by silicon phthalocyanine or zinc phthalocyanine compounds.

[0011] In one embodiment of the present invention, each targeting ligand in the above-mentioned conjugate compound can independently bind to any one of the following cell surface proteins: PSMA, FOLR1 and TRPV6.

[0012] Furthermore, when the above-mentioned conjugate compound contains 2 or 4 targeting ligands, the above-mentioned conjugate compound can bind to any one of the following cell surface protein combinations: PSMA / FOLR1, FOLR1 / TRPV6 and PSMA / TRPV6, preferably PSMA / FOLR1 and FOLR1 / TRPV6, and more preferably PSMA / FOLR1.

[0013] In one embodiment of the present invention, each targeting ligand in the above-mentioned conjugate compound can be independently formed by a polypeptide, an antibody or a small molecule.

[0014] The present invention also provides a coupled compound, which may include a photosensitizer, a targeting ligand and a connecting portion; the connecting portion may be used to connect the photosensitizer and the targeting ligand and / or connect a plurality of the targeting ligands.

[0015] The present invention also provides a coupled compound, which may include a photosensitizer, a targeting ligand, a linker and a spacer; the linker may be used to connect the photosensitizer and the targeting ligand; the spacer may be used to connect a plurality of the targeting ligands.

[0016] In one embodiment of the present invention, the above-mentioned coupled compound may have a structure as shown in Formula I,

[0017]

[0018] in,

[0019] LG indicates targeting ligand;

[0020] S represents a spacer;

[0021] L represents a linker;

[0022] PS indicates photosensitizer;

[0023] n is 1 or 2;

[0024] and

[0025] The two LGs are identical to or different from each other;

[0026] When n is 2, two The same as or different from each other.

[0027] Furthermore, the compound of formula I may have a structure as shown in formula IA,

[0028]

[0029] wherein LG, S, L and PS are as defined in Formula I.

[0030] Alternatively, further, the compound of formula I may have a structure as shown in formula IB,

[0031]

[0032] wherein LG, S, L and PS are as defined in Formula I.

[0033] In one embodiment of the present invention, each LG in the above-mentioned compound of Formula I, Formula IA or Formula IB can independently bind to any one of the following cell surface proteins: PSMA, FOLR1 and TRPV6.

[0034] Further, when containing 2 or 4 LGs, the above-mentioned compounds of Formula I, Formula IA or Formula IB can bind to any one of the following cell surface protein combinations: PSMA / FOLR1, FOLR1 / TRPV6 and PSMA / TRPV6, preferably PSMA / FOLR1 and FOLR1 / TRPV6, and more preferably PSMA / FOLR1.

[0035] In one embodiment of the present invention, each LG in the above-mentioned compound of Formula I, Formula IA or Formula IB can be independently formed by a polypeptide, an antibody or a small molecule.

[0036] In one embodiment of the present invention, each LG in the above-mentioned compound of formula I, formula IA or formula IB can be independently formed by any one of the ligand compounds of formula LG-I, LG-II and LG-III.

[0037]

[0038] In formula LG-I,

[0039] R LG1 For hydroxyl,

[0040] Preferably, R LG1 For hydroxyl,

[0041] Formula LG-II is pteroic acid, folic acid or an analog thereof; preferably, the folic acid analog is selected from 5-methyltetrahydrofolate, 5-formyltetrahydrofolate, 10-formylfolate, methotrexate, aminopterin and raltitrexed;

[0042] Formula LG-III comprises all or part of the amino acids in the polypeptide EGKLSSNDTEGGLCKEFLHPSKVDLPR; preferably, formula LG-III comprises 9 to 27 amino acids in the above polypeptide; more preferably, formula LG-III has at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity with the polypeptide KEFLHPSKVDLPR; further preferably, formula LG-III is the polypeptide KEFLHPSKVDLPR.

[0043] In one embodiment of the present invention, each LG in the above-mentioned compounds of formula I, formula IA or formula IB can be independently a targeting ligand formed by a ligand compound of formula LG-I or an optical isomer thereof and binding to the cell surface protein PSMA; a targeting ligand formed by a ligand compound of formula LG-II or an optical isomer thereof and binding to the cell surface protein FORL1; or, a targeting ligand formed by a ligand compound of formula LG-III or an optical isomer thereof and binding to the cell surface protein TRPV6.

[0044] In one embodiment of the present invention, each LG in the compound of formula I, formula IA or formula IB can be independently formed by any one of the following ligand compounds or their optical isomers.

[0045]

[0046] In the present invention, the LG formed by the above-mentioned ligand compound can be various, and the present invention does not make specific restrictions on this. For example, it can be a monovalent group obtained by losing a hydrogen atom from a primary amino group in the structure (preferably a primary amino group located at the end of the main chain) (i.e., a secondary amino group, which can be connected with a carboxyl group of other fragments (e.g., S) in the conjugate to form an amide group), or a monovalent group obtained by losing a hydroxyl group in the structure (preferably a carboxyl group located at the end of the main chain) (i.e., a carboxyl group, which can be connected with a secondary amino group of other fragments in the conjugate to form an amide group), or a monovalent group obtained by losing a hydrogen atom from a hydroxyl group in the structure (preferably a hydroxyl group located at the end of the main chain) (e.g., an alkoxy group or an aryloxy group, which can be connected with a carboxyl group of other fragments in the conjugate to form an ester group).

[0047] In the present invention, each ligand compound can independently form any one of the following fragments, wherein the end marked by * is connected to S in the conjugate.

[0048]

[0049] In the present invention, the above-mentioned compounds of formula IA or formula IB are relatively preferred because the structures of these conjugates contain at least two targeting ligands, which can act in multiple ways to improve the therapeutic effect and reduce toxic side effects; at the same time, the two targeting molecules enhance the affinity of the conjugate to the target cells and reduce off-target toxicity.

[0050] Furthermore, when the conjugate compound of the present invention contains at least two targeting ligands, a connecting portion (e.g., a spacer) can be used to connect the two targeting ligands. In the present invention, the spacer can be either cleavable (or degradable) or non-cleavable (or non-degradable), and the present invention has no special requirements for the specific structure of the spacer.

[0051] In one embodiment of the present invention, each S in the compound of formula I, formula IA or formula IB can be independently formed by any one of the following spacer compounds or optical isomers thereof.

[0052]

[0053] In the present invention, each spacer compound can independently form any one of the following fragments, wherein the end marked by * is connected to one LG in the conjugate; the end marked by ** is connected to another LG in the conjugate; the group marked by *** can react with the corresponding group in the linker compound (for example, the cycloaddition reaction of the alkynyl group and the azide group, or the double bond addition reaction of the alkenyl group in the maleimide ring and the thiol group), thereby completing the connection between S and L.

[0054]

[0055] In one embodiment of the present invention, each S in the above-mentioned compounds of formula I, formula IA or formula IB can be independently formed by any one of S-1, S-2, S-3 and S-4 spacer compounds or their optical isomers and additional amino acids or amino acid combinations or their optical isomers, wherein the additional amino acids or amino acid combinations can be selected from Lys, Cys, Glu, Lys-Cys, Cys-Cys, Lys-Cys-Lys, Arg-Arg, Ala-Ser-Asn, Ala-Ala-Ala, Ser-Ser-Arg, Pro-Arg, Asp-Asp-Lys-Cys and Pro-Leu-Gly, and the additional amino acids or amino acid combinations can be optionally amidated (for example, Lys is amidated to L-2,6-diaminohexanamide).

