Near-infrared lymphography composition and its dosage and administration

Abstract

A contrast agent for imaging lymph nodes or lymphatic vessels with near-infrared light that can selectively image primary lymph nodes relative to secondary or higher lymph nodes. A dose of the composition suitable for imaging lymph nodes or lymphatic vessels.

Description

[Technical Field]

[0001]

[0001] The present invention relates to a composition for imaging lymph nodes or lymphatic vessels with near-infrared light, which can selectively image primary lymph nodes over secondary and subsequent lymph nodes, and to the dosage and administration method of the composition suitable for imaging lymph nodes or lymphatic vessels. [Background technology]

[0002]

[0002] The lymphatic system, along with the blood system, is an important part of the body's circulatory system, transporting fluids from peripheral tissues and playing a role in the immune system. In the lymphatic system, lymph is generated when tissue fluid derived from blood plasma flows into lymphatic capillaries. Lymph then flows from the lymphatic vessels through the thoracic duct and lymph nodes to the blood system. Lymph nodes function as a filter in the lymphatic system and also provide immune function through immune cells against non-self antigenic targets such as foreign substances, pathogens, and cancer cells. Lymphatic system disorders can lead to bodily dysfunction, such as idiopathic or surgically-induced secondary lymphedema. When cancer metastasizes to other organs in the body, it travels through the dominant lymphatic system and distant sites. The lymph node that first passes through from the cancer site is known as the sentinel lymph node. Sentinel lymph node biopsy, which detects systemic metastasis via the lymphatic system, is one of the most important diagnostic methods in modern cancer treatment. Because the lymphatic system is difficult to visualize with the naked eye, various techniques for detecting it have been devised and put to practical use. Indigo carmine, a slightly purplish blue pigment long known as Blue No. 2, is widely used as a food additive. Medically, it is administered intravenously for renal function testing and intracellularly as a contrast agent. Indigo carmine is also used in sentinel lymph node biopsy, but identification can be difficult, and improving the identification rate remains a challenge. Techniques using the radioisotope technetium-99m have also become important tools for sentinel lymph node biopsy, but they have drawbacks, including the complexity of administering it to multiple sites on the patient one day before surgery, inaccurate localization due to radioactivity rather than visual detection, the need for a nuclear license, and the preparation, handling, and disposal of nuclear waste. Radiation exposure to patients and medical personnel is also an issue.

[0003]

[0003] In recent years, fluorescence imaging techniques using near-infrared wavelengths, which easily penetrate biological components, have become widespread, and medical applications of fluorescence imaging are increasing. Near-infrared wavelengths are longer than the visible light wavelength range that can be seen by the human eye, and are therefore invisible to the naked eye. However, because they can be detected with high sensitivity using, for example, a CCD camera, they are an effective means of assisting visualization during surgery. Indocyanine green is known as a fluorescent reagent that uses near-infrared wavelengths, and is commercially available in Japan as a drug for liver and circulatory function tests, a fluorescent angiography agent, and a drug for identifying sentinel lymph nodes.

[0004]

[0004] When administered around a tumor, indocyanine green tends to migrate to the sentinel lymph node and then to downstream lymph nodes. This migration can result in bright green lymph nodes that appear to be sentinel lymph nodes but are actually secondary or tertiary lymph nodes. Also, dilated lymphatic vessels can produce extremely bright signals that can be mistaken for lymph nodes. As a result, the use of indocyanine green carries the risk of misidentifying unnecessary lymph nodes for removal or removing lymphatic vessels mistaken for lymph nodes. Currently, there are no fluorescent agents in the near-infrared region that can selectively and sensitively identify sentinel lymph nodes.

[0005]

[0005] Patent Document 1 describes cyclodextrin-bound indocyanine compounds represented by Chemical Formula 15, Chemical Formula 16, Chemical Formula 19, or Chemical Formula 20, including the compound represented by Formula I below (pudexacyaninium), as fluorescent reagents utilizing near-infrared wavelengths. Diagnostic compositions containing these compounds are applicable to sentinel lymph node identification, lymphedema evaluation, intraoperative cholangiography, tumor marking, coronary angiography, abdominal angiography (hepatic artery, abdominal aorta, gastrointestinal blood flow, etc.) for cancer (breast cancer, esophageal cancer, gastric cancer, colon cancer, prostate cancer, skin cancer, etc.), fundus angiography, cerebral circulation evaluation, intraoperative imaging in neurosurgery, etc.

[0006] [ka]

[0007]

[0006] According to Non-Patent Documents 1 and 2, the compound represented by formula I is a compound used as a ureteral imaging agent during surgery. [Prior art documents] [Patent documents]

[0008] [Patent Document 1] U.S. Patent Publication No. 2012 / 0302881 [Non-patent literature]

[0009] [Non-Patent Document 1] Mol Imaging Biol (2021), published online: 11 May 2021, DOI: 10.1007 / s11307-021-01613-0 [Non-patent document 2] Clinical Pharmacology in Drug Development, 2021, 10(12) 1460-1468 Summary of the Invention

[0010]

[0007] A composition for use in imaging lymph nodes or lymphatic vessels with near-infrared light is provided. The composition according to the present specification contains a compound represented by the following formula I or a pharmaceutically acceptable salt thereof, and is characterized by being capable of imaging primary lymph nodes selectively over secondary and subsequent lymph nodes.

