method

Abstract

The present invention provides endoplasmic reticulum-targeted chemiluminescent agents and their use in methods of photodynamic therapy (PDT).In particular, the present invention provides compounds and pharma-ceutically acceptable salts of general formula (I), in which A represents a chemiluminescent moiety, each L, which may be the same or different, is either a direct bond or a linker, each B, which may be the same or different, represents an endoplasmic reticulum-targeting moiety, n is an integer between 1 and 3, preferably 1, and x is an integer between 1 and 3, preferably 1.Such compounds, when used in combination with a photosensitizer or photosensitizer precursor, find particular use in the treatment of deep tumors, such as glioblastoma multiforme (GBM). [Formula 1] JPEG2024536927000054.jpg18170

Description

[Technical field]

[0001] The present invention relates to improvements in and relating to methods of photodynamic therapy (PDT), in particular for the targeted treatment of diseases and conditions characterized by hyperproliferative and / or abnormal cells without the need for an external light source, and more particularly, to such methods for treating tumors, in particular tumors that are inaccessible using existing PDT methods.

[0002] The invention further relates to novel endoplasmic reticulum-targeted chemiluminescent agents, methods for their preparation, and their use as intracellular light sources in methods of PDT using photosensitizers or photosensitizer precursors. [Background technology]

[0003] Conventional treatment of internal tumors typically involves invasive surgery, radiation therapy, non-curative chemotherapy, or a combination of these. Intracranial tumors such as glioblastoma multiforme (GBM) are an example of deep tumors that are very difficult to treat due to their location and aggressive characteristics. Approximately 28,000 new cases of malignant gliomas such as GBM are diagnosed annually in the EU and the US, with 240,000 patients diagnosed annually worldwide. The current standard of care consists of highly invasive (craniotomy) surgery that removes about 99% of the tumor, but leaves behind about 1 billion cells, leading to recurrence. Radiation therapy may be used as an adjunct to surgery (at 60-65 Gy) and, when combined with surgery, can reduce the number of cancer cells left behind to a few million, but does not have a significant effect on cancers such as GBM, which tend to metastasize to several locations that also have radiation-resistant cancer cells inside. Furthermore, radiation therapy is not specific for the destruction of cancerous tissue versus normal tissue. In addition to radiation therapy, chemotherapy with temozolomide can also be used. However, these therapies limit the patient's overall survival, do not result in a curative outcome, and are primarily cytostatic, as the cells eventually (within about one year of treatment) develop resistance and the treatment is no longer effective. The combination of surgery and radiation therapy increases median survival from 4.5 months (untreated) to 12.1 months. Additional chemotherapy with temozolomide extends survival to 14.6 months. The relative survival rate for adults diagnosed with GBM is less than 30% within one year of diagnosis, and only 3% of patients survive more than five years after initial diagnosis.

[0004] Therefore, deep-seated and hard-to-reach tumors such as GBM remain extremely difficult to treat, and existing therapies offer only modest increases in survival rates.Therefore, there is an urgent need to develop more targeted, minimally invasive therapeutic approaches with improved efficacy.

[0005] Other methods known to be used to treat tumors include PDT, which involves administering a photosensitizer either locally or systemically, followed by exposure of the affected area to photoactivatable light that interacts with ambient oxygen to produce cytotoxic intermediates, resulting in the destruction of cells and the shutdown of the tumor vasculature.

[0006] PDT provides cancer therapy through the synergistic effect of three essential but individually non-chemotoxic components: (i) a photoactivatable drug, a photosensitizer (PS), (ii) light of the appropriate wavelength to activate the PS, and (iii) the presence of oxygen, the final generator of toxic species. The antitumor effects of PDT can be classified into three main interrelated effects: (i) a direct cytotoxic effect, mainly mediated via either type I or type II mechanisms (the former generates reactive oxygen species (ROS) and ultimately hydroxyl radicals, while the type II mechanism, which is prominent in the majority of PS, generates harmful singlet oxygen [O( 1 Δ g ) or 1 O2]), (ii) damage to the tumor vasculature, and (iii) induction of an inflammatory response that may lead to the development of systemic immunity as a result of PDT-induced oxidative stress.

[0007] Photosensitizers currently approved for use in methods of photodynamic therapy and diagnosis include protoporphyrin IX (PpIX), which is generated from its biosynthetic non-photosensitive precursor, 5-aminolevulinic acid (5-ALA). Following exogenous administration of 5-ALA, the heme biosynthetic cycle promotes its conversion to the active photosensitizer PpIX in cellular mitochondria. Cancer cells treated with 5-ALA accumulate large amounts of PpIX, mainly due to higher amounts of PpIX synthesis-promoting enzymes and / or significantly lower amounts of ferrochelatase (which catalyzes the chelation of iron with PpIX in the mitochondrial matrix to generate heme, which is then transported out of the mitochondria). Subsequent exposure to light excites PpIX from its ground singlet state to its excited singlet state. It then undergoes intersystem crossing to a longer-lived excited triplet state. When triplet-state PpIX interacts with molecular oxygen (which is in its ground triplet state), energy transfer occurs from PpIX to oxygen, which results in a spin flip between the two molecules, forming a cytotoxic excited singlet state of molecular oxygen while PpIX relaxes back to its ground singlet state.

[0008] Photosensitizers are also known for use in methods of photodynamic diagnosis of cancerous cells and tissues, and can also be used to guide surgical resection of tumor masses. For example, PDT is used as an adjunct to surgery in the treatment of bladder cancer. PpIX fluorescence induced by 5-ALA is also currently used intraoperatively for fluorescence-guided resection in the treatment of GBM (see Stummer et al., Lancet Oncol., 2006, 7(5):392-401). However, due to limitations of conventional PDT procedures (e.g., light reach and light penetration into tissue), they currently cannot provide any significant benefit in treating this invasive condition without the need for surgical intervention.

[0009] The main limitations of existing methods of PDT as anticancer treatment are poor judgment of the treatment area by the clinician, poor definition of the tumor volume to be treated, and limited depth of penetration of photoactivating light into tissue (approximately 1.5 cm). This leads to ineffective treatment and residual viable cancer cells. For example, 5-ALA-based PDT is specific and efficient with very high cure rates for the treatment of actinic keratosis and basal cell carcinoma, but only for lesions thinner than 2 mm. For thicker lesions or non-superficial cancers, 5-ALA PDT cannot guarantee cure of the patient and is only palliative for large tumors. This is due to the limited tissue penetration of light at the activation wavelength of PpIX (635 nm).

[0010] In some cases, PDT can be used to treat tumors located deeper in solid organs or hollow organs such as the esophagus, but this usually involves the use of devices such as optical fibers to guide catheters or endoscopes for photoactivation of photosensitizers. Not only is this a complex procedure, it also prevents access to certain areas of the body, making the treatment somewhat invasive. It also cannot eradicate the entire cancer cell and cannot be applied to multifocal disease (e.g., glioma) or multifocal metastases. Thus, although suitable for treating superficial tumors, the use of existing PDT methods in treating deep tumor cells and anatomically difficult-to-reach lesions is very limited.

[0011] Since the main limitation of PDT is the reach of light to the lesion, especially when the cancerous lesion is located in deep organs such as the brain, liver, or pancreas, several efforts have been made to utilize bioluminescence or chemiluminescence as an intracellular light source to provide the light necessary for photodynamic cell suicide after administration of a photosensitizer. One such treatment, first developed in 2013 (Theodossou et al., Cancer Research, 2013, 63:1818-21), is BLADe (BioLuminescence Activated Destruction). BLADe relies on intracellular transfection with firefly luciferase enzyme, followed by administration of a photosensitizer and luciferin, the natural substrate of luciferase. The main drawback of this method is the need to genetically modify cells to produce luciferase and the need for colocalization of the above three factors with ATP. Moreover, this colocalization must be in close proximity to the vulnerable intracellular singlet oxygen target.

[0012] Since then, there have been several attempts to utilize luminescence to achieve the desired PDT effect (see, for example, Hsu et al., Biomaterials, 2013, 34(4):1204-12; and Baacirova et al., Luminescence, 2011, 26(6):410-5). Laptev et al. (Br. J. Cancer, 2006, 95(2):189-96) have previously proposed the use of luminol (5-amino-2,3-dihydro-1,4-phthalazinedione) together with transferrin-hematoporphyrin conjugates to kill cells by intracellular luminescence. Although they provided sufficient proof of concept (95% cytotoxicity), the following drawbacks are associated with these methods, making them unfeasible in the clinic: (i) non-specific intracellular targeting, (ii) the need for transferrin as an iron source in the study by Laptev et al., and (iii) lack of design for proximity to intracellular ROS.

[0013] Most recently, in WO 2019 / 243757, the inventors proposed the use of mitochondria-targeted chemiluminescent agents in methods of PDT to treat tumors such as GBM. This initial work involves chemical modification of luminol to attach groups that target luminol to cell mitochondria and efficiently transport it across the mitochondrial membrane. Mitochondrial respiration provides the reactive oxygen species (ROS) and transition metal catalysts required for luminol luminescence. Furthermore, PpIX is biosynthesized in mitochondria and is therefore in close proximity to luminol for efficient activation and harmful singlet oxygen generation. In this way, mitochondrial ROS that pose no direct threat to cell survival are "upgraded" from within the cell to highly cytotoxic ROS that inflict lethal cell damage. This action is specific to cancerous lesions because high levels of PpIX are produced at the target site. Thus, chemically modified luminol ("mitochondrially-targeted luminol") is used as a self-sustaining intracellular light source, and the target cell mitochondria are used as the power source to "switch on the light." This in turn activates the cytotoxic activity of the photosensitizer (eg, PpIX) within the tumor cells.

[0014] In an extension of previous work, we propose here an alternative, non-invasive method of PDT that involves the use of chemically modified chemiluminescent agents, such as luminol, that can accumulate in the endoplasmic reticulum (ER) of target tumor cells.

