Luminescent dyes having a narrow range of emission wavelengths, compositions containing the same and methods of making and using the same

Abstract

To provide a chlorin derivative having a narrow emission band.SOLUTION: The compound is represented by formula (I). (M: metal, H). R5, R10, R15:H, alkoxy, etc. R2, R3, R12, R13: H, cyanos, carboxylic acids, formyls, acetyls, etc. ) In some embodiments, the chlorin derivative is PEGylated. In some embodiments, the chlorin derivative has high water solubility (e.g., 10mg / mL or more). The chlorin derivatives can include bioconjugatable groups to form conjugates with, for example, antibodies and nanoparticles.SELECTED DRAWING: Figure 1

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This invention claims priority to U.S. Provisional Patent Application No. 62 / 850,442, filed May 20, 2019. and is incorporated herein by reference in its entirety. [Technical Field]

[0002] This invention relates generally to chlorin derivatives, and in some embodiments, to narrow scope The present invention relates to water-soluble chlorin derivatives having long emission wavelengths, and further to their conjugates and their chlorin derivatives. The present invention relates to methods for making and using phosphorus derivatives and conjugates thereof. [Background technology]

[0003] Fluorescent dyes that are water-soluble and suitable for conjugation with other substances ranging from nanoparticles to biological targeting agents There has been a huge expansion of applications requiring this. These include imaging, sensing, and photodynamic therapy of living cells, cells, and whole organisms. Typically, fluoresceins have absorption in the red (600-700 nm) spectral region, Chlorin molecules are one of the few chromophores available for photochemical studies, making them ideal for these applications. There is particular interest in The success of the above applications is due to (1) significant solubility in aqueous saline, which allows for intermolecular aggregation; (2) Non-specific binding to cellular components is minimized. (3) incorporating a single reactive group for conjugation, thereby producing the product and (4) providing sufficient quantities for the experiment. However, the large solubility of chlorins is a major factor in determining their synthesis. Aqueous surfaces present a challenge to water solubility. Summary of the Invention [Problem to be solved by the invention]

[0004] Therefore, there is a continuing need to provide new chlorin derivatives. , those with improved water solubility (e.g., water solubility greater than 1 mg / mL), especially and / or have a narrow range of absorption and emission wavelengths. These include, but are not limited to, those characterized by emission bands.

[0005] It should be noted that this summary lists several embodiments of the invention and in many cases, This summary merely lists many different embodiments. In the embodiments listed, one or more representative features may be mentioned. , which are also exemplary. Such embodiments generally have the same advantages as those described above. Similarly, these characteristics are not listed in this overview. This is applicable to other embodiments of the present invention, whether or not they are excessively repetitive. To avoid this, this summary does not list or attempt to list all possible combinations of such features. I'm not suggesting it. [Means for solving the problem]

[0006] In some embodiments, the present invention provides a compound represented by formula (I): [ka] wherein M is a metal or two hydrogen atoms; R5, R 10 and R 15 are each independently a hydrogen atom, an alkoxy group, or a group represented by the following formula: a linker group selected from the group consisting of: -L1-(X1-L2) p -G wherein p is 0 or 1, L1 is alkylidene, and X1 is -C(=O)NH- or -NHC(=O)- and L2 is -(CH2CH2O) q -alkylene- (wherein q is 1 to 24), alkylene, or substituted alkylene (optionally, the substituted alkylene The alkylene is bound to one or more groups containing a polyoxyethylene chain and / or an amide group. G is a bioconjugable group; and R2, R3, R 12 and R 13 are each independently a hydrogen atom, cyano, halo, or perhydrogen atom. alkyl, sulfonate, sulfonamide, ester, carboxylic acid, formyl, acetyl L1-(X1-L2) p a linker group and a solubilizing group represented by G; (wherein the solubilizing group is -aryl-(R s ) w and -alkynyl-aryl-(R s ) w wherein w is an integer of 0 to 5, and R s is a group represented by the following formula: . -X2-(L3) z -R 17 , where z is 0 or 1, and X2 is —CH2NHC(═O)—, —C(═O)NH-alkyl L3 is -C(=O)-alkylene-C(=O)-NH - and R 17 Ha-(C2H4O) m -R 18 , -C(=O)C2H4-(OC2H4) m OR 18and -(C2H4O) n -C2H4-C(=O)NH-C(R 19 )3 are selected where m is an integer of 4 or greater (optionally, m is an integer of 8 or greater), and n is a number from 1 to 5. is an integer, R 18 is lower alkyl (optionally methyl), and R 19 -CH2O-C2 H4-C(=O)NH-(C2H4O) m R 18 It is.) However, R2, R3, R 12 and R 13 At least one of the -aryl-(R s ) w or -alkynyl-aryl-(R s ) w )

[0007] In some embodiments, R, R 10 and R 15 is a hydrogen atom, methoxy and the formula -L1-(X1-L2) p In some embodiments, the linker group is selected from the group represented by -G. Hey, R 10 is the formula -L1-(X1-L2) p -G is a linker group. In an embodiment of the present invention, R 10 is the formula -L1-(X1-L2) p -G is a linker group wherein L1 is phenylene, p is 1, X1 is -C(=O)NH-, and L2 is alkyl. alkylene, and G is selected from a carboxylic acid or an activated ester. In some embodiments, R 10 is expressed by the following formula: [ka] In some embodiments, R and R 13are -aryl-(R s ) w Yes (Optionally, R3 and R 13 are -phenyl-(R s )2). Some real In the embodiment, each R s is a group represented by the following formula: -X2-(L3)zR 17 (Wherein z is 0, X2 is —CH2NHC(═O)—, R 17 Ha-(C2H4O) m -R 18 (wherein m is an integer of 12 to 24.) .) In some embodiments, R and R 13 are expressed by the following equations, respectively. [ka]

[0008] In some embodiments, the compound has the formula: [ka] The present invention, in some embodiments, provides a covalently bonded conjugate formed between: A composition comprising: (a) R2, R3, R5, R 10 , R 12 , R 13 and R 15 At least one of the A compound represented by formula (I) according to claim 1, wherein (b) Small molecules, antigens, microparticles, nanoparticles, polymers, peptides, proteins, antibodies or one or more of the group consisting of antibody fragments, nucleic acids, hormones and growth factors .

[0009] In some embodiments, the present invention provides a compound represented by formula (I), or the covalent conjugate formed, (a) R2, R3, R5, R 10 , R 12 , R 13 and R 15 At least one of the A compound represented by formula (I) according to claim 1, wherein (b) Small molecules, antigens, microparticles, nanoparticles, polymers, peptides, proteins, antibodies or one or more of the group consisting of antibody fragments, nucleic acids, hormones and growth factors and a pharmaceutically acceptable carrier. In some embodiments, the present invention provides methods for detecting a target. In an embodiment, the target is a compound, a cell, or a particle, and the method comprises forming a This involves labeling the target with a covalently bound conjugate formed therefrom. (a) R2, R3, R5, R 10 , R 12 , R 13 and R 15 At least one of the A compound represented by formula (I) according to claim 1, wherein (b) Small molecules, antigens, microparticles, nanoparticles, polymers, peptides, proteins, antibodies or one or more of the group consisting of antibody fragments, nucleic acids, hormones and growth factors In some embodiments, the method comprises the use of flow cytometry.

[0010] The present invention provides, in some embodiments, a method for imaging a cell, tissue, or organism. In some embodiments, the method comprises administering to a patient a compound of formula (I), This includes the use of covalently bonded conjugates formed between: (a) R2, R3, R5, R 10 , R12 , R 13 and R 15 At least one of the A compound represented by formula (I) according to claim 1, wherein (b) Small molecules, antigens, microparticles, nanoparticles, polymers, peptides, proteins, antibodies or one or more of the group consisting of antibody fragments, nucleic acids, hormones and growth factors The present invention, in some embodiments, provides a method for treating a disease in a patient in need thereof. In some embodiments, the method comprises the steps of: a compound of formula (I) or a covalent conjugate formed between: (a) R2, R3, R5, R 10 , R 12 , R 13 and R 15 At least one of the A compound represented by formula (I) according to claim 1, wherein (b) Small molecules, antigens, microparticles, nanoparticles, polymers, peptides, proteins, antibodies or one or more of the group consisting of antibody fragments, nucleic acids, hormones and growth factors or administering to a patient a pharmaceutical composition comprising the compound or a conjugate thereof; and - irradiating at least a portion of the patient with light (optionally, the disease is a hyperproliferative disease; Further optionally, the disease is cancer. In some embodiments, the present invention provides a compound having a solubility of greater than about 1 mg / ml in aqueous solution. (optionally, having a solubility of about 2.5 mg / ml or more in aqueous solution, and optionally, a water-soluble chloride (having a solubility of about 10 mg / ml or more in aqueous solution); We provide dyes.

[0011] In some embodiments, the present invention provides a synthetic intermediate for a compound represented by the following formula (I): A method of manufacturing, [ka] wherein M is a metal or two hydrogen atoms; R5, R 10 and R 15 are each independently a hydrogen atom, an alkoxy group, or a group represented by the following formula: a linker group selected from the group consisting of: -L1-(X1-L2) p -G wherein p is 0 or 1, L1 is alkylidene, and X1 is -C(=O)NH- or -NHC(=O)- and L2 is -(CH2CH2O) q -alkylene- (wherein q is 1 to 24), alkylene, or substituted alkylene (optionally, the substituted alkylene The alkylene is bound to one or more groups containing a polyoxyethylene chain and / or an amide group. G is a bioconjugable group; and R2, R3, R 12 and R 13 are each independently a hydrogen atom, cyano, halo, or perhydrogen atom. alkyl, sulfonate, sulfonamide, ester, carboxylic acid, formyl, acetyl L1-(X1-L2) p a linker group and a solubilizing group represented by G; (wherein the solubilizing group is -aryl-(R s ) w and -alkynyl-aryl-(R s ) w wherein w is an integer of 0 to 5, and R s is a group represented by the following formula: . -X2-(L3)zR 17 , where z is 0 or 1, and X2 is —CH2NHC(═O)—, —C(═O)NH-alkyl L3 is -C(=O)-alkylene-C(=O)-NH - and R 17 Ha-(C2H4O) m -R 18 , -C(=O)C2H4-(OC2H4) m OR 18 and -(C2H4O) n -C2H4-C(=O)NH-C(R 19 )3 are selected where m is an integer of 4 or greater (optionally, m is an integer of 8 or greater), and n is a number from 1 to 5. is an integer, R 18 is lower alkyl (optionally methyl), and R 19 -CH2O-C2 H4-C(=O)NH-(C2H4O) m R 18 It is.) However, R2, R3, R 12 and R 13 At least one of the -aryl-(R s ) w or -alkynyl-aryl-(R s ) w ) A method is provided comprising the steps of: (a) providing a compound represented by formula (I'): [ka] wherein M is a metal or two hydrogen atoms; R5',R 10 ' and R 15 ' are each independently a hydrogen atom, an alkoxy, [ka] Selected from; R2',R3',R 12 ' and R 13 ' are each independently a hydrogen atom, an ester, a carboxylic acid, or a carboxylic acid, formyl, acetyl, [ka] is selected from However, R2', R3', R 12 ' and R 13 At least one of the ' [ka] ) (b) The compound provided in step (a) is dissolved in a solution containing 4 molar (M) HCl in dioxane. with a solution to provide a compound of formula (I"): [ka] wherein M is a metal or two hydrogen atoms; R5'',R 10 '' and R 15 '' are each independently a hydrogen atom, an alkoxy, [ka] Selected from; R2, R3, R 12 '' and R 13 '' each independently represent a hydrogen atom, an ester, Carboxylic acid, formyl, acetyl, [ka] is selected from However, R2'', R3'', R 12 '' and R 13 At least one of the [ka] )

[0012] In some embodiments, the compound of the present invention is a compound represented by the following formula: or a conjugate thereof. [ka] In some embodiments, the compound is a small molecule, an antigen, a microparticle, a nanoparticle, or , polymers, peptides, proteins, antibodies or antibody fragments, nucleic acids, hormones and compounds The nucleotide sequence is conjugated to one or more of the group consisting of:

[0013] It is therefore an object of the present invention to provide chlorin derivatives, in some embodiments, water-soluble Chlorin derivatives, conjugates thereof, pharmaceutical compositions containing said derivatives and / or said conjugates, and methods of using and making them. These and other objects are achieved in whole or in part by the present invention. The above-mentioned objects, other objects, and advantages of the present invention will become more apparent from the following description, drawings, and examples. It will be apparent to those skilled in the art upon review of the above. [Brief explanation of the drawings]

[0014] [Figure 1] Scheme 1, an exemplary scheme for synthesizing representative WH building blocks of dibromochlorin derivatives of the present invention. [Figure 2] Scheme 2, an exemplary scheme for synthesizing representative EH building blocks of dibromochlorin derivatives of the present invention. [Figure 3] 1 is an exemplary scheme for synthesizing compound CP-1, Scheme 3. [Figure 4] 1 is an exemplary scheme for synthesizing the western half of compound CP-1, Scheme 4. [Figure 5] 1 is an exemplary scheme for synthesizing the eastern half of compound CP-1 and the cyclized intermediate compound C-11, Scheme 5. [Figure 6]6 is an exemplary scheme of the final step in the synthesis of compound CP-1, Scheme 6. [Figure 7] Scheme 7 is an exemplary scheme for the final step in the synthesis of compound 4. BEST MODE FOR CARRYING OUT THE INVENTION

[0015] The present invention is described more fully below. However, there are several embodiments of the invention. Although some, but not all, of the invention may be embodied in many different forms. These embodiments should not be construed as being limited to the embodiments set forth herein. The disclosure is provided so that the disclosure satisfies applicable legal requirements.

[0016] I. Definition The terminology used herein is for the purpose of describing particular embodiments only and does not limit the scope of the present invention. It is not intended to be limiting. All technical and scientific terms used herein, unless otherwise defined below, It is intended to have the same meaning as commonly understood by one of ordinary skill in the art. References to techniques employed in the specification may include variations or equivalents of those techniques that would be apparent to those skilled in the art. These are references to techniques commonly understood in the art, including permutations of techniques such as: Although the following terms are believed to be well understood by those of ordinary skill in the art, the following definitions are provided for purposes of the present invention. are described to facilitate the explanation of It will be understood that in describing the present invention, numerous techniques and steps are disclosed. Each of these has its own advantages and each may incorporate one or more of the techniques disclosed in the others. It may also be used with any or all of the above. Therefore, for the sake of clarity, this description does not unnecessarily include all possible combinations of the individual steps. I will refrain from repeating all of the above. Nevertheless, the specification and claims The combination should be read with the understanding that it is within the full scope of the present invention and claims. It is.

[0017] In accordance with long-standing patent law practice, "one" as used herein, including the claims, The words "a," "an," and "the," when used, refer to "one or more." For example, the phrase "fluorescent microparticles and / or nanoparticles" refers to multiple identical fluorescent particles. one or more fluorescent microparticles and / or nanoparticles, including optical microparticles and / or nanoparticles; Similarly, the phrase "at least one" refers to an entity. As used herein, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 75, 100 or more, including 1 to 100 and greater than 100 Including, but not limited to, integer values.

[0018] Unless otherwise indicated, ingredients, reaction conditions, etc., used in the specification and claims All numbers expressing quantities are to be deemed to be modified in all instances by the term "about." A measurable value such as mass, weight, time, volume, concentration or percentage (%) As used herein, the term "about" refers to a range of some magnitude from the stated value. In some embodiments, ±20%, in some embodiments, ±10%, in some embodiments In some embodiments, ±5%, in some embodiments ±1%, in some embodiments ±0.5%, In some embodiments, variations of ±0.1% are included, and these variations are within the scope of the present invention for carrying out the disclosed methods. Accordingly, unless indicated to the contrary, the present specification and accompanying patents The numerical parameters set forth in the claims are sought to be obtained by the present invention. These are approximations that may vary depending on the desired properties. As used herein, when used in the context of listing entities, the term "and / or" refers to The terms "A, B, C, The phrase "and / or D" includes A, B, C, and D individually, but also A, B, C, and D.

[0019] Synonymous with "including," "containing," or "characterized by" The term "comprising" is inclusive or open-ended and is not intended to be a citation. It does not exclude additional elements and / or method steps. "Comprising" is used in claim language. is a term used in a methodology where the named elements and / or steps are present but other elements and steps are not. Additional steps and / or methods may be added and still be within the scope of the claims. As used herein, the phrase "consisting only of" refers to the The phrase "consisting only of" excludes any element, step, or ingredient that is not included in the invention. , appears in a clause in the body of a claim, rather than immediately following the preamble, it The scope of the claims is limited to only the elements recited within, and no other elements are excluded from the scope of the entire claims. As used herein, the phrase "consisting essentially of" extends a claim to: The materials and / or steps specified and the basic and novel features of the disclosure and claims (multiple possibilities) is limited to the range that does not substantially affect the The polymer and / or nanoparticles are bonded to a polymer matrix and at least one polymer bonded thereto. When referring to "consisting essentially of" a polymer matrix, the polymer matrix being referred to is a fluorescent matrix. The polymer matrix is ​​the only one present in the microparticles and / or nanoparticles. This means:

[0020] With respect to the terms "comprising," "consisting of," and "consisting essentially of," When one of the terms is used herein, the claimed subject matter of this disclosure includes the other. For example, in some embodiments, Thus, the present invention relates to fluorescent microparticles and / or nanoparticles. After that, the present invention provides a polymer matrix of the present invention and at least one polymer bonded thereto. Fluorescent microparticles and / or nanoparticles consisting essentially of chlorins of the present invention, Fluorescent materials consisting only of a polymer matrix and at least one chlorin bound to it It will be understood to include microparticles and / or nanoparticles.

[0021] As used herein, "halo" refers to any suitable halo, including -F, -Cl, -Br, and -I. He points to Gen. As used herein, "mercapto" refers to an --SH group. As used herein, "azido" refers to the group --N3. As used herein, "cyano" refers to a -CN group. As used herein, "hydroxyl" refers to an --OH group. As used herein, "nitro" refers to the group --NO.sub.2.

