WO2024084287A1

WO2024084287A1 - Composition for near infrared lymphatic imaging and uses and dosages therefor

Info

Publication number: WO2024084287A1
Application number: PCT/IB2023/000642
Authority: WO
Inventors: Sadaaki Kimura; Hiroshi FUSHIKI; Leticia DELGODO-HERRERA; Jeffrey RAIZER
Original assignee: Astellas Pharma Inc.
Priority date: 2022-10-21
Filing date: 2023-10-20
Publication date: 2024-04-25

Abstract

Disclosed are imaging agents for near-infrared imaging of lymph nodes or lymphatic vessels that can be used to selectively image the primary lymph node relative to secondary and subsequent lymph nodes and dosages for this composition suitable for imaging lymph nodes or lymphatic vessels.

Description

COMPOSITION FOR NEAR-INFRARED LYMPHATIC IMAGING AND USES AND DOSAGES THEREFOR

FIELD

[0001] The present disclosure relates to a composition for near-infrared imaging of lymph nodes or lymphatic vessels in which the primary lymph node can be imaged selectively relative to secondary and subsequent lymph nodes, and to uses and dosages of this composition suitable for imaging lymph nodes or lymphatic vessels.

BACKGROUND

[0002] The lymphatic system, along with the blood system, is an important circulatory system in the human body that transports fluids from peripheral tissues and forms part of the immune system. In the lymphatic system, lymphatic fluid is produced by interstitial fluid derived from blood plasma components flowing into the capillary lymphatic vessels and joins the blood system from the lymphatic vessels via the thoracic duct or lymph nodes. Lymph nodes perform a filtering role in the lymphatic system and an immune function from immune cells opposing non-autoantigen targets such as foreign substances, pathogens, and cancer cells. Disorders of the lymphatic system induce biodysfunction, such as idiopathic or surgically associated secondary lymphedema. When cancer develops in a body and metastasizes to other organs, it passes through the dominant lymphatic system to metastasize in distant locations. The lymph node that is passed first after the cancer site is known as the sentinel lymph node, and a biopsy of the sentinel lymph node to check for systemic metastasis via the lymphatic system has become one of the most important diagnostic methods in modern cancer treatments. Because it is difficult to check the lymphatic system with the naked eye, various methods of lymphatic system detection have been devised and put into practical use. Indigo carmine is a slightly purplish blue pigment known as Blue No. 2 that has been widely used as a food additive for a long time. In medical applications, it is used in renal function tests by intravenous administration and as an imaging agent by intra-tissue administration. Indigo carmine is also used in sentinel lymph node biopsies, but identification can sometimes be difficult and improving the identification rate is a challenge. A technique using the radioisotope technetium-99m has also become an important tool in sentinel lymph node biopsies, but is hampered by complications in administration to multiple sites on a patient a day prior to surgery, inexact location identification due to radioactivity detection rather than visual detection, as well as the requirements for nuclear licensing as well as preparing, handling and disposing of nuclear waste. Moreover, exposure of patients and medical staff to radiation is a drawback.

[0003] In recent years, fluorescence imaging techniques using near-infrared wavelengths that easily pass through biological components have become popular, and there have been increased medical applications of such imaging. Near-infrared wavelengths are on the longer wavelength side of the visible light wavelength range, and cannot be visually perceived. However, because these wavelengths can be detected with high sensitivity using, for example, a CCD camera, they are an effective means of enhanced visual recognition during surgery. Indocyanine green is a known fluorescent reagent that uses near-infrared wavelengths, and is marketed in Japan as an agent for hepatic/circul atory function testing, a fluorescent angiography agent, and a sentinel lymph node identifying agent. [0004] Indocyanine green tends to migrate when administered around a tumor to the sentinel lymph node and then to lymph nodes further downstream. This migration can led to bright green nodes that appear to be sentinel nodes but are in fact second and third tier nodes, and can lead to very bright signals that get misidentified as nodes when they are, in fact, dilated lymphatic channels. As a result, use of indocyanine green presents a risk of misidentifying and removing lymph nodes that do not require resection, and excision of channels misidentified as nodes. At the present time, there are no fluorescent agents that use the near-infrared region for selective and highly sensitive identification of sentinel lymph nodes.