[0011] [ka]

[0012]

[0008] Also provided is a composition for imaging lymphatic vessels and lymph nodes with near-infrared light, which contains a compound represented by formula I or a pharmaceutically acceptable salt thereof, and this composition is used so that the compound of formula I or a pharmaceutically acceptable salt thereof is administered locally in an amount of 0.005 to 10 mg per dose, calculated as the weight of the free form of the compound of formula I.

[0013]

[0009] Further provided is a method for imaging a patient's primary lymph nodes, the method comprising administering to the patient a composition containing an effective amount of a compound represented by formula I or a pharmaceutically acceptable salt thereof, and irradiating the patient with near-infrared light to image the patient's primary lymph nodes, wherein the primary lymph nodes are imaged selectively over secondary and subsequent lymph nodes. [Brief explanation of the drawings]

[0014] [Figure 1]

[0010] Near-infrared fluorescence images of the area around the groin 10 minutes (left), 20 minutes (middle), and 30 minutes (right) after subcutaneous administration of ICG (R: right) and compound (I) (pudexacyaninium chloride) (L: left) near the second nipple of a miniature pig. [Figure 2]

[0011] Near-infrared fluorescence images of the area around the administration site within 10 seconds after subcutaneous administration of 0.1 mL of aqueous solutions of compound (I) at (1) 1 mg / mL, (2) 0.1 mg / mL, (3) 0.01 mg / mL, and (4) 0.001 mg / mL to a minipig. [Figure 3]

[0012] Near-infrared fluorescence images (Head: head side, Tail: tail side) of the inguinal region and excised inguinal lymph nodes at the time of maximum tendency after abdominal subcutaneous administration of 0.1 mL of an aqueous solution of Compound (I) and ICG to a miniature pig. (1) 1 mg / mL aqueous solution of Compound (I) (percutaneous observation of inguinal lymph nodes) (2) 1 mg / mL aqueous solution of Compound (I) (observation at the time of excision of inguinal lymph nodes) (3) 0.1 mg / mL aqueous solution of Compound (I) (percutaneous observation of inguinal lymph nodes) (4) 0.1 mg / mL aqueous solution of Compound (I) (observation at the time of excision of inguinal lymph nodes) (5) 0.01 mg / mL aqueous solution of Compound (I) (inguinal lymph nodes) (6) 0.01 mg / mL aqueous solution of Compound (I) (observation of inguinal lymph nodes during excision) (7) 5 mg / mL aqueous solution of ICG (observation of inguinal lymph nodes during excision) (8) 5 mg / mL aqueous solution of ICG (observation of inguinal lymph nodes during excision) (9) 2.5 mg / mL aqueous solution of ICG (observation of inguinal lymph nodes during excision) (10) 2.5 mg / mL aqueous solution of ICG (observation of inguinal lymph nodes during excision) [Figure 4]

[0013] Near-infrared fluorescence and bright-field images were taken 10 minutes after subcutaneous administration of compound (I) (ASP5354, 1 mg / mL) and ICG (2.5 mg / mL) to a beagle dog, and near-infrared fluorescence and bright-field images were taken before and after exposure of the axillary lymph nodes. (1) is a percutaneous observation of the thoracic lymphatic vessels. (2) Images of the axillary lymph nodes were taken at the time of exposure and after excision. [Figure 5]

[0014] Near-infrared fluorescence images (1, 2) of the area around the popliteal lymph node 20 minutes after subcutaneous administration of Compound (I) and ICG into the interdigital space of the hind limb of a beagle dog, near-infrared fluorescence images (3, 4) of the area around the internal iliac lymph node 90 to 100 minutes after subcutaneous administration, and photographs and near-infrared fluorescence images (5 to 12) of the lymph node after excision. More specifically, the images in Figure 5 show the following: (1) In vivo near-infrared fluorescence image of the area around the right below-the-knee lymph node 20 minutes after administration of compound (I). (2) In vivo near-infrared fluorescence image of the area around the left below-the-knee lymph node 20 minutes after administration of ICG. (3) In vivo near-infrared fluorescence image of the area around the right internal iliac lymph node 100 minutes after administration of compound (I). (The arrow indicates the ureter, and the dotted line indicates the internal iliac lymph node.) (4) In vivo near-infrared fluorescence image of the area around the left internal iliac lymph node 93 minutes after administration of ICG. (The arrow indicates the ureter, and the dotted line indicates the internal iliac lymph node.) (5) Right below-the-knee lymph node excised after administration of compound (I). (6) Ex vivo near-infrared fluorescence image of right subpopliteal lymph node excised after administration of compound (I). (7) Photograph of left subpopliteal lymph node excised after administration of ICG. (8) Ex vivo near-infrared fluorescence image of left subpopliteal lymph node excised after administration of ICG. (9) Photograph of right internal iliac lymph node excised after administration of compound (I). (10) Ex vivo near-infrared fluorescence image of right internal iliac lymph node excised after administration of compound (I). (11) Photograph of left internal iliac lymph node excised after administration of ICG. (12) Ex vivo near-infrared fluorescence image of left internal iliac lymph node excised after administration of ICG. DETAILED DESCRIPTION OF THE INVENTION

[0015]

[0015] The objective of the present invention is to provide a composition for imaging lymph nodes or lymphatic vessels with near-infrared light, which can selectively image primary lymph nodes over secondary or higher lymph nodes, and to establish a dosage regimen suitable for imaging lymph nodes or lymphatic vessels using the composition. Detection of sentinel lymph nodes (SLNs) in patients with cancer (e.g., localized breast cancer, melanoma, cervical cancer, head and neck malignancies, or gastrointestinal malignancies) can diagnose the presence of lymphatic spread and avoid complications associated with extensive lymph node removal. This is important because the degree of lymphatic complications has been shown to be related to the number of lymph nodes removed.