[0022] As used herein, "alkyl" as used alone or as part of another group means 1 or 2 to 10, It is a straight or branched chain hydrocarbon containing 20 or 50 carbon atoms (e.g., C1-C4 Alkyl, C4-C 10 Alkyl, C 11 ~C 50 alkyl). Representative examples of alkyl are are methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, and isobutyl. butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3- Methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, n-heptyl, Examples include, but are not limited to, n-octyl, n-nonyl, and n-decyl. As used herein, "lower alkyl" is a subset of alkyl and in some embodiments In some cases, straight or branched chain hydrocarbons containing 1 to 4 carbon atoms are preferred. Representative examples of lower alkyl include methyl, ethyl, n-propyl, isopropyl, Examples include, but are not limited to, n-butyl, isobutyl, tert-butyl, and the like. The term "alkyl" or "lower alkyl" includes, unless otherwise indicated, substituted and unsubstituted alkyl. and both substituted and unsubstituted lower alkyl, these groups being selected from the group consisting of: may be substituted with a group selected from: halo, alkyl, haloalkyl, alkenyl, alkyn ... quinyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heptyl cyclo, heterocycloalkyl, hydroxyl, alkoxy, alkenyloxy, alkoxy quinyloxy, haloalkoxy, cycloalkoxy, cycloalkylalkyloxy, a Aryloxy, arylalkyloxy, heterocycloalkoxy, heterocycloalkoxy S(O)m, alkynyl-S(O)m, mercapto, alkyl-S(O)m, alkenyl-S(O)m, alkynyl-S(O)m (O)m, cycloalkyl-S(O)m, haloalkyl-S(O)m, cycloalkylal alkyl-S(O)m, aryl-S(O)m, arylalkyl-S(O)m, heterocyclo- S(O)m, heterocycloalkyl-S(O)m, amino, carboxy, alkylamino , alkenylamino, alkynylamino, haloalkylamino, cycloalkylamino, Cycloalkylamino, arylamino, arylalkylamino, heterocycloamino , heterocycloalkylamino, disubstituted amino, acylamino, acyloxy, ester , amide, sulfonamide, urea, alkoxyacylamino, aminoacyloxy, nitrite b or cyano (wherein m is 0, 1, 2 or 3).

[0023] As used herein, "alkylene" refers to a difunctional alkyl group which may be substituted or unsubstituted. refers to a linear, branched or cyclic alkyl group, where "alkyl" is as defined above. . As used herein, "alkenyl" as used alone or as part of another group means 1 or 2 to 1 A straight or branched chain hydrocarbon containing 0, 20 or 50 carbon atoms (e.g., C1-C 4 Alkenyl, C4-C 10 Alkenyl, C 50 Alkenyl or lower alkyl having 1 to 4 carbon atoms Alkenyl), usually containing 1 to 4 double bonds in the chain. Representative examples of alkenyl include vinyl, 2-propenyl, 3-butenyl, 2-butenyl, 4-pentenyl, 3-pentenyl, 2-hexenyl Examples include, but are not limited to, 3-hexenyl, 2,4-heptadienyl, and the like. stomach. The term "alkenyl" or "lower alkenyl" includes both substituted and unsubstituted alkenyl, unless otherwise indicated. These groups include both the alkyl and lower alkenyl groups listed above. It may be substituted with groups as described in relation to alkyl. As used herein, "alkenylene" refers to a difunctional linear alkyl group that may be substituted or unsubstituted. "Alkenyl" refers to a substituted or unsubstituted aryl, branched or cyclic alkenyl group, wherein "alkenyl" is defined above.

[0024] As used herein, "alkynyl" as used alone or as part of another group means an alkyl group having 1 or 20 to 10 carbon atoms. A straight or branched chain hydrocarbon containing 10, 20 or 50 carbon atoms (e.g., C1 to C4 alkynyl; C4~C 10 Alkynyl; C 11 ~C 50 Alkynyl, or 1 carbon atom ~4 lower alkynyl), which usually contains one triple bond in the chain. Examples include 2-propynyl, 3-butynyl, 2-butynyl, 4-pentynyl, 3-pentynyl, etc. The term "alkynyl" or "lower alkynyl" includes, but is not limited to, the following: The term, unless otherwise indicated, includes substituted and unsubstituted alkynyl or substituted and unsubstituted lower alkynyl. These groups are intended to include both alkyl and lower alkyl groups as defined above. may be substituted with the same groups as described in connection with As used herein, "alkynylene" refers to a difunctional alkyl group that may be substituted or unsubstituted. refers to a straight-chain, branched, or cyclic alkynyl group, where "alkynyl" is defined above . As used herein, an "alkylidene chain" may be substituted or unsubstituted, saturated or refers to an optionally unsaturated, difunctional linear, branched and / or cyclic organic group, optionally It may contain 1, 2 or 3 heteroatoms selected from the group consisting of N, O and S. Examples include alkylene, alkenylene, alkynylene, arylene, alkarylene, and These include, but are not limited to, aryl, arylalkylene, and aralkylene. See US Pat. No. 5,533. The alkylidene chain may contain any suitable number of carbon atoms. Good (e.g., C1~C 4; C4~C 10 ;C 10 ~C 20 ;C 20 ~C 50 ).

[0025] As used herein, "alkoxy" alone or as part of another group refers to an oxy group (- an alkyl or lower alkyl group, as defined herein, attached to the parent molecular moiety through an alkyl group (O-), Representative examples of alkoxy include methoxy, ethoxy, propoxy, 2-propoxy, butoxy, t-butoxy, pentyloxy, hexyloxy, etc. Not limited. As used herein, "acyl" alone or as part of another group refers to a -C(O)R group ( wherein R is aryl, alkyl, alkenyl, alkynyl, cycloalkyl or other suitable The substituents are any suitable substituents described herein, such as substituents. As used herein, "haloalkyl" as used alone or as part of another group means any alkyl group of the present invention. at least one alkyl group, as defined herein, attached to the parent molecular moiety through an alkyl group, as defined herein; Both refer to a single halogen. Representative examples of haloalkyl are chloromethyl, 2-fluoromethyl, and 2-fluoromethyl. trifluoromethyl, pentafluoroethyl, 2-chloro-3-fluoropentyl Examples of suitable materials include, but are not limited to, fluorine. As used herein, "perhaloalkyl" alone or as part of another group refers to an alkyl In some embodiments, each hydrogen atom in the alkyl group is replaced with a halo. The perhaloalkyl is a perfluoroalkyl group, and each hydrogen atom of the alkyl group is replaced by fluoro. A typical perhaloalkyl group is trifluoro. It is methyl (i.e., -CF3). As used herein, "alkylthio" as used alone or as part of another group means an alkyl, as defined herein, attached to the parent molecular moiety through a thio moiety, as defined herein; Representative examples of alkylthio include methylthio, ethylthio, t-butylthio, hexylthio, These include, but are not limited to, Sirthio.

[0026] As used herein, "aryl" alone or as part of another group refers to a monocyclic carbon atom. Aromatic ring systems or bicyclic carbocyclic fused ring systems containing one or more aromatic rings. Representative examples of aryl include azulenyl, indanyl, indenyl, naphthyl, phenyl, tetrahydrofuran, and the like. The term "aryl" includes substituted and unsubstituted aryl groups unless otherwise indicated. and unsubstituted aryl, which groups are intended to include both alkyl and lower alkyl groups as defined above. It may be substituted with the same groups as described in connection with alkyl. As used herein, "arylene" refers to a bifunctional arylene group that may be substituted or unsubstituted. "Aryl" refers to an aryl group, where "aryl" is defined above. As used herein, "arylalkyl" alone or as part of another group refers to any group within the scope of this specification. An aryl group, as defined herein, appended to the parent molecular moiety through an alkyl group, as defined herein. Representative examples of arylalkyl include benzyl, 2-phenylethyl, and 3-phenyl. These include, but are not limited to, 2-naphth-2-ylpropyl, 2-naphth-2-ylethyl, and the like. The terms "alkarylene" and "aralkyl" as used herein alone or as part of another group are intended to mean "Arylene" refers to at least one arylene group and at least one It refers to a difunctional group containing an alkyl, alkenyl, or alkynyl group. As used herein, "amino" refers to the group --NH.sub.2. As used herein, "alkylamino" alone or as part of another group means - It means the group NHR, where R is an alkyl group. As used herein alone or as part of another group, "arylalkylamino" refers to " refers to the group --NHR, where R is an arylalkyl group.

[0027] As used herein, "disubstituted amino" alone or as part of another group refers to -NR a R b group (in the formula, R a and R b are independently alkyl, haloalkyl, alkenyl, alkynyl, quinyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heptyl and cycloalkyl, cycloaliphatic ... As used herein, "acylamino" alone or as part of another group refers to -NR a R b group (in the formula, R a is an acyl group as defined herein, and R b is a hydrogen atom, alkyl, Haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, a aryl, arylalkyl, heterocyclo, heterocycloalkyl ) means. As used herein, "acyloxy" alone or as part of another group refers to an -OR group ( wherein R is an acyl group as defined herein. "Ester" as used herein alone or as part of another group refers to an ester of -C(O)OR R is a group such as alkyl, cycloalkyl, alkenyl, alkynyl, or aryl. is any suitable substituent. As used herein, "formyl" refers to a -C(O)H group. As used herein, "carboxylic acid" refers to the group --C(O)OH.

[0028] As used herein, "sulfoxyl" refers to a group of the formula -S(O)R, where R is alkyl, silyl, It may be any suitable substituent such as chloroalkyl, alkenyl, alkynyl or aryl. ) refers to a compound represented by the formula: As used herein, "sulfonyl" refers to a group of the formula -S(O)(O)R, where R is alkyl , cycloalkyl, alkenyl, alkynyl, or aryl. It refers to a compound represented by the formula: As used herein, a "sulfonate" refers to a group of the formula -S(O)(O)OR, where R is an alkyl group. any suitable substituents such as alkyl, cycloalkyl, alkenyl, alkynyl or aryl It refers to a compound represented by the formula: As used herein, "sulfonic acid" refers to a compound of the formula -S(O)(O)OH. Point. "Amido" as used herein alone or as part of another group refers to -C(O)NR a R b group (in the formula, R a and R brepresents a hydrogen atom, alkyl, cycloalkyl, alkenyl, or alkoxy group. is any suitable substituent such as quinyl or aryl. As used herein, "sulfonamide" alone or as part of another group refers to a group having an amino acid sequence similar to -S(O )2NR a R b group (in the formula, R a and R b represents a hydrogen atom, alkyl, cycloalkyl, or alkene atom. is any suitable substituent such as aryl, alkynyl, or aryl.

[0029] "Urea" as used herein alone or as part of another group refers to -N(R c )C( O)NR a R b group (in the formula, R a , R b and R c is a hydrogen atom, alkyl, cycloalkyl, is any suitable substituent such as alkenyl, alkynyl, or aryl. As used herein, "alkoxyacylamino" alone or as part of another group means -N(R a )C(O)OR b group (in the formula, R a , R b is a hydrogen atom, alkyl, cycloalkyl The term "aryl" refers to any suitable substituent such as aryl, alkenyl, alkynyl, or aryl. "Aminoacyloxy" as used herein alone or as part of another group means - OC(O)NR a R b group (in the formula, R a and R b is a hydrogen atom, alkyl, cycloalkyl, is any suitable substituent such as alkenyl, alkynyl, or aryl. As used herein alone or as part of another group, "cycloalkyl" refers to 3, 4 or has 5 to 6, 7, or 8 carbon atoms (the carbon atoms are substituted with heterocyclic groups as discussed below) Cycloalkane refers to a saturated or partially unsaturated cyclic hydrocarbon group containing a cyclic alkyl group, which may be substituted. Representative examples of alkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, These rings may optionally be joined by halo or lower alkyl groups. The term "cycloalkyl" may be substituted with additional substituents as described herein, such as alkyl. "Alkyl" is generic and, unless otherwise specified, includes the heterocyclic groups described below. It is intended to

[0030] As used herein, the term "polyoxyethylene chain" refers to a poly(ethylene glycol) (PEG) groups, such as those of the formula -(C2H4O) n - (wherein n is 2 or more (e.g., 3, 4, 5 , 6, 7, 8, 9, or 10 or more). In some embodiments, n refers to a moiety that comprises or consists of: An integer between 4 and 5000, 4 and 1000, 4 and 100, 4 and 50, 4 and 28, or 4 and 25 As used herein, the term "polyoxyethylene chain" refers to a monodisperse or polydisperse PE. The term "monodisperse" may refer to the PEG chains and linear or branched PEG chains. "Polydisperse" refers to PEG with a PDI greater than 1, and Here, the PEG includes those with a Gaussian distribution of chain length and molecular weight.

[0031] As used herein, the term "bioconjugable group" refers to a group that is capable of being attached to another entity (e.g., a protein; Peptides; targeting agents such as antibodies or antibody fragments; polymers; nanoparticles, organic, poly and bonding (e.g., covalent bonding) with groups on a particle such as a polymer or inorganic beads; another solid support surface, etc. to form a bond between one of the chlorin or bacteriochlorin compounds of the invention and another entity. This refers to a reactive chemical functional group that can form a conjugate. For example, aldehydes (via reductive amination) can be covalently bonded to amino groups on amino-substituted biomolecules. ) or carboxylic acid (via carbodiimide activation) The bioconjugable group may be an isoform capable of binding to a recombinant biomolecule. Socyanates, isothiocyanates, iodoacetamides, azides, diazonium salts, etc. amines (including amine derivatives); N-hydroxysuccinimide (NHS) esters ( More commonly, activated esters derived from carboxylic acids; e.g., p-nitrophenyl esters), carboxylic acids or acid derivatives such as acid hydrazides; aldehydes, sulfonyl chlorides , sulfonyl hydrazide, epoxide, hydroxyl group, thiol group, maleimide, azide Other groups include lysine, acryloyl, halo groups, biotin, and 2-iminobiotin However, the present invention is not limited to these.

[0032] The term "microparticle" refers to particles having dimensions (e.g., length, width, diameter, etc.) of about 1,000 μm. refers to a structure having at least one region that is less than about 1000 nm but greater than about 1000 nm. In some embodiments, the dimension is less than about 500 μm, and in some embodiments, Less than about 250 μm, in some embodiments less than about 200 μm, in some embodiments in some embodiments, less than about 150 μm, in some embodiments, less than about 125 μm, In embodiments, less than about 100 μm, in some embodiments, less than about 80 μm, In some embodiments, less than about 70 μm, and in some embodiments, less than about 60 μm; In some embodiments, less than about 50 μm, in some embodiments, about 40 μm; In some embodiments, less than about 30 μm, in some embodiments, less than about 20 μm in some embodiments less than about 10 μm, and in some embodiments less than about 5 μm In some embodiments, this dimension may be less than about 1 μm to about 2 μm. Between 50 μm (e.g., about 5, 10, 15, 20, 30, 40, 50, 60, 70, 80 , 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, or 250 μm). Similarly, the term "nanoparticle" refers to a particle having dimensions (e.g., length, width, diameter, etc.) of approximately 1,000 In some embodiments, the term "structure" refers to a structure having at least one region that is less than 1 nm. Therefore, this dimension is smaller (e.g., less than about 500 nm, less than about 250 nm, less than about 200 less than about 150 nm, less than about 125 nm, less than about 100 nm, less than about 100 nm , less than about 80 nm, less than about 70 nm, less than about 60 nm, less than about 50 nm, less than about 40 nm , less than about 30 nm, or less than about 20 nm). In some embodiments, this dimension is Between about 5 nm and about 250 nm (e.g., about 1, 5, 10, 15, 20, 30, 40, 5 0, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 16 0, 170, 180, 190, 200, 210, 220, 230, 240, or 250n m).

[0033] In some embodiments, the microparticles or nanoparticles are approximately spherical. If the microparticle or nanoparticle is approximately spherical, the characteristic dimension may correspond to the diameter of the sphere. The microparticles or nanoparticles can be in the shape of a disk or plate (e.g., They may be hexagonal, rectangular, polyhedral, rod-shaped, cubic, or irregularly shaped. The microparticle or nanoparticle has a core region (i.e., the space between the outer dimensions of the particle) and outer surface (i.e., the surface that defines the outer dimensions of the particle). In an embodiment, the microparticle or nanoparticle comprises a core of the microparticle or nanoparticle. May have one or more coating layers surrounding or partially surrounding it Thus, for example, a spherical microparticle or nanoparticle may have one or more concentric coatings. Each successive layer may have smaller layers closer to the center of the particle. spreads over the outer surface.

[0034] The terms "polymer" and "polymeric" refer to a polymer that is made up of repeating units (i.e., a given chemical substructure). A polymer is a chemical structure that has multiple copies of a molecule. A polymer is formed from polymerizable monomers. Polymerizable monomers can react to bond with moieties on other molecules of the polymerizable monomer (e.g., A molecule containing one or more moieties that can form bonds (e.g., covalent or coordinate bonds) In some embodiments, each polymerizable monomer molecule is In some cases, the polymerizable monomer can be bonded to one other molecule / moiety. It only bonds to molecules and forms the ends of polymeric materials. The polymer may be organic, inorganic, or a combination thereof. The term "inorganic" as used herein means any material containing one or more of carbon, hydrogen, nitrogen, oxygen, sulfur, phosphorus, or halogens. It refers to a compound or composition that contains at least some atoms other than The inorganic compound or composition may contain one or more silicon atoms and / or one or more In some embodiments, the polymer may contain more than one metal atom. is polystyrene, and the microparticles and / or nanoparticles are made of polystyrene. In some embodiments, the microparticles and / or nanoparticles are made of polystyrene. It's beads.

[0035] As used herein, the term "porphyrin" refers to a compound that typically contains four nitrogen atoms and various It consists of four pyrrole rings with two substitutable hydrogen atoms that can easily replace metal atoms. A typical porphyrin is hemin. As used herein, "chlorin" refers to a compound having one partially saturated pyrrole ring. It is different from porphyrins, and is the basic product of chlorophyll, the green pigment in plant photosynthesis. The chromophore is a chlorin. The terms "chlorin" and "chlorin derivative" are used herein. are used interchangeably.

[0036] The phrase "combine" means any interaction between two entities, e.g. This refers to any interaction between the polymer matrix and the chlorin. In this case, the polymer matrix and chlorin are bonded non-covalently (hydrophobic, electrostatic and foundation interactions, such as, but not limited to, one or more of the following: In some embodiments, the polymer matrix (e.g., nanoparticles) , microparticles, beads, etc.) and chlorine, which exists within the polymer matrix. As a result of the polymer matrix incorporating the chlorins, they bond together. In such an embodiment, the polymer matrix is ​​"doped" with chlorine. The chlorine is also called "doped" or "doped" with a polymer matrix. In some embodiments, the polymer may be considered "embedded" within the polymer. The polymer matrix and chlorin are covalently bonded to the surface of the polymer matrix. They are bonded to each other by bonds.

[0037] As used herein, "treatment" refers to the amelioration of one or more symptoms of a disease or disorder. "Treatment" also means any manner in which the condition or symptoms of a disease or condition are improved or beneficially altered. or hyperproliferative tissue or tissues mediating the disorder To treat angiogenesis or a disease or disorder involving hyperproliferative tissue or angiogenesis. The present invention also encompasses any pharmaceutical method of use of the compositions of the present invention, such as a method of use for the treatment of a rheumatoid arthritis. In this study, the improvement of symptoms of a particular disorder by administering a particular compound or pharmaceutical composition is Any permanent or temporary, persistent or temporary effect that may result from or be associated with the administration of the composition. It refers to any relief, whether temporary or not. As used herein, a "prodrug" refers to a compound that, when administered in vivo, produces one or more metabolized by the above steps or processes or biologically, pharmacologically or therapeutically It is a compound that is converted into an active form of the compound.

