EP4149925A1 - Novel fluorescent compounds for labelling tumour tissue - Google Patents
Novel fluorescent compounds for labelling tumour tissueInfo
- Publication number
- EP4149925A1 EP4149925A1 EP21732450.8A EP21732450A EP4149925A1 EP 4149925 A1 EP4149925 A1 EP 4149925A1 EP 21732450 A EP21732450 A EP 21732450A EP 4149925 A1 EP4149925 A1 EP 4149925A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- chem
- compounds
- compound
- tumor
- alkyl
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 6
- 125000004432 carbon atom Chemical group C* 0.000 description 6
- 239000002244 precipitate Substances 0.000 description 6
- 239000012429 reaction media Substances 0.000 description 6
- -1 2,2-dimethylbutyl Chemical group 0.000 description 5
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- 238000009826 distribution Methods 0.000 description 4
- 239000000975 dye Substances 0.000 description 4
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- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 3
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 3
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 3
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- 239000000090 biomarker Substances 0.000 description 3
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- 239000001632 sodium acetate Substances 0.000 description 3
- 235000017281 sodium acetate Nutrition 0.000 description 3
- 238000012800 visualization Methods 0.000 description 3
- JWUQFQYYMGMPKE-UHFFFAOYSA-N 2-chloro-3-(hydroxymethylidene)cyclohexene-1-carbaldehyde Chemical compound OC=C1CCCC(C=O)=C1Cl JWUQFQYYMGMPKE-UHFFFAOYSA-N 0.000 description 2
- 206010006187 Breast cancer Diseases 0.000 description 2
- WQDUMFSSJAZKTM-UHFFFAOYSA-N Sodium methoxide Chemical compound [Na+].[O-]C WQDUMFSSJAZKTM-UHFFFAOYSA-N 0.000 description 2
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- 208000015486 malignant pancreatic neoplasm Diseases 0.000 description 2
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- 125000001971 neopentyl group Chemical group [H]C([*])([H])C(C([H])([H])[H])(C([H])([H])[H])C([H])([H])[H] 0.000 description 2
- 208000008443 pancreatic carcinoma Diseases 0.000 description 2
- NMHMNPHRMNGLLB-UHFFFAOYSA-N phloretic acid Chemical compound OC(=O)CCC1=CC=C(O)C=C1 NMHMNPHRMNGLLB-UHFFFAOYSA-N 0.000 description 2
- 238000002271 resection Methods 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- MEKOFIRRDATTAG-UHFFFAOYSA-N 2,2,5,8-tetramethyl-3,4-dihydrochromen-6-ol Chemical compound C1CC(C)(C)OC2=C1C(C)=C(O)C=C2C MEKOFIRRDATTAG-UHFFFAOYSA-N 0.000 description 1
- 125000005916 2-methylpentyl group Chemical group 0.000 description 1
- YQWPHBFLHAJVCG-UHFFFAOYSA-N 3-(4-sulfanylphenyl)propanoic acid Chemical compound OC(=O)CCC1=CC=C(S)C=C1 YQWPHBFLHAJVCG-UHFFFAOYSA-N 0.000 description 1
- 125000005917 3-methylpentyl group Chemical group 0.000 description 1
- WXOHKMNWMKZMND-UHFFFAOYSA-N 4-aminohydrocinnamic acid Chemical compound NC1=CC=C(CCC(O)=O)C=C1 WXOHKMNWMKZMND-UHFFFAOYSA-N 0.000 description 1
- 206010052747 Adenocarcinoma pancreas Diseases 0.000 description 1
- 238000011729 BALB/c nude mouse Methods 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- FXVMRKBTKKLDFH-UHFFFAOYSA-N CC1=C(CCCCS(O)(=O)=O)C(C2=C(C=C3)C(S([O-])(=O)=O)=CC=C2)=C3[N+]1(C)C Chemical compound CC1=C(CCCCS(O)(=O)=O)C(C2=C(C=C3)C(S([O-])(=O)=O)=CC=C2)=C3[N+]1(C)C FXVMRKBTKKLDFH-UHFFFAOYSA-N 0.000 description 1
- JKNCPHZDTACFRM-UHFFFAOYSA-N CC1=C(CCCCS(O)(=O)=O)C(C2=C(C=C3OCCCCCC([O-])=O)C(S(O)(=O)=O)=CC=C2)=C3[N+]1(C)C Chemical compound CC1=C(CCCCS(O)(=O)=O)C(C2=C(C=C3OCCCCCC([O-])=O)C(S(O)(=O)=O)=CC=C2)=C3[N+]1(C)C JKNCPHZDTACFRM-UHFFFAOYSA-N 0.000 description 1
- XBPCUCUWBYBCDP-UHFFFAOYSA-N Dicyclohexylamine Chemical compound C1CCCCC1NC1CCCCC1 XBPCUCUWBYBCDP-UHFFFAOYSA-N 0.000 description 1
- SNRUBQQJIBEYMU-UHFFFAOYSA-N Dodecane Natural products CCCCCCCCCCCC SNRUBQQJIBEYMU-UHFFFAOYSA-N 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- XOGTZOOQQBDUSI-UHFFFAOYSA-M Mesna Chemical compound [Na+].[O-]S(=O)(=O)CCS XOGTZOOQQBDUSI-UHFFFAOYSA-M 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 description 1
- 238000011579 SCID mouse model Methods 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 125000003710 aryl alkyl group Chemical group 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 125000002704 decyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
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- 125000003438 dodecyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
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- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
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- 229960000304 folic acid Drugs 0.000 description 1
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- 238000010438 heat treatment Methods 0.000 description 1
- 125000003187 heptyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 125000004446 heteroarylalkyl group Chemical group 0.000 description 1
- 125000005842 heteroatom Chemical group 0.000 description 1
- 125000004051 hexyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
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- 125000001183 hydrocarbyl group Chemical group 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
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- 230000002601 intratumoral effect Effects 0.000 description 1
- 125000000959 isobutyl group Chemical group [H]C([H])([H])C([H])(C([H])([H])[H])C([H])([H])* 0.000 description 1
- 125000001972 isopentyl group Chemical group [H]C([H])([H])C([H])(C([H])([H])[H])C([H])([H])C([H])([H])* 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
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- 239000000463 material Substances 0.000 description 1
- 229960004635 mesna Drugs 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 125000004108 n-butyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 125000000740 n-pentyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 125000004123 n-propyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 238000003333 near-infrared imaging Methods 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 125000001400 nonyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 125000002347 octyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 125000000913 palmityl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 201000002094 pancreatic adenocarcinoma Diseases 0.000 description 1
- 210000004923 pancreatic tissue Anatomy 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000004286 retinal pathology Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 125000002914 sec-butyl group Chemical group [H]C([H])([H])C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 125000003548 sec-pentyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
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- 239000011593 sulfur Substances 0.000 description 1
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- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 230000005748 tumor development Effects 0.000 description 1
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- 238000001429 visible spectrum Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D209/56—Ring systems containing three or more rings
- C07D209/58—[b]- or [c]-condensed
- C07D209/60—Naphtho [b] pyrroles; Hydrogenated naphtho [b] pyrroles
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
- C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
- C07D403/08—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a carbon chain containing alicyclic rings
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
- A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
- A61K49/0032—Methine dyes, e.g. cyanine dyes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/006—Biological staining of tissues in vivo, e.g. methylene blue or toluidine blue O administered in the buccal area to detect epithelial cancer cells, dyes used for delineating tissues during surgery
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
- G01N33/582—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with fluorescent label
Definitions
- the present invention relates to novel fluorescent compounds which can be used for labeling tumor tissue, their method of preparation, as well as their application as a tool for monitoring, diagnosing or assisting in cancer surgery.
- Fluorescent compounds have been used for over fifty years in medicine as markers in non-invasive imaging techniques for monitoring and / or diagnosis.
- Certain existing fluorescent markers have a limited persistence of fluorescence, which makes it necessary to operate on the patient following the injection of the marker and does not make it possible to obtain a satisfactory delineation of the tissue. tumor. In other cases, the accumulation of the label in the tissues is not high enough, which leads to poor labeling and therefore detection problems. The localization of lesions or tumors then their elimination, for example by surgery, is then not complete.
- ICG indocyanine green
- Another major drawback of existing compounds is that they cannot be used directly. In fact, in order to be used, they need to be coupled with other targeting molecules such as antibodies, proteins, molecules specific to tumor tissue, folic acid, or even steroids.
- the present invention overcomes the problems of the prior art explained above by providing fluorescent molecules having a preferential distribution in tumor tissues over healthy tissues and sufficient persistence for their use in techniques of. imaging for monitoring, diagnosis, and / or surgical assistance.
- These new molecules have the major advantage of being able to be used alone and directly, without prior coupling, due to their specific affinity for tumor tissue. They also have, compared to the molecules of the prior art, a much higher remanence in the tumor tissues, going up to a duration of several days, which allows a more extensive elimination of these fluorescent molecules circulating outside the tumor tissues, and thus improved visualization thanks to a better signal-to-noise ratio.