[0015] Although efficient in tumor cell killing, mitochondria-targeted compounds in WO2019 / 243757 may have a limited therapeutic window before they reach toxic concentrations. This is due to the loss of mitochondrial membrane potential as a result of the accumulation of cationic charge of the compound in the mitochondrial matrix. However, in the case of ER targeting, there is no such effect, so resistance is much higher. At the same time, the endoplasmic reticulum is one of the vulnerable intracellular PDT targets, providing access to mitochondria-generated ROS due to its numerous contact sites with mitochondria (Vance, Biochim. Biophys. Acta, 2014, 1841:595-609). Furthermore, agents currently proposed for use in PDT may also take advantage of ER-generated ROS [doi:10.1089 / ars.2014.5851], particularly ROS derived from the unfolded protein response (UPR) [doi:10.1111 / boc.201800050], which are crucial for cancer progression and survival. It is therefore anticipated that these agents may be able to kill tumor cells more efficiently, thus providing an improved treatment over that described in WO 2019 / 243757.

[0016] Although particularly suitable for the treatment of internal tumor masses that cannot be accessed using conventional PDT techniques, the methods of PDT described herein find use in the treatment of all tumor types and any condition involving cellular hyperproliferation. Summary of the Invention

[0017] The present inventors describe a method of PDT that involves the combination of a photosensitizer, or a precursor of a photosensitizer (e.g., 5-ALA), with a chemiluminescent agent capable of targeting the endoplasmic reticulum ("ER"). Affinity for the endoplasmic reticulum is achieved through the use of "modified" chemiluminescent agents, specifically, chemiluminescent agent "conjugates" in which at least one chemiluminescent moiety is bound to at least one endoplasmic reticulum targeting moiety (also referred to herein as an "ER targeting moiety").

[0018] Examples of chemiluminescent agent "conjugates" are chemically modified luminol and acridinium esters, whose chemical modification involves the attachment of one or more chemical groups that target them to the endoplasmic reticulum.

[0019] Thus, broadly speaking, the present invention includes the modification of known chemiluminescent agents so that they are ER-targeted for PDT, particularly on hyperproliferative and / or abnormal cells, e.g., cancer cells. As a result of this modification, the luminescence required to activate the photosensitizer, such as cercosporin, hypericin and PpIX (after its translocation from mitochondria), that accumulates at the target site is "automatic" and often even stronger in cancer cells, which exhibit higher mitochondrial ROS and ER ROS formation.

[0020] For example, the formation of 5-ALA-derived PpIX is highly specific for cancerous GBM lesions. This high specificity allows the possibility of GBM treatment without the need for invasive procedures, but systemic administration of ER-targeted chemiluminescent agents, such as ER-targeted luminol, with 5-ALA is sufficient to eradicate all GMB lesions in the brain. This is particularly important because GBM is highly mobile in the brain after initial development and resurfaces at various brain locations. When used to treat GBM, the present invention also takes advantage of GBM-induced disruption of the blood-brain barrier, so that both the modified chemiluminescent agent (e.g., luminol or acridinium ester) and the photosensitizer or its precursor (e.g., cercosporin or 5-ALA) efficiently reach the GMB lesions in the brain.

[0021] In the present invention, PDT is effectively applied to individual tumor cells without the need for external light sources such as lamps or lasers that are traditionally used in PDT (i.e., the PDT effect is at the single cell level, not mass lesions). In this therapeutic approach to PDT, the depth of light penetration into tissue is no longer a limitation. This establishes the basis for an alternative treatment of cancer, which, although photochemical, involves the administration of two individual non-chemotherapeutic drugs. Thus, the treatment can be repeated multiple times without the risk of adverse side effects. This minimizes the risk of metastasis and maximizes the curative potential of the treatment.

[0022] The present invention further addresses the need for effective treatment of internal cancers such as GBM, which are currently virtually incurable due to their location and invasive nature, without the need for invasive surgery, ionizing radiation, or non-curative chemotherapy. Such treatments can be either as primary treatments and / or as photochemotherapeutic agents that can effectively control disease progression over a lifetime through repeated non-invasive treatments. This approach to the treatment of inaccessible cancers extends to the treatment of other diseases and conditions characterized by hyperproliferative and / or abnormal cells, particularly other cancers, including those that are shallow or superficial. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0023] definition As used herein, the term "chemiluminescent agent" is intended to encompass any of a variety of agents that can emit light as a result of a chemical reaction occurring within the endoplasmic reticulum (ER) of a cell, i.e., any agent that can be "activated" upon localization to the ER. More specifically, a chemiluminescent agent is an agent that emits light after reaction with a substance, such as a reactive oxygen species or a reactive nitrogen species, that is present in or generated in close proximity to the endoplasmic reticulum of a cell (e.g., in adjacent cell mitochondria or other intracellular sources). Such oxygen species include, for example, any reactive oxygen species (ROS), such as oxygen radicals, oxygen superoxide anion, hydroxyl radicals, and hydrogen peroxide. Nitrogen species include, for example, nitric oxide, peroxynitrite, nitric oxide, and the like. As will be understood, any chemiluminescent agent for use in the present invention will be physiologically tolerated.

[0024] As used herein, the term "chemiluminescent moiety" encompasses any chemiluminescent agent, or any moiety derived from a chemiluminescent agent, i.e., a derivative thereof. Any derivative should retain the luminescent properties of the parent molecule, as described above. It should also meet the requirement of physiological tolerability in vivo. Examples of derivatives include chemiluminescent agents bearing one or more additional functional or non-functional groups (e.g., substituents). The term "derivative" also extends to fragments or residues of a chemiluminescent agent.

[0025] The terms "endoplasmic reticulum targeting moiety" and "ER targeting moiety" are used interchangeably herein and are intended to include any physiologically acceptable agent or moiety that can target and thus accumulate in the endoplasmic reticulum. It also includes derivatives of such agents that retain the endoplasmic reticulum targeting properties of the parent molecule. The term "derivative" extends to fragments or residues of endoplasmic reticulum targeting agents.

[0026] As used herein, the term "photosensitizer precursor" is intended to encompass any compound that is metabolically converted to a photosensitizer and is thus essentially equivalent thereto.

[0027] The term "pharmaceutically acceptable salt" as used herein refers to any pharmaceutically acceptable organic or inorganic salt of any of the compounds described herein. A pharmaceutically acceptable salt may contain one or more additional molecules, such as a counterion. The counterion may be any organic or inorganic group that stabilizes the charge of the parent compound. If the compound is a base, a suitable pharmaceutically acceptable salt may be prepared by reacting the free base with an organic or inorganic acid. If the compound is an acid, a suitable pharmaceutically acceptable salt may be prepared by reacting the free acid with an organic or inorganic base.

[0028] The term "pharmacologically acceptable" means that a compound or composition is chemically and / or toxicologically compatible with other ingredients of a formulation or with the patient (e.g., a human) being treated.

[0029] By "pharmaceutical composition" is meant a composition in any form suitable for use for medical purposes.

[0030] As used herein, the term "treatment" includes any therapeutic indication that can benefit a human or a non-human animal (e.g., a non-human mammal). Although both human and animal treatments are within the scope of the present invention, primarily, the present invention is directed to human treatment. The term "treatment" or "therapeutic method" encompasses curative and prophylactic treatment or therapy.

[0031] The term "alkyl" as used herein refers to a univalent saturated, straight or branched carbon chain. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, n-pentyl, iso-pentyl, neo-pentyl, n-hexyl, and the like. The alkyl group preferably contains 1 to 6 carbon atoms, e.g., 1 to 4 carbon atoms.

[0032] The term "alkoxy" as used herein refers to an -O-alkyl group, where alkyl is as defined herein. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propyloxy, and the like.

[0033] The term "aryl" as used herein refers to an aromatic ring system. Such ring systems may be monocyclic or bicyclic and may contain at least one unsaturated aromatic ring. When they contain bicyclic rings, they may be fused. Preferably, such systems contain 6 to 20 carbon atoms, for example, either 6 or 10 carbon atoms. Examples of such groups include phenyl, 1-naphthyl, and 2-naphthyl. A preferred aryl group is phenyl. Unless otherwise specified, any aryl group may be selected from the group consisting of hydroxy, C 1~6 Alkyl, C 1~6 It may be substituted with one or more substituents selected from alkoxy, amino, cyano, and nitro groups, or halogen atoms (e.g., F, Cl, or Br). When two or more substituents are present, they may be the same or different.

[0034] The term "halogen atom" refers to F, Cl, Br or I.

[0035] The term "haloalkyl," as used herein, refers to an alkyl group, as defined herein, in which at least one of the hydrogen atoms of the alkyl group is replaced by a halogen atom, preferably F, Cl, or Br. Examples of such groups include -CHF, -CHF, -CF, -CCl, -CHCl, -CHCF, and the like.

[0036] The term "heterocycle" as used herein refers to a saturated or partially unsaturated 4-6 membered (preferably 5 or 6 membered) carbocyclic ring system in which at least one ring atom is a heteroatom selected from nitrogen, oxygen, and sulfur, and the remaining ring atoms are carbon. The heterocyclic ring structure may be linked to the remainder of the molecule through a carbon or nitrogen atom. Unless otherwise specified, any heterocycle described herein may contain one or more groups, which may be the same or different, such as hydroxy, C 1~6 Alkyl, C 1~6 It may be optionally substituted by alkoxy, amino, cyano, or nitro groups, or halogen atoms (F, Cl, or Br).

[0037] Unless otherwise specified, all substituents are independent of each other.

[0038] When the subscript is the integer 0 (ie, zero), it is intended that the group to which the subscript refers is absent.

[0039] In one aspect, the invention provides an endoplasmic reticulum-targeted chemiluminescent agent for use in a method of photodynamic therapy.

[0040] In one embodiment, the endoplasmic reticulum-targeted chemiluminescent agent for use in the present invention is a chemiluminescent agent "conjugate" that comprises at least one chemiluminescent moiety bound or otherwise associated with at least one endoplasmic reticulum targeting moiety ("ER targeting moiety") that selectively targets the endoplasmic reticulum. When the conjugate comprises two or more chemiluminescent moieties, they may be the same or different. However, they are generally identical. When the chemiluminescent moiety is bound to two or more ER targeting moieties, the ER targeting moieties may be the same or different, but are preferably the same. In one embodiment, the conjugate comprises a single chemiluminescent moiety bound or otherwise associated with a single ER targeting moiety.