[0038] As used herein, an "antibody" generally refers to an antibody that specifically binds to an antigen to form an immune complex. The term "antibody" refers to an immunoglobulin or fragment thereof that binds to any class of immunoglobulin. IgA, IgD, IgE, dual or multiple antigen or epitope specific It may be a chimeric or hybrid antibody having the same properties as a polyclonal antibody, Preferably obtained from a human or suitable animal (e.g., a primate, goat, rabbit, mouse, etc.) The antibody may be affinity-purified. Monoclonal antibodies may also be used in the present invention. These are suitable for use and may be preferred due to their high specificity. Immunization of a mammal with an antigen preparation, immunosuppression of lymphocytes or spleen cells with an immortal myeloma cell line The fusion of the chromosomes and the isolation of specific chromosome clones, which are now considered conventional procedures, Monoclonal antibodies are readily prepared by conventional methods. Other factors, such as interspecies hybridization and genetic engineering of hypervariable regions, may also affect their usefulness. The reason for this is mainly the antigen specificity of the antibody, so it is not excluded. Newer technologies for detecting antibodies (e.g., human monoclonal antibodies, interspecies monoclonal antibodies) , chimeric (e.g., human / mouse) monoclonal antibodies, genetically engineered antibodies, etc. May be used.

[0039] Thus, the terms "antibody" and "antibodies" refer to antibodies produced by immunoglobulin genes or fragments thereof. Immunoglobulin refers to a protein comprising one or more polypeptides substantially encoded by a Globulin genes are typically kappa (κ), lambda (λ), alpha (α), and gamma (γ ), delta (δ), epsilon (ε), and mu (μ) constant region genes, as well as numerous immune The immunoglobulin variable region genes are contained in the light chains. The light chains are classified as kappa or lambda. In immunoglobulins, heavy chains are classified as gamma, mu, alpha, delta, or epsilon, which correspond to the immunoglobulin classes, Ig IgE defines IgA, IgM, IgA, IgD, and IgE. Other species have other light and heavy chain genes (e.g., For example, certain birds have a type of immunoglobulin called IgY that chickens deposit in the yolk of their eggs. ), which are also included in the present invention.

[0040] A typical immunoglobulin (antibody) structural unit is known to comprise a tetramer. A dimer consists of two identical pairs of polypeptide chains, each pair consisting of one "light" chain (an average molecular weight It has two polypeptide chains: one "heavy" chain (average molecular weight about 50-70 kDa) and one "heavy" chain (average molecular weight about 25 kDa). Identical pairs of these are held together in a dimer by disulfide bonds present within the heavy chain region. The N-terminus of each chain contains approximately 100 to 110 or more amino acids that are primarily involved in antigen recognition. The variable light chain (V L ) and variable heavy chain (V H ) are the terms The term refers to the light and heavy chains of these proteins. Antibodies are typically produced as intact immunoglobulins or as fragments purified by various peptidases. It exists as a number of well-characterized fragments that can be generated by digestion with When an antibody molecule is digested with papain, the antibody is cleaved at the N-terminal position of the disulfide bond. This generates three fragments: two identical fragments containing the N-termini of the light and heavy chains. It consists of one "Fab" fragment and one "Fc" fragment containing the C-terminus of the heavy chains linked by disulfide bonds. On the other hand, pepsin digests antibodies at the C-terminus of the disulfide bond in the hinge region to form "F (ab) produces a single fragment known as the '2' fragment, which is linked by a disulfide bond. This F(ab)'2 fragment can be reduced under mild conditions to form a dimer of two Fab fragments. The disulfide bond in the di-region is disrupted, converting the F(ab')2 dimer into two Fab' monomers. The Fab' monomer is essentially a Fab fragment with part of the hinge region. With respect to the fragments, Fab, F(ab)'2 and Fab fragments contain at least one intact antigen-binding domain. (called a "paratope") and thus capable of binding to an antigen.

[0041] Various antibody fragments have been defined in terms of the digestion of intact antibodies, and those skilled in the art will recognize these fragments. Various fragments of the Fab' fragments (including but not limited to Fab' fragments) can be prepared chemically or by recombinant DNA technology. It will be understood that the compound can be synthesized de novo using Method A. The term "antibody" as used herein includes both antibody fragments produced by the modification of whole antibodies and antibody fragments produced using recombinant DNA methodologies. In some embodiments, the term "antibody fragment" refers to both newly synthesized and newly synthesized antibody fragments. "Antibody" includes fragments having at least one antigen-binding domain.

[0042] The antibodies, fragments and derivatives of the present invention also include chimeric antibodies. The term "chimeric" and its grammatical variants used in the context of antibodies in this context refer to certain antibody invariant domains. constant regions derived substantially or exclusively from the variable regions of other species; refers to antibody derivatives having variable regions derived exclusively from the antibody. A particular type of chimeric antibody is A "humanized" antibody is an antibody in which the CDRs of the human antibody have been replaced with those of a mouse antibody, e.g. Antibodies are produced by substituting the complementarity determining regions (CDRs) of antibodies (e.g., See publication WO 1992 / 22653.) Thus, in some embodiments, humanized The antibody does not have any determining regions (CDRs) derived substantially or exclusively from corresponding human antibody regions. A determinant having constant and variable regions and derived substantially or exclusively from a mammal other than a human. (CDR). The antibodies, fragments and derivatives of the present invention may be single chain antibodies and single chain antibody fragments. The antibody fragments comprise at least the variable regions and / or critical determinants (CDRs) of the whole antibody described herein. containing an amino acid sequence having one of the constant domains of the antibody, but lacking some or all of the constant domains of those antibodies. These constant domains are not required for antigen binding but make up the majority of the overall antibody structure. is doing.

[0043] Single-chain antibody fragments avoid many of the problems associated with the use of antibodies containing some or all of the constant domain. For example, single-chain antibody fragments contain a biomolecule and a heavy chain constant region. tend to be free of undesirable interactions between the Furthermore, single-chain antibody fragments are significantly smaller than whole antibodies and have a higher capillary permeability than whole antibodies. Their high transmissivity allows them to more efficiently localize and bind to the target antigen-binding site. Additionally, antibody fragments can be produced on a relatively large scale in prokaryotic cells, facilitating their manufacture. Furthermore, the relatively small size of single-chain antibody fragments may result in greater immunogenicity in recipients than whole antibodies. The single chain antibody fragments of the present invention are less likely to induce a response. , single-chain fragment variable (scFv) antibodies and their derivatives, such as tandem di-scFv, tandem Demtri scFv, diabodies, and further bispecific diabodies, triabodies Di, tetrabody, miniantibody, minibody, tetravalent bispecific molecule, bispecific F(a b')2 fragments and the like.

[0044] As used herein, "infectious agent" means an invading microorganism or parasite. The term "microorganisms" used in the book refers to viruses, bacteria, rickettsia, mycoplasma, and protozoa. "Parasite" refers to an organism that is infectious, generally microscopic or very small. Small multicellular invertebrates or their eggs or juveniles that exhibit antibody-induced clearance or are susceptible to lytic or phagocytic destruction (e.g., malaria parasites, spirochetes, This refers to a company (such as a company office). As used herein, "tumor" means a neoplasm and includes both benign and malignant tumors. The term is used to specifically refer to solid (such as breast, liver, or prostate cancer) or non-solid (such as leukemia) cancers. This includes malignant tumors, which may be any of the above. In addition, this tumor further includes adenocarcinoma (breast, prostate, They can also be divided into subtypes, such as:

[0045] As used herein, a "target" is a molecule that can be detected, diagnosed, damaged, or destroyed by the methods of the present invention. "Target" means a target object to which the target is intended to be administered, including target cells, target tissues, and target compositions. As used herein, "target tissue" and "target cells" refer to tissues that are injured by the therapeutic method. These are the tissues that are intended to be damaged or destroyed. Photosensitizing compounds target these tissues. They bind to or collect in tissues or target cells and then, when exposed to sufficient radiation, destroy these tissues. The tissue or cells are injured or destroyed. The target cells are cells within the target tissue. Target tissues include vascular endothelial tissue, abnormal blood vessel walls in tumors, and solid tumors such as those of the head and neck. (including but not limited to), eye tumors, gastrointestinal tumors, liver tumors, breast tumors, prostate tumors tumors of the glands, tumors of the lungs, non-solid tumors and malignant cells of the hematopoietic and lymphoid tissues, vascularized tissues, autologous Other lesions of the vasculature, bone marrow, tissues, or cells associated with autoimmune diseases, including, but not limited to: The target cells undergo substantially more rapid division than non-target cells. Also included are cells in which

[0046] As used herein, "non-target tissue" refers to tissue that is injured or destroyed by the treatment method. These non-target tissues include all tissues in the subject that are not intended to be affected by the drug. Healthy blood cells and other normal tissues, including those not otherwise identified as targets However, the present invention is not limited to these. As used herein, a "targeted composition" refers to a compound that is not damaged or destroyed by the treatment method. Compositions intended to combat pathogens (bacteria, viruses, fungi, protozoa, toxins, , and cells and tissues infected or infiltrated by them). "Targeting compositions" include prions, toxins, peptides, polymers, and other compounds targeted by this method of treatment. Selectively and specifically identify the organic target intended to be damaged or destroyed. Other compounds that can cause infection include, but are not limited to, infectious organic particles. As used herein, "hyperproliferative tissue" refers to tissue that grows uncontrollably and includes tumors. Inflammatory tissue, tumors, and uncontrolled blood vessel growth (as seen in age-related macular degeneration and glaucoma) These include blood vessel growths that often occur after surgery. As used herein, a "hyperproliferative disorder" refers to a disorder caused by unregulated or abnormal cell proliferation. These disorders share as their underlying pathology excessive cell proliferation caused by uncontrolled Examples of hyperproliferative disorders include cancer or carcinoma, acute and membranous angiogenesis. Proliferative glomerulonephritis, myeloma, psoriasis, atherosclerosis, psoriatic arthritis, rheumatoid arthritis Recurrence of diabetic retinopathy, macular degeneration, corneal neovascularization, choroidal hemangioma, and pterygium This includes, but is not limited to, scarring from glaucoma, excimer laser surgery, and glaucoma filtering surgery. do not have.

[0047] As used herein, a "therapeutically effective dose" is a dose sufficient to prevent progression or cause regression of the disease. or a dose sufficient to alleviate the symptoms caused by the disease. . As used herein, "biological material" refers to both tissue (such as biopsy tissue) and cells, etc. It also refers to bodily fluids such as blood, urine, plasma, cerebrospinal fluid, mucus, and phlegm.

[0048] As used herein, "illumination" and "irradiation" refer to exposing a subject to light of all wavelengths. The irradiation wavelength is preferably selected to match the wavelength that excites the photosensitive compound. The emission wavelength is preferably selected to match the excitation wavelength of the photosensitive compound and to be suitable for detecting the blood Low absorption by non-target tissues of interest, including proteins. The illumination can be coherent (laser) or non-coherent (non-laser), as well as optical The therapeutic effect of the present invention is further defined by the intensity, duration, and timing of administration of the sensitizing compound. The intensity or fluence rate must be sufficient to allow the light to reach the target tissue. The duration or total fluence dose is determined so that the photosensitizing compound has sufficient effect on the target tissue. The timing of administration of the photosensitizing compound must be sufficient. This is important because: 1) the administered photosensitizing compound takes some time to return to the target tissue; and 2) the blood levels of many photosensitizing compounds decrease over time. The radiation energy is provided by an energy source. external to the subject, or implanted in or introduced into the subject (e.g., catheter) by means of a terahertz or optical fiber, or in the form of a capsule or pill (see, for example, U.S. Pat. No. 6,629,492). 273,904) by ingesting a light source, such as a laser or cold cathode light source.

[0049] Some embodiments of the present invention utilize photodynamic therapy (PD) to destroy tumors. Although the present invention is directed to the use of light energy to administer T), other forms of energy may also be used. It is within the scope of the present invention and will be understood by those skilled in the art. - Includes thermal, sonic, ultrasonic, chemical, light, microwave, ionization (such as X-rays and gamma rays), These include, but are not limited to, mechanical and electrical, for example, acoustically induced or activated The drugs that can be activated include gallium-porphyrin complexes (Yumita et al. (1997) Cancer Lett. ters 112:79-86), other porphyrins such as protoporphyrin and hematoporphyrin complex (Umemura et al. (1996) Ultrasonics Sonochemistry 3:S187-S191), ultrasound therapy Other anticancer drugs such as daunorubicin and adriamycin used in the presence of (1987) Japanese Journal of Hyperthermic Oncology 3(2):175-182) However, the present invention is not limited to these. As used herein, a "coupling agent" refers to a compound that couples a photosensitizer to a targeting agent. This refers to a reagent that can A "targeting agent" is an agent that targets a specific tissue, receptor, infectious agent, or other area of ​​the subject's body to be treated. To home to or preferentially bind or associate with (e.g., a target tissue or target composition) Examples of targeting agents include antibodies, ligands, and one member of a ligand-receptor binding pair. Bars, nucleic acids, peptide-nucleic acids (PNAs), aptamers, proteins and peptides, and liposome suspensions (including tissue-targeted liposomes) do not have.

[0050] As used herein, "specific binding pair" and "ligand-receptor binding pair" refer to two different refers to two molecules, one of which specifically confers a particular spatial or polar organization on the other molecule. They have areas on their surface or within their cavities that attract or bind to each other, making both molecules parents to each other. The two members of this specific binding pair are called a ligand and a receptor. The terms ligand and receptor refer to the relationship between a ligand and a receptor. It includes the entire ligand or receptor, or a portion thereof, sufficient for binding to occur between them. Examples of endonucleaser-receptor binding pairs include hormones and hormone receptors (e.g., epidermal growth factor and epidermal growth factor receptors). Growth factor receptors, tumor necrosis factor-α and tumor necrosis factor-receptor, and interferon and interferon- receptors, avidin and biotin or antibiotin); antibody-antigen pairs; enzymes Substrates and drugs, drug receptors; cell surface antigens and lectins; two complementary nucleic acid strands; nucleic acid strands and complementary oligonucleotides; interleukins and interleukin receptors; and stimulation Factors and their receptors (granulocyte-macrophage colony-stimulating factor (GMCSF) and GMCSF receptor, and macrophage colony-stimulating factor (MCSF) and MCSF receptors), which Not limited to:

[0051] The "linker" may be a bioconjugable group, a cross-coupling group, a surface-attaching group, a hydrophilic group, etc. , an aromatic or aliphatic group (substituted or unsubstituted, optionally , which may contain heteroatoms such as N, O or S. Examples of such linkers include: Aryl, alkyl, heteroaryl, heteroalkyl (e.g., oligoethyleneglycol, Linkers include, but are not limited to, linkers such as hydroxyl groups, peptides, and polysaccharides. Patients (subjects) to be treated by the methods of the present invention for diagnostic or therapeutic purposes include human patients and Other animal patients for veterinary purposes (especially mammals such as dogs, cats, horses, monkeys, chimpanzees, etc.) This includes both patients with and without diabetes. More specifically, the terms "patient," "subject," and "recipient" as used herein The terms can be used interchangeably and refer to members of any invertebrate or vertebrate species. Thus, the term "patient" is meant to encompass any member of the animal kingdom. The diagram shows that the phylum Chordata (e.g., Osteichthyes (bony fish), Amphibians (amphibians), and Reptiles These include, but are not limited to, reptiles (reptiles), avian species (birds), and mammalia (mammals).

[0052] The compositions and methods of the present invention are particularly useful for warm-blooded vertebrates. The invention relates to mammals and birds, more particularly to mammalian-derived and / or mammalian Compositions and methods are provided for use in animals, including mammals such as humans and It includes other primates, as well as important mammals that are endangered (such as the Siberian tiger). mammals of economic importance to humans (animals raised on farms for human consumption) ) and / or socially important mammals (pets or animals kept in zoos) Also provided are uses of the compositions and methods of the present invention in birds, including birds that: Endangered birds, birds kept in zoos, birds kept as pets ( parrots, cockatiels, etc.), specifically, for example, turkeys, chickens, ducks, geese, Poultry such as rofowl are also included as they are also economically important to humans. Accordingly, there is also provided a method for using the compositions and methods of the present invention in livestock. Including, but not limited to, domesticated pigs (pigs and hogs), ruminants, horses, and poultry It will not be done.

[0053] II. Chlorin Compounds In some embodiments, the present invention provides chlorin derivatives. In an embodiment, the chlorin derivative of the present invention is a water-soluble chlorin derivative, wherein The chlorin derivatives are dissolved in an aqueous solution (e.g., water, saline, PBS, etc.) at a concentration of approximately 1 mg / mL. The water solubility is in the milliliter (mg / mL) range or higher. This is provided by adding a solubilizing group containing a PEG chain to the position of the chain. In embodiments, the PEG chain is attached to the chlorin through a different group and / or longer than the PEG chain attached to the PEGylated chlorin compound. Therefore, the solubility of the chlorins of the present invention in aqueous solutions is about 2.5 mg / mL or more. In some embodiments, the chlorin has a solubility in aqueous solution of about 5.0 mg / mL or greater. In some embodiments, the chlorin has a concentration of about 10 mg / mL or more in water. In some embodiments, the chlorin has a solubility in aqueous solution of about In some embodiments, the solubility is about 700, about 800, about 900 μM or greater. wherein the chlorin has a solubility of about 1, 1.5, 2, 2.5, or 3 mM or more in aqueous solution. It has. In some embodiments, the chlorin derivatives (including water-soluble chlorin derivatives) The bioconjugable group may be a linker moiety containing a bioconjugable group, and the bioconjugable group may be a linker moiety containing a bioconjugable group. The functional group allows the chlorin derivative to act, for example, as a targeting agent or a substance to be detected. In some embodiments, the conjugated molecule can be used to conjugate another substance that can Substances to which this chlorin derivative can be conjugated include small molecules (e.g., about 900 Da). Non-polymeric synthetic molecules with molecular weights of 1000 Da or less), antigens, microparticles , nanoparticles, polymers, peptides, proteins, antibodies or antibody fragments, nucleic acids, hormones, or The bioconjugable group can be, for example, a carboxylic acid or an active ester, In some embodiments, the compound may comprise a hydroxyl, an amine, a thiol, or an aldehyde. In some embodiments, the linker moiety further comprises both an arylene group and an alkylene group. In some embodiments, the linker moiety is an arylene group proximal to the main chlorin structure. and / or alkynylene groups, provided that the alkylene groups are not proximal to the bioconjugable group. be.