[0056] Furthermore, a double ligand compound (BLG) can be formed by the spacer compound and the two ligand compounds, and the latter also falls within the protection scope of the present invention.

[0057] In the present invention, each BLG can independently be any one of the following compounds, wherein the alkynyl group or thiol group can react with the corresponding group in the linker compound (for example, the azido group or the olefinic group in the maleimide ring) to complete the connection between the biligand fragment and the linker fragment.

[0058]

[0059]

[0060] In one embodiment of the present invention, each BLG can be independently any one of the following compounds or their optical isomers.

[0061]

[0062]

[0063] Furthermore, when the conjugate compound of the present invention contains at least two targeting ligands, a connecting part (e.g., a linker) can be used to connect the photosensitizer (PS) fragment and the biligand (BLG) fragment. In the present invention, the linker can be either cleavable (or degradable) or non-cleavable (or non-degradable), and the present invention has no special requirements for the specific structure of the linker.

[0064] In one embodiment of the present invention, each L in the above-mentioned compound of formula I, formula IA or formula IB can be independently formed by any one of the linker compounds of formula LI and L-II,

[0065]

[0066] In formula LI,

[0067] s is any integer from 1 to 8, preferably any integer from 3 to 5, more preferably 4 or 5;

[0068] R L1 is amino, carboxyl or Where W is and the two ends are connected to other fragments through amide bonds; t is 0 or 1; u is any integer from 1 to 4, preferably 2 or 3, more preferably 3; v is any integer from 1 to 4, preferably 1 or 2, more preferably 2; preferably, R L1 For amino, carboxyl,

[0069]

[0070] In Formula L-II,

[0071] p is any integer from 1 to 4, preferably any integer from 1 to 3, more preferably 2 or 3;

[0072] q is any integer from 0 to 3, preferably any integer from 0 to 2, more preferably 0 or 1;

[0073] r is 0 or 1;

[0074] Preferably, when p is 2, q is 1, and r is 1; or

[0075] Preferably, when p is 3, q ​​is 0 or 1, and r is 0.

[0076] In one embodiment of the present invention, each L in the compound of formula I, formula IA or formula IB can be independently formed by any one of the following linker compounds or optical isomers thereof.

[0077]

[0078] In the present invention, each linker compound can independently form any one of the following fragments, wherein the end marked by * is connected to PS in the conjugate; the group marked by *** can react with the corresponding group in the spacer compound (for example, the cycloaddition reaction of the alkynyl group and the azide group, or the double bond addition reaction of the alkenyl group in the maleimide ring and the thiol group), thereby completing the connection between L and S.

[0079]

[0080]

[0081] In one embodiment of the present invention, in the above-mentioned compounds of formula I, formula IA or formula IB, when S can be formed by any one of the spacer compounds S-1 and S-2, L can be formed by any one of the linker compounds L-2A, L-2B and L-2C, and the two are connected to each other through a cycloaddition reaction of an alkynyl group and an azide group (e.g., [3+2] cycloaddition reaction).

[0082] In one embodiment of the present invention, in the above-mentioned compounds of formula I, formula IA or formula IB, when S can be formed by any one of the spacer compounds S-3 and S-4, L can be formed by any one of the linker compounds L-1A, L-1B, L-1C, L-1D, L-1E and L-1F, and the two are connected to each other through the double bond addition reaction of the alkenyl group in the maleimide ring and the thiol group (for example, thiol-Michael reaction).

[0083] Furthermore, the photosensitizer (PS) contained in the coupled compound of the present invention can undergo a photochemical reaction after absorbing light, thereby generating reactive oxygen species that induce cancer cell death, thereby achieving a therapeutic effect.

[0084] In one embodiment of the present invention, the above-mentioned photosensitizer can be formed by porphyrins, phthalocyanines (for example, silicon phthalocyanine or zinc phthalocyanine), cyanines or Bodipy compounds, preferably formed by phthalocyanine compounds, more preferably formed by silicon phthalocyanine or zinc phthalocyanine compounds.

[0085] In one embodiment of the present invention, the PS in the compound of formula I, formula IA or formula IB can be formed by any one of the following photosensitizer compounds or their optical isomers.

[0086]

[0087] In the present invention, the photosensitizer compound may form any of the following fragments, wherein the end marked by * is connected to L in the conjugate.

[0088]

[0089]

[0090] In one embodiment of the present invention, in the above-mentioned compounds of formula I, formula IA or formula IB, when PS can be formed by any one of the photosensitizer compounds PS-1 and PS-3, L can be formed by any one of the linker compounds L-1A and L-2B, and the two are connected to each other through an amidation reaction of a carboxyl group and a primary amino group.

[0091] In one embodiment of the present invention, in the above-mentioned compounds of formula I, formula IA or formula IB, when PS can be formed by a PS-2 photosensitizer compound, L can be formed by an L-2A linker compound, and the two are connected to each other through an etherification reaction of two hydroxyl groups.

[0092] In one embodiment of the present invention, in the above-mentioned compounds of formula I, formula IA or formula IB, when PS can be formed by any one of the photosensitizer compounds PS-4, PS-7 and PS-8, L can be formed by any one of the linker compounds L-1B, L-1F and L-2C, and the two are connected to each other through an esterification reaction of a hydroxyl group and a carboxyl group.

[0093] In one embodiment of the present invention, in the above-mentioned compounds of formula I, formula IA or formula IB, when PS can be formed by any one of the photosensitizer compounds PS-5 and PS-6, L can be formed by any one of the linker compounds L-1C, L-1D and L-1E, and the two are interconnected through a nucleophilic substitution reaction between a silanol group and a silyl ether group.

[0094] Furthermore, the coupled compound intermediate (CM) formed by the linker compound and the photosensitizer compound also falls within the protection scope of the present invention.

[0095] In the present invention, each CM can be independently any one of the following compounds, wherein the alkenyl group in the azide group or maleimide ring can react with the corresponding group (e.g., alkynyl group or thiol group) in the spacer compound to complete the connection between the coupled compound intermediate and the spacer fragment.

[0096]

[0097]

[0098]

[0099]

[0100] In one embodiment of the present invention, each CM may be independently any one of the following compounds or their optical isomers.

[0101]

[0102]

[0103]

[0104] In the present invention, when different fragments used to construct the coupled compound are connected in series via an amide group, the amide group can be formed by a condensation reaction of a primary amino group (e.g., an amino group in a compound forming L, S or LG) and a carboxyl group (e.g., a carboxyl group in a compound forming PS). Generally, the above condensation reaction can be carried out in the presence of a coupling agent so as to activate the carboxylic acid as a preferred electrophilic reagent, thereby promoting the forward progress of the reaction. Exemplary coupling agents include, but are not limited to, EDC (1-ethyl-3-(3-dimethylaminopropyl) carbodiimide), DCC (dicyclohexylcarbodiimide), HOBt (1-hydroxybenzotriazole), HATU (O-(7-azabenzotriazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate), etc. In addition, the formation of an amide bond can also be achieved by activating a carboxylic acid with N-hydroxysuccinimide (NHS) to form a succinimide ester, which can be further reacted with an amine in the absence of any other coupling agent.