- the present invention relates to a compound of formula (I) [Chem. 1] where n1 and n2 are each an integer from 0 to 15,
- Ri, R2, R3, R4, R5 and He are each independently selected from H, OH, SH, NH 2 , SO3R10 and X-R11-Y,
- R11, R'11, R ”n being independently selected from C1 to C15 alkyl, aryl, heteroaryl, (C1 to Cis alkyl) aryl, (C1 to Cis alkyl) heteroaryl, aryl (C1 to C15 alkyl) and heteroaryl (C1-C15 alkyl);
- Y, Y ’, Y being independently selected from H, halogen, COOR’10 or amide; R7 and Re being each selected from H, OH, SH, NH2, C1-C15 alkyl, and X'-R'n-Y ';
- R 9 being chosen from H, OH, SH, NH2 and X ”-R” nY ”, said compound comprising at least one group X-R11-Y, X'-R'n-Y 'or X” -R ”n - Y ”with Y, Y ', and / or Y” which is COOR'10.
- the present invention also relates to the process for preparing the compounds of formula (I) according to the invention, as well as the process for labeling tumor tissue with one of the compounds according to the invention or prepared according to the process of the invention. .
- the first subject of the present invention relates to a compound of formula (I)
- ni and n2 are each an integer from 0 to 15, Ri, R2, R3, R4, R6 and R6 are each independently selected from H, OH, SH, NH 2 , SO3R10 and X-R11-Y,
- X, X ’, X being independently O, S or NH,
- R11, R'11, R ”H being independently selected from C1 to C15 alkyl, aryl, heteroaryl, (C1 to C18 alkyl) aryl, (C1 to Cis alkyl) heteroaryl, aryl (C1 to C15 alkyl) and heteroaryl (C1-C15 alkyl);
- Y, Y ’, Y being independently selected from H, halogen, COOR’10 or amide;
- R7 and R8 being each selected from H, OH, SH, NH2, C1-C15 alkyl, and X'-R'n-Y '; R 9 being chosen from H, OH, SH, NH2 and X ”-R” nY ”, said compound comprising at least one X-R11-Y, X'-R'n-Y 'or X” -R ”n group - Y ”with Y, Y ', and / or Y” which is COOR'10.
- C 1 to C 15 alkyl means a hydrocarbon chain, cyclic, linear or branched, containing from 1 to 15 carbon atoms, preferably from 2 to 6 carbon atoms and more preferably still from 4 to 6 carbon atoms, in particular 5 carbon atoms and possibly being in particular a methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, tert-butyl chain, n-pentyl, 1-methylbutyl, 2,2-dimethylbutyl, 2-methylpentyl, 2,2-dimethylpropyl, isopentyl, neopentyl, 2-pentyl, hexyl, 2-hexyl, 3-hexyl, 3-methylpentyl, heptyl, octyl, nonyl, decyl, dodecyl, palmityl.
- heptyl octyl
- heteroaryl means an aromatic group, containing one or more aromatic rings, optionally substituted, and comprising at least one heteroatom other than carbon and hydrogen.
- arylalkyl is understood to mean an aryl group substituted by one or more alkyl groups, said alkyl groups possibly being C1 to C15 alkyl groups, preferably containing from 1 to 15 carbon atoms. carbon.
- heteroarylalkyls is understood to mean a heteroaryl group substituted by one or more alkyl groups, said alkyl groups possibly being C 1 to C 15 alkyl groups, preferably containing from 1 to 15 carbon atoms. carbon.
- m or n2 are independently of one another, equal to 1, 2, 3, 4 or 5, more preferably still 3 or 4.
- n-i n2 and is preferably equal to 1, 2, 3, 4 or 5, more preferably still 3 or 4.
- the molecule is symmetrical.
- it comprises a single group X ”-R” nY ”with Y” which is COOR'10 which is carried by R9 and / or 2 groups X-R11-Y with Y which is COOR'10, one carried by one of Ri, R2, R30U Rz, preferably one of Ri, R2 or R3 and the other carried by one of R4, Rs, R6 or Rs, preferably one of R4, Rs or R6.
- R10, and / or R'10 may be identical.
- X, X ', and / or X may be the same, Y, Y', and / or Y” may be the same, Ru, R'11, and / or R ”n may be the same.
- the compounds according to the invention can in particular be chosen from the compounds of the following general formula: for which X ”can be O, S or NH, then corresponding to the formulas below:
- the compounds of formula (I) according to the invention can in particular be chosen from the compounds of formula (I) for which Ri, R2, R3, R4, Rs and Re ne are not all simultaneously H, which in this case excludes the compounds according to formula (II) below:
- the compound of formula (I) according to the invention can preferably be chosen from the compounds of the following general formulas:
- the compounds according to the invention can be chosen from the compounds of the following formulas [Chem. 5] [Chem. 7] in which R1, R2, R3, R4, Rs, Re, R7, Rs and R9 are as defined above. According to a preferred embodiment, the compounds according to the invention can be chosen from the following compounds:
- the second object of the present invention relates to the process for preparing the compounds of formula (I) according to the invention comprising a reaction step between:
- the third object of the invention relates to the method of labeling tumor tissue with one of the compounds according to the invention or prepared according to the method of the invention.
- tumor tissue is understood to mean the tissue consisting of tumor cells which are abnormal proliferative cells, and supporting tissue, also called tumor stroma or interstitial tissue, made up of cells and extra-cellular material in which the tumor vasculature is located.
- the fluorescent compounds according to the invention have the particularity, after their diffusion in the body, of being trapped in the tumor tissue, while they are eliminated from healthy tissue. This feature makes it possible to use these fluorescent compounds directly, without prior coupling to another labeling molecule, thus making their use simpler, faster and more efficient than that of the compounds of the prior art. It has been observed that this elimination from healthy tissue increases over time. In general, between 24 and 72 hours, preferably between 36 and 60 hours and more preferably 48 hours after the administration of these compounds, their elimination from healthy tissues is complete. However, they remain trapped in tumor tissue. This property makes it possible to have a clear differentiation of tumor tissues from healthy tissues and thus the use of these compounds in monitoring, diagnostic and / or surgical assistance applications in the context of cancerous diseases. This differentiation lasts for 6 to 48 hours, preferably 12 to 36 hours, making it possible to program the diagnosis or the surgery in a targeted manner.
- the compounds according to the invention can thus in particular be used in the context of cancers, for example hormone-dependent, such as breast cancer or digestive cancers, such as pancreatic cancer.
- hormone-dependent such as breast cancer
- digestive cancers such as pancreatic cancer.
- pancreatic cancer tumors are particularly difficult to remove in their entirety by surgery because they are not easily delimited.
- the use of the compounds according to the invention makes it possible to obtain a better visualization of the contours of the tumors thanks to the differentiation of labeling between the tumor tissue and the healthy tissue, and thus a more efficient tumor resection by surgery.
- the invention also relates to the use of one of the compounds according to the invention or prepared according to the method of the invention in a method of labeling tumor tissue.
- This method of labeling tissues requires the administration of the compounds intravenously, intraarterially, or into another vessel, in particular a lymphatic vessel, either by local injection or by local application, preferably intravenously.
- Another object of the invention relates to a composition
- a composition comprising one of the compounds according to the invention or prepared according to the process of the invention and at least one pharmaceutically acceptable adjuvant.
- the invention also relates to one of the compounds according to the invention or prepared according to the process of the invention or a composition comprising one of the compounds according to the invention or prepared according to the process of the invention for its use. in a method of labeling and / or detecting tumor tissue, and / or in the surgical treatment of tumors.
- the invention also relates to a method of detecting tumor tissue comprising a step of labeling tumor tissue with one of the compounds according to the invention or prepared according to the method of the invention, and a step of detection by medical imaging in fluorescence or fluorescence spectrometry.
- FIG. 1 shows the median values and standard deviations of the tumor / abdomen intensity ratios as a function of post-injection times of Compound 2 (CJ215) and ICG.
- FIG. 2 shows the results of ex vivo imaging of pancreatic tumors after injection of two compounds according to the invention and a fluorescent agent of the prior art (ICG).
- the reaction medium is cooled to room temperature and the precipitate is separated by filtration and washed with diethyl ether to provide 4.33 g (yield: 43.9%) of a green solid.
- the crude product is purified by flash column chromatography (reverse phase C18 silica gel, 0-25% acetonitrile / water).
- Example 8 Comparison of a compound according to the invention and of a fluorescent agent of the prior art (ICG) for in vivo imaging of mammary tumors
- ICG fluorescent agent of the prior art
- ICG or indocyanine green / Infracyanine
- ICG is a fluorescent agent of the prior art, already approved for use in humans for the evaluation of cardiac and hepatic functions, as well as in ophthalmology, for retinal pathologies. It is also being evaluated in numerous clinical trials around the world for guiding the surgical procedure during tumor resection, or the mapping of lymph nodes draining tumors, by near infrared imaging.
- the ICG was compared with the compound (2) according to the invention, the synthesis of which is described in the preceding example 2, this compound is called CJ215 in this study.
- the study included a total of 30 mice divided into three groups.
- the tumor grafts were all carried out with 50,000 4T1-Dendra2 / 20 ⁇ l cells injected into 2 mammary glands contralateral for each of the mice.
- the injections of biomarkers (compound 2 called CJ215 in this study and ICG) were carried out on D9 post-tumor grafting (in order to limit the appearance of necrosis in tumors).
- the change in the intensity of the fluorescence signals observed for each of the biomarkers over time was evaluated by microscopic image.
- the ability of the two markers to produce a signal specifically localized to the tumor was assessed quantitatively by calculating the ratio of the specific signal bound to the tumor to the non-specific signal in the surrounding tissues.
- the imaging protocol was carried out at times 2h, 24h, 48h, 4 and 6 days post-injection for all the mice. All the images produced at each acquisition time were acquired on an I VIS Spectrum imager (Perkin Elmer) with the following parameters:
- the quantitative measurements were carried out on the non-deconvoluted raw images.