[0054] The chlorin derivatives of the present invention may contain at least one solubilizing group. In embodiments, the solubilizing group comprises one or more polyoxyethylene chains (PEG chains). In some embodiments, the solubilizing group comprises at least two PEG chains. In some embodiments, the PEG chains are monodisperse and have at least four -C In some embodiments, the PEG chain comprises at least one repeating H2CH2O- unit. It contains at least 6, 8, 10 or 12 -CH2CH2O- repeating units. The solubilizing group may comprise two PEG6, PEG8, PEG10 or PEG12 groups. In some embodiments, the PEG chain has 12 or more -CH2CH2O- repeating units (e.g., from about 12 to about 24 or about 28 -CH2CH2O- repeating units) In some embodiments, the compound comprises two different pyrrolic carbon atoms. In some embodiments, the two solubilizing groups are: Each contains two PEG chains. In some embodiments, the solubilizing group is a β-pyrrole substituent, The group contains an arylene or alkynyl-arylene group attached directly to the β-pyrrole carbon atom. However, this group does not contain a direct oxo linker between the PEG chain and the aryl. In an embodiment, the solubilizing group is one or more aryl groups between the aryl group and the PEG chain. In some embodiments, the amide bond further comprises one or more of the following: The above alkylene spacers (eg, ethylene, propylene, etc.) are included.

[0055] In some embodiments, the chlorin derivative is a compound of Formula (I): [ka] wherein M is a metal or two hydrogen atoms; R5, R 10 and R 15 are each independently a hydrogen atom, an alkoxy group, or a group represented by the following formula: a linker group selected from the group consisting of: -L1-(X1-L2) p -G wherein p is 0 or 1, L1 is alkylidene, and X1 is -C(=O)NH- or -NHC(=O)- and L2 is -(CH2CH2O) q -alkylene- (wherein q is 1 to 24), alkylene, or substituted alkylene (for example, the substituted alkylene The alkylene is bound to one or more groups containing a polyoxyethylene chain and / or an amide group. G is a bioconjugable group; and R2, R3, R 12 and R 13 are each independently a hydrogen atom, cyano, halo, or perhydrogen atom. perhaloalkyl (e.g., including, but not limited to, trifluoromethyl); alkyl), sulfonates, sulfonamides, esters, carboxylic acids, formyl, acetyl, Formula-L1-(X1-L2) p a linker group and a solubilizing group represented by G; (wherein the solubilizing group is -aryl-(R s ) w and -alkynyl-aryl-(R s ) w wherein w is an integer of 0 to 5, and R s is a group represented by the following formula: . -X2-(L3) z -R 17 , where z is 0 or 1, and X2 is —CH2NHC(═O)—, —C(═O)NH-alkyl L3 is -C(=O)-alkylene-C(=O)-NH - and R 17 Ha-(C2H4O) m -R 18 , -C(=O)C2H4-(OC2H4) m OR 18 and -(C2H4O) n -C2H4-C(=O)NH-C(R 19 )3 are selected , where m is 4 or more (e.g., at least 8, at least 10 or more, or n is an integer of 1 to 5, and R 18 is a low-level arch (e.g., methyl), and R 19 -CH2O-C2H4-C(=O)NH-(C2H4 O) m R 18 It is.) However, R2, R3, R 12 and R 13 At least one of the -aryl-(R s ) w or -alkynyl-aryl-(R s ) w )

[0056] M is any suitable metal ion (e.g., Pd, Pt, Mg, Al, Ga, In, Sn , Au, Ni, Cu, Co, Fe or Zn) or none (e.g., In the case of , it can be replaced by two hydrogens (-H, -H). The two nitrogen atoms of the phosphorus ring are protonated. In some embodiments, M is Zn or is substituted with two hydrogen atoms (-H, H). Therefore, The compounds include metallochlorin derivatives and free base chlorin derivatives. In some embodiments, R, R 10 and R 15 are each independently hydrogen atoms. methyl, methoxy and the formula -L1-(X1-L2) p -G. In some embodiments, R, R 10 and R 15 At least one of the formula -L 1-(X1-L2) p In some embodiments, R 10 is the formula: -L1-(X1-L2) p -G is a linker group. In some embodiments, L1 is arylene or alkarylene (e.g., arylene In some embodiments, the alkyl group is an alkyl or alkynyl group bonded to an alkyl group. and L1 is phenylene or C≡C-phenyl-. In some embodiments, p is 1, which linker group may comprise a polyoxyethylene chain to improve solubility and / or The reactivity of G compared to the reactivity of G in an equivalent chlorin where G is directly bound to L1 It may contain a spacer (e.g., an alkylene spacer) to improve In some embodiments, the linker group (R, R 10 and R 15 in , formula:-L1-(X1-L2) p -G (wherein L1 is phenylene, p is 1, and X1 is -C(=O)NH-, L2 is alkylene, and G is a carboxylic acid or an activated ester. (e.g., NHS-ester). In some embodiments, L2 is ethylene. The group contains a β-alanine spacer to improve the reactivity of G.

[0057] In some embodiments, the linker group (e.g., R, R10 and R 15 Inoke ) is expressed by the following formula: [ka] In some embodiments, at least one of R2 and R3, and R 12 and R 13 One of the groups is -aryl-(R s ) w or -alkynyl-aryl-(R s ) w in Thus, in some embodiments, the compound of formula (I) comprises at least two For example, in some embodiments, R and R 12 are both -aryl-(R s ) w or -alkynyl-aryl-(R s ) w Some In the embodiment, R and R 13 are -aryl-(Rs)w or -alkyl- It is aryl-(Rs)w. In some embodiments, R and R 13 are -aryl-(R s ) w in In some embodiments, R and R 13 are -phenyl-(R s ) It is 2. In some embodiments, each R s is -X2-(L3) z -R 17 where , z is 0, X2 is -CH2NHC(=O)-, and R 17 Ha-(C2H4O) m -R 18where m is an integer between 12 and 24 (i.e., 12, 13, 14, 15, 16, 1 7, 18, 19, 20, 21, 22, 23 or 24). In some embodiments, m is 12 and R 18 is methyl. In an embodiment, R and R 13 are expressed by the following equations, respectively. [ka]

[0058] In some embodiments, the compound is C-1 having the structure: [ka] In some embodiments, the compound is Compound 4, which has the structure: [ka]

[0059] In some embodiments, the present invention provides a covalently bonded conjugate formed between: (a) R2, R3, R5, R 10 , R 12 , R 13 and R 15 At least one of (b) a compound of formula (I) according to claim 1, wherein one of the groups is a linker group; and (c) a small molecule , antigens, microparticles, nanoparticles, polymers, peptides, proteins, antibodies or antibody fragments one or more of the group consisting of: fragments, nucleic acids, hormones and growth factors. In an embodiment, for example, the conjugate may comprise (i.e., as the bioconjugable group G) Compounds of formula (I) containing a linking group comprising a carboxylic acid or an active ester can be used to treat small molecules, peptides, It can be formed by reacting with the amino groups of a protein, antibody, or polymer. do.

[0060] III.Synthesis method Methods for synthesizing chlorins that may be suitable for use in the present invention are described, for example, in U.S. Pat. Nos. 212,055 and 8,980,565, each of which is incorporated by reference in its entirety. In some embodiments, the compounds of the present invention represented by formula (I) are incorporated herein by reference. The compound is a suitable trans-beta-substituted chlorin (where the two beta-chlorin substituents are halo( For example, providing a chlorine (Br) substituent, and then a beta-substituent reacting it to replace a suitable water-soluble group Methods for synthesizing trans-beta substituted chlorins are already known, for example, No. 6,559,372, which is incorporated herein by reference in its entirety. can be. For example, in some embodiments, the chlorin of formula (I) is The dihydrodipyrrin building block representing "stanhalf" (WH) is used in this chlorin. by condensing it with a building block representing the "eastern half" (EH) of In some embodiments, the WH building block can be prepared by , may have the following structure: [ka] In the formula, S1, S2, S7, S9, S12 and S13 each independently represent a hydrogen atom, Aryl, substituted aryl, phenyl, cycloalkyl, alkyl, substituted alkyl, alkene Nyl, alkynyl, halo, alkoxy, alkylthio, perfluoroalkyl, perfluoro Aryl, pyridyl, cyano, thiocyano, nitro, amino, alkylamino, acyl , sulfoxyl, sulfonyl, imido, amido and carbamoyl; or S7 and S 13 together form =O, or S1 and S2 together are substituted or unsubstituted alkylene In some embodiments, S 12 is a hydrogen atom be.

[0061] The EH building block compound may have the structure: [ka] In the formula, S3, S4, S5, S6 and S 11 are each independently a hydrogen atom, an aryl , substituted aryl, phenyl, cycloalkyl, alkyl, substituted alkyl, alkenyl, Alkynyl, halo, alkoxy, alkylthio, perfluoroalkyl, perfluoroaryl aryl, pyridyl, cyano, thiocyano, nitro, amino, alkylamino, acyl, sulfo S6 and S7 are selected from the group consisting of xyl, sulfonyl, imido, amido and carbamoyl. 5 together form a substituted or unsubstituted alkylene or arylene group. In embodiments, Z is halo (eg, Br, Cl, or I).

[0062] More specifically, dibromo-substituted chlorins and the corresponding bromo-substituted dihydropyrins. Methods for preparing WH and EH building blocks are already known. For example, Krayer et al. (2009) Journal of Porphyrins and Phthalocyanines 13(10):1098-1110; Jiang See, e.g., J.M. et al. (2014) Organic & Biomolecular Chemistry 12:86-103. Chlorin derivatives containing two bromo substituents at the 3- and 13-positions of the chlorin are shown, for example, in FIG. As shown in Scheme 1, WH-bis(2-formylpyrrole) was prepared from 2-formylpyrrole. It can be prepared from a bonding block. As shown in Scheme 1 of Figure 1, 2-formylpyrrole (pyrrole-2-carboxylate) The brominated aldehyde is first brominated using N-bromosuccinimide (NBS) to form the bromo-substituted aldehyde. The nitrogen atom of pyrrole a is then protected with a suitable protecting group Pg to give the protected pyrrole a. In some embodiments, the protecting group is a bromo-substituted pyrrole b. The nitrogen atom of roll a was deprotonated, and then the deprotonated compound was treated with p-toluenesulfonyl ether. The compound is installed by contacting the compound with phenylsulfonyl chloride to provide N-tosylpyrrole. The protected pyrrole b may be a tosyl group. (e.g., in the presence of potassium acetate and a slight excess of methylamine hydrochloride) to form the aldo The carbon-carbon double bond in this product c is converted into a condensation product c by a suitable reducing agent (e.g., LiBH4) to give compound d, which can be reacted with a non-nucleophilic base (e.g., diazabicyclo[4.2.1]cyclopentasiloxane). In the presence of lowundecene (DBU), treatment with mesityl oxide leads to a Michael addition reaction. The Michael addition product e was then treated with formamide and zinc powder to undergo reductive cyclization. , to give the protected dihydropyrin f, which is deprotected to give the chlorin WH.

[0063] Suitable EH chlorin building blocks can be prepared as shown in Scheme 2 in Figure 2. Ru. Laha et al. (2003) Organic Process Research & Development 7(6):799-812; t al. (2013) Journal of Organic Chemistry 78(21):10678-10691; and Liu et al. (201 6) See also New Journal of Chemistry 40(9):7721-7740. Phosphorus building blocks can be used as linker groups (e.g., methyl esters of after hydrolysis), or further (e.g., further function as a bioconjugable group or react an amino-containing alkyl group containing a second functional group capable of reacting to form a bioconjugable group; (via reaction with a chlorine compound) to provide a long linker group in the corresponding chlorin. The alkyl group may contain aryl substituents. As shown in Scheme 2 of Figure 2, pyrrole was reacted with methyl-4-formylbenzoate to give dipyrromethane g can be produced by the addition of methyl-4-formylbenzoyl methyl methyl methyl methyl methyl benzoyl ... When aldehydes other than benzoic acid methyl esters are used, dipyrromethanes substituted with groups other than methyl benzoate can be obtained. Dipyrromethane g can be converted to aldehyde h by Vilsmeier formylation. Aldehyde h is dibrominated using N-bromosuccinimide (NBS) to give Dibromodipyrromethane EH can be obtained. The WH and EH building blocks are prepared by the addition of an acid (e.g., trifluoroacetic acid or tosyl Condensation in the presence of an acid (Brønsted or Lewis acid, such as TsOH) to form a condensation product Then, in an organic solvent, a base (e.g., 2,2,6,6-tetramethylpiperidine), an oxidizing agent (e.g., trimethylpiperidine), In the presence of silver fluoromethanesulfonate and a metal salt (zinc acetate), the compound is oxidatively cyclized to form methyl Optionally, the metal (e.g., Zn) can be reacted with an acid (e.g., thiazolinone) to obtain thiazolinone. Trifluoroacetic acid) to provide the chlorin free base. In embodiments, the chlorin free base can have the structure: [ka]

[0064] The halo substituents of dihalochlorins, such as the dibromochlorin free bases described above, can further be selected from those skilled in the art. Coupling reactions known in the field (Stillet coupling, Hiyama coupling, Suzuki coupling) Coupling, Negishi coupling, Sonogashira coupling, Kumada coupling, etc. (including but not limited to) can be used to provide solubilizing groups. Dihalochlorins can be reacted with boronic acids in the presence of a Pd(0) catalyst; organotination in the presence of a Pd catalyst with pseudohalides or organosilanes in the presence of Pd catalysts; with pseudohalides or organosilanes in the presence of Ni or Pd catalysts with an organozinc compound in the presence of Zn, or with a Grignard reagent in the presence of Ni or Pd catalyst. In some embodiments, the dihalochlorine (e.g., The dibromochlorin free base is a suitable arylboronic acid Suzuki coupling reaction partner. Jiang et al. (2015) New Journal of Chemistry 39(7):5694 -5714; and Zhang et al.(2016) New Journal of Chemistry 40(9):7750-7767. In some embodiments, the arylboronic acid is preferably a boronic acid that is capable of reacting with the arylboronic acid in a Suzuki coupling reaction. It may contain additional or protected chemical functionalities that can be further elaborated later. For example, in some embodiments, the Suzuki coupling reaction can be carried out by reacting 1 or Further protected amino groups may be included, which, after deprotection, can be reacted with an appropriate PEG reagent. (e.g., activated PEG esters) can be reacted with Suzuki coupling reactions. Suitable catalysts for the reaction include, but are not limited to, Pd(PPh3)4 and PdCl2(PPh3)4. Suitable solvents include water, toluene, THF, dioxane, DMF, and combinations thereof. Suitable bases for use in Suzuki coupling reactions include Cs2CO3, K3PO4, NaOH, NaOEt , NaOtBu, Na2CO3, K2CO3, alkyllithium compounds, and trialkylamines (e.g. , triethylamine), and the like. In some embodiments, the compounds of the present invention may be used in combination with Suzuki or other types of coupling. t-butyloxycarbonyl (BOC) protecting groups (optional) for amino groups present on the reagent in combination with the use of t-butyl ester protection of the carboxylic acid, for example, in the linker group. Surprisingly, trifluoroacetic acid (TFA) can be used. While this is a common method for BOC deprotection, the use of TFA during the preparation of the compounds of the present invention Deprotection of the BOC group, especially in compounds containing, for example, alkyne bonds, can result in significant degradation. It has been found that BOC deprotection results in: This has been carried out using other conditions, e.g., 4M HCl in dioxane, which is not possible using TFA. This avoids the decomposition seen when using

[0065] IV. Pharmaceutical Compositions The compounds of the present invention can be provided as pharmaceutically acceptable salts. Contains N,N'-dibenzylethylenediamine, chloroprocaine, choline, ammonia, Diethanolamine and other hydroxyalkylamines, ethylenediamine, N-methyl Tilglucamine, procaine, N-benzylphenethylamine, 1-para-chlorobenzyl- 2-pyrrolidin-1'-ylmethyl-benzimidazole, diethylamine and other amines methylamine, piperazine and tris(hydroxymethyl)aminomethane; lithium, potassium alkali metal salts such as, but not limited to, sodium and barium; alkaline earth metal salts such as, but not limited to, sodium and magnesium; zinc transition metal salts; and sodium hydrogen phosphate and disodium phosphate, etc. Other metal salts include, but are not limited to, hydrochlorides and sulfates. Salts of mineral acids, including but not limited to acetate, lactate, malate, tartrate, citrate, ascorbate, including, but not limited to, benzoate, succinate, butyrate, valerate, and fumarate Pharmaceutically acceptable salts include, but are not limited to, salts of organic acids that are not soluble in water. Ternates include those with acidic groups (carboxylic, phosphoric, phosphinic, sulfonic, sulfinic, and and boronic acid. Aryl, heteroaryl, aralkyl, heteroaralkyl, cycloalkyl and hetero This includes, but is not limited to, esters of cyclyl.

[0066] The compounds of the present invention may also include prodrugs of the compounds disclosed herein. As noted above, a "prodrug" is a compound that, upon in vivo administration, undergoes one or more steps or metabolized by a process or otherwise converted into a biologically, pharmaceutically or therapeutically active form To generate a prodrug, a compound that is converted into a pharmaceutically active compound is The compound is modified so that the active compound is regenerated by metabolic processes. The drugs are used to alter the metabolic stability or transport properties of drugs, to mask side effects or toxicity. , designed to improve the taste of the drug or to modify other properties or characteristics of the drug. Knowledge of in vivo pharmacodynamic processes and drug metabolism has made this possible. Those skilled in the art, once a pharmaceutically active compound is known, can design a prodrug of that compound. (See, for example, Nogrady (1985) Medicinal Chemistry: A Biochemical Approach) , Oxford University Press, New York, New York, USA, pages 388-392 .)

[0067] utility The methods and intermediates of the present invention are useful for the synthesis of compounds of formula (I) as described Such compounds may be used as such or in further modified forms (e.g., salts, metallated compounds, etc.). Pandey et al., U.S. Patent Application Publication No. 2004 / 0044197, in a manner similar to other compounds described for photodynamic therapy, such as those described in , are useful for diagnostic and therapeutic purposes, as described in more detail below.

[0068] stability An advantage of some embodiments of the chlorin compounds of the present invention is their stability and absorption characteristics. Therefore, the present invention provides an active compound of the present invention (e.g., a compound represented by formula (I) or its pharmaceutically acceptable salts, prodrugs or conjugates (e.g., proteins, peptides or or a conjugate with a targeting agent such as an antibody), and has a wavelength of about 600 to about 800 nm and a wavelength of about 10,0 00~300,000M -1 cm -1 having or characterized by a peak molar absorption coefficient in solution of (a) providing a composition to be applied, which has a peak molar absorption coefficient at a specified wavelength; that (a) the active compound must be placed in solution to determine May exhibit additional peaks outside this range or multiple peaks within this range , hope you understand.) Furthermore, the present invention provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof in a solvent. salts, prodrugs or conjugates (e.g., with targeting agents such as proteins, peptides or antibodies) The amount of solvent is not critical. The composition may comprise from 0.01 (or 1) to 99 (or 99.99) weight percent of the composition. Approximately 10,000 to 300,000 M at wavelengths of 600 to approximately 800 nm -1 cm -1 Peak mole absorption in the above solutions Before measuring the molar absorption, the agglomerated particles are It is preferable to stir as needed to bring the molecules back into solution, but it is best to stir as soon as possible after actually using the composition. It should be understood that some degree of aggregation may be desirable when dissolving. Depending on the compound and the intended use of the compound, either organic solvents or aqueous solvents may be used. It may also be a combination of these. The composition may be mixed with the chlorin compound or compounds in "neat" form or with a solvent. Whether the chlorin compound or compounds are mixed, they can be stored in a sealed container (e.g., a flask). Store in the original container (samples or vials) at room temperature and away from ambient light for at least 3 or 4 months. When the chlorin compound of the present invention is present in an amount of more than about 10, 15, or 20 weight percent of the chlorin compound of the present invention, This decomposition can be detected by spectroscopy, thin layer chromatography, can be determined according to the prior art by NMR spectroscopy and / or mass spectrometry. do.