[0105] <Second Aspect>

[0106] The present invention provides the following specific presentation forms of the conjugate compound.

[0107]

[0108]

[0109]

[0110]

[0111]

[0112]

[0113]

[0114] Preferably, the present invention provides the following specific forms of the coupled compound or optical isomers thereof.

[0115]

[0116]

[0117]

[0118]

[0119]

[0120]

[0121]

[0122] <Third Aspect>

[0123] The present invention provides a pharmaceutical composition comprising the conjugate compound described in <the first aspect> or <the second aspect>.

[0124] Preferably, the pharmaceutical composition further comprises at least one pharmaceutically acceptable excipient.

[0125] <Fourth Aspect>

[0126] The present invention provides use of the conjugate compound described in <the first aspect> or <the second aspect> or the pharmaceutical composition described in <the third aspect> in the preparation of a drug for diagnosing, preventing and / or treating a disease or condition.

[0127] <Fifth Aspect>

[0128] The present invention provides the conjugate compound described in <the first aspect> or <the second aspect> or the pharmaceutical composition described in <the third aspect>, which is used for diagnosing, preventing and / or treating a disease or condition.

[0129] <Sixth Aspect>

[0130] The present invention provides a method for diagnosing, preventing and / or treating a disease or condition, comprising: administering a diagnostically, preventively and / or therapeutically effective amount of the conjugate compound described in <the first aspect> or <the second aspect> or the pharmaceutical composition described in <the third aspect> to an individual in need thereof.

[0131] Preferably, in the <Fourth Aspect>, <Fifth Aspect> and / or <Sixth Aspect>, the disease or condition is cancer or a precancerous lesion; preferably, the cancer or precancerous lesion occurs in a cavity, a lumen or a body surface; more preferably, the cancer is selected from skin cancer, esophageal cancer, colorectal cancer, lung cancer, cervical cancer and bladder cancer.

[0132] Effects of the Invention

[0133] The conjugate compound of the present invention (or its pharmaceutically acceptable derivative form, such as an addition salt) combines the advantages of ligand-drug conjugates and photodynamic technology. By coupling a photosensitizer to a ligand with a targeting effect, while utilizing the advantages of the photosensitizer drug itself, such as small side effects and low drug resistance, the targeting of the photosensitizer is improved, thereby providing a possible drug-making method for the photosensitizer drug. BRIEF DESCRIPTION OF THE DRAWINGS

[0134] Figure 1 This is the MS spectrum of intermediate 1 in Example 3.

[0135] Figure 2 This is the MS spectrum of compound CM-5' in Example 3.

[0136] Figure 3 This is the MS spectrum of compound 9' in Example 3.

[0137] Figure 4 This is the MS spectrum of compound 10' in Example 4.

[0138] Figure 5 This is the MS spectrum of compound CM-4' in Example 5.

[0139] Figure 6 This is the MS spectrum of compound 7' in Example 5.

[0140] Figure 7 is the MS spectrum of compound 8' in Example 6.

[0141] Figure 8 This is the MS spectrum of compound BLG-2' (compound NC) in Example 7.

[0142] Fig. 9 This is the MS spectrum of compound 11′ in Example 7.

[0143] Fig.10 The graph shows the effect of compound 8' on the body weight of mice in each group of SKOV-3 model (mean ± standard error).

[0144] Fig.11 The graph shows the effect of compound 8' on the body weight changes of mice in each group of the SKOV-3 model (mean ± standard error).

[0145] Fig.12 The graph shows the effect of compound 8' on the tumor volume of each group of mice in the SKOV-3 model (mean ± standard error).

[0146] Fig.13 The graph shows the effect of compound 8' on the changes in tumor volume in each group of mice in the SKOV-3 model (mean ± standard error). DETAILED DESCRIPTION

[0147] General Terms and Definitions

[0148] Unless otherwise stated, the terms used in the present invention have the following meanings.

[0149] In the present invention, "target" or "drug target" refers to the binding site between drugs and biological macromolecules of the body, mainly involving receptors, enzymes, ion channels, transporters, immune system, genes, etc. Among existing drugs, more than 50% of drugs target receptors, which become the main and most important targets; more than 20% of drugs target enzymes, especially enzyme inhibitors, which have a special status in clinical applications; about 6% of drugs target ion channels; 3% of drugs target nucleic acids; and the targets of 20% of drugs are still to be further studied.

[0150] In the present invention, "conjugate compound", "conjugate", "conjugated drug" or "drug conjugate" refers to a pharmaceutically acceptable compound formed by connecting fragments of different functions through optional connecting fragments and capable of exerting corresponding activities, such as antibody-drug conjugates (ADC), polypeptide-drug conjugates (PDC), small molecule-drug conjugates (SMDC), radionuclide-drug conjugates (RDC), ligand-photosensitizer conjugates (LPC), etc.

[0151] In the present invention, "photosensitizer" refers to a drug that can trigger a photodynamic reaction to destroy cell structures.

[0152] In the present invention, "ligand-photosensitizer conjugate" or "ligand-coupled photosensitizer" refers to a pharmaceutically acceptable conjugate formed by connecting fragments such as a photosensitizer and a targeting ligand for at least one target through a linker and a spacer, wherein the photosensitizer portion can exert a photodynamic effect, the targeting ligand portion is used to exert targeted positioning for one or more specific targets (such as receptors), and the linker portion and the spacer portion are used to connect the photosensitizer portion and the targeting ligand portion to each other to form the complete structure of the ligand-photosensitizer conjugate.

[0153] In the present invention, "targeting ligand" refers to any molecule or part that can be targeted to a target site, target tissue, target organ, target cell or target cell intraregional area. In some embodiments, the targeting ligand makes the part connected to the targeting ligand more distributed in the target site, target tissue, target organ, target cell or target cell intraregional area compared to the non-target site, non-target tissue, non-target organ, non-target cell or non-target cell intraregional area, for example, at least 10%, 20%, 50%, 80%, 100%, 150%, 200%, 300%, 400%, 500% or higher. In some embodiments, compared to the non-targeting ligand, the conjugate compound or reagent with the targeting ligand is distributed more in the target site, target tissue, target organ, target cell or target cell intraregional area, for example, at least 10%, 20%, 50%, 80%, 100%, 150%, 200%, 300%, 400%, 500% or higher. In some embodiments, the targeting ligand is capable of triggering or promoting the specific binding of a conjugate compound containing such a targeting ligand to a target molecule, triggering or promoting the endocytosis of the conjugate compound by target cells, triggering or promoting the enrichment of the conjugate compound around target cells and / or entering target cells.