[0005] US 2012/0302881 A1 mentions that cyclodextrin-bonded indocyanine compounds represented by Chemical Formula 15, Chemical Formula 16, Chemical Formula 19, or Chemical Formula 20, which include a compound represented by the Formula (I) (Pudexacianinium), are fluorescent reagents that utilize near-infrared wavelengths. Diagnostic compositions containing this compound can be applied to sentinel lymph node identification, evaluation of lymphedema, intraoperative bile ductography, 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 evaluations, and intraoperative angiography in brain surgery.

[0006] According to Mol Imaging Biol (2021), published online: May 11, 2021 (Non-Patent Document 1), DOI: 10.1007/s11307-021-01613-0 and Clinical Pharmacology in Drug Development, 2021, 10 (12) 1460-1468 (NonPatent Document 2), the compounds represented by Formula (I) are compounds used as ureteral imaging agents during surgery.

SUMMARY

[0007] Provided herein are compositions for use in near-infrared imaging of lymph nodes or lymphatic vessels. The compositions disclosed herein comprise a compound of Formula (I) or a pharmaceutically acceptable salt thereof,

and the compositions are characterized in that a primary lymph node is selectively imaged relative to secondary and subsequent lymph nodes.

[0008] Also provided is a composition for near-infrared imaging of lymph nodes or lymphatic vessels comprising the compound of Formula (I) or a pharmaceutically acceptable salt thereof, wherein the composition is used such 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, based upon the compound of Formula (I) weight in free form.

[0009] Further provided herein is a method of imaging a primary lymph node in a patient comprising administering to the patient an effective amount of a composition comprising a compound of Formula (I) or a pharmaceutically acceptable salt thereof, and exposing the subject to near infrared light to image the primary lymph node in the subject, wherein the primary lymph node is selectively imaged relative to a secondary lymph node and subsequent lymph nodes.

BRIEF DESCRIPTION OF THE FIGURES

[0010] Fig. 1 shows near-infrared fluorescence images captured near the inguinal region 10 minutes (left), 20 minutes (center), and 30 minutes (right) after subcutaneous administration of ICG (R: right) and Compound (I) (Pudexacianinium Chloride) (L: left) near the second nipple of a mini pig.

[0011] Fig. 2 shows near-infrared fluorescence images captured near the administration site within 10 seconds of subcutaneous abdominal administration of 0.1 mL aqueous solutions containing (1) 1 mg/mL, (2) 0.1 mg/mL, (3) 0.01 mg/mL, and (4) 0.001 mg/mL of Compound (I) to a mini pig.

[0012] Fig. 3 shows near-infrared fluorescence images taken near the inguinal region and after removal of inguinal lymph nodes at the time the maximum tendency was shown after subcutaneous abdominal administration of 0.1 mL aqueous solutions containing 1 mg/mL ((1) - transdermal observation of inguinal lymph node; (2)- observation of inguinal lymph node at time of extraction), 0.1 mg/mL ((3) - transdermal observation of inguinal lymph node; (4)- observation of inguinal lymph node at time of extraction), and 0.01 mg/mL ((5) - transdermal observation of inguinal lymph node; (6)- observation of inguinal lymph node at time of extraction), of Compound (I) and 5 mg/ mL ((7) - transdermal observation of inguinal lymph node; (8)- observation of inguinal lymph node at time of extraction), and 2.5 mg/mL ((9) - transdermal observation of inguinal lymph nodes; (10)- observation of inguinal lymph node at time of extraction) of ICG to a mini pig. [0013] Fig. 4 shows near-infrared fluorescence images and bright-field images 10 minutes after subcutaneous chest administration of Compound (I) (ASP5354 at 1 mg/mL) and ICG (at 2.5 mg/mL) to beagles and nearinfrared fluorescence images and bright-field images of the exposed and removed axillary lymph nodes. The observation of (1) was a transdermal observation of thoracic lymphatic vessels. The observation of the axillary lymph nodes in (2) was obtained at time of exposure and at time of extraction.