- the acquisition time was set in automatic mode. In this mode, the system determines the acquisition time necessary to reach the imposed target value (6000 counts) within the allotted time (fixed at 2 min).
- Figure 1 reports the median values and standard deviations of the tumor / abdomen intensity ratios as a function of post-injection times of CJ215 and ICG.
- Example 9 Comparison of two compounds according to the invention and of a fluorescent agent of the prior art (ICG) in ex vivo imaging of pancreatic tumors.
- a model of orthotopic pancreatic adenocarcinoma in mice has been developed. Tumor cells were amplified subcutaneously in SCID mice and the resulting fragments were then surgically implanted into the pancreas of irradiated BALB / c nude mice.
- MRI 4.7T, PharmaScan, Bruker Biospin
- Animals were subjected to weak fluorescence in order to minimize autofluorescence. Fluorescent imaging was performed with a charge coupled device (CCD) camera (PhotonRT, BiospaceLab) with excitation at 700 nm and emission filter at 770 nm.
- CCD charge coupled device
- ex vivo fluorescent images were acquired.
- the fluorescent compounds according to the invention 2 (CJ215) and CJ319 (the structure of which is detailed below) were injected intravenously at 2 mg / kg, 39 days after implantation of the tumor fragments, while the volumes average tumors were about 70 mm 3 .
- Indocyanine green (ICG) a dye widely used in intraoperative tumor imaging, was included as a control.
Abstract
The subject matter of the present invention is novel fluorescent compounds that can be used for labelling tumour tissue, the method for preparing same, and also the application thereof as a monitoring or diagnostic tool or a tool for assisting with cancer surgery.
Description
Description Description
Titre : Nouveaux composés fluorescents pour le marquage de tissu tumoral Title: New Fluorescent Compounds for Tumor Tissue Labeling
[0001] La présente invention a pour objet de nouveaux composés fluorescents utilisables pour le marquage de tissu tumoral, leur procédé de préparation, ainsi que leur application comme outil de surveillance, de diagnostic ou d’aide à la chirurgie de cancers. [0001] The present invention relates to novel fluorescent compounds which can be used for labeling tumor tissue, their method of preparation, as well as their application as a tool for monitoring, diagnosing or assisting in cancer surgery.
Domaine technique Technical area
[0002] Le marquage de tissu tumoral par des composés fluorescents présente un grand intérêt dans le domaine de l'imagerie médicale, car il permet entre autres la localisation de tumeurs. [0002] The labeling of tumor tissue with fluorescent compounds is of great interest in the field of medical imaging, since it allows, among other things, the localization of tumors.
Technique antérieure Prior art
[0003] Des composés fluorescents sont utilisés depuis plus de cinquante ans en médecine comme marqueurs dans des techniques d'imagerie non invasives pour la surveillance et/ou le diagnostic. Fluorescent compounds have been used for over fifty years in medicine as markers in non-invasive imaging techniques for monitoring and / or diagnosis.
Problème technique Technical problem
[0004] L’émergence de nouvelles technologies d’imagerie de fluorescence au service de la chirurgie, nécessitant une sensibilité et une précision améliorées conduit à la recherche de nouvelles molécules fluorescentes toujours plus performantes. [0004] The emergence of new fluorescence imaging technologies in the service of surgery, requiring improved sensitivity and precision, has led to the search for new fluorescent molecules that are always more efficient.
[0005] Dans le contexte de maladies telles que le cancer, il est notamment nécessaire d’avoir une distribution préférentielle des molécules fluorescentes dans les tissus tumoraux par rapport aux tissus sains, ainsi qu’une persistance suffisamment longue de la fluorescence pour permettre une meilleure spécificité de marquage et fournir une aide à l’acte chirurgical, par exemple pour délimiter des zones de tumeurs à éliminer. [0005] In the context of diseases such as cancer, it is in particular necessary to have a preferential distribution of the fluorescent molecules in the tumor tissues relative to the healthy tissues, as well as a sufficiently long persistence of the fluorescence to allow better specificity of marking and provide an aid to the surgical act, for example to delimit areas of tumors to be eliminated.
[0006] Certains marqueurs fluorescents existants présentent une persistance de fluorescence limitée, ce qui oblige à opérer le patient dans la foulée de l’injection du marqueur et ne permet pas d’obtenir une délimitation satisfaisante du tissu
tumoral. Dans d’autres cas, l’accumulation du marqueur dans les tissus n’est pas suffisamment importante, ce qui entraîne un faible marquage et donc des problèmes de détection. La localisation de lésions ou de tumeurs puis leur élimination par exemple par chirurgie n’est alors pas complète. [0006] Certain existing fluorescent markers have a limited persistence of fluorescence, which makes it necessary to operate on the patient following the injection of the marker and does not make it possible to obtain a satisfactory delineation of the tissue. tumor. In other cases, the accumulation of the label in the tissues is not high enough, which leads to poor labeling and therefore detection problems. The localization of lesions or tumors then their elimination, for example by surgery, is then not complete.
[0007] Un autre problème présenté par les marqueurs fluorescents existants, notamment le vert d’indocyanine (ICG), qui est l’un des seuls colorants utilisés pour le marquage des tumeurs au cours de l’acte chirurgical, est la nécessité de la présence de néo-vaisseaux tumoraux pour obtenir un marquage du tissu tumoral. Par ailleurs, le vert d’indocyanine, à l’image des autres colorants existants dans l’art antérieur, n’est visible dans les tissus tumoraux que jusqu’à 24h après son injection. Cette durée courte ne permet pas d’avoir une bonne élimination du colorant circulant hors des tissus tumoraux, ce qui provoque une mauvaise visualisation car le rapport signal sur bruit est faible. [0007] Another problem presented by existing fluorescent markers, in particular indocyanine green (ICG), which is one of the only dyes used for the labeling of tumors during the surgical act, is the need for it. presence of tumor neo-vessels to obtain labeling of the tumor tissue. Furthermore, indocyanine green, like other dyes existing in the prior art, is only visible in tumor tissue for up to 24 hours after its injection. This short duration does not allow good elimination of the dye circulating outside the tumor tissues, which causes poor visualization because the signal to noise ratio is low.
[0008] Un autre inconvénient majeur des composés existants est qu’ils ne peuvent pas être utilisés directement. En effet, ils nécessitent pour être utilisés d’être couplés avec d’autres molécules de ciblage telles que des anticorps, des protéines, des molécules spécifiques du tissu tumoral, l’acide folique, ou encore des stéroïdes. Another major drawback of existing compounds is that they cannot be used directly. In fact, in order to be used, they need to be coupled with other targeting molecules such as antibodies, proteins, molecules specific to tumor tissue, folic acid, or even steroids.
[0009] La présente invention permet de s’affranchir des problèmes de l’art antérieur précédemment explicités en fournissant des molécules fluorescentes ayant une distribution préférentielle dans les tissus tumoraux par rapport aux tissus sains et une persistance suffisante pour leur utilisation dans des techniques d'imagerie pour la surveillance, le diagnostic, et/ou l’assistance à la chirurgie. Ces nouvelles molécules présentent l’avantage majeur de pouvoir être utilisées seules et directement, sans couplage préalable, du fait de leur affinité spécifique pour le tissu tumoral. Elles ont par ailleurs par rapport aux molécules de l’art antérieur une rémanence dans les tissus tumoraux beaucoup plus élevée, allant jusqu’à une durée de plusieurs jours, ce qui permet une élimination plus poussée de ces molécules fluorescentes circulant hors des tissus tumoraux, et ainsi une visualisation améliorée grâce à un meilleur rapport signal sur bruit. The present invention overcomes the problems of the prior art explained above by providing fluorescent molecules having a preferential distribution in tumor tissues over healthy tissues and sufficient persistence for their use in techniques of. imaging for monitoring, diagnosis, and / or surgical assistance. These new molecules have the major advantage of being able to be used alone and directly, without prior coupling, due to their specific affinity for tumor tissue. They also have, compared to the molecules of the prior art, a much higher remanence in the tumor tissues, going up to a duration of several days, which allows a more extensive elimination of these fluorescent molecules circulating outside the tumor tissues, and thus improved visualization thanks to a better signal-to-noise ratio.