[0154] In the present invention, "ligand" may include a variety of chemical molecules or polypeptides that have specific binding affinity for a selected target, and the selected target may be a cell surface protein (e.g., a cell surface receptor or a cell surface antigen), a specific protein, a cell, a tissue, an organ, etc. In some embodiments, the ligand may specifically bind to a cell surface receptor. In some embodiments, the ligand may specifically bind to a cell surface antigen. In some embodiments, the ligand may specifically bind to a specific protein, and the specific protein may be a disease-causing protein. In some embodiments, the overexpression of the specific protein may cause a disease or the specific protein may be a mutant protein that causes a disease. In some embodiments, the ligand of the present application may be expressed as 10 -6 ~10 -11 M(K d In some embodiments, the ligand of the present application binds to the target with an affinity of at least 10 -6 , at least 10 -7 , at least 10 -8 or at least 10 -9 M(K dIn some embodiments, the ligand of the present application binds to the target with an affinity of at least two, three, four, five, six, eight, ten, twenty, fifty, one hundred or more times higher than the affinity of the non-target (e.g., other cell surface receptors, cell surface antigens or specific proteins, etc.). In some embodiments, the expression of the cell surface receptors, cell surface antigens, and specific proteins of the present application on the surface of target cells (e.g., cancer cells or cells with abnormal physiological functions) or in target cells is significantly higher than that in normal cells. The term "significant" used in the present application refers to a statistically significant difference, or a significant difference that can be recognized by a person skilled in the art.

[0155] In the present invention, "FOLR1" refers to folate receptor 1, a glycosylphosphatidylinositol (GPI)-anchored glycoprotein that binds folate with nanomolar affinity, thereby promoting receptor-mediated endocytosis. Fast-growing solid malignancies, including ovarian and lung cancers, rely on folate for metabolism and nucleic acid synthesis.

[0156] In the present invention, "TRPV6" is transient receptor potential cation channel subfamily V member 6, which is a highly selective calcium ion transmembrane transport channel that mediates the active transport of calcium ions from the extracellular to the intracellular. TRPV6 is expressed in normal human kidneys, gastrointestinal tract, pancreas, mammary glands, salivary glands, etc., but is mainly expressed in intestinal epithelial cells, which participate in the transport of calcium ions into cells. Therefore, when the number or function of TRPV6 channels changes, changes in calcium ion regulation may be caused, further leading to structural or functional abnormalities of related tissues and organs. Compared with normal tissues, TRPV6 expression is significantly increased in malignant tumors such as breast cancer, bile duct cancer, ovarian cancer, squamous cell lung cancer, and prostate cancer, and its abnormal expression may be related to the formation and progression of tumors.

[0157] In the present invention, "PSMA" is prostate-specific membrane antigen, which refers to a type II transmembrane glycoprotein present in the membrane of prostate epithelial cells, consisting of 750 amino acids, which have 19 intracellular amino acids, 24 transmembrane amino acids and 707 extracellular amino acids. Prostate-specific membrane antigen is expressed in normal prostate epithelial cells, but its expression level in prostate cancer cells is much higher. Compared with the prostate-specific antigen traditionally used for clinical detection, prostate-specific membrane antigen is a more sensitive and specific prostate cancer tumor marker, especially in hormone-refractory prostate cancer and prostate cancer metastasis, which are highly expressed, and has high sensitivity and specificity in distinguishing prostate cancer from other types of malignant tumors. At the same time, in a variety of non-prostate-derived solid tumors (such as lung cancer, bladder cancer, gastric cancer, pancreatic cancer, kidney cancer and colorectal cancer, etc.), prostate-specific membrane antigen is also highly specifically expressed on tumor vascular endothelial cells.

[0158] In the present invention, "pharmaceutical composition" refers to a composition for pharmaceutical use, which contains a small molecule drug, a polypeptide, an antibody (or an antibody-like ligand) or a conjugate thereof as an active pharmaceutical ingredient (API), and other components (e.g., pharmaceutically acceptable excipients). The pharmaceutical composition can be prepared using any method known to those skilled in the art. For example, conventional mixing, dissolving, granulating, emulsifying, grinding, encapsulating, embedding and / or lyophilizing processes.

[0159] In the present invention, "pharmaceutically acceptable excipients" refer to auxiliary materials widely used in the field of drug production. The main purpose of using excipients is to provide a pharmaceutical composition that is safe to use, stable in nature and / or has specific functionality, and also to provide a method so that after the drug is administered to the subject, the active ingredient can be dissolved at a desired rate, or to promote the active ingredient to be effectively absorbed in the body of the subject receiving the drug. Pharmaceutically acceptable excipients can be inert fillers, or they can be functional ingredients that provide a certain function to the pharmaceutical composition (for example, stabilizing the overall pH value of the composition or preventing the degradation of the active ingredient in the composition). Examples of "pharmaceutically acceptable excipients" include, but are not limited to, adhesives, suspending agents, emulsifiers, diluents (or fillers), granulating agents, adhesives, disintegrants, lubricants, anti-adhesive agents, glidants, wetting agents, gelling agents, absorption delay agents, dissolution inhibitors, enhancers, adsorbents, buffers, photosensitizers, preservatives, colorants, flavoring agents, sweeteners, etc.

[0160] The technical scheme of the present invention will be described below in conjunction with specific embodiments. The following embodiments are provided to further illustrate the present invention, but are not intended to limit the scope of the present invention. It will be apparent to those skilled in the art that various changes and improvements can be made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention.

[0161] The starting materials used in the present invention can be synthesized by methods known in the art, or purchased by conventional commercial means. The separation and purification of the compounds of the present invention can be achieved by methods well known to those skilled in the art, including but not limited to column chromatography (CC), high performance liquid chromatography (HPLC), ultra-high performance liquid chromatography (UPLC), etc. The structural identification of the compounds of the present invention can be achieved by methods well known to those skilled in the art, including but not limited to nuclear magnetic resonance (NMR), mass spectrometry (MS), etc.

[0162] Abbreviations

[0163]

[0164] Preparation method of double ligand compound (BLG)

[0165]

[0166] Load the resin into a solid phase reaction column, add DMF, and bubble nitrogen into the solvent to swell the resin for 30 minutes. Add Fmoc-Arg-OH, DCC, and DMAP, react at 25°C for 3 hours, then cap with acetic anhydride and pyridine for 1 hour, wash with DMF 3 times, remove the Fmoc protecting group on the resin with DBLK, and then wash with DMF 5 times. Weigh Fmoc-Pro-OH and HOBt, dissolve them with DMF, add DIC to the above solution in an ice water bath at 0°C, and mix to activate it for 5 minutes. Add the solution to the above reaction column, react for 3 hours, drain the solvent, wash the resin in the reaction column 3 times, and then remove the Fmoc protecting group with DBLK. Repeat the above operation, and sequentially couple Fmoc-Leu-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Val-OH, Fmoc-Lys(Boc)OH, Fmoc-Ser(tBu)-OH, Fmoc-Pro-OH, Fmoc-His(Trt)-OH, Fmoc-Leu-OH, Fmoc-Phe-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Lys(Boc)-OH, Fmoc-Cys(Trt)-OH, Fmoc-Glu-OtBu and pteroic acid according to the structure to obtain a peptide resin. After the peptide resin is cleaved with a cleavage solution, it is filtered, the cleavage solution is collected and poured into MTBE, the solid is precipitated, and it is washed with MTBE to obtain a crude product, which is prepared by preparative high performance liquid chromatography (Pre-HPLC) to obtain BLG-4'.