[0014] Fig. 5 shows near-infrared fluorescence images near the lymph nodes below the kneecaps 20 minutes after subcutaneous administration of Compound (1) and ICG to the interdigital portion of the hind limb of a beagle (1, 2), near-infrared fluorescence images near the internal iliac lymph nodes 90 to 100 minutes after subcutaneous administration (3, 4), and photographs and near-infrared fluorescence images after removal (5- 12). More specifically, the pictures of Fig. 5 show: (1) An in vivo near-infrared fluorescence image near a lymph node below the right kneecap 20 minutes after administration of Compound (I); (2) An in vivo near-infrared fluorescence image near a lymph node below the left kneecap 20 minutes after administration of ICG; (3) An in vivo near-infrared fluorescence image near a right internal iliac lymph node 100 minutes after administration of Compound (I) where the arrow indicates the urinary tract and the dotted-line circle indicates the internal iliac lymph node; (4) An in vivo near-infrared fluorescence image near a left internal iliac lymph node 93 minutes after administration of ICG where the arrow indicates the urinary tract and the dotted-line circle indicates the internal iliac lymph node; (5) A photograph of a lymph node below the right kneecap extracted after administration of Compound (I); (6) An ex vivo near-infrared fluorescence image of a lymph node below the right kneecap extracted after administration of Compound (I); (7) A photograph of a lymph node below the left kneecap extracted after administration of ICG; (8) An ex vivo near-infrared fluorescence image of a lymph node below the left kneecap extracted after administration of ICG (9) A photograph of a right internal iliac lymph node after administration of Compound (I) (10) An ex vivo near-infrared fluorescence image of a right internal iliac lymph node after administration of Compound (I); (11) A photograph of a left internal iliac lymph node after administration of ICG; and (12) An ex vivo near-infrared fluorescence image of a left internal iliac lymph node after administration of ICG.

DETAILED DESCRIPTION

[0015] The object of the present disclosure is to provide a composition for near-infrared imaging of lymph nodes or lymphatic vessels in which the primary lymph node can be imaged selectively relative to secondary and subsequent lymph nodes, and to establish uses and dosages for this composition suitable for imaging lymph nodes or lymphatic vessels. Sentinel lymph node (SLN) detection in patients with cancer (e.g., localized breast cancer, melanoma, cervical, head and neck malignancy, or gastrointestinal malignancy) allows for diagnosing the presence of lymphatic spread and avoids the complications related to extended removal of the lymph nodes. This is important as it has been shown that the degree of lymphatic complications is related to the number of lymph nodes removed.

[0016] Indocyanine green, which has been used as a sentinel lymph node identification agent, tends to spread beyond the primary lymph node to secondary and subsequent lymph nodes. This trait is probably due to indocyanine green having high hydrophobicity and a tendency to become adsorbed to lipids. Therefore, a study of compounds represented by Formula (I) (Pudexacianinium, sometimes referred to as "ASP5354” in the present specification) was conducted, which have higher water solubility.

[0017] The compound represented by Formula (I) is an iodine-free inodcyanine compound which shows hydrophilic properties by conjugation with cyclodextrin while maintaining the near infrared fluorescence (NIR-F) property. The compound represented by Formula (I) is almost entirely excreted into the urine through the kidneys after intravenous administration due to its hydrophilic nature.

[0018] It was discovered that compositions containing a compound represented by Formula (I) or a pharmaceutically acceptable salt thereof do not spread to secondary and subsequent lymph nodes as much as ICG, and, therefore, can be used to selectively image primary lymph nodes relative to secondary and subsequent lymph nodes. The compound's nonclinical data also indicates that it traverses the lymphatic vessels into the lymph nodes at an increased rate over ICG, due to its physical and chemical properties. It was also discovered that the local 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.

[0019] The "primary lymph node" refers to the lymph node that the imaging agent (e.g., a compound of Formula (I)) first reaches via lymphatic vessels after the imaging agent has been administered locally near a lesion, such as a tumor. The sentinel lymph node is the lymph node that cancer cells first reach from the primary tumor of the cancer via lymph vessels. "Secondary and subsequent lymph nodes" refers to lymph nodes located farther than the primary lymph node from the administration site of the imaging agent via lymph vessels. The ability to "selectively" image a primary lymph node relative to the secondary and subsequent lymph nodes means that when exposed to near-infrared light (around 800 to 830 nm), the fluorescence emitted at the primary lymph node is identifiably stronger than the fluorescence emitted at the secondary and subsequent lymph nodes.