[0069] solubility An advantage of some embodiments of the compounds of the present invention is their water solubility. The present invention provides a method for preparing a liquid containing (a) an aqueous solvent (e.g., distilled water, saline, buffer solution), and (b) about 1, 2 , from 5 or 10 μM to 200, 300 or 500 mM, solubilized in aqueous solvent. Pharmaceutical formulations comprising, consisting of, or consisting essentially of the active compounds of the invention are also included. The present invention provides compositions including, but not limited to,

[0070] Formulation of pharmaceutical compositions The pharmaceutical compositions of the present invention comprise one or more compounds of the present invention in a pharmaceutically acceptable carrier. The composition comprises a therapeutically effective amount of the above, which is used to treat hyperproliferative tissue or angiogenesis-related or Prevention of one or more symptoms of a disease or disorder in which hyperproliferative tissue or angiogenesis is implicated These compounds are useful in the treatment or amelioration of diseases or disorders associated with hyperproliferative tissue or angiogenesis. These include cancer, psoriasis, atherosclerosis, heart disease and age-related macular degeneration. Suitable pharmaceutical carriers for administration of the compounds of the present invention include those suitable for particular modes of administration. Any carrier known to those skilled in the art to be suitable is included. The pharmaceutical composition preferably exhibits the absorption characteristics and storage or stability properties described above. Additionally, the compound may be formulated as the sole pharmaceutically active ingredient in the composition. or can be combined with other active ingredients. The composition may comprise one or more compounds of the present invention (e.g., a compound of formula (I) In some embodiments, the compound is administered as a solution, suspension, tablet, or the like for oral administration. Suitable formulations such as tablets, dispersible tablets, pills, capsules, powders, sustained release formulations or elixirs or sterile solutions or suspensions for parenteral administration, as well as transdermal patch formulations and dry powder formulations. In some embodiments, the compounds are formulated into inhalers and the like. The pharmaceutical compositions may be formulated using techniques and procedures well known in the art (see, for example, Ansel (1985) In troduction to Pharmaceutical Dosage Forms, Fourth Edition, Lea & Febiger, Philad Elphia, Pennsylvania, USA, page 126.)

[0071] In this composition, an effective concentration of one or more compounds or their pharmaceutically acceptable salts is The derivative is mixed with a suitable pharmaceutical carrier. The compound may be prepared as described above prior to formulation. The corresponding salts, esters, enol ethers or esters, acetals, ketals, octadecyl ether ... ester, hemiacetal, hemiketal, acid, base, solvate, hydrate or prodrug The concentration of the compound in the composition is determined by the amount of the compound administered. When hyperproliferative tissue or angiogenesis is involved, or when hyperproliferative tissue or angiogenesis is involved, delivers an amount that treats, prevents, or ameliorates one or more symptoms of a disease or disorder This is an effective concentration. In some embodiments, the composition is formulated for single administration. To prescribe, a drug is prescribed to treat a condition with an active ingredient that is intended to alleviate, prevent, or improve one or more symptoms. The compound is dissolved, suspended, dispersed or otherwise mixed in a selected carrier at an effective concentration by weight fraction. Combine. The active compound (i.e., a compound of formula (I) or a pharmaceutically acceptable salt thereof) The compound (salt, prodrug or conjugate) is intended to provide a therapeutic effect without undesirable side effects to the patient receiving the treatment. In this therapeutically effective amount, the compound is present in a pharmaceutically acceptable carrier in an amount sufficient to exert a beneficial effect. Effective concentrations are determined by the inclusion of the inclusions described herein and in U.S. Pat. No. 5,952,366 (Pandey et al.). This can be determined empirically by testing the compound in vitro and in vivo systems. From this, dosages for humans can be extrapolated.

[0072] The concentration of the active compound in the pharmaceutical composition determines the rate of absorption, inactivation and excretion of the active compound. The excretion rate, as well as the physicochemical properties of the compound, the administration schedule and dosage, and other factors will be apparent to those skilled in the art. For example, the amount delivered will depend on the amount of the drug administered, as described herein, and other factors known to those skilled in the art. Associated with or involving hyperproliferative tissue or angiogenesis , sufficient to ameliorate one or more symptoms of the disease or disorder. In some embodiments, the therapeutically effective dosage is about 0.1 ng / ml of active ingredient. should produce a serum concentration of about 50-100 μg / ml. The therapeutically effective dose is 0.5 mg of active compound per kilogram of body weight per day. 001 (or 0.01, 0.1) mg to 10 (or 100, 1000) mg. Dosage unit forms contain about 0.0 mg of the active ingredient or combination of essential ingredients per dosage unit form. 1 mg (or 0.1 mg, 1 mg) to about 500 mg (or 1000 mg, 2000 mg) ), and in some embodiments, prepared to provide about 10 mg to about 500 mg. It is manufactured. The active ingredient may be administered at once or in several smaller doses spaced apart over time. The exact dosage and duration of treatment are a function of the condition being treated. and using known test protocols or externally from in vivo or in vitro test data. It is understood that concentration and dosage values ​​may also be determined empirically by interpolation. Please note that this may vary depending on the severity of the symptoms to be alleviated. The particular dosing regimen for a patient will depend on their individual needs and the type of drug or drug to be administered or administered. The concentration ranges shown here should be adjusted over time according to the professional judgment of the supervisor. are merely exemplary and are not intended to limit the scope or practice of the compositions of the present invention. stomach.

[0073] If the compound exhibits insufficient solubility, methods for solubilizing the compound may be used. Such methods are known to those skilled in the art and include dimethyl sulfoxide. Use of cosolvents such as DMSO, polyoxyethylene sorbitol esters Use of surfactants such as acetaldehyde (e.g., sold under the trade name TWEEN®) can or by dissolving in aqueous sodium bicarbonate. Derivatives of this compound, such as prodrugs of the compound, may also be used to formulate effective pharmaceutical compositions. It can be used in the event. When the compound(s) are mixed or added, the resulting mixture may be a solution, a suspension, or a mixture of both. The form of the resulting mixture may be a liquid, such as a pharmaceutical, a pharmaceutical preparation ... The effective concentration will depend on many factors, including the solubility of the compound in the selected carrier or vehicle. The dose is an amount sufficient to improve the symptoms of the disease, disorder, or condition being treated, and is empirically can be determined.

[0074] The pharmaceutical composition comprises a tablet containing an appropriate amount of the compound or a pharmaceutically acceptable derivative thereof. , capsules, pills, powders, granules, sterile parenteral solutions or suspensions and oral solutions or suspensions It is provided for administration to humans and animals in unit dosage forms such as liquids and oil-in-water emulsions. can be. The pharmaceutically therapeutically active compounds and derivatives thereof, in some embodiments, comprise: The term "unit dosage form" as used herein refers to a dosage form that is formulated and administered in a unit dosage form or multiple dosage form. As known in the art, individually packaged, physically suitable for human and animal subjects Each unit dose refers to a discrete unit in combination with the required pharmaceutical carrier, vehicle or diluent. Each unit dosage form contains a predetermined amount of a therapeutically active compound sufficient to produce the desired therapeutic effect. Unit dose forms include ampoules and syringes and individually packaged tablets or capsules. may be administered in fractions or multiples thereof. Multiple dosage forms may be administered in separate unit dosage forms. Multiple unit dosage forms are identical and packaged in a single container. Examples include vials, tablet or capsule bottles, or pint or gallon bottles. Thus, multiple-dosage form is a multiple of unit doses that are not segregated in packaging. do.

[0075] Liquid pharmaceutically administrable compositions can be, for example, liquids containing an active compound as defined above, e.g. A compound of formula (I) or a pharmaceutically acceptable salt, prodrug or conjugate thereof and any pharmaceutical adjuvants in a carrier (e.g., water, saline, aqueous dextrose, Dissolve, disperse, or otherwise mix in glycerol, glycol, ethanol, etc. The dosage can be adjusted by forming a solution or suspension. The pharmaceutical composition may also contain additives such as wetting agents, emulsifying agents, solubilizing agents, pH buffering agents, and the like (e.g., acetic acid, Sodium, Sodium Citrate, Cyclodextrin Derivatives, Sorbita Monolaurate triethanolamine acetate, triethanolamine oleate, and other Such agents may contain small amounts of non-toxic auxiliary substances. Actual methods for preparing such dosage forms are known to those skilled in the art or are currently being developed. See, for example, Remington's Pharmaceutical Sciences, 15th Edition. ion, 1975, Mack Publishing Company, Easton, Pennsylvania, USA. A dosage form or composition containing 0.005% to 100% of the active ingredient, with the remainder consisting of a non-toxic carrier. Methods for preparing these compositions are well known to those skilled in the art. Contemplated compositions contain from 0.001% to 100% of the active ingredient, in some embodiments from 0.1 to 95%, in others In this embodiment, the content can be 75 to 85%.

[0076] Compositions for oral administration Oral pharmaceutical dosage forms may be solid, gel, or liquid. Solid dosage forms include tablets, capsules, and Oral tablet types include compressed, chewable tablets, granules, and bulk powders. These include oats and tablets, which may be enteric coated, sugar coated, or The capsule may be a hard or soft gelatin capsule. However, granules and powders may be combined with other ingredients known to those skilled in the art to produce non-effervescent or may be provided in effervescent form.

[0077] Solid Composition for Oral Administration In some embodiments, the pharmaceutical formulation is a solid dosage form, and in some embodiments, In this case, the tablets, pills, capsules, lozenges, etc. The following ingredients (e.g., binders; lubricants; diluents; glidants; disintegrants; colorants; sweeteners) ; flavorings; humectants; emetic coatings; film coatings) or similar substances The binder may include one or more compounds. Examples of binders include microcrystalline cellulose. , tragacanth gum, glucose solution, acacia mucus, gelatin solution, molasses, polyynyl These include pyrrolidine, povidone, crospovidone, sucrose and starch paste. Lubricants include talc, starch, magnesium or calcium stearate, lycopodine Diluents include, for example, lactose, sucrose, starch, and stearic acid. Glidants include methylcellulose, kaolin, salt, mannitol, and dicalcium phosphate. Disintegrants include, but are not limited to, colloidal silicon dioxide. Sodium sucrose, sodium starch glycolate, alginic acid, corn starch, di Potato starch, bentonite, methylcellulose, agar and carboxymethylcellulose Coloring agents include, for example, any of the approved certified water-soluble FD and C dyes. or mixtures thereof, and water-insoluble FD and C dyes suspended in alumina hydrate. Sweeteners include sucrose, lactose, mannitol, and artificial sweeteners such as saccharin. Flavoring agents include those extracted from plants such as fruits, as well as any number of spray-dried flavors. Natural flavors and other ingredients, including but not limited to peppermint and methyl salicylate The humectants include synthetic blends of compounds that produce a pleasant sensation of moisturizing (not moisturizing). Propylene glycol monostearate, sorbitan monooleate, diethyl monolaurate Contains ethylene glycol and polyoxyethylene laural ether. The coatings include fatty acids, fats, waxes, shellac, ammoniated shellac and acetic acid phthalates. Film coatings include hydroxyethyl cellulose, Gellan gum, sodium carboxymethylcellulose, polyethylene glycol 4000 ( PEG 4000) and cellulose acetate phthalate.

[0078] This compound, or a pharmaceutically acceptable derivative thereof, protects it from the acidic environment of the stomach. For example, the composition can be provided in a composition that maintains its integrity in the stomach and in the intestine. The composition may be formulated with an enteric coating that releases the active compound. It may be formulated in combination with an acidic agent or other such ingredient. If the cell is a cell, it may contain, in addition to the above types of materials, a liquid carrier such as a fatty oil. In addition, dosage unit forms can contain various other materials which modify the physical form of the dosage unit. The compound may also contain coatings of, for example, sugars and other enteric agents. , elixirs, suspensions, syrups, wafers, sprinkles, chewing gum, etc. Syrups contain, in addition to the active compounds, sugar as a sweetener. They may contain cellulose, as well as certain preservatives, dyes, colorings and / or flavors. In addition, this active substance may be used in combination with other active substances that do not impair the desired effect, such as antacids, H2 blockers, etc. It can be mixed with other ingredients that supplement the desired action, such as medicines and diuretics. The compound of the present invention or a pharmaceutically acceptable derivative thereof is an active ingredient having a molecular weight of about 98. It can be included in concentrations up to 100% by volume. In some embodiments, tablet and capsule formulations modify the dissolution of the active ingredient. or may be coated, as known by those skilled in the art, to maintain Thus, for example, these include phenyl salicylate, wax and acetate phthalate. They can be coated with a conventional enteric coating such as cellulose.

[0079] Liquid compositions for oral administration Liquid oral dosage forms include aqueous solutions, emulsions, suspensions, and formulations reconstituted from non-effervescent granules. Includes solutions and / or suspensions, as well as effervescent preparations reconstituted from effervescent granules. Solutions include, for example, elixirs and syrups. Emulsions include oil-in-water or It is either water-in-oil type. Elixirs are clear, sweetened, hydroalcoholic medicinal formulations. The pharmaceutically acceptable carriers used include solvents. Syrups are made from sugars such as sucrose. Emulsions are concentrated aqueous solutions of one liquid mixed with another. It is a two-phase system in which small globules are dispersed throughout the emulsion. Pharmaceutically acceptable carriers are non-aqueous liquids, emulsifying agents and preservatives. Use suspending agents and preservatives that are acceptable for use in non-effervescent granules to be reconstituted into a liquid oral dosage form. Pharmaceutically acceptable substances that may be used include diluents, sweeteners and wetting agents. Pharmaceutically acceptable substances used in effervescent granules to be reconstituted into dosage forms include organic acids and diglycerides of ... A carbon dioxide source is included. Coloring agents and flavoring agents are used in all of the above formulations. Solvents include Examples of preservatives include glycerin, sorbitol, ethyl alcohol, and syrup. , glycerin, methyl and propylparaben, benzoic acid, sodium benzoate and alcohol Examples of non-aqueous liquids utilized in emulsions include mineral oil and cottonseed oil. Examples of emulsifying agents include gelatin, acacia, tragacanth, bentonite, and polyoxyethylene. Suspending agents include surfactants such as diethylene sorbitan monooleate. Sodium hydroxymethylcellulose, pectin, tragacanth, xanthan gum, Beegum Sweeteners include sucrose, syrup, glycerin, and saccharin. Contains artificial sweeteners such as propylene glycol monostearate, monostearate, Sorbitan oleate, diethylene glycol monolaurate and polyoxyethylene laurate Organic acids include citric acid and tartaric acid. Carbon dioxide sources include Colorants include sodium bicarbonate and sodium carbonate. Approved certified water-soluble colorants are Contains any of the FD and C dyes and their mixtures. Flavoring agents include those derived from fruits and other plants. Contains extracted natural flavors and a synthetic blend of compounds that produce a pleasant taste sensation. For dosage forms, the formulation may be in solution (e.g., in propylene carbonate, vegetable oils, and / or triglycerides) or In some embodiments, the suspension is encapsulated in a gelatin capsule. The solution, and its preparation and encapsulation, are described in U.S. Pat. No. 4,328,245, U.S. Pat. No. 4,409,222, and U.S. Pat. No. 4,409,222. 39 and U.S. Pat. No. 4,410,545, which are incorporated herein in their entireties. In the case of a liquid dosage form, for example, a solution in polyethylene glycol may be suitable for administration. The composition can be diluted with a sufficient amount of a pharmaceutically acceptable liquid carrier (e.g., water).

[0080] Alternatively, liquid or semisolid oral formulations may be prepared by dissolving the active compound or salt in a liquid or semisolid carrier such as vegetable oil, glucose, or the like. triglycerides, propylene glycol esters (e.g., propylene carbonate) and others The solution or suspension is then dissolved or dispersed in such a carrier, and the solution or suspension is then placed in a hard or soft gel. It can be prepared by encapsulating it in a lattice capsule shell. Useful formulations include those described in U.S. Patent Nos. RE28,819 and 4,358,603. These include, but are not limited to, those incorporated herein in their entirety. The formulation may contain a compound of the present invention, a dialkylated mono- or polyalkylene glycol (1,2-dialkylene glycol), or Methoxymethane, diglyme, triglyme, tetraglyme, polyethylene glycol-3 50-dimethyl ether, polyethylene glycol-550-dimethyl ether, polyethylene glycol Alcohol-750-dimethyl ether (where 350, 550 and 750 are polyethylene ) refers to the approximate average molecular weight of the glycol. One or more antioxidants (butylated hydroxytoluene (BHT), butylated hydroxy Anisole (BHA), propyl gallate, vitamin E, hydroquinone, hydroxycoumarin , ethanolamine, lecithin, cephalin, ascorbic acid, malic acid, sorbitol , phosphoric acid, thiodipropionic acid and its esters, and dithiocarbamates) These include, but are not limited to: Other formulations include aqueous alcohol solutions containing pharmaceutically acceptable acetals. The alcohols used in these formulations include, but are not limited to, one or more Any pharmaceutically acceptable water-miscible solvent having the above hydroxyl group, Acetals include, but are not limited to, glycol and ethanol. However, di(low) alkyl aldehydes such as acetaldehyde diethyl acetal are not (methylene alkyl) acetals.

[0081] Injections, solutions and emulsions In some embodiments featuring injection, either subcutaneously, intramuscularly, or intravenously, Parenteral administration is also contemplated herein. Injectables are liquid solutions or suspensions, liquids that are prepared prior to injection. In conventional forms, either as solid forms suitable for solution or suspension in the body, or as emulsions. The injections, solutions and emulsions may contain one or more Suitable excipients include, for example, water, saline, dextrose, glycerin, PEG-40 ... Furthermore, if necessary, the pharmaceutical composition to be administered may also , wetting or emulsifying agents, pH buffering agents, stabilizers, solubility enhancers, and other such agents (e.g., , sodium acetate, sorbitan monolaurate, triethanolamine oleate, and It may also contain small amounts of non-toxic auxiliary substances, such as cyclodextrins.

