Photodynamic diagnosis and treatment integrated method

Abstract

The present invention provides a method of determining the presence and / or location of a tumor, and / or the recurrence or metastasis of said tumor, comprising a photodynamic diagnosis based on a porphyrin dimer salt photosensitizer, and optionally a method of performing a photodynamic therapy based on a porphyrin dimer salt photosensitizer on said determined tumor, the invention also relates to application of the compound in diagnosis and treatment of tumors, especially tumors suitable for interventional diagnosis and treatment.

Description

Field of the Invention

[0001] The present application relates to a method for photodynamic diagnosis, positioning and treatment of tumors based on porphyrin dimer salt photosensitizer. Background Art

[0002] Photodynamic therapy (PDT) is a new method of treating diseases by using the photodynamic effect of photosensitizers. It is currently mainly used to treat malignant tumors and certain precancerous lesions and benign lesions. After the photosensitizer is injected into the body, it will form a relatively high concentration of accumulation in the tumor tissue over a period of time. When irradiated with light of a specific wavelength, a strong photodynamic effect will occur, causing the diseased tissue to be fully destroyed. PDT has a certain selectivity for target tissues and good controllability of killing. Therefore, it can minimize the damage to normal tissues while ensuring the goal of tumor inactivation. It is a minimally invasive, low-toxic, non-thermogenic local treatment method, and therefore has broad application prospects.

[0003] The present invention provides a method for photodynamic diagnosis, positioning and treatment of tumors based on porphyrin dimer salt photosensitizer, which is used for treating tumors suitable for interventional diagnosis and treatment. Summary of the invention

[0004] The inventors of the present application unexpectedly found that porphyrin dimer salt photosensitizers can show different effects under laser irradiation with different wavelengths and powers. In particular, the inventors of the present application unexpectedly found that when irradiated with a laser of a specific wavelength (e.g., wavelength 365nm or 400nm) far lower than the light dose of photodynamic therapy (e.g., light power below 30mw), the porphyrin dimer salt photosensitizer can emit fluorescence, so that the lesion site enriched with the photosensitizer shows a fluorescence intensity significantly different from that of the non-lesion site. For such lesions identified by the photosensitizer, a laser of a photodynamic therapy wavelength (e.g., wavelength 630nm) can be further used to irradiate with an appropriate dose (e.g., light power 150-800mw), so that the porphyrin dimer salt photosensitizer has a therapeutic photodynamic effect, thereby destroying the lesion tissue with a relatively high accumulation of photosensitizer, and achieving the effect of treating tumors. Therefore, porphyrin dimer salt photosensitizers can achieve the effect of integrated diagnosis and treatment, greatly improve the efficiency of diagnosis and treatment, and reduce the burden on patients.

[0005] Thus, in one aspect, the present application provides a method for determining the presence and / or location of a tumor in an individual suspected of having a tumor, comprising:

[0006] a) administering to the individual a photosensitizer comprising a porphyrin dimer salt so that the photosensitizer is distributed at a body site of the individual suspected of having a tumor;

[0007] b) irradiating the body part with laser light of a first wavelength to excite the porphyrin dimer salt distributed in the body part to emit fluorescence, wherein the first wavelength is lower than the wavelength of photodynamic therapy using the porphyrin dimer salt;

[0008] c) detecting whether the fluorescence at the body part shows a fluorescence intensity difference at a certain position that is different from that at surrounding positions;

[0009] The presence of the fluorescence intensity difference reflects the presence of a tumor, and / or the location showing the fluorescence intensity difference reflects the location of the tumor.

[0010] The present application also provides a method for determining the presence and / or location of a tumor in an individual suspected of having a tumor and treating the determined tumor, comprising:

[0011] a) administering to the individual a photosensitizer comprising a porphyrin dimer salt so that the photosensitizer is distributed at a body site of the individual suspected of having a tumor;

[0012] b) irradiating the body part with laser light of a first wavelength to excite the porphyrin dimer salt distributed in the body part to emit fluorescence, wherein the first wavelength is lower than the wavelength of photodynamic therapy using the porphyrin dimer salt;

[0013] c) detecting whether the fluorescence at the body part shows a difference in fluorescence intensity at a certain position that is different from that at surrounding positions, the presence of the fluorescence intensity difference reflecting the presence of a tumor, and / or the position showing the fluorescence intensity difference reflecting the position of the tumor; and

[0014] d) irradiating the location showing the fluorescence intensity difference with laser light of a second wavelength to perform photodynamic therapy.

[0015] In certain embodiments, step d) includes irradiating the body part with laser light of a first wavelength and irradiating the location in the body part exhibiting the fluorescence intensity difference with laser light of a second wavelength to perform the photodynamic therapy.

[0016] The present application also provides a method for determining the effect of tumor treatment and / or tumor recurrence or metastasis in an individual suffering from a tumor and undergoing treatment, comprising:

[0017] a) administering to the individual a photosensitizer comprising a porphyrin dimer salt so that the photosensitizer is distributed at a part of the body of the individual having a tumor;

[0018] b) irradiating the body part with laser light of a first wavelength to excite the porphyrin dimer salt distributed in the body part to emit fluorescence, wherein the first wavelength is lower than the wavelength of photodynamic therapy using the porphyrin dimer salt;

[0019] c) detecting whether the fluorescence at the body part shows a difference in fluorescence intensity at a certain position that is different from that at surrounding positions, and obtaining a post-treatment fluorescence result;

[0020] The existence of the fluorescence intensity difference reflects the existence of a tumor, and / or the location showing the fluorescence intensity difference reflects the location of the tumor, and the existence and location of the tumor reflect the treatment effect of the tumor and / or the recurrence or metastasis of the tumor.

[0021] In certain embodiments, the method further comprises step d): irradiating the location showing the fluorescence intensity difference with a laser of a second wavelength to perform photodynamic therapy.

[0022] In certain embodiments, step d) includes irradiating the body part with laser light of a first wavelength and irradiating the location in the body part exhibiting the fluorescence intensity difference with laser light of a second wavelength to perform the photodynamic therapy.