[0020] A composition containing a compound represented by Formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient is effective as a composition for near-infrared light imaging of lymph nodes or lymph vessels that can selectively image the primary lymph node relative to the secondary and subsequent lymph nodes. A compound represented by Formula (I) or a pharmaceutically acceptable salt thereof locally administered to a patient once or multiple times, if desired, in an amount of from 0.005 to 10 mg is useful in nearinfrared light imaging of lymph nodes or lymph vessels. [0021] The compound represented by Formula (I) may be a salt formed with anions which is generated by releasing one or more protons from an acid. Specific examples include salts formed with anions which is generated by releasing protons from inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid and phosphoric acid, as well as from organic acids such as formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, mandelic acid, tartaric acid, dibenzoyl tartaric acid, ditoluoyl tartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid, benzene sulfonic acid, p-toluene sulfonic acid, aspartic acid, and glutamic acid. One embodiment of a pharmaceutically acceptable salt of a compound of Formula (I) is a chloride of a compound of Formula (I) (Pudexacianinium chloride), which is a salt with a chloride anion (Cl ).

[0022] As long as no conflicts arise, a combination of these embodiments may be used.

[0023] A compound represented by Formula (I) or a pharmaceutically acceptable salt thereof can be produced using the production methods described in Non-Patent Document 1, WO2011/093098 A1 or WO2021/105888 A1, or using a similar production method.

[0024] Compounds represented by Formula (I) and pharmaceutically acceptable salts thereof also include hydrates and solvates. Compounds represented by Formula (I) and pharmaceutically acceptable salts thereof may be amorphous or crystalline (including crystal polymorphs). Amorphous compounds represented by Formula (I) and pharmaceutically acceptable salts thereof can be produced using the production method described in the Journal of Biomedical Optics 21 (8), 086009 (August 2016) or similar production methods. Crystalline compounds represented by Formula (I) and pharmaceutically acceptable salts thereof can be produced using the method described in WC2021/105888 A1.

[0025] Compounds represented by Formula (I) and pharmaceutically acceptable salts thereof also include compounds labeled with radioactive or non-radioactive isotopes.

[0026] Compounds represented by Formula (I) and pharmaceutically acceptable salts thereof may have tautomers. In the present specification, compounds represented by Formula (I) and pharmaceutically acceptable salts thereof may be described in only one tautomer form, but these may include other tautomers, as well as isolated isomers or mixtures thereof.

[0027] A composition for imaging as disclosed herein can be prepared by a method common in the art using excipients, carriers, and other additives common in the art.

[0028] There are no particular restrictions on how a composition for imaging as disclosed herein is administered. Examples include parenteral administration, such as local administration of an injectable.

[0029] "Local administration" refers to local administration of the composition at a site where biological action is desired. "Local administration” and "administered locally” may also be described as "topical administration” or "administered topically,” respectively. Embodiments include intradermal, subcutaneous, intramuscular, interstitial, subdermal, and subareolar administration. Other embodiments include intradermal and subcutaneous administration. Another embodiment is intramuscular administration. [0030] Injectables may contain a sterile aqueous or non-aqueous solution, suspension, or emulsion. Aqueous solvents include, for example, distilled water for injection or saline. Non-aqueous solvents include alcohols such as ethanol. These compositions may also include excipients, buffers, isotonic agents, antioxidants, surfactants, preservatives, wetting agents, emulsifiers, dispersants, stabilizers, and solubilizing agents. These compositions are also sterilized, for example, by filtration through a bacterial retention filter, or by fungicide or irradiation. These can also be obtained by producing a sterile solid composition and dissolving or suspending this composition in sterile water or a sterile injectable solvent prior to use. One embodiment of a dosage form of a composition for imaging as disclosed herein is an aqueous solution. Examples of solid compositions that can be dissolved prior to use include lyophilized preparations and spray-dried preparations. One embodiment is a lyophilized preparation.