[0041] The chemiluminescent moiety (or moieties) may be attached to the ER targeting moiety (or moieties) via covalent or non-covalent means. It may be attached, for example, via electrostatic interactions, van der Waals forces, and / or hydrogen bonds. Typically, the chemiluminescent moiety (or moieties) and the ER targeting moiety (or moieties) are covalently attached to each other, for example, via one or more covalent bonds. In some cases, the chemiluminescent moiety (or moieties) may be covalently attached to the ER targeting moiety (or each thereof) via a linking group (or "spacer").

[0042] A chemiluminescent agent "conjugate" for use in the present invention may be a compound having the following general formula (I), or a pharma- ceutically acceptable salt thereof:

[0043] [ka] where A represents a chemiluminescent moiety; each L may be the same or different and is either a direct bond or a linker; each B may be the same or different and represents an endoplasmic reticulum targeting moiety; n is an integer from 1 to 3, preferably 1; x is an integer of 1 to 3, and preferably 1.

[0044] In one embodiment of formula (I), both n and x are 1. Thus, a chemiluminescent agent "conjugate" for use in the present invention may be a compound of formula (II), or a pharma- ceutically acceptable salt thereof:

[0045] [ka] wherein A, L, and B are as defined herein.

[0046] Chemiluminescent agents suitable for use in the present invention are known in the art and include, for example, luminol, isoluminol, lucigenin, acridinium esters, oxalate esters, and known analogs and derivatives thereof. Any known chemiluminescent agent or derivative thereof may be used. Chemiluminescent moieties for use in the present invention may be "derived" from any of these agents. Suitable derivatives may include one or more additional functional or non-functional groups (e.g., substituents), or they may include fragments or residues of such agents that retain chemiluminescent activity.

[0047] Preferred for use in the present invention are chemiluminescent moieties derived from luminol, isoluminol, and acridinium esters. Such compounds may contain one or more additional substituents, e.g., C 1~6 Alkyl, C 1~6 It may be substituted by alkoxy, amino, cyano, nitro and aryl (e.g., phenyl) groups, or halogen atoms. For example, the phenyl ring of luminol or isoluminol may be substituted by halogen atoms, C 1~6 It may be substituted with one or more (eg, 1 or 2) groups selected from alkyl and phenyl groups.

[0048] As will be appreciated, in order to be bound to the ER targeting moiety (or moieties) via a covalent bond or linker, the chemiluminescent moiety is typically a "derivative" of the parent chemiluminescent agent. For example, it may lack one or more terminal atoms or groups following the formation of a covalent bond either to the linker or directly to the ER targeting agent, and thus may be considered a "residue" of the original molecule. For example, in the case of luminol, the primary amine group is "derivatized" by the loss of a single hydrogen atom (i.e., -NH2 to -NH-), since it may form the point of attachment to either the linker or the ER targeting moiety. Other forms of derivatization may be envisioned, including the introduction of functional groups that may react to form a covalent bond with the ER targeting moiety (or moieties). The particular form of "derivatization" required for any given chemiluminescent agent depends on its structure and can be readily determined by any skilled chemist.

[0049] In formula (I) or (II), the chemiluminescent moiety has the following structure: * indicates the point (or points) of attachment to the linker L or directly to the ER targeting moiety;

[0050] [ka] During the ceremony, R 1 is hydrogen or C 1~6 Alkyl, preferably C 1~3 an alkyl group such as alkyl (e.g., methyl); Each R 2 teeth, - C 1~6 Alkyl, - C 1~6 Alkoxy, - -NR 5 R 6 (In the formula, R 5 R 6 H and C 1~6 Alkyl, preferably H and C 1~3 alkyl (e.g., -CH3), - optionally substituted aryl (e.g., optionally substituted phenyl); n is an integer of 0 to 3, preferably 0, 1 or 2, for example, 0 or 1; m is an integer of 0 to 2, for example, 0 or 1; R is hydrogen or C 1~6 Alkyl, preferably C 1~3 alkyl (e.g., methyl); X is a monovalent anion, such as Cl, Br, I, OTos, ClO4, NO3, PF6, or BF4 anion; Y is an optionally substituted aryl (or arylene) group, for example, an optionally substituted phenyl (or phenylene).

[0051] In one embodiment, R 1is hydrogen or methyl. Preferably, R 1 is hydrogen.

[0052] In one embodiment, each R 2 is C 1~6 alkyl and unsubstituted phenyl. For example, each R 2 may be independently selected from methyl, ethyl, n-propyl, iso-propyl, n-butyl and phenyl.

[0053] When Y is a substituted aryl or arylene group, examples of suitable substituents include one or more halogen atoms (e.g., F, Cl, Br, I), 1~6 Alkyl (e.g., tert.butyl, propyl, ethyl or methyl), -COOC 1~6 Examples include alkyl (eg, -COOCH3), nitro or cyano groups.

[0054] Different substitution patterns on the aromatic ring of luminol or isoluminol may be selected by one of skill in the art, and in one embodiment, the substitution pattern may be selected to enhance chemiluminescence. In one embodiment, the chemiluminescent moiety for use in the present invention is luminol or a substituted derivative thereof, for example, which has the following structure: * indicates a point of attachment to a linker L or directly to an ER targeting moiety,

[0055] [ka] During the ceremony, R 17 is hydrogen, C 1~6 Alkyl (preferably C 1~4 Alkyl, more preferably C 1~3 alkyl (e.g., methyl), or optionally substituted aryl (e.g., optionally substituted phenyl); R 18 is hydrogen or C 1~6 Alkyl (preferably C 1~4 Alkyl, more preferably C1~3 alkyl, for example methyl); R 19 is hydrogen or C 1~6 Alkyl (preferably C 1~4 Alkyl, more preferably C 1~3 alkyl, for example, methyl).

[0056] In one embodiment, R 17 and R 18 are both hydrogen, and R 19 is other than hydrogen, for example, R 19 is C 1~3 Alkyl, for example methyl.

[0057] In another embodiment, R 18 is hydrogen and R 17 and R 19 Each of is other than hydrogen, e.g., R 17 is optionally substituted phenyl, R 19 is C 1~6 Alkyl (eg, methyl).

[0058] Non-limiting examples of chemiluminescent moieties for use in the present invention include the following, wherein: * indicates the point of attachment to a linker L or directly to the ER targeting moiety).

[0059] [ka]

[0060] Any agent known to be capable of selectively targeting the endoplasmic reticulum may be used to provide the "ER targeting moiety" in the conjugate described herein. Such agents are typically hydrophobic in nature to make them suitable for distribution to the ER membrane environment. Particularly suitable for use in the present invention are sulfonamide agents due to their ability to localize to the endoplasmic reticulum. Such agents have one or more sulfonamide groups. Typically, they contain one sulfonamide group.

[0061] As will be appreciated, in order to be linked to the chemiluminescent moiety (or moieties) via a covalent bond or linker, the ER targeting moiety may in some cases be a "derivative" of the parent drug. For example, it may lack one or more terminal atoms or groups following the formation of a covalent bond, either to the linker or directly to the chemiluminescent agent (or agents), and thus may be considered a "residue" of the original molecule. The particular form of "derivatization" required for any given ER targeting moiety will depend on its structure and can be readily determined by any skilled chemist.

[0062] Sulfonamide ER targeting moieties for use in the present invention include the following: * indicates the point of attachment to a linker L or directly to the chemiluminescent moiety,

[0063] [ka] During the ceremony, R 14 is C 1~6 is an alkyl or an optionally substituted aryl group; R 15 is hydrogen or C 1~6 is alkyl,

[0064] [ka] represents an optionally substituted nitrogen-containing heterocycle.

[0065] In one embodiment, R 14 is C 1~6 Alkyl, preferably C 1~3 Alkyl, for example, -CH3.

[0066] In one embodiment, R 14 is an optionally substituted aryl group. For example, R 14 may be an optionally substituted phenyl or naphthyl.

[0067] R 14 If is replaced, C 1~3 Alkyl (e.g., methyl), C 1~3 Alkoxy (e.g., methoxy), C 1~3 Haloalkyl (e.g., -CF3), -OC 1~3 haloalkyl (e.g., -OCF), cyano, nitro, -NR' (wherein each R' is independently H or C 1~3 alkyl), halogen (e.g., F, Cl, or Br), -COR" (wherein R" is H or C 1~3 alkyl), and -COOR" (wherein R" is H or C 1~3 The preferred substituents are C 1~3 It is an alkyl (e.g., methyl). Typically, there may be one or two substituents, e.g., a single substituent. When a single substituent is present on a phenyl ring, the substituent may be in the ortho, meta, or para position. In one embodiment, the substituent is in the para position. For example, a methyl group may be present in the para position of the phenyl ring.

[0068] In one embodiment, R 14 is an unsubstituted aryl group, for example, unsubstituted phenyl or naphthyl.

[0069] In one embodiment, R 15 is hydrogen or -CH3, for example, hydrogen.

[0070] In one embodiment,

[0071] [ka] represents a piperazinyl ring, which may be linked to the chemiluminescent moiety via a linker L, or directly via a ring carbon or via an additional ring nitrogen atom, but is preferably linked via an additional ring nitrogen atom. In a preferred embodiment, the ER targeting moiety can be represented as follows:

[0072] [ka] During the ceremony, R 14 is as defined herein; * indicates the point of attachment to the linker, L, or directly to the chemiluminescent moiety.

[0073] In one embodiment, the ER targeting agent is a group having one of the following structures:

[0074] [ka] During the ceremony, R 15 is as defined herein; Each R 16 is independently 1~3 Alkyl (e.g., methyl), C 1~3 represents an alkoxy (e.g., methoxy) or a halogen atom (e.g., Cl or Br); u is an integer of 0 to 5, preferably 1 to 3, for example, 1; * indicates the point of attachment to the linker, L, or directly to the chemiluminescent moiety.

[0075] Non-limiting examples of ER targeting moieties for use in the present invention include the following, wherein: * indicates the point of attachment to a linker L, or directly to the chemiluminescent moiety.

[0076] [ka]

[0077] In the above general formulas (I) and (II), the exact nature of the linker L is not considered to be important for the performance of the present invention, as long as it serves the intended function of linking the chemiluminescent moiety to the ER targeting moiety (or moieties), thus enabling targeted delivery of the chemiluminescent moiety to the endoplasmic reticulum. The linker may be rigid or flexible, and may be cleavable (e.g., photocleavable) in vivo at the desired target site. Generally, it comprises an organic group.