[0167] BLG-1', BLG-2' and BLG-3' were obtained by steps similar to the above method.

[0168] Example 1: Synthesis of Compound 5'

[0169]

[0170] Step 1: Synthesis of CM-3

[0171] Under nitrogen protection, PS-3 (0.60 g, 0.799 mmol) was added to DMF (9 ml), and then EDC·HCl (0.32 g), HOSU (0.19 g) and DIPEA (0.4 ml) were added, and the reaction was carried out at room temperature for 17 h. After the reaction was completed, the reaction solution was poured into MTBE (30 ml), and a solid was precipitated. The mixture was stirred for 10 min and filtered. The filter cake was washed with MTBE and dried to obtain an intermediate. The intermediate was dissolved in DMF (9 ml), DIPEA (0.3 ml) was added, and then raw material 1 (0.17 g) was added. The reaction was carried out at room temperature for 2 h. After the reaction was completed, the reaction solution was purified by preparative chromatography (acetonitrile / water = 5%-80%, v / v) to obtain CM-3 (0.50 g, yield: 65.8%).

[0172] Step 2: Synthesis of compound 5'

[0173] Under nitrogen atmosphere, CM-3 (9.8 mg, 6.09 mmol) was added to DMF (0.8 ml), and purified water (0.17 ml) was added, and the mixture was stirred at 25°C. BLG-1' (19.2 mg) was weighed and added to the reaction solution. The pH value of the reaction system was adjusted to about 8 with DIPEA, and cuprous bromide (2.7 mg) was added. The reaction was allowed to react for 1 h. After the reaction was completed, the mixture was purified by preparative chromatography (acetonitrile / water = 5%-80%, v / v) to give compound 5' (16.2 mg, yield: 55.2%).

[0174] Example 2: Synthesis of Compound 6'

[0175]

[0176] Compound 6' was synthesized by referring to the method of compound 5'.

[0177] Example 3: Synthesis of Compound 9'

[0178]

[0179] Step 1: Synthesis of intermediate 1

[0180] PS-5 (3.0 g, 5.2 mmol) was dispersed in pyridine (30 ml), (3-aminopropyl)dimethylethoxysilane (6.7 g, 41.6 mmol) was added dropwise, the mixture was refluxed for 4 h under argon atmosphere, and most of the pyridine was removed by concentration under reduced pressure; the residue was filtered, washed with water-ethanol solution (v:v=2:1), and then vacuum dried to obtain intermediate 1 (2.8 g, 4.06 mmol, yield: 78%); MS (ESI-): m / z 803.9 [MH] - (like Figure 1 shown).

[0181] Step 2: Synthesis of CM-5'

[0182] Under nitrogen atmosphere, intermediate 1 (2.7 g, 3.357 mmol) and 6-maleimidocaproic acid (1.4 g, 6.71 mmol) were dissolved in DMF (50 ml), HATU (3.8 g, 10.07 mmol) was added, and then DIPEA (2.6 g, 20.14 mmol) was added dropwise, and the mixture was reacted at room temperature for 1.5 h. Water (150 ml) was added to quench the reaction, and the mixture was extracted with dichloromethane (50 ml*3). The organic phases were combined, washed with water, washed with saturated brine, dried and concentrated under reduced pressure, and purified by preparative chromatography (acetonitrile / water = 5%-80%, v / v) to obtain CM-5' (1.9 g, 2.15 mmol, yield: 64.2%); MS (ESI+): m / z 885.6 [M+H] + (like Figure 2 shown).

[0183] Step 3: Synthesis of compound 9'

[0184] Under nitrogen atmosphere, CM-5' (50 mg, 0.0566 mmol) was dissolved in DMF (7.5 mL), and an aqueous solution (3.0 mL) of BLG-4' (165.6 mg, 0.079 mmol) was added dropwise. The mixture was reacted at room temperature for 1 h. After dissolution with PBS solution, the mixture was purified by preparative chromatography (acetonitrile / water = 5%-80%, v / v) to give compound 9' (83.0 mg, yield: 49.5%); MS (ESI+): m / z 991.6 [M-3H] 3- (like Figure 3 shown).

[0185] Example 4: Synthesis of Compound 10'

[0186]

[0187] Referring to the method of compound 9', compound 10' was synthesized (yield: 14.5%); MS (ESI-): m / z 801.2 [M-3H]3- (like Figure 4 shown).

[0188] Example 5: Synthesis of Compound 7'

[0189]

[0190] Step 1: Synthesis of intermediate 1

[0191] PS-5 (3.0 g, 5.2 mmol) was dispersed in pyridine (30 ml), (3-aminopropyl)dimethylethoxysilane (6.7 g, 41.6 mmol) was added dropwise, and the mixture was refluxed for 4 h under argon atmosphere and concentrated under reduced pressure to remove most of the pyridine; the residue was filtered, washed with water-ethanol solution (v:v=2:1), and then dried in vacuo to obtain intermediate 1 (2.8 g, 4.06 mmol, yield: 78%).

[0192] Step 2: Synthesis of CM-4'

[0193] Under nitrogen atmosphere, intermediate 1 (4.90 g, 6.09 mmol) and 6-maleimidocaproic acid (3.08 g, 14.61 mmol) were dissolved in DMF (100 mL), HATU (6.94 g, 18.28 mmol) was added, and DIPEA (4.73 g, 36.56 mmol) was added dropwise, and the mixture was reacted at room temperature for 1.5 h. Water (300 ml) was added to quench the reaction, and the solid was precipitated by stirring, filtered, and the filter cake was washed with water, dried, and purified by silica gel column chromatography (dichloromethane / methanol = 100 / 1-10 / 1, v / v) to obtain CM-4' (1.9 g, 1.60 mmol, yield: 26%); MS (ESI+): m / z 1192.7 [M+H] + (like Figure 5 shown).

[0194] Step 3: Synthesis of compound 7'

[0195] Under nitrogen atmosphere, CM-4' (20 mg, 0.0168 mmol) was dissolved in DMF (3 mL), and an aqueous solution (1.2 ml) of BLG-4' (84.31 mg, 0.0403 mmol) was added dropwise. The pH value was adjusted to 6-7 with PBS solution. The mixture was reacted at room temperature for 1 h. After dissolution with PBS solution, the mixture was purified to obtain compound 7' (33.6 mg, 0.0063 mmol, yield: 37.5%); MS (ESI-): m / z 1342.3 [M-4H] 4- (like Figure 6 shown).

[0196] Example 6: Synthesis of Compound 8'

[0197]

[0198] Referring to the method of compound 7', compound 8' was synthesized (yield: 54.4%); MS (ESI-): m / z 1411.8 [M-3H] 3- (like Figure 7 shown).

[0199] Example 7: Synthesis of Compound 11'

[0200]

[0201] Step 1: Synthesis of intermediate 1

[0202] PS-5 (3.0 g, 5.2 mmol) was dispersed in pyridine (30 ml), (3-aminopropyl)dimethylethoxysilane (6.7 g, 41.6 mmol) was added dropwise, and the mixture was refluxed for 4 h under argon atmosphere and concentrated under reduced pressure to remove most of the pyridine; the residue was filtered, washed with water-ethanol solution (v:v=2:1), and then dried in vacuo to obtain intermediate 1 (2.8 g, 4.06 mmol, yield: 78%).