[0023] In some embodiments, the tumor is suitable for interventional diagnosis and treatment, and optionally, is suitable for endoscopic examination. In some embodiments, the tumor is a digestive tract tumor (e.g., esophageal cancer, gastric cancer, gastrointestinal stromal tumor, rectal cancer), a urinary tract tumor (e.g., bladder tumor), a reproductive system tumor (e.g., vaginal tumor, cervical cancer, ovarian cancer, endometrial cancer, prostate cancer) or an intra-abdominal tumor (e.g., liver tumor, pancreatic tumor, kidney tumor).

[0024] In certain embodiments, the administration comprises administration by injection, infusion or transdermal administration.

[0025] In certain embodiments, the porphyrin dimer salt has a structure shown in formula (I), formula (II) or formula (III):

[0026]

[0027]

[0028] in

[0029] R is independently selected from C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy and C 1-6acyl, which is optionally substituted by one or more substituents selected from halogen and hydroxy,

[0030] M is an alkali metal, an alkaline earth metal or NH4 + ,

[0031] p is the reciprocal of the valence of M,

[0032] and solvates thereof, especially hydrates.

[0033] In certain embodiments, wherein R is independently selected from C optionally substituted with one or more hydroxyl groups 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl and C 1-6 Acyl, M is an alkali metal or NH4 + , and p is 1, and solvates thereof, especially hydrates.

[0034] In certain embodiments, wherein R is independently selected from -CH=CH2, -CH(OH)CH3, and -COCH3, M is an alkali metal or NH4 + , and p is 1, and solvates thereof, especially hydrates.

[0035] In certain embodiments, wherein R is independently selected from -CH=CH2 and -CH(OH)CH3, M is an alkali metal or NH4 + , and p is 1, and solvates thereof, especially hydrates.

[0036] In certain embodiments, wherein M is an alkali metal, such as Na or K.

[0037] In certain embodiments, the porphyrin dimer salt bound by an ether bond is selected from:

[0038] Bis[1-[sodium 6,7-dipropionate-1,3,5,8-tetramethyl-4-vinyl-2-porphine]ethyl]ether (DVDMS-1),

[0039] Bis[1-[sodium 6,7-dipropionate-1,3,5,8-tetramethyl-2-vinyl-4-porphine]ethyl]ether (DVDMS-2),

[0040] 1-[6,7-dipropanoic acid sodium-1,3,5,8-tetramethyl-2-vinyl-4-porphine]ethyl 1-[6',7'-dipropanoic acid sodium-1',3',5',8'-tetramethyl-4-vinyl-2-porphine]ethyl ether (DVDMS-3),

[0041] Bis[1-[6,7-dipropanoic acid potassium-1,3,5,8-tetramethyl-4-vinyl-2-porphine]ethyl]ether,

[0042] Bis[1-[6,7-dipropanoic acid potassium-1,3,5,8-tetramethyl-2-vinyl-4-porphine]ethyl]ether,

[0043] 1-[6,7-dipropionate potassium-1,3,5,8-tetramethyl-2-vinyl-4-porphine]ethyl 1-[6',7'-dipropionate potassium-1',3',5',8'-tetramethyl-4-vinyl-2-porphine]ethyl ether,

[0044] Bis[1-[6,7-dipropionate ammonium-1,3,5,8-tetramethyl-4-vinyl-2-porphine]ethyl]ether,

[0045] Bis[1-[6,7-dipropionate ammonium-1,3,5,8-tetramethyl-2-vinyl-4-porphine]ethyl]ether, and

[0046] 1-[6,7-dipropionate ammonium-1,3,5,8-tetramethyl-2-vinyl-4-porphine]ethyl 1-[6',7'-dipropionate ammonium-1',3',5',8'-tetramethyl-4-vinyl-2-porphine]ethyl ether.

[0047] In some embodiments, the porphyrin dimer salt is sodium porphyrin (DVDMS), which is a sodium salt of porphyrin dimer bonded by an ether bond. In some embodiments, the sodium porphyrin has the structure shown in the above formula (I), formula (II), or formula (III). In some embodiments, the porphyrin dimer salt is DVDMS-1, DVDMS-2, DVDMS-3 or DVDMS-M (a mixture of DVDMS-1, DVDMS-2, and DVDMS-3).

[0048] In certain embodiments, the porphyrin dimer salt is administered to a subject at a dose of 0.1-1 mg / kg body weight.

[0049] In certain embodiments, the porphyrin dimer salt is administered by perfusion or transdermally, and step b) is performed half an hour to one hour after the administration of the porphyrin dimer salt.

[0050] In certain embodiments, the porphyrin dimer salt is administered by intravenous injection, and step b) is performed 24 hours after the administration of the porphyrin dimer salt.

[0051] In some embodiments, the first wavelength is in the range of 365-405 nm. In some embodiments, the first wavelength is 405 nm.

[0052] In some embodiments, the laser power of the first wavelength is in the range of 10 mw-30 mw.

[0053] In certain embodiments, the fluorescence can be observed by naked eye, or an image of the fluorescence can be collected at a wavelength of 620 nm.

[0054] In certain embodiments, the second wavelength is a wavelength of 630±3 nm, such as 630 nm, 631 nm or 632 nm.

[0055] In some embodiments, the laser power of the second wavelength is in the range of 150-800 mw.

[0056] On the other hand, the present application provides a use of a porphyrin dimer salt for preparing an integrated diagnostic and therapeutic reagent, wherein the integrated diagnostic and therapeutic reagent is used to determine the presence and / or location of a tumor in an individual suspected of having a tumor, to determine the efficacy of tumor treatment and / or tumor recurrence in an individual suffering from a tumor and undergoing treatment, or to determine the presence and / or location of a tumor in an individual suspected of having a tumor and treating the determined tumor.

[0057] In yet another aspect, the present application also provides a photosensitizer suitable for diagnosing and treating cancer in an individual, wherein the photosensitizer comprises the porphyrin dimer salt as described above.