Résumé de l’invention
[0010] La présente invention concerne un composé de formule (I) [Chem. 1]
dans laquelle n1 et n2 sont chacun un entier de 0 à 15, Summary of the invention The present invention relates to a compound of formula (I) [Chem. 1] where n1 and n2 are each an integer from 0 to 15,
Ri, R2, R3, R4, R5 et He sont chacun indépendamment choisis parmi H, OH, SH, NH2, SO3R10 et X-R11-Y, Ri, R2, R3, R4, R5 and He are each independently selected from H, OH, SH, NH 2 , SO3R10 and X-R11-Y,
R10, R’10, étant indépendamment H, Na ou K, X, X’, X” étant indépendamment O, S ou NH, R10, R'10, being independently H, Na or K, X, X ’, X” being independently O, S or NH,
R11, R’11, R”n, étant indépendamment choisis parmi alkyle en Ci à C15, aryle, hétéroaryle, (alkyl en Ci à Cis)aryle, (alkyl en Ci à Cis)hétéroaryle, aryl(alkyle en Ci à C15) et hétéroaryl(alkyle en Ci à C15); R11, R'11, R ”n, being independently selected from C1 to C15 alkyl, aryl, heteroaryl, (C1 to Cis alkyl) aryl, (C1 to Cis alkyl) heteroaryl, aryl (C1 to C15 alkyl) and heteroaryl (C1-C15 alkyl);
Y, Y’, Y” étant indépendamment choisis parmi H, halogène, COOR’10 ou amide ; R7 et Re étant chacun choisis parmi H, OH, SH, NH2, alkyle en Ci à C15, et X’-R’n- Y’; Y, Y ’, Y” being independently selected from H, halogen, COOR’10 or amide; R7 and Re being each selected from H, OH, SH, NH2, C1-C15 alkyl, and X'-R'n-Y ';
R9 étant choisi parmi H, OH, SH, NH2 et X”-R”n-Y”, ledit composé comportant au moins un groupement X-R11-Y, X’-R’n-Y’ ou X”-R”n- Y” avec Y, Y’, et/ou Y” qui est COOR’10. [0011] La présente invention concerne également le procédé de préparation des composés de formule (I) selon l’invention, ainsi que le procédé de marquage de tissu tumoral avec un des composés selon l’invention ou préparé selon le procédé de l’invention. R 9 being chosen from H, OH, SH, NH2 and X ”-R” nY ”, said compound comprising at least one group X-R11-Y, X'-R'n-Y 'or X” -R ”n - Y ”with Y, Y ', and / or Y” which is COOR'10. The present invention also relates to the process for preparing the compounds of formula (I) according to the invention, as well as the process for labeling tumor tissue with one of the compounds according to the invention or prepared according to the process of the invention. .
Exposé de l’invention [0012] Le premier objet de la présente invention concerne un composé de formule (I) Disclosure of the invention [0012] The first subject of the present invention relates to a compound of formula (I)
[Chem. 2]
dans laquelle ni et n2 sont chacun un entier de 0 à 15, Ri, R2, R3, R4, RÔ et R6 sont chacun indépendamment choisis parmi H, OH, SH, NH2, SO3R10 et X-R11-Y, [Chem. 2] wherein ni and n2 are each an integer from 0 to 15, Ri, R2, R3, R4, R6 and R6 are each independently selected from H, OH, SH, NH 2 , SO3R10 and X-R11-Y,
Rio, R’10, étant indépendamment H, Na ou K, Rio, R'10, being independently H, Na or K,
X, X’, X” étant indépendamment O, S ou NH, X, X ’, X” being independently O, S or NH,
R11, R’11, R”H étant indépendamment choisis parmi alkyle en Ci à C15, aryle, hétéroaryle, (alkyl en Ci à Cis)aryle, (alkyl en Ci à Cis)hétéroaryle, aryl(alkyle en Ci à C15) et hétéroaryl(alkyle en Ci à C15) ; R11, R'11, R ”H being independently selected from C1 to C15 alkyl, aryl, heteroaryl, (C1 to C18 alkyl) aryl, (C1 to Cis alkyl) heteroaryl, aryl (C1 to C15 alkyl) and heteroaryl (C1-C15 alkyl);
Y, Y’, Y” étant indépendamment choisis parmi H, halogène, COOR’10 ou amide ;Y, Y ’, Y” being independently selected from H, halogen, COOR’10 or amide;
R7 et R8 étant chacun choisis parmi H, OH, SH, NH2, alkyle en Ci à C15, et X’-R’n- Y’; R9 étant choisi parmi H, OH, SH, NH2 et X”-R”n-Y”, ledit composé comportant au moins un groupement X-R11-Y, X’-R’n-Y’ ou X”-R”n- Y” avec Y, Y’, et/ou Y” qui est COOR’10. R7 and R8 being each selected from H, OH, SH, NH2, C1-C15 alkyl, and X'-R'n-Y '; R 9 being chosen from H, OH, SH, NH2 and X ”-R” nY ”, said compound comprising at least one X-R11-Y, X'-R'n-Y 'or X” -R ”n group - Y ”with Y, Y ', and / or Y” which is COOR'10.
[0013] Au sens de la présente invention, on entend par « alkyles en Ci à C15», une chaîne hydrocarbonée, cyclique, linéaire ou ramifiée, contenant de 1 à 15 atomes de carbone, de préférence de 2 à 6 atomes de carbone et plus préférentiellement encore de 4 à 6 atomes de carbone, notamment 5 atomes de carbone et pouvant être notamment une chaîne méthyle, éthyle, n-propyle, iso- propyle, n-butyle, iso-butyle, sec-butyle, tert-butyle, n-pentyle, 1-méthylbutyle, 2,2- diméthylbutyle, 2-méthylpentyle, 2,2-diméthylpropyle, isopentyle, néopentyle, 2- pentyle, hexyle, 2-hexyle, 3-hexyle, 3-méthylpentyle, heptyle, octyle, nonyle, décyle, dodécyle, palmityle.
[0014] Au sens de la présente invention, on entend par « aryles » un groupement aromatique, contenant un ou plusieurs cycles aromatiques, éventuellement substitué. For the purposes of the present invention, the term "C 1 to C 15 alkyl" means a hydrocarbon chain, cyclic, linear or branched, containing from 1 to 15 carbon atoms, preferably from 2 to 6 carbon atoms and more preferably still from 4 to 6 carbon atoms, in particular 5 carbon atoms and possibly being in particular a methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, tert-butyl chain, n-pentyl, 1-methylbutyl, 2,2-dimethylbutyl, 2-methylpentyl, 2,2-dimethylpropyl, isopentyl, neopentyl, 2-pentyl, hexyl, 2-hexyl, 3-hexyl, 3-methylpentyl, heptyl, octyl, nonyl, decyl, dodecyl, palmityl. For the purposes of the present invention, the term “aryl” is understood to mean an aromatic group, containing one or more aromatic rings, optionally substituted.
[0015] Au sens de la présente invention, on entend par « hétéroaryles » un groupement aromatique, contenant un ou plusieurs cycles aromatiques, éventuellement substitué, et comportant au moins un hétéroatome différent du carbone et de l’hydrogène. [0015] For the purposes of the present invention, the term "heteroaryl" means an aromatic group, containing one or more aromatic rings, optionally substituted, and comprising at least one heteroatom other than carbon and hydrogen.
[0016] Au sens de la présente invention, on entend par « arylalkyles », un groupement aryle substitué par un ou plusieurs groupements alkyles, lesdits groupements alkyles pouvant être des groupements alkyles en Ci à C15, contenant de préférence de 1 à 15 atomes de carbone. For the purposes of the present invention, the term “arylalkyl” is understood to mean an aryl group substituted by one or more alkyl groups, said alkyl groups possibly being C1 to C15 alkyl groups, preferably containing from 1 to 15 carbon atoms. carbon.
[0017] Au sens de la présente invention, on entend par « hétéroarylalkyles », un groupement hétéroaryle substitué par un ou plusieurs groupements alkyles, lesdits groupements alkyles pouvant être des groupements alkyles en Ci à C15, contenant de préférence de 1 à 15 atomes de carbone. For the purposes of the present invention, the term “heteroarylalkyls” is understood to mean a heteroaryl group substituted by one or more alkyl groups, said alkyl groups possibly being C 1 to C 15 alkyl groups, preferably containing from 1 to 15 carbon atoms. carbon.
[0018] Selon un mode de réalisation particulier, dans la formule ci-dessus, m ou n2 sont indépendamment l’un de l’autre, égaux à 1 , 2, 3, 4 ou 5, plus préférentiellement encore 3 ou 4. [0018] According to a particular embodiment, in the above formula, m or n2 are independently of one another, equal to 1, 2, 3, 4 or 5, more preferably still 3 or 4.
[0019] Selon un autre mode de réalisation particulier, dans la formule ci-dessus n-i=n2 et est de préférence égal à 1 , 2, 3, 4 ou 5, plus préférentiellement encore 3 ou 4. According to another particular embodiment, in the above formula n-i = n2 and is preferably equal to 1, 2, 3, 4 or 5, more preferably still 3 or 4.
[0020] Selon un autre mode de réalisation particulier, la molécule est symétrique. Dans ce cas, elle comprend un seul groupe X”-R”n-Y” avec Y” qui est COOR’10 qui est porté par R9 et/ou 2 groupes X-R11-Y avec Y qui est COOR’10, l’un porté par l’un de Ri, R2, R30U Rz, de préférence l’un de Ri, R2 ou R3 et l’autre porté par l’un de R4, Rs, R6 ou Rs, de préférence l’un de R4, Rs ou R6. [0020] According to another particular embodiment, the molecule is symmetrical. In this case, it comprises a single group X ”-R” nY ”with Y” which is COOR'10 which is carried by R9 and / or 2 groups X-R11-Y with Y which is COOR'10, one carried by one of Ri, R2, R30U Rz, preferably one of Ri, R2 or R3 and the other carried by one of R4, Rs, R6 or Rs, preferably one of R4, Rs or R6.
[0021] Selon un mode de réalisation particulier, dans la formule ci-dessus, R10, et/ou R’10, peuvent être identiques. De la même façon, X, X’, et/ou X” peuvent être identiques, Y, Y’, et/ou Y”peuvent être identiques, Ru, R’11, et/ou R”n peuvent être identiques.