[0016]

[0016] Indocyanine green, which has been used as a drug for identifying sentinel lymph nodes, has a tendency to leak beyond the primary lymph nodes into secondary and subsequent lymph nodes. This characteristic is probably due to the highly hydrophobic nature of indocyanine green, which has the property of being adsorbed to lipids. Therefore, the present inventors investigated a compound with higher water solubility represented by the following formula I (pudexacyaninium, also referred to as "ASP5354" in this specification).

[0017] [ka]

[0018]

[0017] The compound represented by formula I is an iodine-free indocyanine compound that exhibits hydrophilicity through binding with cyclodextrin while maintaining near-infrared fluorescence (NIR-F) properties. Due to its hydrophilicity, the compound represented by formula (I) is excreted almost completely in the urine via the kidney after intravenous administration.

[0019] It has been found that a composition containing a compound represented by formula (I) or a pharmaceutically acceptable salt thereof can selectively image primary lymph nodes over secondary lymph nodes because it does not leak into secondary or higher lymph nodes as does ICG. Furthermore, non-clinical data on this compound has shown that due to its physical and chemical properties, it travels through lymphatic vessels to reach lymph nodes at a faster rate than ICG. It has also been found that topical administration of a predetermined dose of a composition containing a compound represented by formula (I) or a pharmaceutically acceptable salt thereof is suitable for near-infrared imaging of lymphatic vessels and lymph nodes.

[0020]

[0019] "Primary lymph node" refers to the lymph node to which the contrast agent of the present invention (e.g., a compound of Formula I) first reaches via lymphatic vessels after local administration of the contrast agent near a lesion such as a tumor. A sentinel lymph node is the lymph node to which cancer cells first reach via lymphatic vessels from the primary cancer lesion. When a contrast agent is locally administered near the primary cancer lesion, the sentinel lymph node is the primary lymph node. "Secondary or later lymph node" refers to a lymph node located farther from the administration site of the contrast agent than the primary lymph node, via lymphatic vessels from the primary lymph node. "Selective imaging of primary lymph nodes over secondary or later lymph nodes" refers to the fact that, when irradiated with near-infrared light (around 800 to 830 nm), the fluorescence from the primary lymph node is distinguishably stronger than the fluorescence emitted from secondary or later lymph nodes.

[0021]

[0020] A composition containing a compound of Formula I or a pharmaceutically acceptable salt thereof as an active ingredient is useful for imaging lymph nodes or lymphatic vessels with near-infrared light, and is capable of imaging primary lymph nodes selectively over secondary or higher lymph nodes. When administered locally to a patient in an amount of 0.005 to 10 mg, either once or multiple times (if necessary), the composition is useful for imaging lymph nodes or lymphatic vessels with near-infrared light.

[0022] The compound represented by formula I may be a salt formed with an anion generated by releasing one or more protons from an acid. Specific examples include salts with an anion generated by releasing a proton from an inorganic acid (hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid, etc.) or an organic acid (formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, mandelic acid, tartaric acid, dibenzoyltartaric acid, ditoluoyltartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, aspartic acid, glutamic acid, etc.). One embodiment of the pharmaceutically acceptable salt of the compound represented by formula I is a salt formed with a chloride anion (Cl -) is a salt of the compound of formula I, such as the chloride salt (pudexacyaninium chloride).

[0023]

[0022] As long as no contradiction occurs, a combination of these aspects may also be used.

[0023] The compound represented by formula I and its pharmaceutically acceptable salts can be prepared, for example, by the method described in Non-Patent Document 1, WO2011 / 093098A1 or WO2021 / 105888A1, or a method similar thereto.

[0024]

[0024] The compound represented by formula I or a pharmaceutically acceptable salt thereof also includes a hydrate or solvate. The compound represented by formula I or a pharmaceutically acceptable salt thereof may be amorphous or crystalline (including crystalline polymorphism). The amorphous form of the compound represented by formula I or a pharmaceutically acceptable salt thereof can be prepared by the method described in Journal of Biomedical Optics 21(8), 086009 (August 2016) or a method similar thereto. Crystals of the compound represented by formula I or a pharmaceutically acceptable salt thereof can be prepared by the method described in WO2021 / 105888A1.

[0025] The compounds of formula I or pharmaceutically acceptable salts thereof include compounds labeled with various radioactive or non-radioactive isotopes.