[0058] In another aspect, the present application also provides a device for integrated diagnosis and treatment applicable to the method as described above, the device comprising:

[0059] a) Interventional devices that can be implanted in the body;

[0060] b) a first device capable of emitting laser light at said first wavelength; and

[0061] c) a second device capable of emitting laser light at said second wavelength;

[0062] The first device and the second device are arranged on the interventional device, and can be controlled to emit the laser after the interventional device is implanted in the body.

[0063] In some embodiments, the first device and the second device can be controlled to emit laser light of the first wavelength and laser light of the second wavelength, respectively, at the same time.

[0064] In some embodiments, the first device and the second device can be manipulated so that the first device emits a laser of the first wavelength at the first time, and the second device emits a laser of the second wavelength at the second time. Without wishing to be bound by theory, the method for photodynamic diagnosis, positioning, and treatment of tumors based on porphyrin dimer salt photosensitizers described in this application has outstanding advantages in some medical application scenarios. Most of the fluorescence diagnostic equipment and photodynamic therapy equipment currently used in clinical practice have only a single function. Fluorescence diagnostic equipment can only be used for diagnosis and surgical navigation, and photodynamic therapy equipment can only be used for treatment. However, this application realizes the integration of photodynamic diagnosis and treatment through the different reactions of porphyrin dimer salt photosensitizers to laser irradiation with different parameters, so that diagnosis and treatment are completed simultaneously in one interventional medical operation (only the laser emission wavelength and control power need to be switched), which is particularly suitable for tumors suitable for interventional diagnosis and treatment, such as tumors suitable for endoscopes. The present invention has at least the following advantages: 1. Treatment can be performed immediately upon discovery of the lesion, reducing the damage caused by repeated endoscopic operation; 2. It is conducive to more targeted treatment of the diseased tissue and minimizes damage to normal tissue; 3. Based on the strength of the fluorescent reaction, the density of the diseased tissue can be preliminarily determined, and the light dose used in the treatment process can be adjusted accordingly to achieve personalized treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0065] Figure 1 Shows the in vivo fluorescence imaging of mice injected with sodium porphyrin (dose: 0.2 mg / kg).

[0066] Figure 2 The in vitro fluorescence imaging of various tissues or organs of mice injected with sodium thiourea (dose: 0.2 mg / kg) at the 32h time point is shown, from top to bottom: tumor, muscle tissue, heart, liver, spleen, lung, kidney, brain, and skin.

[0067] Figure 3 The fluorescence imaging data of various tissues or organs of mice injected with sodium porphyrin (dose: 0.2 mg / kg) are shown. The tissue fluorescence intensity of the target region (ROI) is calculated by Living Image software, and after deducting the background of each tissue or organ, a curve graph of the relative fluorescence intensity (ratio of tumor to normal tissue) changing with time is drawn.

[0068] Figure 4 Endoscopic NIR imaging and section staining of a bladder cancer patient are shown. DETAILED DESCRIPTION

[0069] The following description of the application is intended only to illustrate various embodiments of the application. Therefore, the specific modifications discussed should not be interpreted as limiting the scope of the application. It will be apparent to those skilled in the art that various equivalents, changes and modifications can be made without departing from the scope of the application, and it should be understood that these equivalent embodiments will be included herein. All references cited in this application, including publications, patents and patent applications, are incorporated into this application by reference in their entirety.

[0070] The present application provides a method for determining the presence and / or location of a tumor in an individual suspected of having a tumor, and optionally a method for treating the determined tumor, as well as a method for determining the effectiveness of tumor treatment and / or tumor recurrence or metastasis in an individual who has a tumor and has undergone treatment.

[0071] In one aspect, the method described herein comprises the following steps:

[0072] a) administering a photosensitizer comprising a porphyrin dimer salt to the individual so that the photosensitizer is distributed at a body site of the individual suspected of having a tumor.

[0073] b) irradiating the body part with a laser of a first wavelength to excite the porphyrin dimer salt distributed in the part to emit fluorescence, wherein the first wavelength is lower than the wavelength of photodynamic therapy using the porphyrin dimer salt. In some embodiments, the above method described in the present application also includes:

[0074] c) detecting whether the fluorescence in the body part shows a fluorescence intensity difference at a certain position that is different from that at surrounding positions, wherein the existence of the fluorescence intensity difference reflects the existence of a tumor, and / or the position showing the fluorescence intensity difference reflects the location of the tumor, optionally, the existence and location of the tumor reflect the treatment effect of the tumor and / or the recurrence or metastasis of the tumor.

[0075] In some embodiments, the above method described in the present application further comprises:

[0076] d) irradiating the location showing the fluorescence intensity difference with laser light of a second wavelength to perform photodynamic therapy.

[0077] In some embodiments, the above method described in the present application further comprises:

[0078] e) while irradiating the body part with laser light of a first wavelength, irradiating the location in the body part showing the fluorescence intensity difference with laser light of a second wavelength to perform photodynamic therapy.

[0079] In this application, the terms "tumor", "cancer", "malignant tumor" and "cancer" are used interchangeably herein to refer to cells that exhibit relatively abnormal, uncontrolled and / or autonomous growth, such that the cells exhibit an abnormal growth phenotype characterized by significantly uncontrolled cell proliferation. As used herein, the terms "tumor", "cancer", "malignant tumor" and "cancer" may include primary, or metastatic. In certain embodiments, the tumor is a tumor suitable for interventional diagnosis and treatment.

[0080] In this application, the term "interventional diagnosis and treatment" refers to making tiny channels in the skin, blood vessels, etc., or delivering diagnostic or therapeutic instruments or equipment to the diagnosis and treatment site for disease diagnosis and treatment through the original channels of the human body without exposing the lesion through surgery. Examples of the interventional diagnosis and treatment include, but are not limited to, endoscopy, interventional ultrasound angiography, etc. In some embodiments, the interventional diagnosis and treatment is endoscopy. Examples of tumors suitable for interventional diagnosis and treatment include, but are not limited to, digestive tract tumors, such as esophageal cancer, gastric cancer, gastrointestinal stromal tumors, and rectal cancer; urinary tract tumors, such as bladder tumors; reproductive system tumors, such as vaginal tumors, cervical cancer, ovarian cancer, endometrial cancer, and prostate cancer; intra-abdominal tumors, such as liver tumors, pancreatic tumors, and kidney tumors.