[0082] Implantation of a sustained-release or sustained-release system (e.g., a rice See US Patent No. 3,710,795, which is incorporated herein in its entirety. Briefly, the compounds of the present invention are distributed in a solid internal matrix. The solid inner matrix may be, for example, polymethyl methacrylate, poly(methyl methacrylate), or Butyl methacrylate, plasticized or unplasticized polyvinyl chloride, plasticized nylon, plasticized Polyethylene terephthalate, natural rubber, polyisoprene, polyisobutylene, polybutylene Polyethylene, ethylene-vinyl acetate copolymer, silicone rubber, poly Dimethylsiloxane, silicone carbonate copolymer, hydrophilic polymer, e.g. For example, hydrogels of acrylic and methacrylic acid esters, collagen, cross-linked polyvinyl alcohol, and cross-linked partially hydrolyzed polyvinyl acetate, which is the outer polymer membrane ( For example, polyethylene, polypropylene, ethylene / propylene copolymer, ethylene / acrylic copolymer Ethyl acrylate copolymer, ethylene / vinyl acetate copolymer, silicone rubber, polydimethyl Siloxane, neoprene rubber, chlorinated polyethylene, polyvinyl chloride, salts with vinyl acetate Vinyl chloride copolymer, vinylidene chloride, ethylene and propylene, ionomer polyethylene Terephthalate, butyl rubber, epichlorohydrin rubber, ethylene / vinyl alcohol copolymer ethylene / vinyl acetate / vinyl alcohol copolymer, and ethylene / vinyloxyethylene They are polyol copolymers, which are insoluble in body fluids. The compound diffuses through the outer polymeric membrane in a release rate controlling step. The percentage of active compound contained in such parenteral compositions will depend on the specific nature of the composition and the amount of the compound. The effectiveness of the treatment will depend largely on the activity of the agent and the needs of the patient.

[0083] Parenteral administration of the composition includes intravenous, subcutaneous, and intramuscular administration. Preparation includes a sterile solution ready for injection; ready to be combined with a solvent immediately before use Sterile dry soluble products (including hypodermic tablets), such as lyophilized powders; preparations for injection Ready to be combined with vehicle and sterile emulsion immediately before use These solutions may be either aqueous or non-aqueous. This may also be the case. If administered intravenously, suitable carriers include physiological saline or phosphate buffered saline (PB S), and thickeners and solubilizers (glucose, polyethylene glycol and polypropylene and mixtures thereof. Pharmaceutically acceptable carriers used in parenteral formulations include aqueous vehicles, non-aqueous vehicles, and antibacterial agents, isotonic agents, buffers, antioxidants, local anesthetics, suspending and dispersing agents, emulsifiers, sequestrants The term "anticoagulant" includes a coagulant or chelating agent, and other pharmaceutically acceptable substances.

[0084] Examples of aqueous vehicles include sodium chloride injection, Ringer's injection, and isotonic dextrose injection. Non-aqueous parenteral vehicles include sterile water injection, dextrose and lactated Ringer's injection. These include fixed oils of vegetable origin, cottonseed oil, corn oil, sesame oil, and peanut oil. or fungistatic concentrations of antimicrobial agents may be added to parenteral preparations packaged in multi-dose containers. This includes phenol or cresol, mercury, benzyl alcohol, chlorobutanoic acid, methyl and propyl p-hydroxybenzoates, thimerosal, benzal chloride Isotonic agents include sodium chloride and dextromethorphan. Buffers include phosphate and citrate. Antioxidants include sulfuric acid. Local anesthetics include procaine hydrochloride. Suspending and dispersing agents Contains sodium carboxymethylcellulose, xanthan gum, hydroxypropyl methylcellulose The emulsifiers include polysorbate 80 ( (sold under the trade name TWEEN® 80). Used as a sequestering or chelating agent for metal ions. Pharmaceutical carriers include ethyl alcohol for water-miscible vehicles, polyethylene glycol for water-miscible vehicles, and EDTA. Ethylene glycol and propylene glycol; and sodium hydroxide and hydrochloric acid for pH adjustment , citric acid or lactic acid. The concentration of the pharmaceutically active compound is sufficient to produce the desired pharmacological effect upon injection. The exact dosage can be adjusted to provide an effective amount. As such, the dosage will depend on the age, weight and condition of the patient or animal.

[0085] Unit-dose parenteral formulations are packaged in ampoules, vials, or syringes with needles. All preparations for parenteral administration are prepared as known and practiced in the art. Therefore, it must be sterile. Illustratively, intravenous or intraarterial infusion of a sterile aqueous solution containing an active compound is an effective mode of administration. Another embodiment is a live agent that is injected as needed to produce the desired pharmaceutical effect. It is a sterile aqueous or oily solution or suspension containing the substance. Injectables are designed for local and systemic administration. Dosage may vary depending on the specific dosage, with concentrations of the active compound in the treated tissue ranging from about 0.1% w / w to about 90% w / w or more. In some embodiments, it is formulated to be greater than 1% w / w. This compound may be used to generate a more soluble active product or to generate a prodrug. To obtain the desired product, the product may be suspended in micronized or other suitable form or derivatized. The form of the mixture to be administered will depend on the intended mode of administration and the composition of the compound in the selected carrier or vehicle. The effective concentration depends on many factors, including the solubility of the drug. and can be determined empirically.

[0086] freeze-dried powder It can be reconstituted for administration as solutions, emulsions and other mixtures. Sintered dry powders can also be used to practice the present invention. can be reconstituted or formulated as a gel. The sterile, lyophilized powder contains the compound of the invention or a pharmaceutically acceptable derivative thereof in a suitable solution. The solvent is prepared by dissolving the drug in a solvent that contains excipients or powders that improve stability. Other pharmacological components of the powder or reconstituted solution prepared from the powder may also be included. Excipients include dextrose, sorbital, fructose, corn syrup, xylitol, Examples include, but are not limited to, glycerin, glucose, sucrose, or other suitable agents. This solvent also contains citrate, sodium phosphate, or potassium phosphate. or other such buffers known to those skilled in the art, and in some embodiments In some cases, it may include a neutral pH. The solution is then sterile filtered and then lyophilized under standard conditions known to those skilled in the art. In one embodiment, the resulting solution is The compound is then distributed into multiple vials for lyophilization. Each vial contains a single dose of the compound. This lyophilized powder can be stored at room temperature, such as at about 4°C to room temperature. It can be stored under appropriate conditions. This lyophilized powder, when reconstituted with water for injection, is formulated for use in parenteral administration. To reconstitute, add this lyophilized powder to sterile water or another suitable carrier. The exact amount will depend on the compound selected; this amount can be determined empirically. Cut.

[0087] Topical administration The topical mixture is prepared as described for local and systemic administration. The mixture may be in the form of a solution, suspension, emulsion, or the like, and may be in the form of a cream, gel, ointment, Emulsions, solutions, elixirs, lotions, suspensions, colorants, pastes, foams, Aerosol, irrigation, spray, suppository, bandage, skin patch, or other formulation suitable for topical administration It is prescribed as a substance. The compound or a pharmaceutically acceptable derivative thereof may be formulated as an educt for topical application, such as by inhalation. and can be formulated as an aerosol (see, e.g., U.S. Pat. Nos. 4,044,126; 4,414,209). and 4,364,923, which are useful for treating inflammatory diseases, particularly asthma. and US Pat. No. 6,629,293, each of which describes an aerosol for delivering steroids, each of which is the subject of the present invention in its entirety. These formulations for administration to the respiratory tract may be administered as aerosols or nebulizers. It may be in the form of a solution for infusion or as a finely divided powder for insufflation; It may be used alone or in combination with an inert carrier such as lactose. In some embodiments, the particle diameter of the object is less than 50 μm, and in some embodiments, In this case, it is less than 10 μm. The compound is administered topically to the skin or mucous membranes (e.g., eyes) in the form of a gel, cream, or lotion. It may also be formulated for topical application, such as for use in the eyes, or for intracisternal or intraspinal application. Topical administration can be by transdermal delivery, and also by administration to the eyes or mucous membranes, or by inhalation therapy. The active compound may be administered alone or in combination with other pharmaceutically acceptable excipients. These solutions, especially ophthalmic solutions, can also be administered in combination with nasal solutions. It can be formulated as a 0.01% to 10% isotonic solution at a pH of about 5 to 7 containing appropriate salts.

[0088] Compositions for other routes of administration Other routes of administration, such as transdermal patches and rectal administration, including iontophoretic and electrophoretic devices, are also possible. That's to be expected here. Transdermal patches, including iontophoretic and electrophoretic devices, are well known to those of skill in the art. For example, such patches are disclosed in U.S. Pat. Nos. 6,267,983, 6,261,595, and 6,261,595. No. 6,256,533, U.S. Patent No. 6,167,301, U.S. Patent No. 6,024,975, U.S. Patent No. 6,010715 , U.S. Patent No. 5,985,317, U.S. Patent No. 5,983,134, U.S. Patent No. 5,948,433, and U.S. No. 5,860,957, which is incorporated herein in its entirety. For example, pharmaceutical dosage forms for rectal administration include rectal suppositories, capsules, and Rectal suppository, as used herein, means a solid substance for insertion into the rectum, which is a tablet. melts or softens at body temperature to release one or more pharmacologically or therapeutically active ingredients Pharmaceutically acceptable substances used in rectal suppositories generally include bases or vehicles to raise the melting point. Examples of bases include cocoa butter (theobroma oil), glycerin, gelatin, carbowax (polyoxyethylene glycol) and fatty acid mono- and di- and triglycerides. That's fine. Agents that raise the melting point of suppositories include spermaceti and wax. Rectal suppositories are prepared by the compressed or molded method. In some embodiments, the weight of a rectal suppository is is about 2 to 3 grams. Tablets and capsules for rectal administration are formulated with the same pharmaceutically acceptable substances as formulations for oral administration. and are produced in the same manner.

[0089] Targeted Formulations The compounds of the present invention or pharmaceutically acceptable derivatives thereof may also be used to treat specific tissues, receptors, infections, and the like. The agent may be formulated to target the agent or other area of ​​the patient's body to be treated. Many such targeting methods are well known to those skilled in the art. Methods are contemplated for use in the present compositions. Non-limiting examples of targeting methods include: For example, U.S. Patent No. 6,316,652, U.S. Patent No. 6,274,552, U.S. Patent No. 6,271,359, U.S. Patent No. US Patent No. 6,253,872, US Patent No. 6,139,865, US Patent No. 6,131,570, US Patent No. 6,1 20,751, U.S. Patent No. 6,071,495, U.S. Patent No. 6,060,082, U.S. Patent No. 6,048,736, U.S. Patent No. 6,039,975, U.S. Patent No. 6,004,534, U.S. Patent No. 5,985,307, U.S. Patent No. 5 ,972,366, U.S. Patent No. 5,900,252, U.S. Patent No. 5,840,674, U.S. Patent No. 5,759,542 and U.S. Patent No. 5,709,874, which are incorporated herein in their entireties. It can be enjoyed.

[0090] Liposomes In some embodiments, the liposomes include tissue-targeted liposomes, such as tumor-targeted liposomes. Liposomal suspensions may also be suitable as pharmaceutically acceptable carriers. They can be prepared according to methods known to those skilled in the art, for example, liposomal pharmaceutical formulations. can be prepared as described in U.S. Pat. No. 4,522,811, the entire contents of which are incorporated herein by reference. Briefly, egg phosphatidylcholine and brain phosphatidylcholine are mixed inside the flask. By drying the mixture of acetylcholine and acetylserine (molar ratio 7:3), it was possible to form lysates such as multilamellar vesicles (MLVs). A flask was filled with phosphate buffered saline lacking divalent cations (PBS). A solution of the compound of the present invention in saline is added and the flask is shaken until the lipid film is dispersed. The resulting vesicles are washed to remove unencapsulated compounds and centrifuged. The cells are then pelleted and resuspended in phosphate buffered saline (PBS).

[0091] Ligand In some embodiments, the compounds disclosed herein are administered to a target tissue or target composition. Using specific ligands (e.g., ligand or ligand-receptor pairs (antibodies and antigens) Anti-tumor agents can be used to target specific target tissues or target compositions. Antibodies against tumors or infectious lesions (viruses, bacteria, fungi, parasitic infections, and antigens and products that bind to such microorganisms Antibodies and antibody fragments that specifically bind to markers that bind to the markers are particularly U.S. Patent No. 3,927,193, U.S. Patent No. 4,331,647, U.S. Patent No. 4,348,376, U.S. Patent No. Patent No. 4,348,376, U.S. Patent No. 4,361,544, U.S. Patent No. 4,468,457, U.S. Patent No. 4,444,7 44, U.S. Pat. No. 4,818,709, and U.S. Pat. No. 4,624,846, which are and the like, which are incorporated herein in their entirety. Antibodies against antigens of tumors such as lymphoma, sarcoma, or melanoma can be used.

[0092] A wide range of monoclonal antibodies against various infectious disease pathogens have been developed, and Pollin (198 4) Summarized in a review by the European Journal of Clinical Microbiology 3(5):387-398 These include: Monoclonal antibodies (MAbs) against these antigens include: Streptococcus aureus Galactia, Legionella pneumophila, Streptococcus pyogenes, Escherichia coli, Neisseria gonorrhoeae, Neisseria meningitidis, Streptococcus pneumoniae, Haemophilus influenzae B, Treponema pallidum, Lyme disease, spirochetes, Pseudomonas aeruginosa, etc. Antibacterial monoclonal antibodies (MAb) against bacteria, Brucella, Mycobacterium tuberculosis, tetanus toxin, etc. Plasmodium falciparum, Plasmodium vivax, Toxoplasma gondii, Trypanosoma langermanii -Ma, Trypanosoma cruzi, Trypanosoma rhododesiensei, Trypanosoma marcesii, Man Schistosoma sonnei, Schistosoma japonicum, Mesocestoides corti, Emelia tenella, Onchocerca volvulus, Leishmania tropica, Trichinella spiralis, Theileria parva, Taenia hydatidi, Taenia saginata, Taenia saginata, etc. Antiprotozoan monoclonal antibodies (MAbs) against HIV-1, -2, and -3, types A, B, C, and D , Ravivirus, influenza virus, cytomegalovirus, simplex virus Viruses I and II, human serum parvo-like virus, respiratory syncytial virus, varicella zoster virus Virus, Hepatitis B virus, Measles virus, Adenovirus, Human T-cell leukemia virus , Epstein-Barr virus, mumps virus, Sindbis virus, mouse mammary tumor Viruses, feline leukemia virus, lymphocytic choriomeningitis virus, wart virus, bull Viruses such as leukemia virus, polio virus, dengue virus, and rubeola virus. Antiviral monoclonal antibodies (MAb) against rabies virus, murine leukemia virus, etc. Acholeplasma laidlowii, Mycoplasma arthritis, Mycoplasma hyorini Antimycoplasma monoclonal antibody against Mycoplasma orreli, M. arginine, M. pneumoniae, etc. Clonal antibodies (MAbs)

[0093] Suitable monoclonal antibodies (MAbs) are the most common antigens responsible for the majority of infections in humans. It has been developed against a wide range of microorganisms (bacteria, viruses, protozoa, and other parasites) and Many of these antibodies have been used in the past for in vitro diagnostic purposes. Newer monoclonal antibodies (MAbs) that can be generated in combination with the compounds of the invention are These compounds are suitable for use as targeted agents. Monoclonal antibodies (MAbs) against malaria parasites have been reported to target sporozoites, merozoites, and cytozoites. Monoclonal antibodies can be directed against the zoont and gametocyte stages. (circumsporozoite antigen) and in vitro and in rodent models. It has been shown to neutralize sporozoites. Yoshida et al. (1980) Science 207:71-73 against T. gondii, the protozoan parasite responsible for toxoplasmosis. A monoclonal antibody was developed. Kasper et al. (1982) Journal of Immunology 12 9:1694-1699. Monoclonal antibodies (MAbs) have been developed to target the Schistosoma surface antigen. It has been developed against Schistosoma and has been shown to be effective against Schistosoma in vitro and in vivo. It is known that Simpson et al.(1981) Parasitology 83:163-177; Smith et al.(1982) P arasitology 84:83-91; Gryzch et al. (1982) Journal of Immunology 129:2739-2743; odda et al. (1982) Journal of Immunology 129:2326-2328; and Dissous et al. (1982) J See the Journal of Immunology 129:2232-2234.

[0094] Mixtures of antibodies and immunoglobulin classes can be used, as can hybrid antibodies. Multispecific, including hybrid, antibodies and antibody fragments can be used in the methods of the invention. Particularly preferred for detecting and treating target tissues are at least two different substantially identical at least two of the antibodies or antibody fragments are monospecific antibodies or antibody fragments At least two different antigens produced or associated with the target lesion, or produced by the target tissue. Specific for at least two different epitopes or molecules of the native or related marker substance. Multispecific antibodies and antibody fragments with dual specificities are described in U.S. Pat. No. 4,361,544. The hybrids can be prepared in the same manner as the anti-tumor marker hybrids disclosed in Other techniques for preparing rDNA antibodies are described, for example, in U.S. Pat. No. 4,474,893 and U.S. Pat. No. 4,479,895, which are incorporated herein in their entireties. This technique is disclosed in Milstein et al. (1984) Immunology Today 5:299.

[0095] Antibody fragments useful in the present invention include F(ab')2, F(ab)2, Fab', including hybrid fragments. Fab, Fv, etc. Preferred fragments are Fab', F(ab')2, Fab, and F(ab)2. It retains the hypervariable antigen-binding region of the immunoglobulin and has a size similar to or smaller than the Fab' fragment. Any subfragment of the same size as the antibody fragment is also useful, including those that incorporate the antigen-binding site and function as targeting vehicles in vivo in substantially the same manner as natural immunoglobulin fragments , genetically engineered and / or recombinant proteins (whether single-chain or multi-chain) Such single-stranded binding molecules are disclosed in U.S. Pat. No. 4,946,778. and is incorporated herein by reference in its entirety. Fab' antibody fragments are derived from the F(ab')2 fragment. F(ab')2 fragments themselves are easily produced by reactive cleavage of intact immunoglobulins. Fab antibody fragments can be generated by cleavage of intact immunoglobulins under reducing conditions. F(ab)2 derived from papain digestion or careful papain digestion of whole immunoglobulins It can be created by cutting fragments. The ligand, or one member of a ligand-receptor binding pair, is used to bind the compound of the invention to a specific target. Ligands can be conjugated to the compounds of the invention for targeting to tissues or target compositions. Examples of endonucleaser-receptor binding pairs are described in U.S. Pat. Nos. 4,374,925 and 3,817,837. , which are incorporated herein in their entireties.