[0026] The compound represented by formula I or a pharmaceutically acceptable salt thereof may exist in the form of a tautomer. In this specification, the compound represented by formula I or a pharmaceutically acceptable salt thereof may be described in only one form of a tautomer, but other isomers are also included, as well as isolated isomers or mixtures thereof.

[0026]

[0027] The imaging compositions of the present invention can be prepared by methods commonly known in the art using excipients, carriers, and other additives commonly known in the art.

[0028] There are no particular limitations on how the imaging composition of the present invention is administered, and examples include parenteral administration such as local administration of an injection.

[0027]

[0029] "Local administration" refers to administering a composition locally to the site where a biological effect is desired. "Local administration" and "administered locally" can also be written as "topical administration" and "administered topically," respectively. Some embodiments include intradermal, subcutaneous, intramuscular, interstitial, subdermal, and subareolar administration. Other embodiments include intradermal and subcutaneous administration. Another embodiment is intramuscular administration.

[0028]

[0030] Injectables contain sterile aqueous or non-aqueous solutions, suspensions, or emulsions. Aqueous solvents include, for example, distilled water for injection or physiological saline. Non-aqueous solvents include alcohols such as ethanol. Such compositions may further contain excipients, buffers, isotonicity agents, antioxidants, surfactants, preservatives, wetting agents, emulsifiers, dispersants, stabilizers, or solubilizers. These compositions can be sterilized, for example, by filtration through a bacteria-retaining filter, sterilizing with a sterilizer, or by irradiation. Alternatively, these compositions can be prepared as sterile solid compositions, which can be dissolved or suspended in sterile water or a sterile injectable solvent before use. In some embodiments, the imaging compositions described herein are in the form of aqueous solutions. Examples of solid compositions that can be dissolved before use include freeze-dried and spray-dried formulations, and in some embodiments, they are freeze-dried formulations.

[0029]

[0031] The compound of Formula I or a pharmaceutically acceptable salt thereof may be administered once, or multiple times as desired. In some embodiments, the administration is once, and in other embodiments, twice. The compound of Formula I or a pharmaceutically acceptable salt thereof is administered before surgery or intraoperatively. In some embodiments, it is administered before surgery, and in other embodiments, it is administered intraoperatively. In some embodiments, it is administered first before surgery, and then again once intraoperatively. Preoperative administration may be within 24 hours, 18 hours, 12 hours, 10 hours, 8 hours, 6 hours, 4 hours, 3 hours, 2 hours, 1.5 hours, 1 hour, 45 minutes, 30 minutes, 15 minutes, or immediately before surgery (e.g., within 5 minutes).

[0030]

[0032] The dose per administration of the compound of formula I or a pharmaceutically acceptable salt thereof may be 0.005 to 10 mg, 0.01 to 10 mg, 0.03 to 10 mg, 0.05 to 10 mg, 0.05 to 5 mg, 0.05 to 4 mg, 0.05 to 2 mg, 0.05 to 1 mg, or 0.1 to 1 mg, calculated as the weight of the free form of the compound of formula I. In some embodiments, it is 0.05 mg, 0.2 mg, 0.6 mg, 1 mg, 2 mg, or 4 mg. In other embodiments, the amounts administered in multiple doses may be the same or different.

[0031]

[0033] The site of administration of the compound of Formula I or a pharmaceutically acceptable salt thereof may be any suitable site in a patient in need thereof. In some cases, the site of administration is the periareolar region. In some cases, the site of administration is near the left chest (left breast). In some cases, the site of administration is near the right chest (left breast). In various cases, after administration to the patient's chest, the patient's chest is massaged for about 5 minutes (which may promote transfer of the imaging compound). In other cases, no massage is performed.

[0032]

[0034] In the case of an aqueous solution (aqueous solution), the concentration of the compound represented by Formula I or a pharmaceutically acceptable salt thereof may be 0.1 to 5 mg / mL, 0.1 to 3 mg / mL, 0.3 to 3 mg / mL, 0.5 to 3 mg / mL, or 1 to 3 mg / mL, calculated as the free form. In one embodiment, it is 0.1 mg / mL, 0.3 mg / mL, 0.5 mg / mL, 1 mg / mL, 2 mg / mL, 3 mg / mL, or 5 mg / mL. In another embodiment, it is 1 mg / mL, 2 mg / mL, or 3 mg / mL. In another embodiment, it is 1 mg / mL. In another embodiment, it is 3 mg / mL.

[0033]

[0035] The total amount of aqueous solution of the compound of Formula I or a pharmaceutically acceptable salt thereof administered topically in a single or multiple doses may be 0.05 mL to 5 mL in terms of the free form. In some embodiments, the amount is 0.05 mL, 0.1 mL, 0.2 mL, 0.3 mL, 0.4 mL, 0.5 mL, 0.6 mL, 0.8 mL, 1.0 mL, 2.0 mL, 3.0 mL, 4.0 mL, or 5.0 mL. In various embodiments, the amount is 0.05 mL, 0.2 mL, 0.6 mL, 1.0 mL, 2.0 mL, or 4.0 mL. In some embodiments, the amount is 1.0 mL. In various embodiments, the amount is 2.0 mL.