[0081] In the present application, the photosensitizer comprising a porphyrin dimer salt refers to a compound structure having two porphyrin structures connected by covalent bonding or a linker. In certain embodiments, the porphyrin dimer salt has a structure shown in the following formula (I), formula (II), or formula (III):

[0082]

[0083]

[0084] in

[0085] R is independently selected from C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy and C 1-6 acyl, which is optionally substituted by one or more substituents selected from halogen and hydroxy,

[0086] M is an alkali metal, an alkaline earth metal or NH4 + ,

[0087] p is the reciprocal of the valence of M,

[0088] and solvates thereof, especially hydrates.

[0089] In this application, “C 1-6"Alkyl" means a branched or straight-chain monovalent saturated hydrocarbon group having 1 to 6 carbon atoms. 1-4 Alkyl. Examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, and n-hexyl.

[0090] In this application, “C 2-6 "Alkenyl" means a straight or branched monovalent unsaturated hydrocarbon group having 2 to 6 carbon atoms and containing one or more double bonds. 2-4 Examples include, but are not limited to, vinyl, prop-1-enyl, allyl, but-1-enyl, pent-1-enyl, pent-1,4-dienyl, and the like.

[0091] In this application, “C 2-6 "Alkynyl" means a straight or branched monovalent unsaturated hydrocarbon group having 2 to 6 carbon atoms and containing one or more triple bonds. 2-4 Examples include, but are not limited to, ethynyl, prop-1-ynyl, prop-2-ynyl, but-1-ynyl, pent-1-ynyl, pent-1,3-diynyl, and the like.

[0092] In this application, “C 1-6 "Alkoxy" means a group R'-O-, wherein R' is a C 1-6 Alkyl. Preferably C1-4 alkoxy. Examples thereof include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, isopentoxy, neopentoxy and n-hexoxy.

[0093] In this application, “C 1-6 "Acyl" means a group R'-C(O)-, wherein R' is H or a branched or straight-chain monovalent saturated hydrocarbon radical having 1 to 5 carbon atoms. Preferably C 1-4 Acyl. Examples include, but are not limited to, formyl, acetyl, propionyl, butyryl, valeryl, hexanoyl, and the like.

[0094] In the present application, "optional" or "optionally" means that the event or situation described later can occur but does not have to occur, and the description includes the situation where the event or situation occurs and the situation where it does not occur. For example, "C1-6 alkyl ..., which is optionally substituted with one or more substituents selected from halogen and hydroxyl" means that C1-6 alkyl can be, but not necessarily, substituted with one or more halogen and / or hydroxyl, and the description includes unsubstituted C1-6 alkyl and C1-6 alkyl substituted with one or more halogen and / or hydroxyl.

[0095] In the present application, "halogen" means fluorine, chlorine, bromine and iodine, preferably fluorine, chlorine and bromine, more preferably fluorine and chlorine.

[0096] In this application, "hydroxy" means the group -OH.

[0097] In this application, "valence" means the number of shared electron pairs formed by one atom of a chemical element. The valence of an ion is equal to its charge. For example, alkali metals and NH4 + The valence number of is 1, and the valence number of alkaline earth metals is 2.

[0098] In the present application, "solvate" means a solvent addition form formed by the addition of solvent molecules to the porphyrin dimer salts described herein which are bonded together by mutual attraction. If the solvent is water, the solvate formed is a hydrate, such as a monohydrate or a dihydrate; if the solvent is an alcohol, the solvate formed is an alcoholate, such as an addition form with methanol or ethanol.

[0099] In certain embodiments, R is independently selected from C optionally substituted with one or more hydroxyl groups. 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl and C 1-6 Acyl, M is an alkali metal or NH4 + , and p is 1.

[0100] In certain embodiments, R is independently selected from -CH=CH2, -CH(OH)CH3, and -COCH3, and M is an alkali metal or NH4 + , and p is 1.

[0101] In certain embodiments, R is independently selected from -CH=CH2 and -CH(OH)CH3, and M is an alkali metal or NH4 + , and p is 1.

[0102] In certain embodiments, M is an alkali metal.

[0103] In certain embodiments, M is Na or K.

[0104] In certain embodiments, the porphyrin dimer salt bound by an ether bond is selected from the group consisting of: bis[1-[sodium 6,7-dipropionate-1,3,5,8-tetramethyl-4-vinyl-2-porphine]ethyl]ether, bis[1-[sodium 6,7-dipropionate-1,3,5,8-tetramethyl-2-vinyl-4-porphine]ethyl]ether, 1-[sodium 6,7-dipropionate-1,3,5,8-tetramethyl-2-vinyl-4-porphine]ethyl1-[6',7'-sodium dipropionate-1',3',5',8'-tetramethyl-4-vinyl-2-porphine]ethylether, bis[1-[potassium 6,7-dipropionate-1,3,5,8-tetramethyl-4-vinyl-2-porphine]ethyl]ether, bis[1-[potassium 6,7-dipropionate-1,3,5,8-tetramethyl-2- Vinyl-4-porphine]ethyl]ether, 1-[6,7-dipropionate potassium-1,3,5,8-tetramethyl-2-vinyl-4-porphine]ethyl 1-[6',7'-dipropionate potassium-1',3',5',8'-tetramethyl-4-vinyl-2-porphine]ethyl ether, di[1-[6,7-dipropionate ammonium-1,3,5,8-tetramethyl-4-vinyl-2-porphine]ethyl 1-[6,7-dipropionate-1,3,5,8-tetramethyl-2-vinyl-4-porphine]ethyl] ether, di[1-[6,7-dipropionate-1,3,5,8-tetramethyl-2-vinyl-4-porphine]ethyl] ether, and 1-[6,7-dipropionate-1,3,5,8-tetramethyl-2-vinyl-4-porphine]ethyl 1-[6',7'-dipropionate-1',3',5',8'-tetramethyl-4-vinyl-2-porphine]ethyl ether.

[0105] In certain embodiments, the porphyrin dimer salt is DVDMS, which is a sodium salt of a porphyrin dimer bonded by an ether bond. In some embodiments, DVDMS has the structure shown in the above formula (I), formula (II), or formula (III).