[0031] A compound represented by Formula (I) or a pharmaceutically acceptable salt thereof may be administered once or, if desired, multiple times. In one embodiment, the number of administrations is once. In another embodiment, it is twice. A compound represented by Formula (I) or a pharmaceutically acceptable salt thereof is administered before or during surgery. In one embodiment, it is administered before surgery. In another embodiment, it is administered during surgery. In one embodiment, it is administered first before surgery and then once again during surgery. Administration before surgery can be up to 24 hours before surgery, up to 18 hours before, up to 12 hours before, up to 10 hours before, up to 8 hours before, up to 6 hours before, up to 4 hours before, up to 3 hours before, up to 2 hours before, up to 1 .5 hours before, up to 1 hour before, up to 45 minutes before, up to 30 minutes before, up to 15 minutes before, or just before (e.g., within 5 minutes).

[0032] The amount of the compound represented by Formula (I) or a pharmaceutically acceptable salt thereof administered per dose (based upon free form weight of Formula (I)) 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 mg to 2 mg, 0.05 to 1 mg, or 0.1 to 1 mg. In one embodiment, it is 0.05, 0.2, 0.6, 1 , 2, or 4 mg. In another embodiment, the amount administered over multiple administrations is the same or different.

[0033] The site of administration of the compound represented by Formula (I) or a pharmaceutically acceptable salt thereof can be any suitable site for the patient in need thereof. In some cases, the administration is periareolar. In some cases, the administration is near the patient's left breast. In some cases, the administration is near the patient's right breast. In various cases, upon administration to a patient's breast, the patient's breast is massaged for about 5 minutes, which can assist in migration of the compound for imaging. In alternate cases, no massaging is performed.

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

[0035] The total volume locally administered over single or multiple administrations of an aqueous solution of the compound represented by Formula (I) or a pharmaceutically acceptable salt thereof in free form may be 0.05 mL to 5 mL. In some embodiments, the volume is 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.8, 1.0, 2.0, 3.0, 4.0 or 5.0 mL. In various embodiments, the volume is 0.05, 0.2, 0.6, 1 .0, 2.0 or 4.0 mL. In some embodiments, the volume is 1 .0 mL. In various embodiments, the volume is 2.0 mL.

EMBODIMENTS

1 . A composition for near-infrared imaging of lymph nodes or lymphatic vessels comprising a compound of Formula (I) or a pharmaceutically acceptable salt thereof,

the composition characterized in that the primary lymph node is selectively imaged relative to secondary and subsequent lymph nodes.

2. The composition according to embodiment 1, wherein the composition is an aqueous solution having a concentration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof of from 0.1 to 5 mg/mL, based upon the compound of Formula (I) weight in free form.

3. The composition according to embodiment 2, wherein the concentration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof is from 0.5 to 3 mg/mL, based upon the compound of Formula (I) weight in free form.

4. The composition according to embodiment 3, wherein the concentration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof is 1 mg/mL, based upon the compound of Formula (I) weight in free form.

5. The composition according to any of embodiments 1 to 3, wherein the compound of Formula (I) or a pharmaceutically acceptable salt thereof is a chloride salt.

6. A composition for near-infrared imaging of lymph nodes or lymphatic vessels comprising a compound of Formula (I) or a pharmaceutically acceptable salt thereof,

wherein the composition is used such 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, based upon the compound of Formula (I) weight in free form.

7. The composition according to embodiment 6, wherein the composition is administered to a patient once.

8. The composition according to embodiment 6, wherein the composition is administered to a patient multiple times.

9. The composition according to any of embodiments 6 to 8, wherein the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof administered per dose is from 0.05 to 5 mg, based upon the compound of Formula (I) weight in free form.

10. The composition according to embodiment 9, wherein the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof administered per dose is from 0.05 to 4 mg, based upon the compound of Formula (I) weight in free form.

11 . The composition according to any of embodiments 6 to 8, wherein the composition is an aqueous solution having a concentration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof of from 0.1 to 5 mg/mL, based upon the compound of Formula (I) weight in free form.

12. The composition according to embodiment 11, wherein the concentration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof is from 0.5 to 3 mg/mL, based upon the compound of Formula (I) weight in free form.

13. The composition according to embodiment 12, wherein the concentration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof is 1 mg/mL, based upon the compound of Formula (I) weight in free form.

14. The composition according to embodiment 10, wherein the concentration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof is 1 mg/mL, based upon the compound of Formula (I) weight in free form.