[0078] The linking group L may be hydrophilic or hydrophobic in nature. It may be either branched (including dendritic) or linear, but is preferably linear. If the linking group is branched, it may, for example, carry two or more ER targeting moieties. The linking group may be aliphatic and / or aromatic and may contain one or more cycloalkyl, heterocyclic, aryl or heteroaryl rings. Thus, the linking group may be aliphatic, (poly)cyclic and / or (poly)aromatic in nature.

[0079] The chain length of the linker may vary, but generally it may comprise a backbone containing 1 to 20 atoms (e.g., 1 to 20 carbon atoms), preferably 2 to 15, e.g., 2 to 12 atoms. In some cases, the length of the linker may be varied to adjust the precise positioning of the chemiluminescent portion of the conjugate relative to the ER targeting moiety.

[0080] The linker L can be, for example, 1~3 Alkyl, -O(C 1~3 ) an alkylene chain (preferably C ) optionally substituted with one or more groups selected from alkyl, -OH, cycloalkyl and aryl groups; 1~15 Alkylene, e.g., C 2~11 alkylene), and one or more of the -CH2- groups in the alkylene chain may be -O-, -CO-, -NR- (wherein R is H or C 1~6 Alkyl, preferably C 1~3The linker L may be replaced by a group independently selected from alkyl (e.g., methyl), cycloalkyl, heterocyclic, aryl and heteroaryl groups. In one embodiment, the -CH2- group of the alkylene chain may be replaced by such a group. For example, the linker L may be a group -CO-.

[0081] Suitable linker groups can be readily determined by one of ordinary skill in the art. Examples of suitable linkers include optionally substituted alkylene groups, preferably unsubstituted linear alkylene groups, such as -C3H6-, -C4H8-, -C6H 12 -, -C8H 16 -, -C 10 H 20 - and -C 11 H 22 In one embodiment, short chain alkylene groups such as -C3H6- and -C4H8- are preferred.

[0082] When one or more -CH2- groups of the alkylene chain are replaced by a group, these may be replaced either by -O- or -CO- groups, or by a heterocycle (e.g., a saturated heterocycle such as a piperazinylene group), or an aryl ring (e.g., phenylene). Examples of such linkers in which one or more -CO- are present include -CO-, -CO-CH2-, -CO-CH6, -CO-CH5H6, -CO-CH 10 -, -CO-C6H 12 - and -CO-C 10 H 20 Other examples of suitable linkers in which more than one -O- group is present include oligo- or polyethylene glycol groups, preferably polyethylene glycol groups containing 1 to 4 ethylene oxide units, for example 2 or 4 ethylene oxide units.

[0083] In certain embodiments, the chemiluminescent agent conjugate for use in the present invention is a compound of formula (III), or a pharma- ceutically acceptable salt thereof:

[0084] [ka] In the formula, L 1 is either a direct bond or any linker described herein, B 1 is any endoplasmic reticulum targeting moiety described herein, R 3 is hydrogen or C 1~3 an alkyl group such as alkyl (e.g., methyl); Each R 4 is C 1~6 Alkyl, and -NR 5 R 6 are independently selected from R 5 and R 6 H and C 1~6 Alkyl, preferably H and C 1~3 alkyl (e.g., -CH3); p is an integer of 0 to 3, preferably 0, 1 or 2, for example, 0 or 1.

[0085] Preferred compounds of formula (III) include the following compounds of formulae (IIIa) and (IIIb):

[0086] [ka] In the formula, L 1 , B 1 , R 3 , R 4 , and p are as defined herein.

[0087] In formulae (III), (IIIa) and (IIIb), L 1 is preferably selected from one of the following:

[0088] [ka] In the formula, a is an integer of 1 to 10, preferably 3 to 10. b is an integer of 1 to 4, for example, 2.

[0089] In one embodiment of Formula (III), (IIIa) and (IIIb), L 1 The basis

[0090] [ka] where a is 2 or 3.

[0091] In one embodiment of formula (III), (IIIa) or (IIIb), B 1 may be one of the following groups:

[0092] [ka] During the ceremony, R 14 , R 15 and

[0093] [ka] is as defined herein.

[0094] In certain embodiments, the chemiluminescent agent conjugate for use in the present invention is a compound of formula (IV), or a pharma- ceutically acceptable salt thereof:

[0095] [ka] In the formula, L 2 is either a direct bond or any linker described herein, B 2 is any endoplasmic reticulum targeting moiety described herein, Each R 6 is halogen (e.g., F, Cl, Br, I) and C 1~6 alkyl (e.g., tert-butyl); q is an integer of 0 to 4, preferably 0 or 2; Z is a monovalent anion, such as Cl, Br, I, or CF3OSO2 anion.

[0096] In one embodiment of formula (IV), L 2 represents one of the following groups:

[0097] [ka] In the formula, a is an integer of 1 to 10, and preferably 3, 4, or 5.

[0098] In one embodiment of formula (IV), L 2 The basis

[0099] [ka] Represents.

[0100] In one embodiment of formula (IV), B 2 may be one of the following groups:

[0101] [ka] During the ceremony, R 14 , R 15 and

[0102] [ka] is as defined herein.

[0103] In certain embodiments, the chemiluminescent agent conjugate for use in the present invention is a compound of formula (V), or a pharma- ceutically acceptable salt thereof:

[0104] [ka] In the formula, L 3is either a direct bond or any linker described herein, B 3 is any endoplasmic reticulum targeting moiety described herein, Each R 7 is halogen (e.g., F, Cl, Br, I), -COR 8 (In the formula, R 8 is hydrogen or C 1~6 alkyl), cyano, and C 1~6 alkyl (e.g., tert-Bu); r is an integer of 0 to 5, preferably 0 or 3; Z is a monovalent anion, such as Cl, Br, I, or CF3OSO2 anion.

[0105] In formula (V), L 3 is preferably C 1~10 Alkylene, e.g., C 1~6 It is alkylene.

[0106] In one embodiment of formula (V), B 3 may be one of the following groups:

[0107] [ka] During the ceremony, R 14 , R 15 and

[0108] [ka] is as defined herein.

[0109] In certain embodiments, the chemiluminescent agent conjugate for use in the present invention is a compound of formula (VIa), (VIb), or a pharma- ceutically acceptable salt thereof:

[0110] [ka] In the formula, L 4 is either a direct bond or any linker described herein, A 1 is any chemiluminescent moiety described herein.

[0111] In formula (VI), (VIa) or (VIb), L 4 may be selected from the following:

[0112] [ka] In the formula, a is an integer of 1 to 10, and preferably 3 to 10.

[0113] In one embodiment of Formula (VI), (VIa) or (VIb), L 4 The basis

[0114] [ka] where a is 2 or 3.

[0115] In one embodiment of formula (VI), (VIa) or (VIb), A 1 is selected from one of the following:

[0116] [ka] In the formula, R 3 is hydrogen or C 1~3 an alkyl group such as alkyl (e.g., methyl); Each R 4 is C 1~6 Alkyl, and -NR 5 R 6 are independently selected from R 5 and R 6 H and C 1~6 Alkyl, preferably H and C 1~3 alkyl (e.g., -CH3); p is an integer of 0 to 3, preferably 0, 1 or 2, for example, 0 or 1; Z is a monovalent anion, such as Cl, Br, I, or CF3OSO2 anion; Each R 9 is halogen (e.g., F, Cl, Br, I) and C 1~6 alkyl (e.g., tert-Bu); s is an integer of 0 to 4, and preferably 0, 2, or 3.

[0117] In certain embodiments, the chemiluminescent agent conjugate for use in the present invention is a compound of formula (VII), or a pharma- ceutically acceptable salt thereof:

[0118] [ka] In the formula, L 5 is either a direct bond or any linker described herein, B 4 is any endoplasmic reticulum targeting moiety described herein, Each R 10 is C 1~6 Alkyl (e.g., methyl), and -NR 11 R 12 are independently selected from R 11 and R 12 H and C 1~6 Alkyl, preferably H and C 1~3 alkyl (e.g., -CH3); t is an integer of 0 to 3, preferably 1 or 2.

[0119] Preferred compounds of formula (VII) include compounds of formula (VIIa):

[0120] [ka] In the formula, L 5 , B 4 , R 11 and R 12is as defined herein, and R 13 is H or C 1~3 It is an alkyl.

[0121] In formulae (VII) and (VIIa), L 5 is preferably C 1~11 Alkylene, more preferably C 2~8 Alkylene, for example propylene.

[0122] The chemiluminescent drug conjugates described herein can be prepared using methods and procedures known in the art.Suitable methods include those described in WO 2019 / 243757, the entire contents of which are incorporated herein by reference.Methods for preparing luminol derivatives for use in the present invention include those described by Mikroulis et al. in J.Org.Chem. (see https: / / doi.org / 10.1021.acs.joc.1c00890), the entire contents of which are incorporated herein by reference.

[0123] Methods that can be used to covalently attach the chemiluminescent agent to the endoplasmic reticulum targeting moiety include known coupling techniques. The exact method used will depend on the exact nature of the chemiluminescent agent, the endoplasmic reticulum targeting moiety, and the linker (if present), particularly the nature of the pendant functional groups involved in the formation of the bond. If pendant functional groups are already present on the binding partner, they can be used in linking the various moieties. If necessary, one or more components of the conjugate (i.e., the chemiluminescent moiety, the linker, and the ER targeting moiety) may be functionalized to include, for example, reactive functional groups that can be used to link the components. Suitable reactive groups include carboxylic acids, hydroxy, thiol, carbonyl, acid halides, primary and secondary amines, aryl halides and pseudoaryl halides, alkyl halides and pseudoalkyl halides, alkenyl halides and pseudoalkenyl halides, terminal alkynes, clickable moieties, and the like. Methods for introducing such functional groups are well known in the art.

[0124] Examples of methods that can be used to covalently attach a chemiluminescent agent to one or more ER targeting moieties include, but are not limited to, amide bond formation, ether bond formation, ester bond formation, thioester bond formation, cross-coupling reactions, olefin metathesis reactions, aromatic electrophilic substitution, click chemistry, nucleophilic substitution reactions, and the like.

[0125] The compounds for use as starting materials in the preparation of the conjugates described herein are known from the literature or can be commercially available. Alternatively, they can be easily obtained by methods known from the literature. A more detailed description of the methods for preparing the compounds for use according to the present invention can be found in the examples.