[0106] In certain embodiments, the porphyrin dimer salt is di[1-[sodium 6,7-dipropionate-1,3,5,8-tetramethyl-4-vinyl-2-porphine]ethyl]ether (DVDMS-1), di[1-[sodium 6,7-dipropionate-1,3,5,8-tetramethyl-2-vinyl-4-porphine]ethyl]ether (DVDMS-2), 1-[sodium 6,7-dipropionate-1,3,5,8-tetramethyl-2-vinyl-4-porphine]ethyl 1-[6',7'-sodium dipropionate-1',3',5',8'-tetramethyl-4-vinyl-2-porphine]ethyl ether (DVDMS-3), or a mixture of the above three (DVDMS-M).

[0107]

[0108] More about the photosensitizer containing porphyrin dimer salt is described in CN102030765B, and the disclosure of the patent is incorporated herein in full. Without wishing to be bound by theory, the photosensitizer containing porphyrin dimer salt has a parent ring porphyrin ring, which belongs to endogenous substances in the human body, so that the metabolites of the photosensitizer in the human body are safe and non-toxic. And the time for the porphyrin ring to avoid direct sunlight is short, only about three days, and the integrated operation of diagnosis and treatment greatly shortens the cycle of treatment and diagnosis, so that the photosensitizer in the patient's body can play a maximum role when performing the integrated operation of diagnosis and treatment and metabolize out of the body quickly after the operation is completed, avoiding adding burden to the patient's body.

[0109] In the present application, the photosensitizer comprising the porphyrin dimer salt can be formulated into any suitable form and can be used by any suitable approach, can be formulated into solution, suspension, emulsion, lyophilized preparation etc. for injection (for example intra-arterial, intravenous, intramuscular, subcutaneous, intraperitoneal injection etc.) or perfusion administration, be formulated into ointment, cream, suppository, patch etc. for transdermal administration. Preferred mode of administration is generally injection, perfusion and transdermal administration. Injection administration can make the photosensitizer of the present invention reach distribution equilibrium within, for example, 24 hours. Perfusion and transdermal administration can make the photosensitizer of the present invention reach distribution equilibrium within, for example, 0.5-1 hour.

[0110] In certain embodiments, the porphyrin dimer salt is administered in a range of 0.1-1 mg / kg body weight, for example, 0.1-0.9 mg / kg body weight, 0.1-0.8 mg / kg body weight, 0.1-0.7 mg / kg body weight, 0.1-0.6 mg / kg body weight, 0.1-0.5 mg / kg body weight, 0.1-0.4 mg / kg body weight, 0.1-0.3 mg / kg body weight, or 0.1-0.2 mg / kg body weight.

[0111] In this application, the term "fluorescence" refers to when a substance is irradiated with incident light of a specific wavelength, it absorbs light energy and enters an excited state, and immediately de-excites and emits output light with a wavelength longer than the incident light. The output light with this property is fluorescence.

[0112] In the present application, the term "fluorescence intensity difference" refers to the difference in the intensity of the light emitting fluorescence when irradiated by the same specific wavelength of incident light due to the difference in the properties of the irradiated substance. In some embodiments, the fluorescence intensity difference refers to the difference in fluorescence intensity between tumor tissue and non-lesion tissue. In some embodiments, the fluorescence intensity difference can be observed by the naked eye. In some embodiments, the dynamic image of the fluorescence intensity difference can be collected at an appropriate wavelength, for example, at a wavelength of 620nm.

[0113] In the present application, the first wavelength is an excitation wavelength, which can excite the photosensitizer containing porphyrin dimer salt distributed in a certain body part to emit fluorescence when irradiating the body part, and its light dose is not enough to induce a photodynamic reaction that is lethal to the human body. In some embodiments, the wavelength range of the first wavelength is 365nm-405nm. In some embodiments, the first wavelength is 405nm. In some embodiments, the range of the laser power of the first wavelength is 10mw-30mw. In some embodiments, the laser power of the first wavelength is 10mw.

[0114] In the present application, the second wavelength is a therapeutic wavelength, and a relatively high concentration of photosensitizer containing porphyrin dimer salt is accumulated in the tumor tissue. When a certain body part is irradiated with the second wavelength, a strong photodynamic effect of the photosensitizer can be caused, so that the diseased tissue is fully destroyed. In some embodiments, the wavelength range of the second wavelength is 630±3nm, such as 630nm, 631nm or 632nm. The light dose (laser power) of the second wavelength is much higher than the first wavelength. In some embodiments, the laser power of the second wavelength ranges from 150-800mw.

[0115] In this application, the term "individual" means an animal, including mammals and non-mammals. Mammals refer to any member of the mammal class, including but not limited to humans, non-human primates (such as chimpanzees and other apes and monkeys), farm animals (such as cattle, horses, sheep, goats and pigs), domestic animals (such as rabbits, dogs and cats), laboratory animals (including rodents, such as rats, mice and guinea pigs), etc. Examples of non-mammals include but are not limited to birds, etc.

[0116] In this application, the term "treatment" means to alleviate one or more disease-related symptoms, delay or avoid disease progression or deterioration, or cure a disease. For cancer, treatment may include curbing cancer development, reducing its severity, delaying its growth, inhibiting its metastasis, and reducing recurrence, etc. However, treatment does not require that the disease or symptoms associated with it be completely eliminated.

[0117] In the present application, the term "tumor recurrence" refers to the reappearance of a tumor of the same type as the primary tumor after the tumor at the primary site regresses after treatment.

[0118] In the present application, the term "tumor metastasis" refers to the metastasis to adjacent sites or distant organs via lymph or blood, in addition to the primary esophageal lesion.

[0119] In the present application, the term "integrated diagnosis and treatment of tumors" refers to treating the determined tumor while determining whether the tumor exists and / or the location of the tumor.