15. The composition according to any of embodiments 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 of embodiments 6 to 15, wherein the composition selectively images the primary lymph node relative to secondary and subsequent lymph nodes. 17. The composition according to any of embodiments 6 to 16, wherein the local administration is intradermal, subcutaneous, intramuscular, interstitial, subdermal, or subareolar administration.

18. A method of imaging a primary lymph node in a patient comprising administering to the patient an effective amount of a composition comprising a compound of Formula (I) or a pharmaceutically acceptable salt thereof,

exposing the patient to near infrared light to image the primary lymph node in the subject, wherein the primary lymph node is selectively imaged relative to a secondary lymph node and subsequent lymph nodes.

19. The method according to embodiment 18, wherein the composition is an aqueous solution having a concentration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof of from 0.1 to 5 mg/mL, based upon the compound of Formula (I) weight in free form.

20. The method according to embodiment 19, wherein the concentration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof is from 0.5 to 3 mg/mL, based upon the compound of Formula (I) weight in free form.

21 . The method according to embodiment 20, wherein the concentration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof is 1 mg/mL, based upon the compound of Formula (I) weight in free form.

22. The method according to embodiment 18, wherein the composition is administered in an amount of 0.005 to 10 mg per dose, based upon the compound of Formula (I) weight in free form.

23. The method according to any of embodiments 18 to 22, wherein the administration is local.

24. The method according to embodiment 23, wherein the local administration is intradermal, subcutaneous, intramuscular, interstitial, subdermal, or subareolar administration.

25. The method according to any of embodiments 18 to 24, wherein the composition is administered to the patient once.

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

27. The method according to any of embodiments 18 to 26, wherein the composition is administered to the patient before surgery. 28. The method according to any of embodiments 18 to 27, wherein the composition is administered to the patient during surgery.

29. The method according to any of embodiments 18 to 28, wherein the compound of Formula (I) or a pharmaceutically acceptable salt thereof is a chloride salt.

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

31 . Use of a composition for the manufacture of an agent for near-infrared imaging of a primary lymph node in a patient, wherein the composition comprises a compound of Formula (I) or a pharmaceutically acceptable salt thereof,

and the near-infrared imaging results in selective imaging of a primary lymph node relative to secondary and subsequent lymph nodes in the patient.

32. The use according to embodiment 31, wherein the composition is an aqueous solution having a concentration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof of from 0.1 to 5 mg/mL, based upon the compound of Formula (I) weight in free form.

33. The use according to embodiment 32, wherein the concentration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof is from 0.5 to 3 mg/mL, based upon the compound of Formula (I) weight in free form.

34. The use according to embodiment 33, wherein the concentration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof is 1 mg/mL, based upon the compound of Formula (I) weight in free form.

35. The use according to embodiment 31 , wherein the composition is for administration in an amount of 0.005 to 10 mg per dose, based upon the compound of Formula (I) weight in free form.

36. The use according to any of embodiments 31 to 34, wherein the composition is for local administration to the patient.

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

38. The use according to any of embodiments 31 to 37, wherein the composition is for administration to the patient once. 39. The use according to any of embodiments 31 to 37, wherein the composition is for administration to the patient multiple times.

40. The use according to any of embodiments 31 to 39, wherein the composition is for administration to the patient before surgery.

41 . The use according to any of embodiments 31 to 40, wherein the composition is for administration to the patient during surgery.

42. The use according to any of embodiments 31 to 41 , wherein the compound of Formula (I) or a pharmaceutically acceptable salt thereof is a chloride salt.

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

44. A method of imaging lymph nodes or lymphatic vessels in a patient comprising

(a) administering locally to the patient an amount of 0.005 to 10 mg per dose of a composition comprising a compound of Formula (I) or a pharmaceutically acceptable salt thereof,

wherein the amount is based upon the compound of Formula (I) weight in free form; and

(b) exposing the patient to near infrared light to image the lymph node or the lymphatic vessels in the patient.

45. Use of a composition for the manufacture of an agent for near-infrared imaging of lymph nodes or lymphatic vessels in a patient, wherein the composition comprises a compound of Formula (I) or a pharmaceutically acceptable salt thereof,

and the composition is for local administration in an amount of 0.005 to 10 mg per dose, based upon the compound of Formula (I) weight in free form. EXAMPLES

[0036] The present disclosure will now be described in greater detail with reference to examples. However, these examples do not limit the scope of the present disclosure.