[0203] Step 2: Synthesis of CM-6'

[0204] Under nitrogen protection, intermediate 1 (200 mg) and raw material 2 (161 mg) were dissolved in DMAc (5 ml), HOBt (33 mg) was added, and DIPEA (64 mg) was added dropwise. The reaction was carried out at room temperature for 0.5 h. Sampling was performed and after the reaction was completed, the reaction solution was poured into water (40 ml), and the solid was precipitated. The solid was filtered, the filter cake was washed with water (10 ml*4), and vacuum dried to obtain a dark green solid CM-6' (144 mg, yield: 31.6%).

[0205] Step 3: Synthesis of compound 11'

[0206] Under nitrogen protection, CM-6' (140 mg) was dissolved in DMF (5 mL), and BLG-2' (ie, compound NC, such as Figure 8 The reaction mixture was added with 1% ethyl acetate (as shown) (300 mg) in an aqueous solution (5 mL); the pH value of the PBS solution was adjusted to about 7, and the reaction was carried out at room temperature; after the intermediate control reaction was completed, the reaction solution was subjected to preparative chromatography purification (acetonitrile / water = 5%-80%, v / v), and lyophilized to obtain compound 11' (110 mg, yield: 29.6%); MS (ESI-): m / z 1217.3 [M-4H] 4- (like Fig. 9 shown).

[0207] Comparative Example: Synthesis of Compound PC

[0208]

[0209] Step 1: Synthesis of Raw Material 3

[0210] Fmoc-Cys(Trt)-OH was immobilized on an amino resin by Fmoc chemical solid phase synthesis, and then the Fmoc protecting group was removed with a 20% piperidine DMF solution, and the mixture was washed with DMF to remove impurities. A DMF solution of Fmoc-Glu-OtBu was added, and HATU and DIPEA were added for condensation reaction. Ninhydrin developed a light yellow color, and the mixture was washed with DMF to remove impurities. The Fmoc removal and condensation steps were repeated to obtain a crude resin. The product was cleaved from the resin with trifluoroacetic acid and the protecting group was removed at the same time to obtain a crude raw material 3, which was purified by preparative chromatography to obtain raw material 3.

[0211] Step 2: Synthesis of compound PC

[0212] Referring to the method of compound 7', compound PC was synthesized (yield: 26.3%).

[0213] Experimental Example 1: Experimental study on the proliferation inhibition of different cells by compounds 8', 9', 10', NC (BLG-2') and PC

[0214] 1. Experimental Materials

[0215] 1.1 Cell line information

[0216] Table 1. Expression of multiple targets in different cells

[0217] Cell Name Chinese name FOLR1 PSMA TRPV6 293T Human renal epithelial cells Not express Not express Not express 293T-FOLR1 / PSMA Human renal epithelial cells High expression High expression Not express SK-OV-3 Human ovarian cancer cells High expression Low expression Low expression HT29 Human colon cancer cells Medium Expression Low expression Low expression

[0218] The results in Table 1 showed that 293T cells did not express FOLR1, PSMA and TRPV6; 293T-FOLR1 / PSMA cells highly expressed FOLR1 and PSMA, but did not express TRPV6; SK-OV-3 cells highly expressed FOLR1, but lowly expressed PSMA and TRPV6; HT29 cells expressed FOLR1, and lowly expressed PSMA and TRPV6.

[0219] 1.2 Main reagents: 1640 folic acid-free culture medium, fetal bovine serum, penicillin-streptomycin solution, L-glutamine and CCK8.

[0220] 2. Experimental Procedure

[0221] 2.1 Cell plating

[0222] Prepare cells in advance, digest with trypsin, collect, and count; dilute cells to 2×10 using complete cell culture medium. 4Cells / ml were plated on a 96-well plate, 100 μl of diluted cell solution was added to each well, negative control (without drug, other conditions were the same) and blank control (containing only cell culture medium) wells were set up on each plate; the 96-well plate with cells was placed in a 37°C, 5% CO2 incubator and cultured overnight.

[0223] 2.2 Dilution and sample loading

[0224] The sample was diluted with culture medium (initial concentration 10 μM, 5-fold gradient dilution), with a total of 9 concentration gradients; the culture medium in the well plate was discarded, and the drug solution was added to a 96-well plate at a volume of 100 μl per well, with 3 replicates, and a negative control, a blank control and a non-illuminated group were set up and placed in a 37°C, 5% CO2 incubator for 24 h; 16 J / cm 2 The well plate was irradiated with light and then placed in a 37°C, 5% CO2 incubator for 24 hours.

[0225] 2.3 Chromogenic plate reading

[0226] Take CCK-8 colorimetric solution, add 10 μl (10% of the liquid volume in the well) to each well, incubate at 37°C for an appropriate time (try to keep the OD value within the range of 1.0-2.5), remove the culture plate cover of the 96-well plate, place it in a microplate reader (Molecular Devices SpectraMax iD5), and read the value at 450 nm.

[0227] 2.4 Data Processing

[0228] GraphPad Prism 8.0.2 was used to perform four-parameter fitting to obtain the IC 50 value.

[0229] 2.5 Experimental Results and Analysis

[0230] Table 2. IC values ​​of compounds under illuminated and non-illuminated conditions 50

[0231]

[0232] Note: N / A means no value was fitted and no obvious inhibitory effect.

[0233] The results in Table 2 show that under illumination, compounds 8', 9' and 10' have a significant killing effect on cells that highly express FOLR1 and PSMA; under non-illumination conditions, compounds 8', 9', 10', PC and NC have no significant inhibitory effect on cells; therefore, the following conclusions can be drawn:

[0234] (1) The ligand-photosensitizer conjugate of the present invention requires light conditions to exert its efficacy, so the compound NC that does not contain a photosensitizer (PS) does not show an inhibitory effect on cells;

[0235] (2) The efficacy of the drug is correlated to a certain extent with the cell expression. When the cells express highly, the killing effect is higher (293T-FOLR1 / PSMA), and when the cells express low or no expression, the killing effect is lower / no killing (293T, HT29). This is also in line with the concept of targeted precision diagnosis and treatment.

[0236] (3) When the dosage concentrations are equivalent, the two-arm dual-ligand conjugate (e.g., compound 8') shows a higher killing effect than the one-arm dual-ligand conjugate (e.g., compound 10') or the two-arm single-ligand conjugate (e.g., compound PC), whether for 293T-FOLR1 / PSMA or SK-OV-3, indicating that the two-arm dual-ligand structure can further enhance the affinity of the conjugate to the target cells and improve the efficacy; moreover, when achieving equivalent efficacy, the actual dosage of the two-arm dual-ligand conjugate can also be reduced, further improving the safety of the drug.

[0237] Although the embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and cannot be understood as limiting the present invention. Without departing from the principles and purpose of the present invention, those skilled in the art may change, modify, replace and modify the above embodiments within the scope of the present invention, and these changes, modifications, replacements and modifications are all within the scope of the present invention.