[0120] Methods for determining the presence and / or location of a tumor

[0121] The present application provides a method for determining the presence and / or location of a tumor in an individual suspected of having a tumor, comprising:

[0122] a) administering to the individual a photosensitizer comprising a porphyrin dimer salt so that the photosensitizer is distributed at a body site of the individual suspected of having a tumor;

[0123] b) irradiating the body part with laser light of a first wavelength to excite the porphyrin dimer salt distributed in the body part to emit fluorescence, wherein the first wavelength is lower than the wavelength of photodynamic therapy using the porphyrin dimer salt;

[0124] c) detecting whether the fluorescence at the body part shows a difference in fluorescence intensity at a certain position that is different from that at surrounding positions;

[0125] The presence of the fluorescence intensity difference reflects the presence of a tumor, and / or the location showing the fluorescence intensity difference reflects the location of the tumor.

[0126] Method for determining the presence and / or location of a tumor and treating the determined tumor

[0127] The present application provides a method for determining the presence and / or location of a tumor in an individual suspected of having a tumor and treating the determined tumor, comprising:

[0128] a) administering to the individual a photosensitizer comprising a porphyrin dimer salt so that the photosensitizer is distributed at a body site of the individual suspected of having a tumor;

[0129] b) irradiating the body part with laser light of a first wavelength to excite the porphyrin dimer salt distributed in the body part to emit fluorescence, wherein the first wavelength is lower than the wavelength of photodynamic therapy using the porphyrin dimer salt;

[0130] c) detecting whether the fluorescence at the body part shows a difference in fluorescence intensity at a certain position that is different from that at surrounding positions, the presence of the fluorescence intensity difference reflecting the presence of a tumor, and / or the position showing the fluorescence intensity difference reflecting the position of the tumor; and

[0131] d) irradiating the location showing the fluorescence intensity difference with laser light of a second wavelength to perform photodynamic therapy.

[0132] The step d) includes irradiating the body part with laser light of the first wavelength and irradiating the location in the body part showing the fluorescence intensity difference with laser light of the second wavelength to perform the photodynamic therapy.

[0133] Methods for determining the effectiveness of tumor treatment and / or tumor recurrence or metastasis

[0134] The present application provides a method for determining the effect of tumor treatment and / or tumor recurrence or metastasis in an individual suffering from a tumor and undergoing treatment, comprising:

[0135] a) administering to the individual a photosensitizer comprising a porphyrin dimer salt so that the photosensitizer is distributed at a part of the body of the individual having a tumor;

[0136] b) irradiating the body part with laser light of a first wavelength to excite the porphyrin dimer salt distributed in the body part to emit fluorescence, wherein the first wavelength is lower than the wavelength of photodynamic therapy using the porphyrin dimer salt;

[0137] c) detecting whether the fluorescence at the body part shows a difference in fluorescence intensity at a certain position that is different from that at surrounding positions, and obtaining a post-treatment fluorescence result;

[0138] The existence of the fluorescence intensity difference reflects the existence of a tumor, and / or the location showing the fluorescence intensity difference reflects the location of the tumor, and the existence and location of the tumor reflect the treatment effect of the tumor and / or the recurrence or metastasis of the tumor.

[0139] In certain embodiments, the method further comprises step d): irradiating the location showing the fluorescence intensity difference with a laser of a second wavelength to perform photodynamic therapy.

[0140] In certain embodiments, step d) includes irradiating the body part with laser light of a first wavelength while irradiating the location in the body part exhibiting the fluorescence intensity difference with laser light of a second wavelength to perform the photodynamic therapy.

[0141] Integrated equipment for tumor diagnosis and treatment

[0142] The present application provides a device for integrated diagnosis and treatment applicable to the method as described above, the device comprising:

[0143] a) Interventional devices that can be implanted in the body;

[0144] b) a first device capable of emitting laser light at said first wavelength; and

[0145] c) a second device capable of emitting laser light at said second wavelength;

[0146] The first device and the second device are arranged on the interventional device, and can be controlled to emit the laser after the interventional device is implanted in the body.

[0147] In some embodiments, the first device and the second device can be controlled to emit laser light of the first wavelength and laser light of the second wavelength, respectively, at the same time.

[0148] In some embodiments, the first device and the second device can be controlled so that the first device emits laser light of the first wavelength at a first time, and the second device emits laser light of the second wavelength at a second time.

[0149] Example

[0150] Material:

[0151] 1. Test drugs and instruments

[0152] 1.1 Test drugs

[0153] DVDMS-2 sodium porphyrin for injection (Shanghai Guangsheng Pharmaceutical Co., Ltd.);

[0154] 1.2 Instruments and Equipment

[0155] Lumina III small animal optical in vivo imaging system (PerkinElmer).

[0156] 1.3 Analysis software

[0157] Living Image software (PerkinElmer).

[0158] 2. Tumor cell lines

[0159] Human esophageal cancer Eca-109 cell line was purchased from China Center for Type Culture Collection (CCTCC) in Wuhan.

[0160] 3. Experimental Animals

[0161] Nu / nu nude mice, weighing 18-22 g, male, were provided by Shanghai Slake Experimental Animal Co., Ltd.

[0162] Example 1. Fluorescence imaging response of sodium chaotoporphyrin-mediated photodynamic diagnosis in the human esophageal cancer Eca-109 mouse model

[0163] Objective: To observe the fluorescence response of sodium chaotoporphyrin at a clinical therapeutic dose when irradiated with 420 nm wavelength laser in the tissue of Eca-109 mouse model of esophageal cancer, and to provide a reference for the clinical application of integrated photodynamic fluorescence diagnosis and treatment.

[0164] plan:

[0165] 1. Preparation of Human Esophageal Cancer Mouse Model

[0166] Human esophageal cancer Eca-109 cells in the logarithmic growth phase were collected and diluted to 5*10 7 The concentration of cells / mL was 0.1 mL per nude mouse, and injected subcutaneously at the base of the right forelimb of the nude mouse. After 7-10 days, a tumor was observed to grow to a diameter of about 8-10 mm at the base of the right thigh of the nude mouse, and then it was used for storage.