[0037] Example 1 : Lymphatic Imaging Test (Mini Pigs)

[0038] Compound (I) and ICG were administered subcutaneously to the chest and abdomen of mini pigs (Gottingen or NIBS), and near-infrared fluorescence images were taken with a near-infrared fluorescence imaging device (PDE-Neo from Hamamatsu Photonics or Fluobeam from Fluoptics). 1 mg/mL of ICG was administered subcutaneously near the right second nipple and 1 mg/mL of Compound (I) was administered subcutaneously near the left second nipple of NIBS mini pigs (body weight 19.74 to 23.13 kg) at volumes of 0.05 mL, and near-infrared fluorescence were observed with a near-infrared fluorescence imaging device 10, 20 and 30 minutes later (Fig. 1). In order to verify the concentration dependence of Compound (I), 0.001, 0.01, 0.1, and 1 mg/mL of Compound (I) were administered subcutaneously to the abdomen of Gottingen minipigs (body weight 6.12 kg) at volumes of 0.1 mL, and near-infrared fluorescence observations were made with a near-infrared fluorescence imaging device immediately after administration as the compound moved from the administration site into the lymphatic vessels (Fig. 2). Also, 0.01, 0.1 and 1 mg/mL of Compound (I) and 2.5 and 5 mg/mL of ICG whose intralymphatic migration had been verified were administered subcutaneously near the second nipple of Gottingen minipigs (body weight 13.7-16.4 kg), and near-infrared fluorescence observations were made with a near-infrared fluorescence imaging device for two hours immediately after administration (Fig. 3). The fluorescent signals of Compound (I) rapidly migrated from the administration site to the inguinal lymph node after administration at concentrations of 0.1 and 1 mg/mL, and the lymph node could be confirmed percutaneously (Fig. 3-(1) and Fig. 3-(3)). Fluorescent signals in the inguinal lymph nodes were percutaneously confirmed with 0.01 mg/mL of Compound (I), but these signals were weak (Fig. 3-(5)). Similarly, fluorescent signals migrated from the administration site to the inguinal lymph node in the case of ICG, and the lymph node could be confirmed percutaneously (Fig. 3-(7), Fig. 3-(9)). At the end of the experiment, the inguinal lymph node was removed and near-infrared fluorescence were observed with the near-infrared fluorescence imaging device at the time of removal. When Compound (I) was administered at concentrations of 0.1 and 1 mg/mL, strong fluorescent signals tended to be observed on the head side (afferent lymphatic vessel side) of the excised lymphatic tissue, and weak fluorescent signals on the tail side (efferent lymphatic vessel side) (Fig. 3-(2) and Fig. 3-(4)). At a 0.01 mg/mL concentration of Compound (I), the fluorescent signals in the lymphatic tissue had disappeared at the time of excision (Fig. 3-(6)). In the case of ICG, strong fluorescent signals were observed throughout the excised lymphatic tissue (Fig. 3-(8), Fig. 3-(10)).

[0039] Example 2: Lymphatic Imaging Test (Dogs)

[0040] 2.5 mg/mL of ICG was administered subcutaneously near the left second nipple and 1 mg/mL of Compound (I) was administered subcutaneously near the right second nipple of a beagle (weight 7.90 kg) at volumes of 0.1 mL, and near-infrared fluorescence observations were made with a near-infrared fluorescence imaging device (Fluobeam from Fluoptics) ten minutes later (Fig. 4-(1 )). Near-infrared fluorescence observations were also made of the axillary lymph nodes with a near-infrared fluorescence imaging device when the lymph nodes were exposed and then after the lymph nodes had been removed (Fig. 4-(2)).

[0041] Example 3: Canine Lymphatic Imaging Test (Dogs, Hind Legs)

[0042] First, 1 mg/mL of Compound (I) (right hind limb) and 2.5 mg/mL of ICG (the left hind limb) were administered to the subcutaneous tissue in the interdigital region of the hind limbs of a beagle dog (male, 7 months old, weight 12.6 kg) at volumes of 0.1 mL, and the region around the gastrocnemius muscle and near the lymph nodes below the kneecaps was imaged with a near-infrared fluorescence imaging device (Fluobeam from Fluoptics) 20 minutes later (Fig. 5-(1) and Fig. 5-(2)). After the abdomen had been opened and the internal iliac lymph nodes exposed, the vicinity of the internal iliac lymph nodes was imaged using the same device (Fig. 5-(3) and Fig. 5-(4)). After euthanasia had been performed about 2.5 hours later, the lymph nodes below the kneecaps and the internal iliac lymph nodes were removed and near-infrared fluorescence images were captured using the same device (Fig. 5-(5) to Fig. 5-(12)).