[0126] The chemiluminescent drug conjugates described herein are themselves novel and form a further aspect of the invention. For example, a method for preparing a chemiluminescent drug conjugate comprising linking one or more chemiluminescent drugs to one or more ER targeting moieties using any of the techniques described herein forms a further aspect of the invention.

[0127] For use in PDT, the ER-targeted chemiluminescent agents described herein are used in combination with a photosensitizer or a precursor of a photosensitizer. The key to the present invention is that they should be in close proximity to each other in the cell ER so that the chemiluminescent agent can "activate" the photosensitizer. These agents can be provided separately for separate, simultaneous, or sequential administration to a patient in a method of PDT. Alternatively, they can be provided as a single formulation in which both the ER-targeted chemiluminescent agent and the photosensitizer (or precursor) are present. Such formulations form part of the present invention.

[0128] For use in the present invention, any photosensitizer (or photosensitizer precursor) must be able to accumulate in the endoplasmic reticulum (or translocate to the ER) of a target cell following its in vivo administration, ensuring that it is in close proximity to the chemiluminescent compound. For example, this may be an ER-localized photosensitizer or its precursor.

[0129] Examples of photosensitizers and precursors that can target the endoplasmic reticulum include 5-ALA and its derivatives (after translocation from mitochondria), mTHPC, temoporfin, chlorin e6, phthalocyanines, anthraquinones and their derivatives (e.g., hypericin, hypocrellins [A, B], cercosporins, calphostin, ersinochromes [A, B, C]), and pharma- ceutically acceptable salts thereof. Other ER-accumulating photosensitizers are known in the art and may also be used in the present invention.

[0130] Other known photosensitizers and precursors may be used in the present invention, subject to appropriate modification to impart the desired targeting properties. For example, they may be encapsulated in suitable nanocarriers with ER targeting capabilities. In these embodiments, a wider range of photosensitizers may be used, and it is envisaged that any known photosensitizer (or precursor) suitable for use in PDT may be used. A range of suitable agents are known in the art, and include, for example, 5-aminolevulinic acid (5-ALA) and derivatives of 5-ALA (leading to the production of protoporphyrin IX), porphyrins, phthalocyanines such as metallated phthalocyanines (e.g., AlPcS) that may be optionally sulfonated, e.g., AlPcS2 or AlPcS 2a di-sulfonated aluminum phthalocyanines such as aluminum phthalocyanine tetra-sulfonate (A1PcS4), sulfonated tetraphenylporphyrins (e.g., TPPS 2a , TPPS4, TPPS1 and TPPS 2o), chlorins such as tetra(m-hydroxyphenyl)chlorin (m-THPC) (e.g., temoporfin, available commercially under the trade name Foscan), chlorin derivatives including bacteriochlorins and ketochlorins, mono-L-aspartyl chlorin e6 (NPe6) or chlorin e6, natural and synthetic porphyrins including hematoporphyrin and benzoporphyrin, anthraquinones and their derivatives (e.g., hypericin, hypocrellins [A, B], cercosporin, calphostin, elsinochrome [A, B, C]).

[0131] Pharmaceutically acceptable salts of any of these photosensitizers (or precursors) may also be used. Such salts include salts with pharma-ceutically acceptable organic or inorganic acids or bases.

[0132] The derivative of 5-ALA that may be used in the present invention includes any derivative of 5-ALA that can form PpIX in vivo.Typically, such derivative is a precursor of PpIX in the heme biosynthetic pathway, and therefore can produce PpIX at target site after administration.Suitable precursor of PpIX includes 5-ALA prodrug, such as 5-ALA ester.

[0133] The following are particularly preferred photosensitizers and precursors for use in the present invention: 5-ALA, mTHPC, temoporfin, chlorin e6, sulfonated aluminum phthalocyanine, anthraquinones and their derivatives (e.g., hypericin, hypocrellin [A, B], cercosporin, calphostin, ersinochrome [A, B, C]), and pharmaceutically acceptable salts thereof. Particularly preferred for use in the present invention are cercosporin, 5-ALA, and its pharmaceutically acceptable derivatives (e.g., pharmaceutically acceptable salts, or methyl or hexyl esters).

[0134] The particular choice of chemiluminescent moiety will depend on a variety of factors, including the nature of the tumor being treated, but can be readily selected by one of ordinary skill in the art. As will be appreciated, the choice of chemiluminescent agent will also depend on the photosensitizer used in the PDT treatment, since the wavelength of light it emits should be suitable for photoactivation of the photosensitizer, either by direct light absorption or by energy transfer mechanisms.

[0135] Examples of suitable chemiluminescent and photosensitizer "pairs" can be easily determined by those skilled in the art. The following are provided as suitable non-limiting examples. When the photosensitizer is PpIX (e.g., generated in vivo after administration of 5-ALA), luminol or isoluminol may be used as the chemiluminescent agent. The photosensitizer hypericin is particularly suitable for use with the chemiluminescent agent lucigenin because these moieties can form a π-stack for highly efficient intramolecular energy transfer, especially in the presence of a coupling agent such as the metal chelators DTPA or EDTA. Luminol and mTHPC are a highly efficient energy transfer pair. Other efficient energy transfer pairs include luminol-erythrosin B, luminol-hypocrelin, luminol-cercosporin, luminol-calphostin, luminol-elsinochrome acridine ester-hypocrelin, lucigenin-hypocrelin, acridine ester-cercosporin, lucigenin-cercosporin, acridine ester-hypericin, and lucigenin-hypericin. Haematoporphyrin derivative (HPD) or sulfonated aluminum phthalocyanine may be used with either luminol or lucigenin. However, these are merely suggestive examples of potential functional pairs, and others can be readily determined by one of skill in the art.

[0136] The ER-targeted chemiluminescent agents described herein are intended for use in methods of photodynamic therapy and are suitable for use in treating disorders or abnormalities of cells or tissues in the body that are responsive to photodynamic therapy. Such methods involve the simultaneous, separate, or sequential use of a photosensitizer or a precursor of a photosensitizer described herein.

[0137] Generally, metabolically active cells respond to photodynamic therapy. Examples of metabolically active cells are cells that undergo abnormal growth, such as increased cell number / cell proliferation, abnormal cell maturation and differentiation, or abnormal cell proliferation. Any condition characterized by such growth patterns can be treated according to the PDT method described herein.

[0138] Disorders or conditions that may be treated include, but are not limited to, malignant and pre-malignant cancerous conditions, such as cancerous growths or tumors and their metastases, tumors, such as sarcomas and carcinomas, particularly solid tumors. The present invention is particularly suitable for the treatment of tumors, particularly tumors located below the surface of the skin, i.e., internal or deep cancers.

[0139] PDT according to the invention may be applied in two ways: (i) as a treatment for malignant or pre-malignant conditions (e.g., gliomas) without the need for an external light source as in conventional PDT, or (ii) as a repeatable, adjunctive, post-operative photochemotherapy to suppress any active neoplastic lesions left behind that may result in either recurrence or dissemination of the disease. The treatment may be effectively used to manage and suppress conditions (e.g., brain cancer) for a lifetime through repeated treatment sessions. Treatment of occult metastases of primary disease may also be performed without the need for prior diagnosis.

[0140] Examples of tumors that can be treated using the present invention are sarcomas, including osteogenic sarcomas and soft tissue sarcomas, carcinomas, such as carcinomas of the breast, lung, brain, bladder, thyroid, prostate, colon, rectum, pancreas, stomach, liver, uterus, liver, kidney, prostate, cervix and ovary, lymphomas, including Hodgkin's lymphoma and non-Hodgkin's lymphoma, neuroblastoma, melanoma, myeloma, Wilms' tumor, leukemias, including acute lymphoblastic leukemia and acute myeloblastic leukemia, astrocytoma, glioma and retinoblastoma, mesothelioma.However, the present invention finds particular value in the treatment of deep cancerous lesions that are difficult to reach non-invasively.The treatment of gliomas (e.g. GMB) forms a preferred embodiment of the present invention.

[0141] Other examples of metabolically active cells are inflamed cells. Thus, inflammatory diseases such as rheumatoid arthritis can also be treated using the PDT method according to the present invention.

[0142] For use in any of the PDT methods described herein, the ER-targeted chemiluminescent agents are generally provided as pharmaceutical compositions containing at least one pharma- ceutically acceptable carrier or excipient. Such compositions form a further aspect of the invention. They may also contain a selected photosensitizer (or precursor), although in most cases it is envisaged that the photosensitizer (or precursor thereof) will be provided in a different formulation for separate administration to the patient.

[0143] The pharmaceutical compositions described herein can be formulated using techniques well known in the art. The route of administration depends on the intended use, particularly the location of the cells or tissues to be treated. Typically, they are administered systemically and therefore may be provided in a form adapted for parenteral administration, for example, by intradermal, subcutaneous, intraperitoneal, intravenous or intratumoral injection, or by infusion by drip infusion. Suitable dosage forms include suspensions and solutions containing the conjugate and / or photosensitizer (or precursor thereof) together with one or more inert carriers or excipients. Suitable carriers include saline, sterile water, phosphate buffered saline, and mixtures thereof. Preferably, the compositions are used in the form of an aqueous suspension or solution in water or saline, for example, phosphate buffered saline.

[0144] The compositions may further contain other agents such as emulsifying agents, suspending agents, dispersing agents, viscosity adjusting agents, solubilizing agents, stabilizers, buffers, preservatives, etc. The compositions may be sterilized by conventional sterilization techniques.

[0145] In one embodiment, the ER-targeted chemiluminescent agent is provided in the form of a solution in water or saline (or any other pharma- ceutical relevant biocompatible vehicle) suitable for intravenous or intratumoral injection, which can be administered in either a single dose or multiple doses.

[0146] In one embodiment, the chemiluminescent drug conjugate can be administered in the form of a sustained release formulation. Suitable delayed release formulations are known in the art and include any formulation that can continuously and sustainably release the drug in vivo. One example of a suitable delayed release formulation is an injectable implant that provides sustained release in vivo. Such an implant can be an in-situ forming implant based on biocompatible and biodegradable polymers that contain nanoparticles of active compounds. These provide sustained delivery of the chemiluminescent drug to ensure long-term light emission, thereby achieving an optimized therapeutic effect of the treatment.