[0167] 2. Experimental process

[0168] Select the qualified tumor-bearing mouse model as described above, and administer sodium porphyrin via tail vein injection at a clinical therapeutic dose of 0.2 mg / kg body weight. Set the following 13 time points after the end of administration: 1h, 4h, 8h, 12h, 16h, 20h, 24h, 28h, 32h, 48h, 72h, 96h, 120h. At different time points after administration, the experimental group mice were irradiated with a laser of 420nm wavelength, and the control group mice were not irradiated. Three experimental group mice and one control group mouse were sacrificed at each time point for in vivo fluorescence imaging of animals, and then important organs such as heart, liver, spleen, lung, and kidney were dissected for organ fluorescence imaging, and muscles, brain, skin, and tumor tissues were used for tissue and organ fluorescence imaging.

[0169] 3. In vivo fluorescence imaging of animals

[0170] The imaging conditions of sodium chloroporphyrin in vivo and tissue were explored and optimized using a small animal optical in vivo imaging system. Based on the naked eye observation and judgment of the fluorescence difference in optical in vivo imaging, the optimal imaging conditions for significant fluorescence difference between tumor tissue and non-lesion tissue were found to be an excitation wavelength of 420nm and an imaging collection wavelength of 620nm (filter), with a light intensity output of 10mW / cm 2 (like Figure 1 shown).

[0171] 4. Fluorescence imaging of sodium porphyrin in animal tissues

[0172] The tissues of the experimental group mice at the same time point were arranged as a group for imaging, and the tissues of the control group mice were used as reference (e.g. Figure 2 shown).

[0173] The fluorescence tissue imaging data were analyzed using Living Image software. The software automatically calculated the tissue fluorescence intensity of the target region (ROI) and plotted the relative fluorescence intensity (ratio of tumor to normal tissue) over time after deducting the background of each tissue or organ (e.g. Figure 3 shown).

[0174] in conclusion:

[0175] Based on the clinical therapeutic dose of sodium thiourea photosensitizer, under the irradiation of low-dose laser light with a wavelength of 420nm, the tumor tissue exhibits a strong fluorescence reaction visible to the naked eye, showing a more obvious fluorescence difference compared with the non-lesioned tissue.

[0176] Example 2. Fluorescence imaging response of sodium porphyrin-mediated photodynamic diagnosis in human patients

[0177] Subjects: Patients suspected of bladder cancer

[0178] Protocol: DVDMS-2 was administered intravenously to the patient at a dose of 0.2 mg / kg body weight, and PDT diagnosis was performed 24 hours later, with a diagnostic light wavelength of 405 nm and a diagnostic light dose of 5-10 J / cm 2 . When a tissue with a significant fluorescence difference compared to the surrounding tissue is observed, the tissue showing a strong fluorescence reaction is sampled and biopsied (e.g. Figure 4 shown).

[0179] Expected results: The biopsy results show that the tissues that show strong fluorescence reactions are cancer cell tissues, and the fluorescence reactions are highly matched with the biopsy results.

[0180] Example 3. Clinical case of integrated photodynamic diagnosis and treatment mediated by sodium chlorophyll

[0181] Subjects: Patients suspected of bladder cancer

[0182] Protocol: DVDMS-2 was intravenously administered to the patient at a dose of 0.2 mg / kg body weight, and PDT diagnosis was performed 24 hours later, with a diagnostic light wavelength of 405 nm and a diagnostic light dose of 5-10 J / cm2. A site with a significant fluorescence difference compared to the surrounding tissue was observed, and the site showing a strong fluorescence reaction was judged to be the tumor site, and the tumor site was irradiated with a laser with a wavelength of 630±3 nm for treatment.

[0183] Expected results: Complete diagnosis and treatment simultaneously in one interventional medical operation. After diagnosis, switch the laser wavelength and power to carry out targeted treatment of the diseased tissue indicated by fluorescence. The light dose of the treatment process can be adjusted according to the intensity of fluorescence to achieve the removal of cancerous tissue.

[0184] While the present application has been particularly shown and described with reference to specific embodiments, some of which are preferred embodiments, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the present application disclosed herein.

Claims

1. A method for determining the presence and / or location of a tumor in an individual suspected of having a tumor and treating the determined tumor, comprising: a) administering to the individual a photosensitizer comprising a porphyrin dimer salt so that the photosensitizer is distributed at a body site of the individual suspected of having a tumor; b) irradiating the body part with laser light of a first wavelength to excite the porphyrin dimer salt distributed in the body part to emit fluorescence, wherein the first wavelength is lower than the wavelength of photodynamic therapy using the porphyrin dimer salt; c) detecting whether the fluorescence at the body part shows a difference in fluorescence intensity at a certain position different from that at surrounding positions, the presence of the fluorescence intensity difference reflecting the presence of a tumor, and / or the position showing the fluorescence intensity difference reflecting the position of the tumor; and d) irradiating the location showing the fluorescence intensity difference with laser light of a second wavelength to perform photodynamic therapy.

2. The method according to claim 1, wherein the step d) comprises irradiating the location in the body part showing the fluorescence intensity difference with laser light of the second wavelength while irradiating the body part with laser light of the first wavelength to perform the photodynamic therapy.

3. A method for determining tumor recurrence or metastasis in an individual suspected of having tumor recurrence or metastasis and treating the determined recurrence or metastasis of the tumor, comprising: a) administering a photosensitizer comprising a porphyrin dimer salt to the individual, so that the photosensitizer is distributed at a body site of the individual where tumor recurrence or metastasis is suspected; b) irradiating the body part with laser light of a first wavelength to excite the porphyrin dimer salt distributed in the body part to emit fluorescence, wherein the first wavelength is lower than the wavelength of photodynamic therapy using the porphyrin dimer salt; c) detecting whether the fluorescence at the body part shows a difference in fluorescence intensity at a certain position that is different from that at surrounding positions; wherein the presence of the fluorescence intensity difference reflects the presence of tumor recurrence or metastasis, and / or the position showing the fluorescence intensity difference reflects the position of tumor recurrence or metastasis; d) irradiating the location showing the fluorescence intensity difference with a laser of a second wavelength to perform photodynamic therapy.

4. The method according to claim 3, wherein the step d) comprises irradiating the location in the body part showing the fluorescence intensity difference with laser light of the second wavelength while irradiating the body part with laser light of the first wavelength to perform the photodynamic therapy.