[0034] Aspects [1] Formula I below:

[0035] [ka]

[0036] or a pharmaceutically acceptable salt thereof, wherein the composition is for imaging lymph nodes or lymphatic vessels with near-infrared light, and characterized in that the composition selectively images primary lymph nodes over secondary and subsequent lymph nodes. [2] The composition according to [1], wherein the composition is an aqueous solution having a concentration of the compound represented by formula I or a pharmaceutically acceptable salt thereof of 0.1 to 5 mg / mL, calculated as the weight of the free form of the compound of formula I. [3] The composition according to [2], wherein the concentration of the compound represented by formula I or a pharmaceutically acceptable salt thereof is 0.5 to 3 mg / mL in terms of the weight of the free form of the compound of formula I. [4] The composition according to [3], wherein the concentration of the compound represented by formula I or a pharmaceutically acceptable salt thereof is 1 mg / mL in terms of the weight of the free form of the compound of formula I. [5] The composition according to any one of [1] to [3], wherein the compound represented by formula I or a pharmaceutically acceptable salt thereof is a chloride salt. [6] Formula I below:

[0037] [ka]

[0038] or a pharmaceutically acceptable salt thereof, wherein the compound is The composition described above, wherein the compound of formula I or a pharmaceutically acceptable salt thereof is used so as to be topically administered in an amount of 0.005 to 10 mg per administration, calculated as the weight of the free form of the compound of formula I. [7] The composition described in [6], which is administered to a patient in a single dose. [8] The composition described in [6], which is administered multiple times to a patient. [9] The composition according to any one of [6] to [8], wherein the amount of the compound represented by formula I or a pharmaceutically acceptable salt thereof per administration is 0.05 to 5 mg, calculated as the weight of the free form of the compound of formula I.

[10] The composition according to [9], wherein the amount of the compound represented by formula I or a pharmaceutically acceptable salt thereof per administration is 0.05 to 4 mg, calculated as the weight of the free form of the compound of formula I.

[11] The composition according to any one of [6] to [8], wherein the composition is an aqueous solution having a concentration of the compound represented by formula I or a pharmaceutically acceptable salt thereof of 0.1 to 5 mg / mL, calculated as the weight of the free form of the compound of formula I.

[12] The composition according to

[11] , wherein the concentration of the compound represented by formula I or a pharmaceutically acceptable salt thereof is 0.5 to 3 mg / mL in terms of the weight of the free form of the compound of formula I.

[13] The composition according to

[12] , wherein the concentration of the compound represented by formula I or a pharmaceutically acceptable salt thereof is 1 mg / mL in terms of the weight of the free form of the compound of formula I.

[14] The composition according to

[10] , wherein the concentration of the compound represented by formula I or a pharmaceutically acceptable salt thereof is 1 mg / mL in terms of the weight of the free form of the compound of formula I.

[15] The composition according to any one of [6] to

[14] , wherein the compound represented by formula I or a pharmaceutically acceptable salt thereof is a chloride salt.

[16] The composition according to any one of [6] to

[15] , which selectively images primary lymph nodes over secondary and subsequent lymph nodes.

[17] The composition according to any one of [6] to

[16] , wherein the local administration is intradermal, subcutaneous, intramuscular, interstitial, subdermal, or subareolar administration.

[18] Formula I below:

[0039] [ka]

[0040] or a pharmaceutically acceptable salt thereof; and irradiating the patient with near-infrared light to image the patient's primary lymph nodes; 1. A method for imaging a patient's primary lymph nodes, comprising: imaging the primary lymph nodes selectively relative to secondary or subsequent lymph nodes.

[19] The method according to

[18] , wherein the composition is an aqueous solution having a concentration of the compound represented by formula I or a pharmaceutically acceptable salt thereof of 0.1 to 5 mg / mL, calculated as the weight of the free form of the compound of formula I.

[20] The method according to

[19] , wherein the concentration of the compound represented by formula I or a pharmaceutically acceptable salt thereof is 0.5 to 3 mg / mL in terms of the weight of the free form of the compound of formula I. [twenty one] The method according to

[20] , wherein the concentration of the compound represented by formula I or a pharmaceutically acceptable salt thereof is 1 mg / mL in terms of the weight of the free form of the compound of formula I. [twenty two] The method according to

[18] , wherein the composition is topically administered in an amount of 0.005 to 10 mg per administration, calculated as the weight of the free form of the compound of formula I. [twenty three] The method according to any one of

[18] to

[22] , wherein the administration is local administration. [twenty four] The method according to

[23] , wherein the local administration is intradermal, subcutaneous, intramuscular, interstitial, subdermal or subareolar administration. [twenty five] The method according to any one of

[18] to

[24] , wherein the composition is administered to a patient in a single dose.

[26] The method according to any one of

[18] to

[24] , wherein the composition is administered to a patient multiple times.

[27] The method according to any one of

[18] to

[26] , wherein the composition is administered to a patient before surgery.

[28] The method according to any one of

[18] to

[27] , wherein the composition is administered to a patient during surgery.

[29] The method according to any one of

[18] to

[28] , wherein the compound represented by formula I or a pharmaceutically acceptable salt thereof is a chloride salt.

[30] The method according to any one of

[18] to

[29] , wherein the patient has localized breast cancer, melanoma, cervical cancer, head and neck malignancy, or gastrointestinal malignancy.