[0147] In another embodiment, the chemiluminescent conjugates can be provided in the form of thermoresponsive formulations that become thermogels at physiological temperatures (i.e., once delivered to the body). They can be formulated to optimally release their load over a period of up to 15 hours, e.g., 10-15 hours. The use of temperature-responsive polymers allows for the formulation of low-viscosity solutions that undergo an in situ phase transition and gel in response to body temperature, suitable for subcutaneous injection. A variety of polymers and their copolymers can be used to optimize the thermosetting properties of the gel network. Such polymeric materials are known and used in the art, and include, for example, poly(lactic-co-glycolic acid) (PLGA), alginate / hyaluronic acid, poly(N-isopropylacrylamide), and poloxamer.

[0148] Nanoparticles and / or microparticles containing chemiluminescent conjugates may also be provided to provide controlled, continuous release of the active agent over an extended period of time, e.g., 10-15 hours. Examples of such carriers include (i) micellar carriers, (ii) liposomes, (iii) dendrimer or polymeric nanocarriers, and (iv) solid lipid nanoparticles. Any such particles may be included in the thermoresponsive formulations described herein to form reservoirs for releasing the active agent in the gel network generated in situ.

[0149] The compositions described herein can be administered systemically (e.g., orally or parenterally) or they can be applied locally (e.g., topically) at or near the affected area. The route of administration depends on the severity, nature, and location of the disease being treated and the photosensitizer (or precursor) used. Compositions that can be administered systemically include plain or coated tablets, capsules, suspensions, and solutions. Compositions that can be administered locally (e.g., topically) include gels, creams, ointments, sprays, lotions, and any other conventional dosage form in the art. Creams, ointments, and gels can be formulated with an aqueous or oily base with the addition of suitable thickening and / or gelling agents.

[0150] Typically, the methods described herein may include an initial step of administration of an effective amount of a composition comprising a photosensitizer, for example, by intravenous injection. The photosensitizer (or precursor) is then distributed to the desired target area of ​​the body, for example, intravenously by either injection and / or infusion, allowing for in situ generation of the active photosensitizer, e.g., PpIX, prior to administration of the ER-targeted chemiluminescent agent. The time profile of PpIX generation in cells following 5-ALA administration may be several hours (typically, this may peak between 2-10 hours after administration), and therefore it is desirable to delay the delivery of the ER-targeted chemiluminescent agent. This may be achieved by delaying its administration or by a delayed release formulation as described herein. It is envisioned that administration of the photosensitizer will typically occur prior to administration of the ER-targeted chemiluminescent agent, although their delivery may nevertheless be simultaneous, for example, when the ER-targeted chemiluminescent agent is provided in the form of a delayed release formulation (e.g., in the form of any of the nanoparticle and / or microparticle carrier systems described herein).

[0151] For example, a patient can receive an initial 5-ALA injection, then a second injection of an ER-targeted chemiluminescent agent within the appropriate therapeutic window at an appropriate time frame, or an infusion containing this agent, for as long as necessary. In this setting, the need to be subject to any post-treatment monitoring is minimized.

[0152] The effective dose of the compositions described herein, the number of doses, and the exact timing of administration will depend on a variety of factors, including the nature of the ER-targeted chemiluminescent agent, the photosensitizer (or precursor), their mode(s) of administration, the condition being treated, the patient, and the like, and can be adjusted accordingly.

[0153] A further aspect of the invention relates to a method of photodynamic therapy of cells or tissue of a patient, the method comprising administering to the cells or tissue: (a) administering, simultaneously, separately or sequentially, an effective amount of an endoplasmic reticulum-targeted chemiluminescent agent as described herein and an effective amount of a photosensitizer or photosensitizer precursor; or (b) administering an effective amount of a pharmaceutical composition comprising an endoplasmic reticulum-targeted chemiluminescent agent described herein and a photosensitizer or photosensitizer precursor.

[0154] In a further aspect, the present invention provides an article of manufacture comprising an endoplasmic reticulum targeted chemiluminescent agent as described herein and a photosensitizer or precursor, for simultaneous, separate or sequential use in a method of photodynamic therapy, e.g., in any of the PDT methods described herein.

[0155] In yet a further aspect, the present invention provides a kit comprising: (i) an endoplasmic reticulum-targeted chemiluminescent agent as described herein, separately (ii) a photosensitizer or photosensitizer precursor, and optionally (iii) instructions for the use of (i) and (ii) in a method of photodynamic therapy. When used, the active components of the kit (i.e., (i) and (ii)) may be administered simultaneously, separately, or sequentially. [Brief description of the drawings]

[0156] The invention will now be further described with reference to the following non-limiting examples and the accompanying drawings. [Figure 1] 1 is a graph of cell viability of LN18 cells in response to compound DZ325+cercosporin. [Diagram 2] 1 is a graph of cell viability of M059K cells in response to compound DZ325+cercosporin. [Diagram 3] 1 is a graph of cell viability of U87, T98G, LN18 and M059K cells in response to compounds EK297 plus cercosporin or 5-ALA. EXAMPLES

[0157] Example 1 - Preparation of N-(3-((1,4-dioxo-1,2,3,4-tetrahydrophthalazin-5-yl)amino)propyl)-4-methylbenzenesulfonamide (DZ325)

[0158] [ka]

[0159] Testing Procedure: Synthesis of N-(3-bromopropyl)-4-methylbenzenesulfonamide 2: Triethylamine (437 mg, 4.32 mmol) was added dropwise to a cooled (0° C.) stirred suspension of 4-toluenesulfonyl chloride (342 mg, 1.79 mmol) and 3-bromopropylamine hydrobromide (453 mg, 2.07 mmol) in dry dichloromethane (10 mL). The resulting mixture was stirred at that temperature for 15 min, then dichloromethane (50 mL) was added, washed with 2N HCl (2×40 mL) and brine (40 mL), dried (Na2SO4) and the solvent was evaporated to give 2 (471 mg, 90%).

[0160] 1H NMR(200MHz,CDCl3)δ:7.76(d,J=8.3Hz,1H,ArH-oS),7.31(d,J=8.5Hz,1H,ArH-oMe),5.01(bs,2H,NH),3 .41(t,J=6.3Hz,1H,CH2Br),3.09(t,J=6.0Hz,1H,CH2N),2.43(s,3H,CH3),2.01(qui,J=6.4Hz,2H,CH2).

[0161] Synthesis of N-(3-((1,4-dioxo-1,2,3,4-tetrahydrophthalazin-5-yl)amino)propyl)-4-methylbenzenesulfonamide DZ325: A solution of luminol (313 mg, 1.77 mmol) and bromide 2 (514 mg, 1.76 mmol) in N-methylpyrrolidone (2 mL) was stirred at 120 °C for 2 days. After cooling, the mixture was poured into stirred ice water (60 mL) and stirred for 15 min. The precipitate formed was filtered, washed with water (3 × 5 mL) and dried in vacuum. The residue was dissolved in 5% THF / dichloromethane and purified by column chromatography (5-10% THF / dichloromethane). The resulting crude DZ325 was triturated with diethyl ether and hexane to give DZ325 as a pale yellow powder (200 mg, 29%).

[0162] 1 H NMR(400MHz,DMSO-d6)δ:11.39(bs,1H,CONH),11.14(bs,1H,CONH),8.97(bs,1H,ArNH),7.66(app d,J=8.3Hz,2H,H-2'),7.59(t,J=5.8Hz,1H,SNH),7.55(t,J=8.1Hz,1H,H-7),7.35(app dd,J=8.5,0.6Hz,2H,H-3'),6.96(d,J=7.3Hz,1H,H-8),6.76(dd,J=8.3,0.5Hz,1H,H-6),3.16(q,J =6.5Hz,2H,ArNCH2),2.85(q,J=6.8Hz,2H,SNCH2),2.35(s,3H,CH3),1.67(qui,J=6.8Hz,2H,CH2).

[0163] 13C NMR(50MHz,DMSO-d6)δ:161.61,151.38,149.90,142.57,137.53,134.51,129.62,126.60,126.53,111.12,110.89,108.72,28.28,20.98,20.96.

[0164] ES-MS C 18 H 19 N4O4S m / z [MH] - : Calculated value 387.1, actual value 387.1.

[0165] Example 2 - Preparation of 9-((2,6-dibromo-4-(4-tosylpiperazine-1-carbonyl)phenoxy)carbonyl)-10-methylacridin-10-ium trifluoromethanesulfonate (EK297).

[0166] [ka]

[0167] Testing Procedure: Synthesis of tert-butyl 4-(3,5-dibromo-4-hydroxybenzoyl)piperazine-1-carboxylate 7: HOBt (259 mg, 1.69 mmol) and EDC (787 mg, 5.07 mmol) were added sequentially to a solution of 3,5-dibromo-4-hydroxybenzoic acid (1 g, 3.38 mmol) and 1-Boc-piperazine (1.89 g, 10.14 mmol) in dry tetrahydrofuran (40 mL) and the resulting mixture was stirred under argon for 7.5 h. The mixture was decanted into ice / water, acidified with 1N HCl, and the precipitate was filtered, washed with water, and dried to give phenol 7 as a white solid (1.35 g, 86%).

[0168] 1 H NMR (200MHz, CDCl3) δ: 7.53 (s, 2H, ArH), 6.32 (bs, 1H, OH), 3.65-3.40 (m, 8H, piperazine), 1.47 (s, 9H, tBu). 13C NMR(50MHz, CDCl3)δ:167.90,154.51,151.52,131.10,129.10,111.04,80.64,47.67(br),43.61(br),28.36.

[0169] ES-HRMS C 16 H 20 m / z of Br2N2NaO4 [M+Na] + : Calculated value 486.9667, actual value 486.9661.

[0170] Synthesis of 2,6-dibromo-4-(4-(tert-butoxycarbonyl)piperazine-1-carbonyl)phenyl acridine-9-carboxylate 8: A suspension of 9-acridine carboxylic acid hydrate (289 mg, 1.29 mmol) in thionyl chloride (3 mL) was stirred at reflux under argon for 5 h. After cooling, excess thionyl chloride was removed under high vacuum and dry dichloromethane (7 mL) was added under argon. To the resulting solution, phenol 7 (600 mg, 1.29 mmol), triethylamine (0.36 mL, 2.58 mmol) and pyridine (0.052 mL, 0.65 mmol) were added successively and the whole was stirred for 18 h. Then dichloromethane (30 mL) was added, the phases were separated and the aqueous phase was washed with dichloromethane (2×30 mL). The organic phases were combined and washed successively with 1N HCl (2×20 mL), aqueous NaHCO3 (20 mL) and brine (20 mL), dried (Na2SO4) and the solvent was evaporated. Column chromatography of the residue (10% MeCN / dichloromethane) afforded 8 as a yellow solid (715 mg, 83%).