[0096] Conjugation to ligands There are many ligand-receptor binding pairs that can serve as targets for antibodies, more specifically Compounds have been identified and are useful for constructing conjugates of such ligands with compounds of formula (I). Techniques for this are well known to those of ordinary skill in the art. For example, Rakestraw et al. Sn(IV) chlorination via covalent attachment to monoclonal antibodies using modified dextran carriers Rakestraw et al. (1990) Proceedings of the National See Academy of Sciences of the USA 87:4217-4221. The compounds of the present invention also , can be conjugated to a ligand such as an antibody by using a coupling agent. The components can be combined in a manner that makes them stable under physiological conditions for the time required for administration and treatment. Any bond that can be formed is suitable, but a covalent bond is preferred. For example, the compound of formula (I) may be directly linked to the targeting agent; or For example, a compound of formula (I) may be linked to an intermediate, which is in turn linked to a targeting agent. It may be indirect, such as: Coupling agents are sensitive to temperature, pH, salt, solvent system, as well as the photosensitizer, backbone (if present). (if present) and other reactants that substantially maintain the chemical stability of the targeting agent. The coupling agent must be able to stably link the component moieties, but the formula ( I) There is a need to minimize or eliminate any denaturation or inactivation of the compound or targeting agent. Many coupling agents react with amines or carboxylic acid salts to form amides or Coupling agents are known in the art for reacting alcohols with carboxylic acid salts to form esters. For example, Bodansky (1993) Principles of Peptide Synthesis, 2nd ed., Springer & Hermanson(1996) Bioconjugate Techniques, 1 st Ed., Academic Press, Ne See www.mhw.nyc.gov.uk / newsroom / newyork / usa.

[0097] Conjugates of the compounds of the invention with a ligand such as an antibody may be conjugated to the antibody via the N-terminus of the peptide. By coupling a carboxylic acid or ester moiety on the compound with a bond, the compound by coupling the compound to a targeting moiety or by other methods known in the art. A variety of coupling agents, including cross-linking agents, can be used for covalent bonding. Examples of such cross-linkers include N,N'-dicyclohexylcarbodiimide (DCC), N-succinimide, N-succinimidyl-5-acetyl-thioacetate (SATA), N-succinimidyl-3-(2-pyridyldithio) e) Propionate (SPDP), ortho-phenylene-dimaleimide (o-PDM) and sulfosulfonate Chuccinimidyl 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (sulfo) -SMCC). For example, Karpovsky et al. (1984) Journal of Experimental M edicine 160(6):1686-1701; and Liu et al. (1985) Proceedings of the National Acad See the Society of Science of the USA 82(24):8648-8652. n et al. (1985) Science 229:81-83 and Glennie et al. (1987) Journal of Immunolog y 139:2367-2375. For example, N,N'-dicyclohexylcarbodiimide (DCC) reacts with the alcohol NHS in DMSO. The active ester can be crosslinked to polylysine by promoting the coupling of carboxyl groups with chlorin. N,N'-dicyclohexyl is a useful coupling agent that can be used to form esters. Carbodiimide (DCC) is commonly used as a coupling agent in peptide synthesis. Another useful crosslinker is N-sulfonyl methylcellulose, a carboxy-reactive crosslinker with a molecular weight of 206.32. Chuccinimidyl-3-(2-pyridyldithio)propionate (SPDP), a primary amine and sulfhydryl groups. The molecular weight of SPDP is 312.4. The length of the sar arm is 6.8 angstroms, and it reacts with NHS ester and pyridyldithio group. It creates a cleavable crosslink that upon further reaction removes the drug and releases the photosensitizer. Other useful conjugates are blocks for two-step crosslinking. N-sequence to introduce a blocked SH group (which is deblocked with hydroxylamine-HCl) Chuccinimidyl-5-acetyl-thioacetate (SATA) and its affinity for amines and sulfhydryls Reactive sulfosuccinimidyl 4-(N-maleimido-methyl)-cyclohexane-1 -carboxylate (sulfo-SMCC). Other crosslinking and coupling agents are also available from Pierce Chem. The proteins can be combined with other proteins or other compositions (e.g., for conjugation to reporter groups or to chelators for metal ion labeling of proteins Additional compounds and processes for the preparation of synthetic derivatives (especially those involving Schiff bases as intermediates) are listed in the European Patent Application No. It is disclosed in EP 0 243 929 B1.

[0098] Photosensitizers containing carboxyl groups are prepared by reacting preformed reactive esters (N-hydroxybenzoates). The imide is reacted with the imide via a carbodiimide-mediated reaction (e.g., NHS ester) or The lysine ε-amino acid of the target polypeptide can be conjugated either in situ or by ester conjugation. The same can be done with sulfones, which can be converted to sulfonyl chlorides that react with amino groups. This also applies to photosensitizers containing acid groups. Chlorins with carboxyl groups are It can be attached to amino groups on polypeptides by the carbodiimide method of The hydroxyl groups of serine or threonine residues or the sulfhydryl groups of cysteine ​​residues bind to the It can also be bonded to a methyl group. A method for linking the components of the conjugate (e.g., attaching a polyamino acid chain that yields a photosensitizer to an antibacterial poly(amino acid) chain) The method of coupling to a peptide involves the use of heterobifunctional cross-linking reagents. These agents bind to a functional group on one chain and another functional group on the second chain. These functional groups are usually amino, carboxyl, sulfhydryl, and aldehyde. There are many permutations of suitable moieties that react with these groups to form It also reacts with differently formulated structures to conjugate them together. Hermanson (1996) B ioconjugate Techniques, 1st Ed., Academic Press, New York, New York, USA; and Me See Rifield et al. (1994) Ciba Foundation Symposium 186:5-20.

[0099] The compound or a pharmaceutically acceptable derivative thereof is contained in (i) packaging material; (ii) within the packaging material; The compounds of the present invention or pharmaceutically acceptable derivatives thereof (which are used to treat hyperproliferative tissue or angiogenesis) to regulate the activity of angiogenesis or to treat hyperproliferative tissue or angiogenesis-mediated diseases or disorders, or hyper Treatment, prevention, or prophylaxis of one or more symptoms of a disease or disorder involving hyperproliferative tissue or angiogenic activity and (iii) the compound or composition, or a pharmaceutical formulation thereof. and the like, and the derivatives thereof are useful for modulating hyperproliferative tissue or angiogenic activity or for treating hyperproliferative Tissue or angiogenesis-mediated diseases or disorders, or diseases involving hyperproliferative tissue or angiogenesis or label indicating that the product is used for the treatment, prevention, or amelioration of one or more symptoms of a disorder. It can be packaged as a product containing The products of the present invention include packaging materials. Packaging materials for use in packaging pharmaceutical products are well known to those skilled in the art. For example, U.S. Patent No. 5,323,907, U.S. Patent No. 5,052,558, and U.S. Patent No. See Patent No. 5,033,252, which is incorporated herein in its entirety. Examples of pharmaceutical packaging materials include blister packs, bottles, tubes, inhalers, pumps, and bags. , vials, containers, syringes, bottles, and suitable and intended administration for the selected formulation. This includes, but is not limited to, any packaging material suitable for the mode of treatment. A wide variety of formulations of compounds and compositions are available to treat hyperproliferative tissue or angiogenesis, as well as to address the symptoms and causes of these conditions. as a broad-spectrum treatment for any disease or disorder in which it is implicated as a mediating or contributing factor It is expected that this will be the case.

[0100] V. Photodynamic Therapy, Diagnostic and Therapeutic Applications In some embodiments, the compounds of formula (II) of the present invention (or their derivatives) The compounds (pharmaceuticals, pharmaceuticals, and pharmaceutical acceptable salts or conjugates thereof) are used in the treatment of diseases (e.g., cancer), including photodynamic therapy (PDT). The compounds can act as photosensitizers in methods for treating diseases such as rheumatoid arthritis (e.g., hyperproliferative diseases). Briefly, the photosensitizing compound, conjugate or pharmaceutical composition generally targets a target tissue, target tissue, or The photosensitizing compound is administered to the patient before the composition or patient is irradiated with light. Administered as described elsewhere. The dose of the photosensitizing compound can be determined clinically. Circulating or locally delivered light should be used to establish an equivalent optimal therapeutic level. A period of time is allowed to pass for the sensitizer to be taken up by the target tissue. The untargeted photosensitizer is cleared from the circulation during this waiting period or, optionally, is transferred to a non-targeted This may provide additional time for the tissue to clear of this unbound compound. This waiting period can be clinically determined and varies from compound to compound. After this waiting period, you can use a laser or non-laser light source (such as fluorescent or incandescent light) using artificial light sources, or natural light sources such as ambient sunlight, The area of ​​illumination is determined by the pathological area to be detected, diagnosed or treated. The length of illumination depends on the location and size of the target. The total or cumulative time may depend on the amount of time spent in the treatment and may be determined empirically. Any time between 1 minute and about 72 hours can be used. The illumination period is between about 60 minutes and 148 hours. The lighting period is between approximately 2 and 24 hours.

[0101] The total fluence or energy of the light used for illumination, measured in joules, is some In some embodiments, the range is between about 10 joules and about 25,000 joules. In embodiments, between about 100 joules and about 20,000 joules, in some embodiments In this case, the range is between about 500 joules and about 10,000 joules. Whether for therapeutic treatment to destroy or injure target tissue or target composition. The wavelength and fluence of light are selected to produce the desired effect. to irradiate at a wavelength that at least partially corresponds to the characteristic light absorption wavelength of the photosensitizer. Preferably, light having the following formula is used. The intensity or power of the light used is measured in watts, with 1 joule equaling 1 watt-second. Therefore, the intensity of the light used for irradiation in the method of the present invention is substantially 500 mW. / cm 2 The total fluence or amount of energy of this light (in joules) is divided by the length of the total exposure time (in seconds), thus increasing the time the target is exposed to radiation. This allows you to increase the amount of total energy or fluence without increasing the intensity of the light you use. In the present invention, the amount of total fluence of irradiation is high enough to activate the photosensitizer. Use.

[0102] In some embodiments of using the compounds of the invention for photodynamic therapy, the compounds The compound is injected into a mammal (e.g., a human) for diagnostic or therapeutic purposes. The dose is usually between about 0.1 and about 0.5 umol / kg body weight. Desired wavelength and energy (e.g., about 10-200 J / cm 2 ) light. For detection, of a wavelength sufficient to cause the compound to fluoresce at a wavelength different from that used to irradiate the compound. When exposed to light, fluorescence is measured. The energy used for detection is The energy required for the treatment is usually significantly lower than that required for therapy. Any one of the photosensitizing compounds of the present invention or pharmaceutically acceptable derivatives thereof may be used in the method of the present invention. The method may be provided in a kit together with instructions for carrying out any of the methods. The instructions may be, for example, printed paper, computer-generated material instructing a person how to perform the method. a disk, video cassette or other device remotely connected to the disk containing instructions on how to carry out the method; computer memory that receives and interprets the data, or (e.g., via the Internet) Any of these instructions may be in any specific form, including but not limited to, any method of providing instructions to a person. using the method described in the classroom or to treat patients using any of the methods of the present invention. In the process, the person can be instructed on how to use the kit.

[0103] Additional and specific examples of methods of using the compounds and compositions of the present invention include: However, it is not limited to these. (i) Treatment of Opportunistic Infections: The compounds, compositions and methods of the present invention are useful for treating opportunistic infections, particularly soft tissue infections. It is useful for photodynamic therapy (PDT) of tissues. For antimicrobial treatment (via PDT), the infecting organism may include (by way of non-limiting example) Staphylococcus aureus Among hospital-acquired infections, Pseudomonas aeruginosa accounts for 8% of surgical wound infections and 1% of blood infections. In some embodiments, the patient has AIDS. immunocompromised patients, such as those with HIV or those receiving immunosuppressant therapy is a person. (ii) Burn treatment: Staphylococcus aureus and gram-positive bacterial infections are common in burns. This multidrug resistance of Staphylococcus aureus presents a serious medical challenge. The compounds, compositions and methods of the present invention are useful for treating opportunistic infections in burns. . (iii) Sepsis: The compounds, compositions and methods of the present invention are effective against Vibrio vulnificus. It is useful in the photodynamic therapy (PDT) treatment of patients suffering from Vibrio vulnificus infection. Cass is a gram-negative bacterium that causes primary sepsis, wound infections, and gastrointestinal disease in humans. . (iv) Ulcers: The compounds, compositions and methods of the present invention are effective against ulcer-causing bacteria (Helicobacter It is useful for photodynamic therapy (PDT) treatment of Helicobacter pylori. (similar to an endoscope, but equipped with a device for shining red or near-infrared light) into the stomach or affected area Treatment can be effectively carried out in an appropriate manner, such as by inserting the device into the

[0104] (v) Periodontal Disease: The compounds, compositions and methods of the present invention are useful for the treatment of periodontal disease, including gingivitis. It is useful in photodynamic therapy (PDT). Periodontal disease is caused by the gram-negative anaerobic bacteria Porphyromonas It is caused by an overgrowth of bacteria such as Solanum gingivalis. Many photodynamic therapy ( As in photodynamic therapy (PDT), the targeting or solubilizing entity combined with the photoactive species is photoactivated. These are essential for proper delivery of the species to the target cells. Actinobacillus actinonzycetemcomitans, Bacterium Rioid forsythus, Campylobacter rectus, Eikenella corrodens, Fusobacter Includes Actinomyces nucleatum subsp. polymorpha, Actinomyces viscosus, and Streptococcus For such uses, the compounds or compositions of the present invention are applied topically (e.g., as a mouthwash or rinse), followed by an external device, oral appliance, or a combination thereof. Light can be emitted by this. (vi) Atherosclerosis: The compounds, compositions and methods of the present invention are useful for treating vulnerable atheroma. It is useful in photodynamic therapy (PDT) for treating atherosclerotic plaques. Without wishing to be bound by any particular theory, it is believed that invading inflammatory macrophages may contribute to the proliferation of coronary arteries. It is thought that they secrete metalloproteinases that break down the thin layer of collagen, leading to thrombosis. This is often fatal. The active compounds are useful for photodynamic therapy (PDT) of vulnerable plaque.

[0105] (vii) Cosmetic and dermatological applications: The compounds, compositions and methods of the present invention are useful in treating hair loss, psoriasis, and other skin conditions. photodynamic therapy for treating a wide range of cosmetic dermatological problems, such as dermatological treatment, or removal of skin discoloration Ruby lasers are currently used for hair removal and are useful in photo-depilatory therapy (PDT). In laser treatments, melanin is the photosensitizing chromophore. Such treatments are effective against dark hair color. The compounds, compositions and methods of the present invention are suitable as near-infrared sensitizers for hair removal. can be used as a chromophore with a more specific and sharper absorption band. It becomes possible to do this. (viii) Acne: The compounds, compositions and methods of the present invention may be used in conjunction with photodynamic therapy to treat acne. Prurigo vulgaris is caused by acne bacteria that infect the sebaceous glands. It is caused by a type of bacterial infection that affects about 80% of young people. Increasing resistance is leading to a surge in acne that is difficult to treat. Photodynamic therapy (PDT) treatment for rheumatoid arthritis usually relies on the addition of aminolevulinic acid. Nolevulinic acid is converted to free base porphyrin in the hair follicle or sebaceous gland. The compositions may be administered to the patient either topically or parenterally (e.g., by subcutaneous injection), depending on the particular condition. It can be administered to a person.

[0106] (ix) Infectious diseases: The compounds, compositions and methods of the present invention may be used in photodynamic therapy to treat infectious diseases. It is useful in PDT, for example, in the treatment of cutaneous ulcerative colitis, which is prevalent in the Mediterranean and Middle Eastern regions. Leishmaniasis and subcutaneous leishmaniasis are currently treated with arsenic-containing compounds Recently, photodynamic therapy (PDT) has shown reasonable efficacy in at least one human patient. The use of the compounds and compositions of the present invention is similarly useful. This potentially offers advantages such as ease of synthesis and better spectral absorption properties. (x) Tissue sealants: The compounds, compositions and methods of the present invention are used to treat tissue sealants in patients in need thereof. As a tissue sealant, it is useful in photodynamic therapy (PDT). They are attractive for wound sealing, tissue joining and closure of tissue defects. The use of such mechanical sealing methods often leads to infection and scarring. It has many uses. (xi) Oncological diseases: The compounds, compositions and methods of the present invention are useful in treating skin cancer, lung cancer, colon cancer, Breast cancer, prostate cancer, cervical cancer, ovarian cancer, basal cell carcinoma, leukemia, lymphoma, squamous cell carcinoma Neoplastic diseases or conditions, including skin cancer, melanoma, plaque-stage cutaneous T-cell lymphoma, and Kaposi's sarcoma is useful in photodynamic therapy (PDT) to treat cancer.

[0107] In addition to photodynamic therapy (PDT), the compositions of the present invention may also be used as image enhancing agents in diagnostic imaging techniques. or to label target tissues or target compositions for radiodiagnosis. In modern medicine, many diagnostic techniques, including magnetic resonance imaging (MRI), are used to diagnose illnesses. There are various treatments available. Detecting cancer at an early stage improves the chances of curing cancerous tissue. Early diagnosis of precancerous lesions and microcancers is an important topic in modern cancer treatment. MRI is a powerful tool in clinical practice because it is non-invasive and provides an accurate three-dimensional representation of the object. As in a typical nuclear magnetic resonance (NMR) experiment, radio frequency pulses are used to stimulate the nuclear spins. Images are produced by exciting and imposing one or more orthogonal magnetic field gradients on the object or specimen. After collecting data at various gradient fields, deconvolution A one-, two-, or three-dimensional image of the specimen / object is generated. Typically, this image is of water. Based on the NMR signal from protons, the signal intensity of a volume element is a function of the water concentration and relaxation time. The local variations of these parameters are the bright contrasts observed in MR images. Provide strikes.

[0108] MRI contrast agents act by increasing the relaxation rate, thereby increasing the enhances the contrast between water molecules in the area being examined and those elsewhere in the body. The effect of this is to decrease both T1 and T2, with the former increasing contrast and the latter decreasing contrast. Therefore, this phenomenon is concentration dependent and usually requires There is an optimal concentration of the paramagnetic species. This optimal concentration depends on the contrast agent used, the location of imaging, Imaging modes (i.e., spin echo, saturation recovery, inversion recovery, and / or various other (e.g., strongly T1- or T2-dependent imaging techniques), and contrast media in which the contrast agent is dissolved or suspended. These factors and their relative importance are well known in the art. For example, Pyket (1982) Scientific American 246:78; and Runge et al. (1983) American Journal of Radiology 141:1209. When used, the contrast agent perfuses the blood vessels, enhancing vascular contrast and detecting organ lesions and infiltration. However, specific tissue labeling for diagnostic radiology is not possible. This remains a challenging task for magnetic resonance imaging (MRI). Efforts to develop cell- and tissue-specific MRI image-enhancing agents by modifying the methodology has been the focus of much research in diagnostic radiology. For example, Antibodies (commonly gadolinium chelate Gd-DTPA) are produced and target tumors and other tissues. has been tested for its effect on MRI contrast agents. See U.S. Patent No. 5,059,415. , which is incorporated herein in its entirety. Unfortunately, the relaxivity of Gd bound to an antibody is The relaxivity of Gd-DTPA has been found to be only slightly better than that of untreated Gd-DTPA. See Janen et al. (1990) Magnetic Resonance in Medicine 13:38-43.