[0022] Les composés selon l’invention peuvent en particulier être choisis parmi les composés de formule générale suivante :
pour laquelle X” peut être O, S ou NH, correspondant alors aux formules ci- dessous :
According to a particular embodiment, in the above formula, R10, and / or R'10, may be identical. Likewise, X, X ', and / or X ”may be the same, Y, Y', and / or Y” may be the same, Ru, R'11, and / or R ”n may be the same. The compounds according to the invention can in particular be chosen from the compounds of the following general formula: for which X ”can be O, S or NH, then corresponding to the formulas below:
[0023] Les composés de formule (I) selon l’invention peuvent en particulier être choisi parmi les composés de formule (I) pour lesquels Ri, R2, R3, R4, Rs et Re ne
sont pas tous simultanément H, ce qui exclut dans ce cas les composés selon la formule (II) ci-dessous :
The compounds of formula (I) according to the invention can in particular be chosen from the compounds of formula (I) for which Ri, R2, R3, R4, Rs and Re ne are not all simultaneously H, which in this case excludes the compounds according to formula (II) below:
[0024] Le composé de formule (I) selon l’invention peut de préférence être choisi parmi les composés de formules générales suivantes : The compound of formula (I) according to the invention can preferably be chosen from the compounds of the following general formulas:
[Chem. 3]
dans lesquelles Ri, R2, R3, R4, Rs, Re, R7, Rs et R9 sont tels que définis précédemment. [Chem. 3] in which R1, R2, R3, R4, Rs, Re, R7, Rs and R9 are as defined above.
[0025] Selon un mode de réalisation particulier, les composés selon l’invention peuvent être choisis parmi les composés de formules suivantes [Chem. 5]
[Chem. 7]
dans lesquelles Ri, R2, R3, R4, Rs, Re, R7, Rs et R9 sont tels que définis précédemment. [0026] Selon un mode de réalisation préféré, les composés selon l’invention peuvent être choisis parmi les composés suivants : According to a particular embodiment, the compounds according to the invention can be chosen from the compounds of the following formulas [Chem. 5] [Chem. 7] in which R1, R2, R3, R4, Rs, Re, R7, Rs and R9 are as defined above. According to a preferred embodiment, the compounds according to the invention can be chosen from the following compounds:
[Chem. 8]
[Chem. 12]
[Chem. 16]
[Chem. 8] [Chem. 12] [Chem. 16]
[0027] Le second objet de la présente invention concerne le procédé de préparation des composés de formule (I) selon l’invention comportant une étape de réaction entre : The second object of the present invention relates to the process for preparing the compounds of formula (I) according to the invention comprising a reaction step between:
[Chem. 17]
et [Chem. 18]
[Chem. 17] and [Chem. 18]
[0028] Cette réaction est effectuée de préférence par chauffage au reflux en présence d’acétate de sodium dans un mélange d’acide acétique et d’anhydride acétique. [0029] Le troisième objet de l’invention concerne le procédé de marquage de tissu tumoral avec un des composés selon l’invention ou préparé selon le procédé de l’invention. This reaction is preferably carried out by heating under reflux in the presence of sodium acetate in a mixture of acetic acid and acetic anhydride. The third object of the invention relates to the method of labeling tumor tissue with one of the compounds according to the invention or prepared according to the method of the invention.
[0030] Au sens de la présente invention, on entend par « tissu tumoral » le tissu constitué de cellules tumorales qui sont des cellules prolifératives anormales, et
d’un tissu de soutien, aussi appelé stroma tumoral ou tissu interstitiel, fait de cellules et de substance extra-cellulaire dans laquelle est située la vascularisation tumorale. For the purposes of the present invention, the term “tumor tissue” is understood to mean the tissue consisting of tumor cells which are abnormal proliferative cells, and supporting tissue, also called tumor stroma or interstitial tissue, made up of cells and extra-cellular material in which the tumor vasculature is located.
[0031] Les composés fluorescents selon l’invention ont la particularité après leur diffusion dans l’organisme d’être piégés dans le tissu tumoral, alors qu’ils sont éliminés des tissus sains. Cette particularité permet d’utiliser ces composés fluorescents directement, sans couplage préalable à une autre molécule de marquage, rendant ainsi leur utilisation plus simple, plus rapide et plus efficace que celle des composés de l’art antérieur. Il a été observé que cette élimination des tissus sains est croissante au cours du temps. En général, entre 24 et 72 heures, de préférence entre 36 et 60 heures et plus préférentiellement 48 heures après l’administration de ces composés, leur élimination des tissus sains est totale. Ils restent cependant piégés dans les tissus tumoraux. Cette propriété permet d’avoir une nette différenciation des tissus tumoraux par rapport aux tissus sains et ainsi l’utilisation de ces composés dans des applications de surveillance, diagnostic et/ou d’assistance à la chirurgie dans un contexte de maladies cancéreuses. Cette différenciation perdure pendant 6 à 48 heures, de préférence 12 à 36 heures, permettant de programmer le diagnostic ou la chirurgie de manière ciblée. [0031] The fluorescent compounds according to the invention have the particularity, after their diffusion in the body, of being trapped in the tumor tissue, while they are eliminated from healthy tissue. This feature makes it possible to use these fluorescent compounds directly, without prior coupling to another labeling molecule, thus making their use simpler, faster and more efficient than that of the compounds of the prior art. It has been observed that this elimination from healthy tissue increases over time. In general, between 24 and 72 hours, preferably between 36 and 60 hours and more preferably 48 hours after the administration of these compounds, their elimination from healthy tissues is complete. However, they remain trapped in tumor tissue. This property makes it possible to have a clear differentiation of tumor tissues from healthy tissues and thus the use of these compounds in monitoring, diagnostic and / or surgical assistance applications in the context of cancerous diseases. This differentiation lasts for 6 to 48 hours, preferably 12 to 36 hours, making it possible to program the diagnosis or the surgery in a targeted manner.
[0032] Les composés selon l’invention peuvent ainsi notamment être utilisés dans le cadre de cancers, par exemple hormonodépendants, tels que le cancer du sein ou de cancers digestifs, tels que le cancer du pancréas. En effet, dans le cancer du pancréas, les tumeurs sont particulièrement difficiles à éliminer dans leur globalité par chirurgie car elles ne sont pas facilement délimitées. L’utilisation des composés selon l’invention permet d’obtenir une meilleure visualisation des contours des tumeurs grâce à la différenciation de marquage entre le tissu tumoral et le tissu sain, et ainsi une résection tumorale plus efficace par chirurgie. The compounds according to the invention can thus in particular be used in the context of cancers, for example hormone-dependent, such as breast cancer or digestive cancers, such as pancreatic cancer. In fact, in pancreatic cancer, tumors are particularly difficult to remove in their entirety by surgery because they are not easily delimited. The use of the compounds according to the invention makes it possible to obtain a better visualization of the contours of the tumors thanks to the differentiation of labeling between the tumor tissue and the healthy tissue, and thus a more efficient tumor resection by surgery.
[0033] L’invention porte également sur l’utilisation d’un des composés selon l’invention ou préparé selon le procédé de l’invention dans une méthode de marquage de tissu tumoral. The invention also relates to the use of one of the compounds according to the invention or prepared according to the method of the invention in a method of labeling tumor tissue.
[0034] Cette méthode de marquage des tissus nécessite l’administration des composés par voie intraveineuse, intra-artérielle, ou dans un autre vaisseau,
notamment un vaisseau lymphatique, soit en injection locale, soit en application locale, préférentiellement par voie intraveineuse. This method of labeling tissues requires the administration of the compounds intravenously, intraarterially, or into another vessel, in particular a lymphatic vessel, either by local injection or by local application, preferably intravenously.
[0035] Un autre objet de l’invention concerne une composition comprenant un des composés selon l’invention ou préparé selon le procédé de l’invention et au moins un adjuvant pharmaceutiquement acceptable. Another object of the invention relates to a composition comprising one of the compounds according to the invention or prepared according to the process of the invention and at least one pharmaceutically acceptable adjuvant.
[0036] L’invention porte également sur l’un des composés selon l’invention ou préparé selon le procédé de l’invention ou une composition comprenant un des composés selon l’invention ou préparé selon le procédé de l’invention pour son utilisation dans une méthode de marquage et/ou de détection de tissu tumoral, et/ou dans le traitement chirurgical de tumeurs. The invention also relates to one of the compounds according to the invention or prepared according to the process of the invention or a composition comprising one of the compounds according to the invention or prepared according to the process of the invention for its use. in a method of labeling and / or detecting tumor tissue, and / or in the surgical treatment of tumors.
[0037] L’invention concerne également une méthode de détection de tissu tumoral comprenant une étape de marquage de tissu tumoral avec un des composés selon l’invention ou préparé selon le procédé de l’invention, et une étape de détection par imagerie médicale en fluorescence ou spectrométrie de fluorescence. The invention also relates to a method of detecting tumor tissue comprising a step of labeling tumor tissue with one of the compounds according to the invention or prepared according to the method of the invention, and a step of detection by medical imaging in fluorescence or fluorescence spectrometry.
Figures Figures
Fig. 1 Fig. 1
[0038] [Fig. 1] montre les valeurs médianes et écarts-types des rapports d’intensité tumeur/abdomen en fonction des temps post-injection du composé 2 (CJ215) et de l’ICG. [0038] [Fig. 1] shows the median values and standard deviations of the tumor / abdomen intensity ratios as a function of post-injection times of Compound 2 (CJ215) and ICG.