[31] 1. Use of a composition in the manufacture of an imaging agent for near-infrared imaging of a primary lymph node in a patient, said composition comprising a compound of Formula I:

[0041] [ka]

[0042] or a pharmaceutically acceptable salt thereof, wherein primary lymph nodes are selectively imaged relative to secondary and subsequent lymph nodes in a patient by imaging with near-infrared light.

[32] The use according to

[31] , wherein the composition is an aqueous solution having a concentration of the compound represented by formula I or a pharmaceutically acceptable salt thereof of 0.1 to 5 mg / mL, calculated as the weight of the free form of the compound of formula I.

[33] The use according to

[32] , wherein the concentration of the compound represented by formula I or a pharmaceutically acceptable salt thereof is 0.5 to 3 mg / mL in terms of the weight of the free form of the compound of formula I.

[34] The use according to

[33] , wherein the concentration of the compound represented by formula I or a pharmaceutically acceptable salt thereof is 1 mg / mL in terms of the weight of the free form of the compound of formula I.

[35] The use according to

[31] , wherein the composition is administered in an amount of 0.005 to 10 mg per administration, calculated as the weight of the free form of the compound of formula I.

[36] The use according to any one of

[31] to

[34] , wherein the composition is administered topically to a patient.

[37] The use according to

[36] , wherein the local administration is intradermal, subcutaneous, intramuscular, interstitial, subdermal or subareolar administration.

[38] The use according to any one of

[31] to

[37] , wherein the composition is administered to a patient in a single dose.

[39] The use according to any one of

[31] to

[37] , wherein the composition is administered to a patient multiple times.

[40] The use according to any one of

[31] to

[39] , wherein the composition is administered to a patient before surgery.

[41] The use according to any one of

[31] to

[40] , wherein the composition is administered to a patient during surgery.

[42] The use according to any one of

[31] to

[41] , wherein the compound represented by formula I or a pharmaceutically acceptable salt thereof is a chloride salt.

[43] The use according to any one of

[31] to

[42] , wherein the patient has localized breast cancer, melanoma, cervical cancer, head and neck malignancy, or gastrointestinal malignancy.

[44] 1. A method of imaging lymph nodes or lymphatic vessels in a patient, comprising: (a) Formula I:

[0043] [ka]

[0044] or a pharmaceutically acceptable salt thereof to a patient in an amount of 0.005 to 10 mg per administration, calculated as the weight of the free form of the compound of formula I; (b) irradiating the patient with near-infrared light to image the patient's lymph nodes or lymphatic vessels; The above method, comprising:

[45] 1. Use of a composition in the manufacture of an imaging agent for imaging lymph nodes or lymphatic vessels in a patient with near-infrared light, said composition comprising a compound of formula I:

[0045] [ka]

[0046] or a pharmaceutically acceptable salt thereof, wherein the composition is topically administered in an amount of 0.005 to 10 mg per administration, calculated as the weight of the free form of the compound of formula I. [Example]

[0047]

[0036] The present invention will be explained in more detail below based on examples, but these examples do not limit the scope of the present invention. Example 1: Lymphatic imaging study (minipigs) Compound (I) and ICG were administered subcutaneously to the thoracic and abdominal tissues of minipigs (Gottingen or NIBS). Near-infrared fluorescence images were captured using a near-infrared fluorescence imaging device (Hamamatsu Photonics PDE-Neo or Fluoptics Fluobeam). 1 mg / mL ICG was administered subcutaneously near the right second nipple of NIBS minipigs (weight 19.74-23.13 kg) in a volume of 0.05 mL, and 1 mg / mL Compound (I) was administered subcutaneously near the left second nipple. Near-infrared fluorescence was observed 10, 20, and 30 minutes later using the near-infrared fluorescence imaging device (Figure 1). To confirm the concentration dependence of Compound (I), 0.001 mg / mL, 0.01 mg / mL, 0.1 mg / mL, and 1 mg / mL of Compound (I) were subcutaneously administered to the abdomen of Göttingen minipigs (6.12 kg) in a 0.1 mL volume. Immediately after administration, near-infrared fluorescence imaging was used to observe the movement of Compound (I) through the lymphatic vessels from the administration site (Figure 2). In addition, 0.01 mg / mL, 0.1 mg / mL, and 1 mg / mL of Compound (I) and 2.5 mg / mL and 5 mg / mL of ICG, which had been confirmed to move through the lymphatic vessels, were subcutaneously administered near the second nipple of Göttingen minipigs (13.7-16.4 kg). Near-infrared fluorescence imaging was used to observe the movement of Compound (I) through the lymphatic vessels for 2 hours (Figure 3). After administration of compound (I), the fluorescent signal rapidly migrated from the administration site to the inguinal lymph nodes (concentrations: 0.1 mg / mL and 1 mg / mL), allowing the lymph nodes to be identified transcutaneously (Figures 3-1 and 3-3). With 0.01 mg / mL of compound (I), a weak fluorescent signal was detected transcutaneously in the inguinal lymph nodes (Figure 3-5). Similarly, ICG also migrated from the administration site to the inguinal lymph nodes, allowing the lymph nodes to be identified transcutaneously (Figures 3-7 and 3-9). At the end of the experiment, the inguinal lymph nodes were excised, and near-infrared fluorescence was observed with a near-infrared fluorescence imaging device. When compound (I) was administered at concentrations of 0.1 mg / mL and 1 mg / mL, a strong fluorescent signal was observed in the cephalad side (afferent lymphatic vessel side) of the excised lymphatic tissue, while a weak fluorescent signal tended to be observed in the caudal side (efferent lymphatic vessel side) (Figures 3-2 and 3-4). With 0.01 mg / mL of Compound (I), the fluorescent signal in lymphatic tissue disappeared at the time of excision (Figures 3-6).With ICG, a strong fluorescent signal was observed throughout the excised lymphoid tissue (Figure 3-8, Figure 3-10).