[0171] 1 H NMR(200MHz,CDCl3)δ:8.74(d,J=8.8Hz,2H,H-4),8.34(d,J=8.8Hz,2H,H-1),7.87(ddd,J=8.8,6.6,1.4Hz,2H ,H-3),7.76(s,2H,H-3'),7.69(ddd,J=8.8,6.7,1.3Hz,2H,H-2),3.80-3.40(m,8H,piperazine),1.49(s,9H,tBu).

[0172] 13 C NMR(50MHz,CDCl3)δ:166.53,163.69,154.43,148.68,147.62,136.32,133.22,131.61,130.36,130 .14,127.69,125.71,122.92,118.29,80.58,47.69(br),43.59(br),43.31(br),42.45(br),28.37.

[0173] ES-HRMS C 30 H 28 m / z[M+H] of Br2N3O5 + : Calculated value 667.0375, actual value 670.0370.

[0174] Synthesis of 4-(4-((acridine-9-carbonyl)oxy)-3,5-dibromobenzoyl)piperazin-1-ium 2,2,2-trifluoroacetate 9: Acridine 8 (162 mg, 0.24 mmol) was added to a mixture of trifluoroacetic acid (2 mL) and chloroform (2 mL) and the solution was stirred for 4 h. The mixture was evaporated to leave salt 9 as a white solid (165 mg, quantitative).

[0175] 1 H NMR (200 MHz, MeOD-d4) δ: 8.84 (d, J = 8.9 Hz, 2H, H-4), 8.36 (d, J = 8.9 Hz, 2H, H-1), 8.12 (ddd, J = 8.8, 6.7, 1.2 Hz, 2H, H-3), 8.02 (s, 2H, H-3'), 7.89 (ddd, J = 8.5, 6.7, 1.1 Hz, 2H, H-2), 4.03-3.83 (m, 4H, piperazine), 3.46-3.33 (m, 4H, piperazine).

[0176] 19 F NMR (188MHz, MeOD-d4) δ: -77.99. ES-HRMS C 25 H 20 m / z[M] of Br2N3O3 + : Calculated value 569.9845, actual value 569.9899.

[0177] Synthesis of 2,6-dibromo-4-(4-tosylpiperazine-1-carbonyl)phenyl acridine-9-carboxylate 10: Tosyl chloride (95 mg, 0.5 mmol) was added to a stirred solution of 9 (310 mg, 0.45 mmol) and DIPEA (0.25 mL, 1.41 mmol) in DMF (8 mL) at 0° C. under argon and the reaction mixture was left stirring for 3 h. Water (30 mL) was added and the precipitate was filtered, washed with water and dried in vacuum to leave the desired product 10 as a yellow solid (235 mg, 73%).

[0178] 1 H NMR(200MHz,CDCl3)δ:8.70(d,J=8.8Hz,2H,H-4),8.32(d,J=8.7Hz,2H,H-1),7.84(ddd,J=8.7,6.7,1.1Hz,2H,H-3),7.71-7.60(m,6H,H-2,3',2 TOS ), 7.35(app d, J=8.1Hz, 2H, H-3 TOS ), 3.74(bs, 4H, piperazine), 3.05(bs, 4H, piperazine), 2.44(s, 3H, CH3).

[0179] 13 C NMR(50MHz,CDCl3)δ:166.47,163.76,148.79,147.90,144.42,135.81,133.25,132.26, 131.71,130.47,130.26,130.09,127.85,127.82,125.76,123.02,118.42,46.05,21.70.

[0180] ES-HRMS C 32 H 26 m / z[M+H] of Br2N3O5S + : Calculated value 723.9939, actual value 723.9998.

[0181] Synthesis of 9-((2,6-dibromo-4-(4-tosylpiperazine-1-carbonyl)phenoxy)carbonyl)-10-methylacridin-10-ium trifluoromethanesulfonate EK297: Methyl triflate (36 mg, 0.22 mmol) was added to a solution of acridine ester 10 (160 mg, 0.22 mmol) in dry dichloromethane (7 mL) and stirred under argon for 24 h. The volatiles were evaporated and the residue was washed with cold EtOAc (4 x 5 mL) (sonication / centrifugation), precipitated with DCM / toluene, and washed with hexane (x2) to leave the acridinium ester EK297 as a yellow powder (48 mg, 25%).

[0182] 1 H NMR(200MHz,CD3CN)δ:8.98(d,J=9.0Hz,2H,H-4),8.72(d,J=9.1Hz,2H,H-1),8.50(ddd,J =9.4,6.9,1.3Hz,2H,H-3),8.14(dd,J=8.7,6.7Hz,2H,H-2),7.78(s,2H,H-3'),7.64(app d,J=8.3Hz,2H,H-2 TOS ), 7.43(d, J=8.2Hz, 2H, H-3 TOS ), 4.90(s,3H,NCH3), 3.95-3.27(m,4H,piperazine), 3.00(app bs,4H,piperazine), 2.44(s,3H,CH3).

[0183] 13 C NMR(50MHz,CD3CN)δ:166.59,162.47,146.98,145.37,143.24,140.59,138.75,133.49, 132.88,130.85,130.64,129.15,128.75,124.42,120.31,118.27,46.72,40.96,21.56. 19 F NMR(188MHz,CD3CN)δ:-78.65.

[0184] ES-HRMS C 33 H 28 m / z[M] of Br2N3O5S +: Calculated value 738.0100, actual value 738.0099.

[0185] Example 3 - Cercosporin or 5-ALA + DZ325 or EK297 in GBM cell lines The efficacy of the chemiluminescent compounds DZ325 and EK297 was tested in cell cultures of human glioblastoma multiforme cell lines (U87, T98G, M059K and LN-18).

[0186] Cells were seeded in 96-well plates and allowed to attach overnight at 37 °C in a humidified atmosphere with 5% CO2. Subsequently, cells were incubated with (i) photosensitizer (cercosporin) or photosensitizer precursor (5-ALA for the generation of photosensitizer PpIX) and (ii) appropriate concentrations of compounds. In the case of 5-ALA, introduction of 5-ALA into the cell culture was performed 1 h before introduction of the chemiluminescent compound to ensure sufficient PpIX production. In the case of cercosporin, it was administered simultaneously with the chemiluminescent compound.

[0187] Chemiluminescent compounds were prepared as 50 mM stock solutions in DMSO (or 1-methyl-2-pyrrolidone) and diluted to their final concentrations in cell culture medium. Cercosporin was prepared as a 4 mM solution in DMSO and appropriately diluted in cell culture medium. 5-ALA was brought to its final concentration in Optimem. In the case of luminol-based compounds (DZ325), in some cases Cu was added to the cell culture medium in the form of CuSO4 at a final concentration of 100 μM.

[0188] 24 hours after introduction of the chemiluminescent compound to the cells, the cell medium was removed and replaced with 100 μL of medium containing 0.5 mg / mL of thiazolyl blue salt for the performance of a standard MTT viability assay. The MTT-containing medium was incubated for 1-3 hours and then removed and replaced with 100 μL of DMSO. End-point absorbance measurements were performed at 561 nm. A medium-only control was used as a 100% cell viability control. The experimental cell viability of the combined treatment (photosensitizer + chemiluminescent compound) was compared to the theoretical additive treatment of photosensitizer + chemiluminescent compound (calculated as follows):

[0189] Cell viability (photosensitizer only)% x cell viability (chemiluminescent part only)% 100% (control)

[0190] A survival rate of the combined treatment (experiment) that is significantly higher than the theoretically calculated additive value is considered evidence of a synergistic effect between the chemiluminescent compound and the photosensitizer.

[0191] The results are presented in Figures 1 and 3. In each figure, an example of synergy is highlighted. For example, in Figure 3, 3 μM cercosporin alone reduced cell viability to 80%, while 400 μM EK297 reduced toxicity to 74.5%. Calculated cell viability is determined according to the following formula (where "PS" = photosensitizer, "CL" = chemiluminescent agent):

[0192]

number

[0193] The calculated cell viability for 3 μM cercosporin + 400 μM EK297 is 59.5%, while the corresponding experimental value of cell viability was found to be 21.5%, suggesting cytotoxic synergy between EK297 and cercosporin. The example of synergy shown in Figures 1 and 2 with DZ325 is calculated in the same way.

Claims

1. A pharmaceutical composition comprising an endoplasmic reticulum-targeted chemiluminescent agent for use in the treatment of cancer by photodynamic therapy, wherein the agent is a conjugate comprising at least one chemiluminescent moiety bound to at least one endoplasmic reticulum-targeting moiety, and the photodynamic therapy comprises the simultaneous or sequential use of a photosensitizer or its precursor; and further; The conjugate is a compound of general formula (I), or a pharmaceutically acceptable salt thereof: 【Chemistry 1】 In the formula, A represents a chemiluminescent moiety selected from the group consisting of luminol, isoluminol, lucigenin, acridinium esters, and oxalate esters; Each L may be the same or different, and is either directly bonded or a linker which is a C1-15 alkylene chain optionally substituted with one or more groups selected from C1-3 alkyl, -O(C1-3) alkyl, -OH, cycloalkyl and aryl groups; and one or more -CH2- groups of the alkylene chain may be replaced with a group independently selected from -O-, -CO-, -NR- (wherein R is H or C1-6 alkyl), cycloalkyl, heterocyclic, aryl and heteroaryl groups; and Each B may be the same or different, and represents an endoplasmic reticulum targeting moiety having one of the following structures: 【Chemistry 2】 During the ceremony, R15 is hydrogen or C1-6 alkyl, Each R16 independently represents a C1-3 alkyl, C1-3 alkoxy, or halogen atom; and u is an integer between 0 and 5; n is an integer from 1 to 3, preferably 1; and A pharmaceutical composition in which x is an integer from 1 to 3, preferably 1.