Fig. 2 Fig. 2
[0039] [Fig. 2] montre les résultats d’imagerie ex vivo de tumeurs pancréatiques après injection de deux composés selon l’invention et d’un agent fluorescent de l’art antérieur (ICG). Exemples [0039] [Fig. 2] shows the results of ex vivo imaging of pancreatic tumors after injection of two compounds according to the invention and a fluorescent agent of the prior art (ICG). Examples
[0040] Exemple 1 : [Chem. 19]
Composé (1) [0040] Example 1: [Chem. 19] Compound (1)
Un mélange de 4-[(5-carboxypentyl)oxy]-6-sulfo-1-(4-sulfobutyl)-2,3,3-triméthyl- benz(e)indolium (sel interne et de disodium) (9 g ; 15 mmol), 2-chloro-1-formyl-3- (hydroxyméthylene)-l-cyclohexene (1 ,30 g ; 7,50 mmol), et acétate de sodium (3g ; 36,6 mmol) dans un mélange acide acétique et anhydride acétique 60/30 est chauffé au reflux pendant 10 minutes. Le milieu réactionnel est refroidi à température ambiante et le précipité est séparé par filtration et lavé avec de l’éther diéthylique pour fournir 4,33 g (rendement : 43,9 %) d’un solide vert. Le produit brut est purifié par chromatographie flash sur colonne (phase inverse gel de silice C18, acétonitrile 0-25% / eau). A mixture of 4 - [(5-carboxypentyl) oxy] -6-sulfo-1- (4-sulfobutyl) -2,3,3-trimethyl-benz (e) indolium (internal and disodium salt) (9 g; 15 mmol), 2-chloro-1-formyl-3- (hydroxymethylene) -l-cyclohexene (1.30 g; 7.50 mmol), and sodium acetate (3g; 36.6 mmol) in a mixture of acetic acid and 60/30 acetic anhydride is heated at reflux for 10 minutes. The reaction medium is cooled to room temperature and the precipitate is separated by filtration and washed with diethyl ether to provide 4.33 g (yield: 43.9%) of a green solid. The crude product is purified by flash column chromatography (reverse phase C18 silica gel, 0-25% acetonitrile / water).
[0041] Exemple 2 : [0041] Example 2:
[Chem. 20]
Composé (2) [Chem. 20] Compound (2)
Au composé (1) (1 g ; 0,76 mmol) en solution dans 500 mL de méthanol, est ajouté du méthylate de sodium (440 mg; 7,6 mmol) . Le milieu réactionnel est chauffé au reflux pendant 16 h et concentré sous vide, puis filtré. Le résidu obtenu est lavé avec du méthanol froid et de l’acétone et séché sous vide pour fournir 450 mg d’un solide vert (rendement : 45%). Le produit brut est purifié par chromatographie flash sur colonne (phase inverse gel de silice C18, acétonitrile 0- 25% / eau). To compound (1) (1 g; 0.76 mmol) dissolved in 500 mL of methanol, is added sodium methoxide (440 mg; 7.6 mmol). The reaction medium is heated at reflux for 16 h and concentrated in vacuo, then filtered. The residue obtained is washed with cold methanol and acetone and dried in vacuo to provide 450 mg of a green solid (yield: 45%). The crude product is purified by flash column chromatography (reverse phase C18 silica gel, 0-25% acetonitrile / water).
[0042] Exemple 3 : [0042] Example 3:
[Chem. 21]
Composé (3) [Chem. 21] Compound (3)
Au composé (1) (400 mg; 0,30 mmol)) en solution dans 20 mL d’un mélange 50/50 méthanol/ NMP (N-méthyl-2-pyrrolidone) est ajouté MeSNa (106 mg; 1 ,5 mmol). Le milieu réactionnel est chauffé au reflux pendant 4 h, puis du diéthyléther (20 mL) est additionné au mélange. Le précipité est filtré et lavé avec le même solvant pour fournir 254 mg de produit brut (rendement : 61% ; odeur de soufre). Le produit brut est purifié par chromatographie flash sur colonne (phase inverse gel de silice C18, acétonitrile 0-25% / eau). [0043] Exemple 4 : To compound (1) (400 mg; 0.30 mmol)) dissolved in 20 mL of a 50/50 methanol / NMP (N-methyl-2-pyrrolidone) mixture is added MeSNa (106 mg; 1.5 mmol) ). The reaction medium is heated at reflux for 4 h, then diethyl ether (20 mL) is added to the mixture. The precipitate is filtered off and washed with the same solvent to provide 254 mg of crude product (yield: 61%; smell of sulfur). The crude product is purified by flash column chromatography (reverse phase C18 silica gel, 0-25% acetonitrile / water). [0043] Example 4:
[Chem. 22]
[Chem. 22]
Composé (4) Compound (4)
Un mélange de 6-sulfo-1-(4-sulfobutyl)-2,3,3-triméthylbenz(e)indolium (sel interne et de DCHA) (2 g ; 4,7 mmol), 2-chloro-1-formyl-3-(hydroxyméthylene)-1- cyclohexene (0,40 g ; 2,35 mmol), et acétate de sodium (0,9 g ; 11 mmol) dans un mélange acide acétique et anhydride acétique 50/20 est chauffé au reflux pendant 15 minutes. Le précipité est séparé par filtration, lavé avec de l’éthanol et de l’acétone et séché sous vide pour fournir 1 ,6 g (rendement : 63,8 %) d’une poudre de couleur brique. Le produit brut est purifié par chromatographie flash sur colonne (phase inverse gel de silice C18, acétonitrile 0-25% / eau). A mixture of 6-sulfo-1- (4-sulfobutyl) -2,3,3-trimethylbenz (e) indolium (internal salt and DCHA) (2 g; 4.7 mmol), 2-chloro-1-formyl -3- (hydroxymethylene) -1- cyclohexene (0.40 g; 2.35 mmol), and sodium acetate (0.9 g; 11 mmol) in a mixture of acetic acid and acetic anhydride 50/20 is heated to reflux for 15 minutes. The precipitate is filtered off, washed with ethanol and acetone, and dried in vacuo to provide 1.6 g (yield: 63.8%) of a brick-colored powder. The crude product is purified by flash column chromatography (reverse phase C18 silica gel, 0-25% acetonitrile / water).
[0044] Exemple 5 : [0044] Example 5:
[Chem. 23]
Composé (5) [Chem. 23] Compound (5)
A de l’acide 3-(4-hydroxyphényl)propionique (660 mg ; 4 mmol) sont ajoutés 8 mL of KOH 1 M méthanolique, 16 mL de DMSO et le composé (4) (500 mg ; 0,25 mmol). Le milieu réactionnel est agité à température ambiante pendant 8h, puis 150 mL d’acétate d’éthyle sont ajoutés goutte-à-goutte. Le précipité est séparé par filtration, lavé avec de l’éthanol et de l’acétone et séché sous vide pour fournir 260 mg (rendement : 45 %) d’une poudre verte. Le produit brut est purifié par chromatographie flash sur colonne (phase inverse gel de silice C18, acétonitrile 0- 25% / eau). To 3- (4-hydroxyphenyl) propionic acid (660 mg; 4 mmol) are added 8 mL of 1 M methanolic KOH, 16 mL of DMSO and compound (4) (500 mg; 0.25 mmol). The reaction medium is stirred at room temperature for 8 hours, then 150 mL of ethyl acetate are added dropwise. The precipitate is filtered off, washed with ethanol and acetone and dried in vacuo to provide 260 mg (yield: 45%) of a green powder. The crude product is purified by flash column chromatography (reverse phase C18 silica gel, 0-25% acetonitrile / water).
[0045] Exemple 6 : [0045] Example 6:
[Chem. 24]
[Chem. 24]
Composé (6) Au compose (4) (520 g ; 0,49 mmol) sont ajoutés de l’acide 4- aminohydrocinnamique (816 mg ; 4,9 mmol), 25 mL de DMSO et de la triéthylamine (500 mg ; 4,9 mmol). Le milieu réactionnel est agité à température ambiante pendant 8h, puis 200 mL d’acétone sont ajoutés goutte-à-goutte. Le précipité est séparé par filtration, lavé avec de l’acétone et séché sous vide pour fournir 430 mg (rendement : 74%) d’une poudre rouge. Le produit brut est purifié par chromatographie flash sur colonne (phase inverse gel de silice C18, acétonitrile 0-25% / eau).
[0046] Exemple 7 : [Chem. 25]
Compound (6) To compound (4) (520 g; 0.49 mmol) are added 4-aminohydrocinnamic acid (816 mg; 4.9 mmol), 25 mL of DMSO and triethylamine (500 mg; 4 , 9 mmol). The reaction medium is stirred at room temperature for 8 h, then 200 ml of acetone are added dropwise. The precipitate is filtered off, washed with acetone and dried in vacuo to provide 430 mg (yield: 74%) of a red powder. The crude product is purified by flash column chromatography (reverse phase C18 silica gel, 0-25% acetonitrile / water). Example 7: [Chem. 25]
Composé (7) A de l’acide 4-mercaptohydrocinnamique (91 mg ; 0,5 mmol) sont ajoutés 1 mL of KOH 1 M méthanolique, 16 mL de DMSO et le composé (4) (500 mg ; 0,25 mmol). Le milieu réactionnel est agité à température ambiante pendant 30 minutes, puis 50 mL d’acétate d’éthyle sont ajoutés goutte-à-goutte. Le précipité est séparé par filtration, lavé avec de l’éthanol et de l’acétone et séché sous vide pour fournir 310 mg (rendement : 54 %) d’une poudre verte. Le produit brut est purifié par chromatographie flash sur colonne (phase inverse gel de silice C18, acétonitrile 0- 25% / eau). Compound (7) A of 4-mercaptohydrocinnamic acid (91 mg; 0.5 mmol) are added 1 mL of 1 M methanolic KOH, 16 mL of DMSO and compound (4) (500 mg; 0.25 mmol) . The reaction medium is stirred at room temperature for 30 minutes, then 50 mL of ethyl acetate are added dropwise. The precipitate is filtered off, washed with ethanol and acetone and dried in vacuo to provide 310 mg (yield: 54%) of a green powder. The crude product is purified by flash column chromatography (reverse phase C18 silica gel, 0-25% acetonitrile / water).