[0048] Example 2: Lymphatic imaging study (dog)

[0040] 2.5 mg / mL ICG was subcutaneously administered to a beagle dog (body weight 7.90 kg) near the left second nipple, and 1 mg / mL Compound (I) was subcutaneously administered to the right second nipple in a volume of 0.1 mL. Ten minutes later, near-infrared fluorescence was observed using a near-infrared fluorescence imaging device (Fluobeam, Fluoptics) (Figure 4-1). Furthermore, the near-infrared fluorescence of the axillary lymph nodes was also observed using the near-infrared fluorescence imaging device after lymph node exposure and subsequent lymph node removal (Figure 4-2).

[0049] Example 3: Canine Lymphatic Imaging Study (Dog, Hind Leg)

[0042] 1 mg / mL of Compound (I) (administered to the right hind paw) and 2.5 mg / mL of ICG (administered to the left hind paw) were administered in 0.1 mL volumes into the subcutaneous tissue between the hind paws of male beagles (7 months old, weighing 12.6 kg). 20 minutes later, images of the area around the gastrocnemius muscle and the area near the popliteal lymph nodes were taken with a near-infrared fluorescence imaging device (Fluoptics Fluobeam) (Figures 5-1 and 5-2). The abdomen was then opened, the internal iliac lymph nodes were exposed, and the area near the internal iliac lymph nodes was photographed with the same device (Figures 5-3 and 5-4). Approximately 2.5 hours later, the animals were euthanized, and the popliteal and internal iliac lymph nodes were removed. Near-infrared fluorescence images were taken with the same device (Figures 5-5 to 5-12).

Claims

1. Formula I below: 【Chemical 1】 or a pharmaceutically acceptable salt thereof, wherein the composition is for imaging lymph nodes or lymphatic vessels with near-infrared light, and characterized in that the composition selectively images primary lymph nodes over secondary and subsequent lymph nodes.

2. The composition according to claim 1, wherein the composition is an aqueous solution having a concentration of the compound represented by formula I or a pharmaceutically acceptable salt thereof of 0.1 to 5 mg / mL in terms of the weight of the free form of the compound of formula I.

3. The composition according to claim 2, wherein the concentration of the compound of formula I or a pharmaceutically acceptable salt thereof is 0.5 to 3 mg / mL in terms of the weight of the free form of the compound of formula I.

4. 4. The composition according to claim 3, wherein the concentration of the compound of formula I or a pharmaceutically acceptable salt thereof is 1 mg / mL in terms of the weight of the free form of the compound of formula I.

5. The composition of any one of claims 1 to 3, wherein the compound of formula I or a pharmaceutically acceptable salt thereof is a chloride salt.

6. Formula I below: 【Chemistry 2】 or a pharmaceutically acceptable salt thereof, wherein the compound is The composition described above, wherein the compound of formula I or a pharmaceutically acceptable salt thereof is used so as to be topically administered in an amount of 0.005 to 10 mg per administration, calculated as the weight of the free form of the compound of formula I.

7. The composition of claim 6 , which is administered to a patient in a single dose.

8. The composition of claim 6 , which is administered multiple times to a patient.

9. The composition according to any one of claims 6 to 8, wherein the dose of the compound represented by formula I or a pharmaceutically acceptable salt thereof per administration is 0.05 to 5 mg, calculated as the weight of the free form of the compound of formula I.

10. The composition according to claim 9, wherein the dose of the compound of formula I or a pharmaceutically acceptable salt thereof per administration is 0.05 to 4 mg, calculated as the weight of the free form of the compound of formula I.

11. The composition according to any one of claims 6 to 8, wherein the composition is an aqueous solution having a concentration of the compound represented by formula I or a pharmaceutically acceptable salt thereof of 0.1 to 5 mg / mL in terms of the weight of the free form of the compound of formula I.

12. The composition according to claim 11, wherein the concentration of the compound of formula I or a pharmaceutically acceptable salt thereof is 0.5 to 3 mg / mL in terms of the weight of the free form of the compound of formula I.

13. The composition according to claim 12, wherein the concentration of the compound of formula I or a pharmaceutically acceptable salt thereof is 1 mg / mL in terms of the weight of the free form of the compound of formula I.

14. The composition according to claim 10, wherein the concentration of the compound represented by formula I or a pharmaceutically acceptable salt thereof is 1 mg / mL in terms of the weight of the free form of the compound of formula I.

15. The composition of any one of claims 6 to 14, wherein the compound of formula I or a pharmaceutically acceptable salt thereof is a chloride salt.