Claims

What is Claimed:

2. The composition according to claim 1, wherein the composition is an aqueous solution having a concentration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof of from 0.1 to 5 mg/mL, based upon the compound of Formula (I) weight in free form.

3. The composition according to claim 2, wherein the concentration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof is from 0.5 to 3 mg/mL, based upon the compound of Formula (I) weight in free form.

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, based upon the compound of Formula (I) weight in free form.

5. The composition according to any of claims 1 to 3, wherein the compound of Formula (I) or a pharmaceutically acceptable salt thereof is a chloride salt.

7. The composition according to claim 6, wherein the composition is administered to a patient once.

8. The composition according to claim 6, wherein the composition is administered to a patient multiple times.

9. The composition according to any of claims 6 to 8, wherein the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof administered per dose is from 0.05 to 5 mg, based upon the compound of Formula (I) weight in free form.

10. The composition according to claim 9, wherein the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof administered per dose is from 0.05 to 4 mg, based upon the compound of Formula (I) weight in free form.

11 . The composition according to any of claims 6 to 8, wherein the composition is an aqueous solution having a concentration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof of from 0.1 to 5 mg/mL, based upon the compound of Formula (I) weight in free form.

12. The composition according to claim 11, wherein the concentration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof is from 0.5 to 3 mg/mL, based upon the compound of Formula (I) weight in free form.

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, based upon the compound of Formula (I) weight in free form.

14. The composition according to claim 10, wherein the concentration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof is 1 mg/mL, based upon the compound of Formula (I) weight in free form.

15. The composition according to any 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 of claims 6 to 15, wherein the composition selectively images the primary lymph node relative to secondary and subsequent lymph nodes.

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

exposing the patient to near infrared light to image the primary lymph node in the patient, wherein the primary lymph node is selectively imaged relative to a secondary lymph node and subsequent lymph nodes.

19. The method according to claim 18, wherein the composition is an aqueous solution having a concentration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof of from 0.1 to 5 mg/mL, based upon the compound of Formula (I) weight in free form.

20. The method according to claim 19, wherein the concentration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof is from 0.5 to 3 mg/mL, based upon the compound of Formula (I) weight in free form.

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, based upon the compound of Formula (I) weight in free form.

22. The method according to claim 18, wherein the composition is administered in an amount of

0.005 to 10 mg per dose, based upon the compound of Formula (I) weight in free form.

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

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

25. The method according to any of claims 18 to 24, wherein the composition is administered to the patient once.

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

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

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

29. The method according to any of claims 18 to 28, wherein the compound of Formula (I) or a pharmaceutically acceptable salt thereof is a chloride salt.

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

32. The use according to claim 31, wherein the composition is an aqueous solution having a concentration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof of from 0.1 to 5 mg/mL, based upon the compound of Formula (I) weight in free form.

33. The use according to claim 32, wherein the concentration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof is from 0.5 to 3 mg/mL, based upon the compound of Formula (I) weight in free form.

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, based upon the compound of Formula (I) weight in free form.

35. The use according to claim 31, wherein the composition is for administration in an amount of 0.005 to 10 mg per dose, based upon the compound of Formula (I) weight in free form.

36. The use according to any of claims 31 to 34, wherein the composition is for local administration to the patient.

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

38. The use according to any of claims 31 to 37, wherein the composition is for administration to the patient once.

39. The use according to any of claims 31 to 37, wherein the composition is for administration to the patient multiple times.

40. The use according to any of claims 31 to 39, wherein the composition is for administration to the patient before surgery.

41 . The use according to any of claims 31 to 40, wherein the composition is for administration to the patient during surgery.

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

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

and the composition is for local administration in an amount of 0.005 to 10 mg per dose, based upon the compound of Formula (I) weight in free form.