[0238] Experimental Example 2: In vivo pharmacodynamics study of compound 8'

[0239] 1. Experimental Methods

[0240] SK-OV-3 cells were cultured in vitro in the presence of McCoy's 5A medium supplemented with 10% fetal bovine serum, 100 U / mL penicillin and 100 μg / mL streptomycin at 37°C in a 5% CO2 incubator. Routine treatment and subculture were performed twice a week. When the cell saturation reached 80%-90% and the number reached the required level, the cells were harvested, counted and inoculated.

[0241] 10*10 6 100 cells (inoculation volume 0.1 mL) were subcutaneously inoculated on the right dorsal flank of each mouse. When the average tumor volume reached 150 mm 3 The day of grouping was set as D0, and the day of first administration was set as D1.

[0242] The animal body weight and tumor size were measured on the day of grouping. According to the size of the animal tumor, 33 animals were randomly divided into 11 groups using Grouping, including a blank control group; a large group of high, medium and low doses of compound 8' (45 mg / kg, 15 mg / kg and 5 mg / kg, respectively), each large group was set up with a light interval of 4, 6 and 8 hours after administration (administered once every two weeks, three times in total) and a 5 mg / kg dose group of compound 8', with a light interval of 4 hours after administration (administered once a week, five times in total). No operation was given to the blank control group. Animal grouping and lighting conditions are shown in Table 3.

[0243] The medication is as follows:

[0244] Route of administration: intravenous administration (iv);

[0245] Dosing frequency: once every two weeks and once a week.

[0246] Table 3. Animal groups and lighting conditions for in vivo pharmacodynamics experiments in mice

[0247]

[0248] 2. Experimental results

[0249] The body weight and body weight changes of experimental mice at different time points are as follows Fig.10 and Fig.11 There was no significant change in the body weight of the animals in each group. After four administrations in group 11, one of the mice was found to be emaciated.

[0250] The tumor volume and tumor volume changes of experimental mice at different time points are shown in Figure 2 Fig.12 and Fig.13 As shown. The animal data of the groups with 4h interval illumination after administration (Groups 1, 4, 7 and 11) were relatively impressive and dose-related, among which 45mg / kg illuminated once every two weeks (Group 7) had a good tumor inhibition effect. 5mg / kg illuminated once a week (Group 11) and 15mg / kg illuminated once every two weeks (Group 4) showed similar tumor inhibition effects. No obvious tumor inhibition effect was observed at 5mg / kg illuminated once every two weeks (Group 1).

Claims

1. A conjugate compound comprising a photosensitizer and 1 to 6 targeting ligands that specifically bind to cell surface proteins.

2. The coupled compound according to claim 1, characterized in that The photosensitizer is formed of porphyrin, phthalocyanine, cyanine dye or Bodipy compound, preferably formed of phthalocyanine compound, more preferably formed of silicon phthalocyanine or zinc phthalocyanine compound.

3. The coupled compound according to claim 2, characterized in that The photosensitizer is formed by any one of the following photosensitizer compounds or optical isomers thereof:

4. The coupled compound according to any one of claims 1 to 3, characterized in that The conjugate compound comprises a photosensitizer and 2 or 4 targeting ligands.

5. The coupled compound according to any one of claims 1 to 4, characterized in that Each targeting ligand in the conjugate compound independently binds to any one of the following cell surface proteins: PSMA, FOLR1 and TRPV6.

6. The coupled compound according to claim 4 or 5, characterized in that When the conjugate compound comprises 2 or 4 targeting ligands, the conjugate compound binds to any one of the following cell surface protein combinations: PSMA / FOLR1, FOLR1 / TRPV6 and PSMA / TRPV6, preferably PSMA / FOLR1 and FOLR1 / TRPV6, and more preferably PSMA / FOLR1.

7. The coupled compound according to any one of claims 1 to 6, characterized in that Each targeting ligand in the conjugate compound is independently formed by a polypeptide, an antibody or a small molecule.

8. The coupled compound according to claim 7, characterized in that Each targeting ligand in the coupled compound is independently formed by any one of the ligand compounds of formula LG-I, LG-II and LG-III, In formula LG-I, R LG1 For hydroxyl, Preferably, R LG1 For hydroxyl, Formula LG-II is pteroic acid, folic acid or an analog thereof; preferably, the folic acid analog is selected from 5-methyltetrahydrofolate, 5-formyltetrahydrofolate, 10-formylfolate, methotrexate, aminopterin and raltitrexed; Formula LG-III comprises all or part of the amino acids in the polypeptide EGKLSSNDTEGGLCKEFLHPSKVDLPR; preferably, formula LG-III comprises 9 to 27 amino acids in the above polypeptide; more preferably, formula LG-III has at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity with the polypeptide KEFLHPSKVDLPR; further preferably, formula LG-III is the polypeptide KEFLHPSKVDLPR.

9. The coupled compound according to claim 8, characterized in that Each targeting ligand in the conjugate compound is independently a targeting ligand formed by a ligand compound of formula LG-I or an optical isomer thereof and binds to a cell surface protein PSMA; A targeting ligand formed by a ligand compound of formula LG-II or an optical isomer thereof and binds to the cell surface protein FORL1; or, a targeting ligand formed by a ligand compound of formula LG-III or an optical isomer thereof and binds to the cell surface protein TRPV6.

10. The coupled compound according to claim 8, characterized in that Each targeting ligand in the conjugate compound is independently formed by any one of the following ligand compounds or their optical isomers:

11. The coupled compound according to any one of claims 1 to 10, characterized in that It also comprises a connecting part; the connecting part is used to connect the photosensitizer and the targeting ligand and / or connect multiple targeting ligands.

12. The coupled compound according to claim 11, characterized in that The connecting part comprises a linker and a spacer; the linker is used to connect the photosensitizer and the targeting ligand; the spacer is used to connect a plurality of the targeting ligands.

13. A coupled compound having a structure as shown in Formula I, in, LG indicates targeting ligand; S represents a spacer; L represents a linker; PS indicates photosensitizer; n is 1 or 2; and The two LGs are identical to or different from each other; When n is 2, two The same as or different from each other.

14. The coupled compound according to claim 13, characterized in that: It has a structure as shown in Formula IA, wherein LG, S, L and PS are as defined in claim 13.

15. The coupled compound according to claim 13, characterized in that: It has a structure as shown in Formula IB, wherein LG, S, L and PS are as defined in claim 13.

16. The coupled compound according to any one of claims 13 to 15, characterized in that Each LG independently binds to any of the following cell surface proteins: PSMA, FOLR1, and TRPV6.

17. The coupled compound according to claim 16, characterized in that: The conjugate compound binds to any one of the following cell surface protein combinations: PSMA / FOLR1, FOLR1 / TRPV6 and PSMA / TRPV6, preferably PSMA / FOLR1 and FOLR1 / TRPV6, and more preferably PSMA / FOLR1.

18. The coupled compound according to any one of claims 13 to 17, characterized in that Each LG is independently formed by a peptide, an antibody or a small molecule.