2. The aforementioned conjugate is a compound of formula (II), or a pharmaceutically acceptable salt thereof: 【Transformation 3】 And, The pharmaceutical composition according to claim 1, wherein A, L, and B are as described in claim 1.

3. The pharmaceutical composition according to claim 1, wherein the chemiluminescent agent or the chemiluminescent portion is luminol, isolminol, or acridinium ester.

4. The linker L is -C ３ H ６ -, -C ４ H ８ -, -C ６ H １２ -, -C ８ H １６ -, -C １０ H ２０ -, -C １１ H ２２ -, -CO-, -CO-CH ２ -, -CO-C ３ H ６ , -CO-C ５ H １０ -, -CO-C ６ H １２ -, -CO-C １０ H ２０ The pharmaceutical composition according to claim 1, which is selected from the group consisting of -, and a polyethylene glycol group containing 1 to 4 ethylene oxide units.

5. The agent is a compound of formula (III), or a pharmaceutically acceptable salt thereof: 【Chemistry 4】 (In the formula, L １ The linker is a direct linker, a linker as defined in claim 1, or a linker selected from the group consisting of -C3H6-, -C4H8-, -C6H12-, -C8H16-, -C10H20-, -C11H22-, -CO-, -CO-CH2-, -CO-C3H6, -CO-C5H10-, -CO-C6H12-, -CO-C10H20-, and polyethylene glycol groups containing 1 to 4 ethylene oxide units. B １ This is the endoplasmic reticulum targeting portion as defined in claim 1, R ３ is hydrogen, or C １～３ Alkyl (for example, methyl) and other alkyl groups, Each R ４ C １～６ Alkyl and -NR ５ R ６ Selected independently from, R ５ and R ６ H and C １～６ From alkyl, preferably H and C １～３ Alkyl (e.g., -CH ３ ) were selected independently of, The pharmaceutical composition according to claim 1, wherein p is an integer from 0 to 3, preferably 0, 1, or 2 (for example, 0 or 1).

6. The agent is a compound of formula (IIIa) or (IIIb): 【Transformation 5】 In the ceremony, L １ , B １ , R ３ , R ４ The pharmaceutical composition according to claim 5, wherein p is as described in claim 5.

7. L １ teeth, 【Transformation 6】 Selected from the group consisting of, In the formula, a is an integer from 1 to 10, preferably from 3 to 10. The pharmaceutical composition according to claim 5 or claim 6, wherein b is an integer from 1 to 4, for example, 2.

8. The agent is a compound of formula (IV), or a pharmaceutically acceptable salt thereof: 【Transformation 7】 (In the formula, L ２ This is a linker as defined in claim 1 or 4, B ２ This is the endoplasmic reticulum targeting portion as defined in claim 1, Each R ６ These are halogens (e.g., F, Cl, Br, I) and C １～６ Selected independently from alkyl groups (e.g., tert-Bu), q is an integer between 0 and 4, preferably 0 or 2. Z is a monovalent anion, such as Cl, Br, I, or CF. ３ OSO ２ The pharmaceutical composition according to claim 1, wherein the composition is an anion.

9. L ２ The following is the basis: 【Transformation 8】 It represents one of the following: The pharmaceutical composition according to claim 8, wherein a is an integer from 1 to 10, preferably 3, 4, or 5.

10. The agent is a compound of formula (V), or a pharmaceutically acceptable salt thereof: 【Chemistry 9】 (In the formula, L ３ This is either a direct linkage or a linker as described in claim 1 or 4. B ３ This is the endoplasmic reticulum targeting portion described in any one of claims 1, Each R ７ These are halogens (e.g., F, Cl, Br, I), -CO ２ R ８ (In the formula, R ８ is hydrogen or C １～６ Alkyl, cyano, and C １～６ Selected independently from alkyl groups (e.g., tert-Bu), r is an integer from 0 to 5, preferably 0 or 3. Z is a monovalent anion, such as Cl, Br, I, or CF. ３ OSO ２ The pharmaceutical composition according to claim 1, wherein the composition is an anion.

11. L ３ C １～１０ Alkylene group, for example, C １～６ The pharmaceutical composition according to claim 10, wherein the component is alkylene.

12. The agent is a compound of formula (VIa), (VIb), or a pharmaceutically acceptable salt thereof: 【Chemistry 10】 And, In the ceremony, L ４ The linker is either directly bonded, the linker described in claim 1, or a linker selected from the group consisting of -C3H6-, -C4H8-, -C6H12-, -C8H16-, -C10H20-, -C11H22-, -CO-, -CO-CH2-, -CO-C3H6, -CO-C5H10-, -CO-C6H12-, -CO-C10H20-, and polyethylene glycol groups containing 1 to 4 ethylene oxide units. A １ The pharmaceutical composition according to claim 1, wherein the chemiluminescent portion is as described in claim 1.

13. L ４ teeth, 【Chemistry 11】 Selected from the group consisting of, The pharmaceutical composition according to claim 12, wherein a is an integer from 1 to 10, preferably from 3 to 10.

14. A １ The following: 【Chemistry 12】 (In the formula, R ３ is hydrogen, or C １～３ Alkyl (for example, methyl) and other alkyl groups, Each R ４ C １～６ Alkyl and -NR ５ R ６ Selected independently from, R ５ and R ６ H and C １～６ From alkyl, preferably H and C １～３ Alkyl (e.g., -CH ３ ) were selected independently of, p is an integer from 0 to 3, preferably 0, 1, or 2, for example, 0 or 1. Z is a monovalent anion, such as Cl, Br, I, or CF. ３ OSO ２ It is an anion, Each R ９ These are halogens (e.g., F, Cl, Br, I) and C １～６ Selected independently from alkyl (e.g., tBu), The pharmaceutical composition according to claim 12 or claim 13, wherein s is selected from any integer between 0 and 4, preferably 0, 2, or 3.

15. The agent is a compound of formula (VII), or a pharmaceutically acceptable salt thereof: 【Chemistry 13】 (In the formula, L ５ The linker is either directly bonded, the linker described in claim 1, or a linker selected from the group consisting of -C3H6-, -C4H8-, -C6H12-, -C8H16-, -C10H20-, -C11H22-, -CO-, -CO-CH2-, -CO-C3H6, -CO-C5H10-, -CO-C6H12-, -CO-C10H20-, and polyethylene glycol groups containing 1 to 4 ethylene oxide units. B ４ This is the endoplasmic reticulum targeting portion described in claim 1, Each R １０ C １～６ Alkyl (e.g., methyl), and -NR １１ R １２ Selected independently from, R １１ and R １２ H and C １～６ From alkyl, preferably H and C １～３ Alkyl (e.g., -CH ３ ) were selected independently of, The pharmaceutical composition according to claim 1, wherein t is an integer from 0 to 3, preferably 1 or 2.

16. The agent is of formula (VIIa): 【Chemistry 14】 (wherein L ５ , B ４ , R １１ and R １２ are as described in claim 15, and R １３ is H or C １～３ alkyl), the pharmaceutical composition according to claim 15.

17. L ５ is C １～１１ alkylene, preferably C ２～８ alkylene, for example, propylene, the pharmaceutical composition according to claim 15 or claim 16.

18. The agent is the following compound: 【Chemistry 15】 A pharmaceutical composition according to claim 1, selected from among.

19. The photosensitizer or precursor is 5-aminolevulinic acid (5-ALA) and derivatives of 5-ALA, protoporphyrin (e.g., protoporphyrin IX), or metallated phthalocyanine (i.e., AlPcS) which can be optionally sulfonated, for example, AlPcS ２ Or AlPcS ２ａ Disulfonated aluminum phthalocyanine, or aluminum phthalocyanine tetrasulfonate (A1PcS ４ Phthalocyanines such as ) and sulfonated tetraphenylporphyrins (e.g., TPPS ２ａ TPPS ４ TPPS １ and TPPS ２ｏ The pharmaceutical composition according to claim 1, selected from chlorine such as tetra(m-hydroxyphenyl)chlorine (m-THPC) (for example, temoporfin, which is commercially available under the trade name Foscan), chlorine derivatives including bacteriochlorine and ketochlorine, mono-L-aspartylchlorine e6 (NPe6) or chlorine e6, natural and synthetic porphyrins including hematoporphyrin and benzoporphyrin, anthraquinones and their derivatives (for example, hypericin, hypocrelin [A, B], cercosporine, carphostin, erusinochrome [A, B, C]).

20. The pharmaceutical composition according to claim 19, wherein the photosensitizer or precursor is selected from: cercosporine, 5-ALA, a derivative of 5-ALA, or a pharmaceutically acceptable salt thereof.

21. The pharmaceutical composition according to claim 19, wherein the photosensitizer is cercosporine.

22. The pharmaceutical composition according to claim 1, wherein the cancer is an internal cancer, for example, a deep-seated cancer.

23. The pharmaceutical composition according to claim 22, wherein the cancer is selected from the group consisting of glioma and other brain cancers, liver cancer and pancreatic cancer, breast cancer, lung cancer and prostate cancer, bile duct cancer, gastric cancer and colon cancer, bladder cancer, cervical cancer, and head and neck cancer.

24. The pharmaceutical composition according to claim 23, wherein the cancer is GBM.

25. A pharmaceutical composition comprising the agent according to any one of claims 1 to 6, 12 to 13, 15 to 16, and 18, together with at least one pharmaceutically acceptable carrier or excipient.

26. A pharmaceutical composition comprising, together with at least one pharmaceutically acceptable carrier or excipient, a drug according to any one of claims 1 to 6, 12 to 13, 15 to 16, and 18, and a photosensitizer or photosensitizer precursor according to any one of claims 1 and 19 to 21.

27. The pharmaceutical composition according to claim 25, for use in the treatment of cancer by photodynamic therapy, preferably for use in the treatment of internal cancer, such as deep-seated cancer.

28. A kit comprising (i) a drug as described in any one of claims 1 to 6, 12 to 13, 15 to 16, and 18; separately (ii) a photosensitizer or photosensitizer precursor as described in any one of claims 1 and 19 to 21; and optionally (iii) instructions for the use of (i) and (ii) in the treatment of cancer by photodynamic therapy.

29. A conjugate according to any one of claims 1 to 6, 12 to 13, 15 to 16, and 18, or a pharmaceutically acceptable salt thereof.

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