16. The composition according to any one of claims 6 to 15, which selectively images primary lymph nodes over secondary and subsequent lymph nodes.

17. The composition according to any one of claims 6 to 16, wherein the topical administration is intradermal, subcutaneous, intramuscular, interstitial, subcutaneous or subareolar administration.

18. 1. A method for imaging primary lymph nodes in a patient, comprising administering to said patient a compound of formula I: 【Chemistry 3】 or a pharmaceutically acceptable salt thereof; and irradiating the patient with near-infrared light to image the patient's primary lymph nodes; The above method, wherein primary lymph nodes are selectively imaged relative to secondary and subsequent lymph nodes.

19. The method according to claim 18, wherein the composition is an aqueous solution having a concentration of the compound represented by formula I or a pharmaceutically acceptable salt thereof of 0.1 to 5 mg / mL in terms of the weight of the free form of the compound of formula I.

20. The method according to claim 19, wherein the concentration of the compound of formula I or a pharmaceutically acceptable salt thereof is 0.5 to 3 mg / mL in terms of the weight of the free form of the compound of formula I.

21. The method according to claim 20, wherein the concentration of the compound of formula I or a pharmaceutically acceptable salt thereof is 1 mg / mL in terms of the weight of the free form of the compound of formula I.

22. 19. The method of claim 18, wherein the composition is topically administered in an amount of 0.005 to 10 mg per administration, calculated as the weight of the free form of the compound of formula I.

23. The method of any of claims 18 to 22, wherein the administration is topical.

24. 24. The method of claim 23, wherein the local administration is intradermal, subcutaneous, intramuscular, interstitial, subcutaneous or subareolar administration.

25. The method of any of claims 18 to 24, wherein the composition is administered to the patient in a single dose.

26. The method of any of claims 18 to 24, wherein the composition is administered to the patient multiple times.

27. The method of any of claims 18 to 26, wherein the composition is administered to the patient preoperatively.

28. The method of any of claims 18 to 27, wherein the composition is administered to a patient during surgery.

29. The method of any one of claims 18 to 28, wherein the compound of formula I or a pharmaceutically acceptable salt thereof is a chloride salt.

30. 30. The method of any of claims 18 to 29, wherein the patient has localized breast cancer, melanoma, cervical cancer, head and neck malignancy or gastrointestinal malignancy.

31. 1. Use of a composition in the manufacture of an imaging agent for near-infrared imaging of a primary lymph node in a patient, said composition comprising a compound of formula I: 【Chemistry 4】 or a pharmaceutically acceptable salt thereof, wherein primary lymph nodes are selectively imaged relative to secondary and subsequent lymph nodes in a patient by imaging with near-infrared light.

32. The use according to claim 31, wherein the composition is an aqueous solution having a concentration of the compound represented by formula I or a pharmaceutically acceptable salt thereof of 0.1 to 5 mg / mL, calculated as the weight of the free form of the compound of formula I.

33. The use according to claim 32, wherein the concentration of the compound of formula I or a pharmaceutically acceptable salt thereof is 0.5 to 3 mg / mL in terms of the weight of the free form of the compound of formula I.

34. The use according to claim 33, wherein the concentration of the compound of formula I or a pharmaceutically acceptable salt thereof is 1 mg / mL in terms of the weight of the free form of the compound of formula I.

35. The use according to claim 31, wherein the composition is administered in an amount of 0.005 to 10 mg per administration in terms of the weight of the free form of the compound of formula I.

36. The use according to any one of claims 31 to 34, wherein the composition is administered topically to a patient.

37. 37. The use according to claim 36, wherein the local administration is intradermal, subcutaneous, intramuscular, interstitial, subcutaneous or subareolar administration.

38. The use according to any one of claims 31 to 37, wherein the composition is administered to a patient in a single dose.

39. The use according to any one of claims 31 to 37, wherein the composition is administered to a patient multiple times.

40. The use according to any one of claims 31 to 39, wherein the composition is administered to a patient before surgery.

41. The use according to any one of claims 31 to 40, wherein the composition is administered to a patient during surgery.

42. 42. The use according to any one of claims 31 to 41, wherein the compound of formula I or a pharmaceutically acceptable salt thereof is a chloride salt.

43. 43. The use according to any one of claims 31 to 42, wherein the patient has localized breast cancer, melanoma, cervical cancer, head and neck malignancy or gastrointestinal malignancy.

44. 1. A method of imaging lymph nodes or lymphatic vessels in a patient, comprising: (a) a compound of formula I: 【Chemistry 5】 or a pharmaceutically acceptable salt thereof to a patient in an amount of 0.005 to 10 mg per administration, calculated as the weight of the free form of the compound of formula I; (b) irradiating the patient with near-infrared light to image the patient's lymph nodes or lymphatic vessels; The above method, comprising:

45. 1. Use of a composition in the manufacture of an imaging agent for near-infrared imaging of lymph nodes or lymphatic vessels in a patient, said composition comprising a compound of formula I: 【Chemistry 6】 or a pharmaceutically acceptable salt thereof, wherein the composition is topically administered in an amount of 0.005 to 10 mg per administration, calculated as the weight of the free form of the compound of formula I.

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