19. The coupled compound according to claim 18, characterized in that: Each LG is independently formed by any one of the ligand compounds of formula LG-I, LG-II and LG-III, In formula LG-I, R LG1 For hydroxyl, Preferably, R LG1 For hydroxyl, Formula LG-II is pteroic acid, folic acid or an analog thereof; preferably, the folic acid analog is selected from 5-methyltetrahydrofolate, 5-formyltetrahydrofolate, 10-formylfolate, methotrexate, aminopterin and raltitrexed; Formula LG-III comprises all or part of the amino acids in the polypeptide EGKLSSNDTEGGLCKEFLHPSKVDLPR; preferably, formula LG-III comprises 9 to 27 amino acids in the above polypeptide; more preferably, formula LG-III has at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity with the polypeptide KEFLHPSKVDLPR; further preferably, formula LG-III is the polypeptide KEFLHPSKVDLPR.

20. The coupled compound according to claim 18, characterized in that Each LG is independently a targeting ligand formed by a ligand compound of formula LG-I or an optical isomer thereof and binds to the cell surface protein PSMA; a targeting ligand formed by a ligand compound of formula LG-II or an optical isomer thereof and binds to the cell surface protein FORL1; or, a targeting ligand formed by a ligand compound of formula LG-III or an optical isomer thereof and binds to the cell surface protein TRPV6.

21. The coupled compound according to claim 18, characterized in that Each LG is independently formed by any of the following ligand compounds or their optical isomers:

22. The coupled compound according to any one of claims 13 to 21, characterized in that Each S is independently formed by any of the following spacer compounds or their optical isomers:

23. The conjugate compound according to claim 22, characterized in that: Each S is independently formed by any one of the spacer compounds of S-1, S-2, S-3 and S-4 or its optical isomers and an additional amino acid or amino acid combination or its optical isomers, wherein the additional amino acid or amino acid combination is selected from Lys, Cys, Glu, Lys-Cys, Cys-Cys, Lys-Cys-Lys, Arg-Arg, Ala-Ser-Asn, Ala-Ala-Ala, Ser-Ser-Arg, Pro-Arg, Asp-Asp-Lys-Cys and Pro-Leu-Gly, and the additional amino acid or amino acid combination is optionally amidated.

24. The conjugate compound according to any one of claims 13 to 23, characterized in that Each L is independently formed by any one of the linker compounds of formula LI and L-II, In formula LI, s is any integer from 1 to 8, preferably any integer from 3 to 5, more preferably 4 or 5; R L1 is amino, carboxyl or Where W is and the two ends are connected to other fragments through amide bonds; t is 0 or 1; u is any integer from 1 to 4, preferably 2 or 3, more preferably 3; v is any integer from 1 to 4, preferably 1 or 2, more preferably 2; preferably, R L1 For amino, carboxyl, In Formula L-II, p is any integer from 1 to 4, preferably any integer from 1 to 3, more preferably 2 or 3; q is any integer from 0 to 3, preferably any integer from 0 to 2, more preferably 0 or 1; r is 0 or 1; Preferably, when p is 2, q is 1, and r is 1; or Preferably, when p is 3, q ​​is 0 or 1, and r is 0.

25. The conjugate compound according to claim 24, characterized in that: Each L is independently formed by any one of the following linker compounds or their optical isomers:

26. The conjugate compound according to claim 25, characterized in that When S is formed by any one of the spacer compounds S-1 and S-2, L is formed by any one of the linker compounds L-2A, L-2B and L-2C, and the two are connected to each other through a cycloaddition reaction between an alkynyl group and an azide group.

27. The conjugate compound according to claim 25, characterized in that When S is formed by any spacer compound among S-3 and S-4, L is formed by any linker compound among L-1A, L-1B, L-1C, L-1D, L-1E and L-1F, and the two are connected to each other through the double bond addition reaction between the alkenyl group in the maleimide ring and the thiol group.

28. The conjugate compound according to any one of claims 13 to 27, characterized in that PS is formed of a porphyrin, phthalocyanine, cyanine dye or Bodipy compound, preferably a phthalocyanine compound, more preferably a silicon phthalocyanine or zinc phthalocyanine compound.

29. The conjugate compound according to claim 28, characterized in that PS is formed by any of the following photosensitizer compounds or their optical isomers:

30. The conjugate compound according to claim 29, characterized in that When PS is formed by any photosensitizer compound of PS-1 and PS-3, L is formed by any linker compound of L-1A and L-2B, and the two are connected to each other through an amidation reaction between a carboxyl group and a primary amino group.

31. The conjugate compound according to claim 29, characterized in that When PS is formed by a PS-2 photosensitizer compound, L is formed by an L-2A linker compound, and the two are connected to each other through an etherification reaction of two hydroxyl groups.

32. The conjugate compound according to claim 29, characterized in that When PS is formed by any photosensitizer compound among PS-4, PS-7 and PS-8, L is formed by any linker compound among L-1B, L-1F and L-2C, and the two are connected to each other through an esterification reaction between a hydroxyl group and a carboxyl group.

33. The conjugate compound according to claim 29, characterized in that When PS is formed by any photosensitizer compound among PS-5 and PS-6, L is formed by any linker compound among L-1C, L-1D and L-1E, and the two are connected to each other through a nucleophilic substitution reaction between a silanol group and a silyl ether group.

34. A conjugate compound, which is any one of the following compounds: Preferably, it is any one of the following compounds or their optical isomers:

35. A pharmaceutical composition comprising the conjugate compound according to any one of claims 1 to 34; Preferably, the pharmaceutical composition further comprises at least one pharmaceutically acceptable excipient.

36. Use of the conjugate compound according to any one of claims 1 to 34 or the pharmaceutical composition according to claim 35 in the preparation of a medicament for diagnosing, preventing and / or treating a disease or condition; Preferably, the disease or condition is cancer or a precancerous lesion; More preferably, the cancer or precancerous lesion occurs in a cavity, lumen or body surface; Further preferably, the cancer is selected from skin cancer, esophageal cancer, colorectal cancer, lung cancer, cervical cancer and bladder cancer.

37. The conjugate compound according to any one of claims 1 to 34 or the pharmaceutical composition according to claim 35 for use in diagnosing, preventing and / or treating a disease or condition; Preferably, the disease or condition is cancer or a precancerous lesion; More preferably, the cancer or precancerous lesion occurs in a cavity, lumen or body surface; Further preferably, the cancer is selected from skin cancer, esophageal cancer, colorectal cancer, lung cancer, cervical cancer and bladder cancer.

38. A method for diagnosing, preventing and / or treating a disease or condition comprising: administering a diagnostically, prophylactically and / or therapeutically effective amount of the conjugate compound according to any one of claims 1 to 34 or the pharmaceutical composition according to claim 35 to an individual in need thereof; Preferably, the disease or condition is cancer or a precancerous lesion; More preferably, the cancer or precancerous lesion occurs in a cavity, lumen or body surface; Further preferably, the cancer is selected from skin cancer, esophageal cancer, colorectal cancer, lung cancer, cervical cancer and bladder cancer.

39. A biligand compound, which is any one of the following compounds: Preferably, it is any one of the following compounds or their optical isomers:

40. A coupled compound intermediate, which is any one of the following compounds: Preferably, it is any one of the following compounds or their optical isomers:

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