5. The method according to any of the preceding claims, wherein the tumor is a tumor suitable for interventional diagnosis and treatment, and optionally, a tumor suitable for endoscopic examination.

6. The method according to any of the preceding claims, wherein the tumor is a digestive tract tumor (e.g., esophageal cancer, gastric cancer, gastrointestinal stromal tumor, rectal cancer), a urinary tract tumor (e.g., bladder tumor), a reproductive system tumor (e.g., vaginal tumor, cervical cancer, ovarian cancer, endometrial cancer, prostate cancer), or an intra-abdominal tumor (e.g., liver tumor, pancreatic tumor, kidney tumor).

7. The method according to any of the preceding claims, wherein the administration comprises administration by injection, infusion or transdermal administration.

8. The method according to any preceding claim, wherein the porphyrin dimer salt is administered to the subject at a dose of 0.1-1 mg / kg body weight.

9. The method according to any one of the preceding claims, wherein the porphyrin dimer salt is administered by intravenous injection and step b) is performed 24 hours after administration of the porphyrin dimer salt.

10. The method according to any one of the preceding claims, wherein the porphyrin dimer salt is administered by perfusion or transdermally, and step b) is performed half an hour to one hour after the administration of the porphyrin dimer salt.

11. The method according to any of the preceding claims, wherein the first wavelength is in the range of 350nm-450nm, optionally, the first wavelength is 405nm.

12. The method according to any of the preceding claims, wherein the power of the laser light at the first wavelength is 10 mw-30 mw.

13. The method according to any of the preceding claims, wherein the fluorescence can be observed by naked eye, or an image of the fluorescence can be collected at a wavelength of 620 nm.

14. A method according to any preceding claim, wherein the second wavelength is a wavelength of 630±3 nm, such as 630 nm, 631 nm or 632 nm.

15. A method according to any preceding claim, wherein the laser power at the second wavelength is 150-800 mw.

16. A method according to any preceding claim, wherein the porphyrin dimer salt has a structure shown in formula (I), formula (II) or formula (III): in R is independently selected from C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy and C 1-6 acyl, which is optionally substituted by one or more substituents selected from halogen and hydroxy, M is an alkali metal, an alkaline earth metal or NH4 + , p is the reciprocal of the valence number of M.

17. The method according to claim 16, wherein R is independently selected from C optionally substituted by one or more hydroxyl groups. 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl and C 1-6 Acyl, M is an alkali metal or NH4 + , and p is 1.

18. The method according to claim 17, wherein R is independently selected from -CH=CH2, -CH(OH)CH3 and -COCH3, and M is an alkali metal or NH4 + , and p is 1.

19. The method according to claim 18, wherein R is independently selected from -CH=CH2 and -CH(OH)CH3, and M is an alkali metal or NH4 + , and p is 1.

20. The process according to any one of claims 16 to 19, wherein M is an alkali metal.

21. The method of claim 20, wherein M is Na or K.

22. The method according to claim 16, wherein the porphyrin dimer salt bound by an ether bond is selected from Bis[1-[sodium 6,7-dipropionate-1,3,5,8-tetramethyl-4-vinyl-2-porphine]ethyl]ether, Bis[1-[sodium 6,7-dipropionate-1,3,5,8-tetramethyl-2-vinyl-4-porphine]ethyl]ether, 1-[6,7-dipropionic acid sodium-1,3,5,8-tetramethyl-2-vinyl-4-porphine]ethyl 1-[6',7'-dipropionic acid sodium-1',3',5',8'-tetramethyl-4-vinyl-2-porphine]ethyl ether, Bis[1-[6,7-dipropanoic acid potassium-1,3,5,8-tetramethyl-4-vinyl-2-porphine]ethyl]ether, Bis[1-[6,7-dipropanoic acid potassium-1,3,5,8-tetramethyl-2-vinyl-4-porphine]ethyl]ether, 1-[6,7-dipropionate potassium-1,3,5,8-tetramethyl-2-vinyl-4-porphine]ethyl 1-[6',7'-dipropionate potassium-1',3',5',8'-tetramethyl-4-vinyl-2-porphine]ethyl ether, Bis[1-[6,7-dipropionate ammonium-1,3,5,8-tetramethyl-4-vinyl-2-porphine]ethyl]ether, Bis[1-[6,7-dipropionate ammonium-1,3,5,8-tetramethyl-2-vinyl-4-porphine]ethyl]ether, and 1-[6,7-dipropionate ammonium-1,3,5,8-tetramethyl-2-vinyl-4-porphine]ethyl 1-[6',7'-dipropionate ammonium-1',3',5',8'-tetramethyl-4-vinyl-2-porphine]ethyl ether.

23. Use of a porphyrin dimer salt as defined in claims 16 to 22 for the preparation of an integrated diagnostic and therapeutic reagent for determining the presence and / or location of a tumor in an individual suspected of having a tumor and for photodynamic therapy after determining the presence and location of the tumor.

24. Use of a porphyrin dimer salt as defined in claims 16 to 22 for the preparation of an integrated diagnostic and therapeutic reagent for determining the presence and / or location of tumor recurrence or metastasis in an individual suspected of having such recurrence or metastasis, and for performing photodynamic therapy after determining the presence and location of the tumor.

25. A method according to any preceding claim, wherein the porphyrin dimer salt is as defined in claims 16 to 22.

26. A device for integrated diagnosis and treatment suitable for the method as claimed in any one of claims 1, 2, 5 to 25, comprising: a) Interventional devices that can be implanted in the body; b) a first device capable of emitting laser light at said first wavelength; and c) a second device capable of emitting laser light at said second wavelength; The first device and the second device are arranged on the interventional device, and can be controlled to emit the laser after the interventional device is implanted in the body.

27. The apparatus of claim 26, wherein the first device and the second device can be controlled to emit the first wavelength laser and the second wavelength laser, respectively, at the same time.

28. The apparatus of claim 26, wherein the first device and the second device are operable such that the first device emits laser light of the first wavelength at a first time and the second device emits laser light of the second wavelength at a second time.

Images