[0047] Exemple 8 : Comparaison d’un composé selon l’invention et d’un agent fluorescent de l’art antérieur (ICG) pour l’imagerie in vivo des tumeurs mammaires L’ICG (ou vert d’indocyanine / Infracyanine) est un agent fluorescent de l’art antérieur, déjà approuvé pour l’utilisation chez l’homme pour l’évaluation des fonctions cardiaques et hépatiques, ainsi qu’en ophtalmologie, pour les pathologies rétiniennes. Il est également en évaluation dans de nombreux essais cliniques à travers le monde pour le guidage du geste chirurgical lors des exérèses tumorales, ou la cartographie des ganglions drainant les tumeurs, par imagerie proche infra-rouge. Example 8: Comparison of a compound according to the invention and of a fluorescent agent of the prior art (ICG) for in vivo imaging of mammary tumors ICG (or indocyanine green / Infracyanine) is a fluorescent agent of the prior art, already approved for use in humans for the evaluation of cardiac and hepatic functions, as well as in ophthalmology, for retinal pathologies. It is also being evaluated in numerous clinical trials around the world for guiding the surgical procedure during tumor resection, or the mapping of lymph nodes draining tumors, by near infrared imaging.
L’ICG a été comparé au composé (2) selon l’invention dont la synthèse est décrite à l’exemple 2 précédent, ce composé est dénommé CJ215 dans cette étude. L’étude a inclus 30 souris au total réparties en trois groupes. Les greffes tumorales ont toutes été réalisées avec 50 000 cellules 4T1-Dendra2 /20 pi injectées dans 2 glandes mammaires en contralatéral pour chacune des souris.
[0048] Les injections des biomarqueurs (composé 2 dénommé CJ215 dans cette étude et ICG) ont été réalisées à J9 post-greffe tumorale (afin de limiter l’apparition de nécrose dans les tumeurs). The ICG was compared with the compound (2) according to the invention, the synthesis of which is described in the preceding example 2, this compound is called CJ215 in this study. The study included a total of 30 mice divided into three groups. The tumor grafts were all carried out with 50,000 4T1-Dendra2 / 20 μl cells injected into 2 mammary glands contralateral for each of the mice. The injections of biomarkers (compound 2 called CJ215 in this study and ICG) were carried out on D9 post-tumor grafting (in order to limit the appearance of necrosis in tumors).
L’évolution de l’intensité des signaux de fluorescence relevée pour chacun des biomarqueurs au cours du temps a été évaluée par image microscopique. La capacité des deux marqueurs à produire un signal spécifiquement localisé à la tumeur a été appréciée quantitativement par le calcul du rapport du signal spécifique lié à la tumeur au signal non spécifique dans les tissus environnants. The change in the intensity of the fluorescence signals observed for each of the biomarkers over time was evaluated by microscopic image. The ability of the two markers to produce a signal specifically localized to the tumor was assessed quantitatively by calculating the ratio of the specific signal bound to the tumor to the non-specific signal in the surrounding tissues.
[0049] Le protocole d’imagerie a été réalisé aux temps 2h, 24h, 48h, 4 et 6 jours post-injection pour toutes les souris. Toutes les images réalisées à chaque temps d’acquisition, ont été acquises sur imageur I VIS Spectrum (Perkin Elmer) avec les paramètres suivants : The imaging protocol was carried out at times 2h, 24h, 48h, 4 and 6 days post-injection for all the mice. All the images produced at each acquisition time were acquired on an I VIS Spectrum imager (Perkin Elmer) with the following parameters:
Pour la détection de la forme GFP de la Dendra2 (détection de la tumeur) : For detection of the GFP form of Dendra2 (tumor detection):
- Excitation à 465 nm - Excitation at 465 nm
- Emission entre 520 et 580 nm Pour la détection des biomarqueurs : - Emission between 520 and 580 nm For the detection of biomarkers:
- Excitation à 745 nm - Excitation at 745 nm
- Emission entre 800 et 840 nm - Emission between 800 and 840 nm
Les mesures quantitatives ont été réalisées sur les images brutes non déconvoluées. Pour les deux fluorophores, la durée d’acquisition a été paramétrée en mode automatique. Dans ce mode, le système détermine le temps d’acquisition nécessaire pour atteindre la valeur cible imposée (6000 counts) dans le temps imparti (fixé à 2 min). The quantitative measurements were carried out on the non-deconvoluted raw images. For the two fluorophores, the acquisition time was set in automatic mode. In this mode, the system determines the acquisition time necessary to reach the imposed target value (6000 counts) within the allotted time (fixed at 2 min).
[0050] La Figure 1 rapporte les valeurs médianes et écarts-types des rapports d’intensité tumeur/abdomen en fonction des temps post-injection du CJ215 et de l’ICG. [0050] Figure 1 reports the median values and standard deviations of the tumor / abdomen intensity ratios as a function of post-injection times of CJ215 and ICG.
La mesure du ratio des intensités tumeur/abdomen illustrée sur cette figure permet de montrer que : The measurement of the ratio of the tumor / abdomen intensities illustrated in this figure makes it possible to show that:
- les rapports d’intensité sont significativement plus élevés pour le composé 2 (CJ215) comparativement à l’ICG, quel que soit le temps post-injection (de 1 ,5 fois
à 2h à plus de 3 fois à J+6) ce qui traduit une capacité à identifier un signal spécifique dans les tumeurs de façon plus précoce et plus spécifique avec le composé 2 (CJ215). Ces résultats indiquent également la possibilité d’améliorer très significativement la spécificité du signal à la tumeur en augmentant le délai entre l’injection du composé 2 (CJ215) et l’imagerie ; - the intensity ratios are significantly higher for compound 2 (CJ215) compared to ICG, regardless of the post-injection time (1.5 times at 2 h to more than 3 times at D + 6) which reflects an ability to identify a specific signal in tumors earlier and more specifically with compound 2 (CJ215). These results also indicate the possibility of very significantly improving the specificity of the signal to the tumor by increasing the time between the injection of compound 2 (CJ215) and imaging;
- le rapport signal sur bruit augmente continuellement pour le composé 2 (CJ215) jusqu’à 6 jours post-injection, dernier jour d’examen considéré dans ce protocole. A ce stade, l’ICG n’est plus observable dans les tumeurs (ceci dès 48h). La grande stabilité du signal intratumoral du composé 2 (CJ215), comparativement aux tissus environnants qui éliminent le produit, offre ainsi une meilleure capacité à identifier les tumeurs et contribue ainsi à améliorer significativement la délimitation fine des marges tumorales, qui reste problématique avec l’ICG. - the signal to noise ratio increases continuously for compound 2 (CJ215) up to 6 days post-injection, the last examination day considered in this protocol. At this stage, ICG is no longer observable in tumors (this from 48 hours). The high stability of the intratumoral signal of compound 2 (CJ215), compared to the surrounding tissues which eliminate the product, thus offers a better capacity to identify tumors and thus contributes to significantly improve the fine delineation of tumor margins, which remains problematic with the ICG.
[0051] Exemple 9 : Comparaison de deux composés selon l’invention et d’un agent fluorescent de l’art antérieur (ICG) en imagerie ex vivo de tumeurs pancréatigues. [0051] Example 9: Comparison of two compounds according to the invention and of a fluorescent agent of the prior art (ICG) in ex vivo imaging of pancreatic tumors.
Un modèle d'adénocarcinome pancréatique orthotopique chez la souris a été développé. Les cellules tumorales ont été amplifiées par voie sous-cutanée chez des souris SCID et les fragments résultants ont ensuite été implantés chirurgicalement dans le pancréas de souris nues BALB/c irradiées. A model of orthotopic pancreatic adenocarcinoma in mice has been developed. Tumor cells were amplified subcutaneously in SCID mice and the resulting fragments were then surgically implanted into the pancreas of irradiated BALB / c nude mice.
Le développement de la tumeur a été suivi in vivo par IRM (4.7T, PharmaScan, Bruker Biospin) à trois moments, D14, 28 et 36. Les animaux ont été soumis à une faible fluorescence afin de minimiser l'autofluorescence. L'imagerie fluorescente a été réalisée avec une caméra à dispositif à couplage de charge (CCD) (PhotonRT, BiospaceLab) avec une excitation à 700 nm et un filtre d'émission à 770 nm. Tumor development was followed in vivo by MRI (4.7T, PharmaScan, Bruker Biospin) at three time points, D14, 28 and 36. Animals were subjected to weak fluorescence in order to minimize autofluorescence. Fluorescent imaging was performed with a charge coupled device (CCD) camera (PhotonRT, BiospaceLab) with excitation at 700 nm and emission filter at 770 nm.
À l'issue de séances d'imagerie in vivo réalisées à 2h, 48h et 164h, des images fluorescentes ex vivo ont été acquises. Les composés fluorescents selon l’invention 2 (CJ215) et CJ319 (dont la structure est détaillée ci-dessous) ont été injectés par voie intraveineuse à 2 mg/kg, 39 jours après l'implantation des fragments de tumeur, alors que les volumes moyens des tumeurs étaient d'environ 70 mm3. Le vert d'indocyanine (ICG), un colorant largement utilisé dans l’imagerie per-opératoire des tumeurs, a été inclus comme témoin. At the end of in vivo imaging sessions carried out at 2h, 48h and 164h, ex vivo fluorescent images were acquired. The fluorescent compounds according to the invention 2 (CJ215) and CJ319 (the structure of which is detailed below) were injected intravenously at 2 mg / kg, 39 days after implantation of the tumor fragments, while the volumes average tumors were about 70 mm 3 . Indocyanine green (ICG), a dye widely used in intraoperative tumor imaging, was included as a control.
[Chem. 26]
(CJ319) [Chem. 26] (CJ319)
[0052] L'imagerie de fluorescence ex vivo décrite sur la Figure 2 a montré que 2 heures après l'injection, les deux composés fluorescents selon l’invention étaient présents dans le pancréas et la tumeur en quantités à peu près équivalentes. Cependant, 48 heures après l'injection, une distribution préférentielle claire a été observée pour la tumeur, les deux composés produisant un signal fluorescent environ quatre fois plus élevé dans la tumeur que dans le tissu pancréatique environnant. Cet effet a persisté six jours après l'injection, bien que le signal ait diminué au fil du temps. En comparaison, le vert d'indocyanine n'a montré aucune accumulation spécifique dans le pancréas ou la tumeur. The ex vivo fluorescence imaging described in Figure 2 showed that 2 hours after injection, the two fluorescent compounds according to the invention were present in the pancreas and the tumor in approximately equivalent amounts. However, 48 hours after injection, a clear preferential distribution was observed for the tumor, with both compounds producing an approximately four-fold higher fluorescent signal in the tumor than in the surrounding pancreatic tissue. This effect persisted for six days after injection, although the signal waned over time. In comparison, indocyanine green did not show any specific accumulation in the pancreas or tumor.
Ces résultats montrent la supériorité des composés de l’invention par rapport à un agent fluorescent de l’art antérieur au niveau de leur distribution spécifique dans un tissu tumoral.
These results show the superiority of the compounds of the invention over a fluorescent agent of the prior art in terms of their specific distribution in tumor tissue.
Claims
Revendications Claims
[Revendication 1] Composé de formule (I) [Chem. 27]
dans laquelle ni et n2 sont chacun un entier de 0 à 15, [Claim 1] Compound of formula (I) [Chem. 27] where ni and n2 are each an integer from 0 to 15,
Ri, R2, R3, R4, R5 et R6 sont chacun indépendamment choisis parmi H, OH, SH, NH2, SO3R10 et X-R11-Y, R10 et R’10 étant indépendamment H, Na ou K, Ri, R2, R3, R4, R5 and R6 are each independently selected from H, OH, SH, NH 2 , SO3R10 and X-R11-Y, R10 and R'10 being independently H, Na or K,
X, X’ et X” étant indépendamment O, S ou NH, X, X ’and X” being independently O, S or NH,
R11, R’11 et R”n étant indépendamment choisis parmi alkyle en Ci à C15, aryle, hétéroaryle, (alkyl en Ci à Cis)aryle, (alkyl en Ci à Ci5)hétéroaryle, aryl(alkyle en Ci à C15) et hétéroaryl(alkyle en Ci à C15) ; Y, Y’ et Y” étant indépendamment choisis parmi H, halogène, COOR’10 ou amide ; R7 et Re étant chacun indépendamment choisis parmi H, OH, SH, NH2, alkyle en Ci à C15, et X’-R’n-Y’; R11, R'11 and R ”n being independently selected from C1 to C15 alkyl, aryl, heteroaryl, (C1 to Cis alkyl) aryl, (C1 to C15 alkyl) heteroaryl, aryl (C1 to C15 alkyl) and heteroaryl (C1-C15 alkyl); Y, Y "and Y" being independently selected from H, halogen, COOR'10 or amide; R7 and Re being each independently selected from H, OH, SH, NH2, C1-C15 alkyl, and X'-R'n-Y ";
Rg étant choisi parmi H, OH, SH, NH2 et X”-R”n-Y”, ledit composé comportant au moins un groupement X-R11-Y, X’-R’n-Y’ ou X”-R”n- Y” avec Y, Y’ et/ou Y” qui est COOR’10. Rg being chosen from H, OH, SH, NH 2 and X ”-R” nY ”, said compound comprising at least one X-R11-Y, X'-R'n-Y 'or X” -R ”n group - Y ”with Y, Y 'and / or Y” which is COOR'10.
[Revendication 2] Composé selon la revendication 1 pour lequel Ri, R2, R3, R4, R5 et R6 ne sont pas tous simultanément H. [Claim 2] A compound according to claim 1 wherein R1, R2, R3, R4, R5 and R6 are not all simultaneously H.
[Revendication 3] Composé selon la revendication 1 ou 2 choisi parmi les composés de formules suivantes : [Chem. 28]
dans lesquelles R-i, R2, R3, R4, Rs, Re, R7, Rs et R9 sont tels que définis à la revendication 1. [Claim 3] A compound according to claim 1 or 2 chosen from the compounds of the following formulas: [Chem. 28] wherein R1, R2, R3, R4, Rs, Re, R7, Rs and R9 are as defined in claim 1.
[Revendication 4] Composé selon l’une des revendications précédentes choisi parmi les composés de formules suivantes [Chem. 30]
[Claim 4] A compound according to one of the preceding claims chosen from the compounds of the following formulas [Chem. 30]
[Chem. 31]
dans lesquelles R-i, R2, R3, R4, Rs, Re, R7, Rs et R9 sont tels que définis à la revendication 1. [Chem. 31] wherein R1, R2, R3, R4, Rs, Re, R7, Rs and R9 are as defined in claim 1.
[Revendication 5] Composé selon l’une des revendications précédentes choisi parmi les composés de formules suivantes [Chem. 33]
[Claim 5] Compound according to one of the preceding claims chosen from the compounds of the following formulas [Chem. 33]
[Chem. 34]
[Chem. 36]
[Chem. 38]
[Chem. 41]
[Chem. 34] [Chem. 36] [Chem. 38] [Chem. 41]
[Revendication 6] Procédé de préparation des composés selon l’une des revendications 1 à 5 comportant une étape de réaction entre : [Chem. 42]
et [Claim 6] Process for preparing the compounds according to one of claims 1 to 5 comprising a reaction step between: [Chem. 42] and
[Chem. 43]
[Revendication 7] Procédé de marquage de tissu tumoral avec un des composés selon l’une des revendications 1 à 5 ou préparé selon la revendication 6. [Chem. 43] [Claim 7] A method of labeling tumor tissue with one of the compounds according to one of claims 1 to 5 or prepared according to claim 6.
[Revendication 8] Utilisation d’un des composés selon l’une des revendications 1 à 5 ou préparé selon la revendication 6 dans une méthode de marquage de tissu tumoral. [Revendication 9] Composé selon l’une des revendications 1 à 5 ou préparé selon la revendication 6 ou composition comprenant un composé selon l’une des revendications 1 à 5 ou préparé selon la revendication 6 pour son utilisation dans une méthode de marquage et/ou de détection de tissu tumoral, et/ou dans le traitement chirurgical de tumeurs.
[Revendication 10] Méthode de détection de tissu tumoral comprenant une étape de marquage de tissu tumoral avec un des composés selon l’une des revendications 1 à 5 ou préparé selon la revendication 6, et une étape de détection par imagerie médicale en fluorescence ou spectrométrie de fluorescence.
[Claim 8] Use of one of the compounds according to one of claims 1 to 5 or prepared according to claim 6 in a method of labeling tumor tissue. [Claim 9] A compound according to one of claims 1 to 5 or prepared according to claim 6 or a composition comprising a compound according to one of claims 1 to 5 or prepared according to claim 6 for its use in a labeling method and / or tumor tissue detection, and / or in the surgical treatment of tumors. [Claim 10] Method for detecting tumor tissue comprising a step of labeling tumor tissue with one of the compounds according to one of claims 1 to 5 or prepared according to claim 6, and a step of detection by medical imaging in fluorescence or spectrometry fluorescence.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| FR2004868A FR3110165B1 (en) | 2020-05-15 | 2020-05-15 | New fluorescent compounds for tumor tissue labeling |
| PCT/FR2021/050832 WO2021229188A1 (en) | 2020-05-15 | 2021-05-12 | Novel fluorescent compounds for labelling tumour tissue |
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| JP (1) | JP2023525601A (en) |
| KR (1) | KR20230010713A (en) |
| CN (1) | CN115605459A (en) |
| BR (1) | BR112022023154A2 (en) |
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| CN103911017B (en) * | 2012-12-28 | 2017-09-15 | 浙江海正药业股份有限公司 | Cyanine dye compound and preparation method thereof, for dual-use function agent of photodynamic therapy and preparation method thereof |
-
2020
- 2020-05-15 FR FR2004868A patent/FR3110165B1/en active Active
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- 2021-05-12 US US17/925,434 patent/US20230203014A1/en active Pending
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| CN115605459A (en) | 2023-01-13 |
| WO2021229188A1 (en) | 2021-11-18 |
| US20230203014A1 (en) | 2023-06-29 |
| FR3110165A1 (en) | 2021-11-19 |
| JP2023525601A (en) | 2023-06-16 |
| FR3110165B1 (en) | 2022-10-28 |
| BR112022023154A2 (en) | 2023-02-07 |
| CA3178232A1 (en) | 2021-11-18 |
| KR20230010713A (en) | 2023-01-19 |
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