[0109] Magnetic resonance imaging (MRI) is generally used to 1 It is used to detect H nuclei, but ,MRI is, 13 C. 15 N, 31 P, 19 NMR spectra of other nuclides such as F can also be detected. 19 F is a living organism is not abundant. 13 C. 15 N, 31 P, or 19 F etc. (especially 19 The present invention provides an isotope useful for MRI of F and administering the compound to a patient, the compound of the present invention accumulates in the target tissue, Then, by NMR imaging, 19 Deposited materials containing MRI-recognizable isotopes such as F NMs with enhanced signal from target tissues or target components due to the presence of compounds IR data is generated. Thus, the compounds of the present invention can be used as image enhancing agents. and specific target tissues or target groups for diagnostic radiology, including magnetic resonance imaging (MRI). Labelling of the composition can be provided. In addition to photodynamic therapy (PDT), the compositions of the present invention can also be used to treat target cells, target tissues, and the like in patients. The compounds of the present invention can be used to detect target tissues or target compositions. When used to detect a target compound, the compound is introduced into a patient and the compound is then applied to the target. Sufficient time is allowed for the compound to accumulate in tissue or bind to the targeting compound. The treatment area is then generally exposed to light of sufficient energy to cause the compound to fluoresce. The energy used here is typically less than that required for photodynamic therapy treatment. Upon exposure to light of the desired wavelength, fluorescence is observed, and The amount of fluorescence of this compound can be measured qualitatively or quantitatively by methods known in the art. It is correlated with the amount of material present.

[0110] The compositions of the present invention may also be used to diagnose the presence of an infectious agent or to identify an infectious agent in a patient. The compounds of the present invention can be used to diagnose infectious diseases (antibodies or antibody fragments). and conjugating the antibody to one or more ligands specific for the infectious agent to selectively associate with the antibody fragment. The targeted compound has sufficient time to bind to the infectious agent and be removed from non-target tissue. After allowing time for the compound to be visualized (e.g., the compound becomes fluorescent). by exposing them to light with sufficient energy to cause a CT scan, or by using diagnostic radiology, including MRI. For example, any one of the compounds of the present invention may be administered to a suitable Helicobacter pylori antigen receptor (e.g., by imaging with a Helicobacter pylori antigen receptor). The antibody can be conjugated to an antibody targeted against a Helicobacter pylori antigen and administered in a pharmaceutical preparation. On the other hand, when introduced into a patient, the conjugated compound is released into the gastric mucus / cortex where the bacteria are found. This compound selectively binds to the target infectious agent, and the unbound compound is not the target. After sufficient time for clearance from the tissue, the patient is examined to determine if Helicobacter pylori is present. This can be done, for example, by 19 MRI due to the presence of F substituents Thus, the accumulated compound can be detected or the fluorescence of this compound can be generated. The energy beam is directed at the target area, for example, using fiber optics, to induce the release of the target compound. This can be done by detecting the fluorescence of the

[0111] In some embodiments, the compounds of the present invention or conjugates thereof are detected by flow cytometry. Flow cytometry is well known and can be useful in, for example, U.S. Patent No. 5,629,666. 167,926, U.S. Patent No. 5,915,925, U.S. Patent No. 6,248,590, U.S. Patent No. 6,589,792, and U.S. Patent No. 6,890,487, which are incorporated herein in their entireties. In some embodiments, the particles to be detected, such as cells, are labeled with a label for detection. The labeling is performed by labeling with a luminescent compound such as a fluorescent substance or fluorophore. Coupling a compound to another compound, such as an antibody (which can be specifically attached to a particle or cell) the uptake or internalization of the luminescent compound into a cell or particle; This can be done by any suitable technique, such as nonspecific adsorption to cells or particles. The active compounds of the present invention are useful in flow cytometry as light-emitting compounds. Flow cytometry techniques (including fluorescence-activated cell sorting or FACS) Implemented according to techniques or modifications thereof that would be apparent to those skilled in the art based on the disclosure of the present invention. can. [Example]

[0112] The following examples provide illustrative embodiments of the present invention and are within the general level of skill of those skilled in the art. In light of this, those skilled in the art will appreciate that the following examples are merely illustrative and do not deviate from the scope of the present invention. It will be understood that many variations, modifications, and variations may be made without departing from the spirit and scope of the invention.

[0113] Example 1 Synthesis of compound CP-1 As shown in Scheme 3 (Figure 3), the Di-BOC protected Suzuki coupling moiety Centaur 1 (CP-1) was prepared. 1-Bromo-3,5-bis(bromomethyl)benzene (CP-1a) N-Bromosuccinimide (NBS, 35.60 g, 200.0 mmol) was added to a flame-dried Add the solution to a three-necked 1 L round-bottom flask (RBF) and equip the flask with a glass stopper, septum-top condenser, and The N-bromosuccinimide (NBS) was heated under high vacuum for 30 minutes. After drying, the flask was flushed with argon and filled with acetonitrile (ACN, 400 mL). Approximately half the volume (~450 mL) was added via a cannula. Cetylbenzene (15.26 g, 80.0 mmol) was added via syringe, followed by short-circuiting the system under a stream of argon. After a time lapse, solid azobisisobutyronitrile (AIBN, 0.670 g, 4.00 mmol) was added. The flask was heated under argon (oil bath set at 90° C.) to a gentle reflux. After 16 h, the reaction mixture was transferred to a single-neck 1 L round-bottom flask (RBF) and concentrated to ACN. The solid residue was further dried under high vacuum and purified by dichloromethane (DCM, 75 mL The mixture was then equilibrated at room temperature and washed with DCM. The resulting filtrate was concentrated, dried under high vacuum, and heated in a water bath set at 65°C. The solid was recrystallized in ethanol (EtOH, 55 mL total) while heating. The resulting solid was filtered and The compound was washed with cold EtOH and then dried under high vacuum. Compound CP-1a (17.40 g, 51%) was obtained. 1 H NMR(400 MHz, CDCl3) δ 4.41(s, 4H), 7.34(s, 1H), 7.47(d, J = 2.0 Hz, 2H).

[0114] 2,2'-((5-bromo-1,3-phenylene)bis(methylene))bis(isoindoline-1,3-dioxide) N) (CP-1b) Compound CP-1a (18.43 g, 53.75 mmol) was placed in a 500 mL round-bottom flask (RBF) equipped with a stir bar. The flask was flushed with argon and filled with dimethylformamide (DMF, 215 mL, 0 0.25 M) was added to the clear, colorless solution, and potassium phthalimide (23.37 g, 123.6 3 mmol) was added little by little. The flask was fitted with a condenser with a drying tube, and the temperature was The mixture was heated in an oil bath set at 0°C. After 16 hours, the mixture was cooled to room temperature, diluted with water (1 L total) and diluted with chloroform (400, The organic layers were combined and extracted with 0.2 N NaOH (500 mL). and water (500 mL). The organic layer was separated, dried over sodium sulfate, and filtered. The resulting solid was further dried under high vacuum and then transferred to a filter. The compound CP-1b was obtained as a white powdery solid after washing with room temperature diethyl ether (Et2O) (3x). Isolated (17.98 g, 70%). 1 H NMR(400 MHz, CDCl3) δ 4.78(s, 4H), 7.42 - 7.47(m, 3H), 7.78 - 7.70(m, 4H), 7 .87 - 7.82(m, 4H).

[0115] (5-Bromo-1,3-phenylene)dimethanamine (CP-1c) Compound CP-1b (7.63 g, 16.06 mmol) was suspended in EtOH (70.0 mL) and heated in an oil bath at 85°C. Add tetrahydrofuran hydrate (4.88 mL, 80.29 mmol) and place a condenser on the flask. The mixture was further heated at reflux for 15 minutes, and then the reaction mixture was slowly cooled to room temperature. . To this was added 6 NaqHCl until the solution was acidic to the litmus test (20 mL total). The resulting mixture was again heated to reflux. The flask was flushed with argon and stirred for 1 hour. The mixture was stirred, then cooled and chilled in an ice bath. The mixture was filtered to give a clear, pale amber solution. The filtrate was cooled in an ice bath and basified with 2N NaOH (30 mL total). The organic layers were combined, washed with brine, and extracted with sodium sulfate. The flask was dried over ice, filtered, and concentrated to a volume of approximately 10 mL. The remaining solution was filtered and the filtrate was concentrated to give 2.3 g of a yellowish oil. This was stored in a refrigerator. The sample was stored at 4°C overnight to solidify, and then further purified. Drying under air gave 2.047 g (59%) of compound CP-1c as an amber semi-solid.

[0116] Di-t-butyl((5-bromo-1,3-phenylene)bis(methylene))dicarbamate (CP-1d) Compound CP-1c (2.00 g, 9.11 mmol) was placed in a flame-dried 250 mL round-bottom flask ( The flask was evacuated and flushed with argon. Tetrahydrofuran (THF, 45 mL) was added and the flask was lowered into a water bath. L, 21.86 mmol) was added and the heterogeneous mixture was cooled in an ice bath. mol) was prepared as a solution in THF (10 mL) and added dropwise in 1 mL portions. The solution was stirred at 0 °C for 1 h. and then allowed to equilibrate to room temperature. The reaction was stirred at room temperature overnight. The mixture was concentrated to give a white solid, which was then purified by ethyl acetate. The organic layer was redissolved in ethyl acetate (70 mL of EtOAc). This organic layer was washed with saturated aqueous NH4Cl, water, and saturated aqueous NaHCO3. The organic layer was then dried over sodium sulfate and filtered. The mixture was filtered and concentrated to a light amber oil, which crystallized on standing. The solid was purified by The fritted filter was washed with 20 / hexane (1:1). The solid was dried under high vacuum. As a result, 3.50 g (93%) of compound CP-1d (white powdery solid) was obtained.

[0117] Coupling Partner 1 (CP-1) Dimethyl sulfoxide (DMSO, reagent grade, 20.0 mL) in a 100 mL round-bottom flask (RBF). The mixture was added to a dry 250 mL round-bottom flask and argon was bubbled through for a total of 45 minutes while stirring. In a RBF solution, compound CP-1d (1.25 g, 3.01 mmol), bis(binacolate)diboron (0.91 7g, 3.61 mmol), potassium acetate (0.886g, 9.03 mmol) and Pd(dppf)Cl2 (0.066g, 0.090 The flask was evacuated for 30 minutes. The flask was flushed with argon. The solution was frozen in a dry ice / acetone bath and placed under vacuum. The reaction mixture was heated in an oil bath at 85°C. After 16 h, the reaction mixture was cooled to room temperature, diluted with EtOAc (100 mL), and washed with brine (3 x 100 mL). The organic layer was dried over sodium sulfate, filtered and concentrated. This concentrate was loaded neat onto a 40 g silica column with minimal DCM rinse and diluted with 0-2% ethanol in DCM. The main product fractions were combined and concentrated to give a clear oil. Further drying under high vacuum solidified the product. Compound CP-1 was obtained as a white waxy solid. and isolated (1.224 g, 88%).

[0118] Example 2 Synthesis of compound C-1 4-Bromopyrrole-2-carboxaldehyde (C-1a) As shown in Scheme 4 (Figure 4), in a 2 L round-bottom flask, stirred HCl (737 mL) in THF (100 mL) was added. Solution Pyrrole-2-carboxaldehyde (70.0 g, 737 mmol) was heated to 0°C under an argon atmosphere. After cooling, N-bromosuccinimide (NBS, 133 g, 737 mmol, reagent grade, recrystallized) was added. The reaction mixture was stirred under argon at 0° C. for 15 min. After the reaction, the solvent was removed by rotary evaporation, and the resulting solid was dried under high vacuum for 2 hours. Water (370 mL) was added to the flask and the suspension was filtered through a Buchner funnel. The filter cake was washed with an additional 370 mL of water. The filtered solid material was placed in a 2 L Erlenmeyer flask. The mixture was transferred to a flask and dissolved in 180 mL of hot ethanol (78°C) while refluxing in a water bath. 00 mL, 100 °C) was added in one go. Upon cooling to room temperature, the product crystallized from the solution. The mixture was further cooled to -10.0°C for 2 to 4 hours to promote crystallization. Filter by air filtration and dry under high vacuum for 24-48 hours to obtain 4-bromopyrrole-2-carboxylate. Light brown crystals of the sulaldehyde (C-1a, 104 g, 81%) were obtained. 1 H NMR (400 MHz, CDCl3) δ 6.91-7.03(m, 1H), 7.05-7.17(m, 1H), 9.48 (s, 1H), 9.6 8 (br s, 1H). 4-Bromo-2-formyl-N-tosylpyrrole (C-1b) In an oven-dried 1 L round-bottom flask, add 90% NaH (10.3 g, 429 mmol) in anhydrous THF (352 mL). The suspension was stirred, evacuated, and cooled to 0° C. under argon. This mixture was treated with Compound C-1 a (62.0 g, 356 mmol) for about 15 minutes. After stirring the mixture at 0° C. for 30 minutes, The reaction was stirred at room temperature for 3 h, and then treated with p-toluenesulfonyl chloride (67.9 g, 356 mmol). The mixture was stirred for 1 hour, and then the reaction was quenched by the slow addition of water (200 mL). Ethyl acetate (200 mL) was added to it, and the organic layer was separated, washed with brine (100 mL), and dried. It was dried (~50 g Na2SO4), filtered, and concentrated on a rotary evaporator to an oily liquid. The liquid was dried overnight under high vacuum in a 1 L round-bottom flask. The resulting crude solid was refluxed in a water bath. The mixture was dissolved in hexane / ethyl acetate (600 mL, 5:1) while cooling to room temperature. The product crystallized from the solution. The mixture was cooled to -10°C for an additional 3 hours to give a further The mixture was filtered by vacuum filtration, and the filtered brown crystals were collected. , and dried under high vacuum to obtain the compound 4-bromo-2-formyl-N-tosylpyrrole (C-1b) (94 g, 81%). 1 H NMR(400 MHz, CDCl3) δ 2.44(s, 3H), 6.77(s, 1H), 7.31(d, J = 13.40 Hz, 1H), 7 .36(d, J = 8.25 Hz, 2H), 7.60(s, 1H), 7.76(d, J = 8.25 Hz, 2H), 8.44(d, J = 13.4 0 Hz, 1H).

[0119] 4-Bromo-2-(2-nitrovinyl)-N-tosylpyrrole (C-1c) In a 500 mL round-bottom flask, 4-bromo-2-formyl-N-tosylpyrrole (C-1b, 84.2 g, 257 mL) was added. mmol), potassium acetate (20.1 g, 205 mmol), methylamine hydrochloride (13.8 g, 205 mmol) and ca. 99. A stirred mixture of acetic acid (1.00 mL) in 5% pure ethanol (90.0 mL) was dissolved in nitromethane (34.6 m The mixture was stirred for 2 hours, then water (200 mL) was added and the resulting The yellow precipitate was filtered by vacuum filtration. The filtered solid material was added to water (500 mL), followed by cold ethanol. The yellow solid was washed with methanol (1.2 L, 0°C) until the eluate was clear. Drying under high vacuum overnight gave 4-bromo-2-(2-nitrovinyl)-N-tosylpyrrole (C-1c, 79g, 83%) was obtained. 1 H NMR (400 MHz, CDCl3) δ2.44(s, 3H), 6.77(s, 1H), 7.31(d, J = 13.40 Hz, 1 H), 7.36(d, J = 8.25 Hz, 2H), 7.60(s, 1H), 7.76(d, J = 8.25 Hz, 2H), 8.4 4(d, J = 13.40 Hz, 1H) 4-Bromo-2-(2-nitroethyl)-N-tosylpyrrole (C-1d) In a 2 L round-bottom flask, a solution of compound C-1c (74.0 g, 199 mmol) in anhydrous THF (1 L) was added to The solution was cooled to -10°C (ice-acetone (1:1)) under vigorously stirring. The reaction mixture was treated with 95% LiBH4 (4.34 g, 199 mmol) in one portion. Stir at -10 °C for 30 min until no material remains. Once this is complete, add cold saturated aqueous NH4Cl ( The reaction mixture was quenched by the slow addition of 340 mL of HCl (at 0°C). The mixture was stirred, extracted with ethyl acetate (340 mL), dried (43 g of anhydrous Na2SO4), and evaporated on a rotary evaporator. The crude solid was concentrated to a dark brown solid and dried under high vacuum in a 2 L round bottom flask for 2 hours. The product was dissolved in isopropyl alcohol (IPA, 1.2 L) under reflux in a water bath. Upon cooling, the product crystallized from the solution. The mixture was further cooled at -10°C for 4 hours. This mixture was filtered by vacuum filtration, and the filtered light brown solid was collected. The crystals were dried under high vacuum overnight to give 4-bromo-2-(2-nitroethyl)-N-tosylpyrrole ( C-1d, 47g, 62%) was obtained. 1 H NMR (400 MHz, CDCl3)δ2.44(s, 3H), 3.39(t, J = 7.01 Hz, 2H), 4.60(t, J = 7.01 Hz, 2H), 6.10(d, J = 1.93 Hz, 1H), 7.32(d, J = 1.93 Hz, 1H), 7.35( d, J = 7.98 Hz, 2H), 7.69(d, J = 7.98 Hz, 2H)

[0120] 6-(4-bromo-1-tosyl-1H-pyrrol-2-yl)-4,4-dimethyl-5-nitrohexan-2-one (C -1e) Compound C-1d (13.3 g, 35.7 mmol) and 1,1-dimethoxy-4-methyl- A mixture of 1,8-diazabicyclo[5.4 The resulting mixture was treated with 1.0]undec-7-ene (DBU, 16 mL, 107.0 mmol, 3.0 equiv.). The mixture was stirred at room temperature for 1 hour and diluted with EtOAc. The organic layer was washed with water (3x, brine) and dried. The excess 1,1-dimethoxy-4-methyl-3-penten-2-one was removed under high vacuum. The resulting crude product was dissolved in a minimum amount of CH2Cl2 (8 mL) and evaporated on a silica cake. The cake was loaded onto a 120 g SiO column containing 25-40% EtOAc in hexane. The major fractions were combined, concentrated, and dried under high vacuum rotor paper. The resulting product was 11.4 g (68%) of 6-(4-bromo-1-tosyl-1H-pyrrol-2-yl)-4,4-dimethyl-5-dimethyl- Trohexen-2-one (C-1e) was obtained as a brown solid. 1 H NMR (400 MHz, CDCl3) δ1.11(s, 3H), 1.24(s, 3H), 2.13(s, 3H), 2.40(AB , J = 17.8 Hz, 1H), 2.43(s, 3H), 2.55(AB, J = 17.8 Hz, 1H), 3.18(AB, J = 16 .2 Hz, 1H), 3.36(ABX, 3J = 16.2 Hz, 2J = 11.8 Hz, 1H), 5.14(AB, J = 11.8 Hz , 1H), 6.00-6.02(m, 1H), 7.22-7.24(m, 1H), 7.34(AB, J = 8.2 Hz, 2H), 7.6 4 (AB, J = 8.2 Hz, 2H) 4-Bromo-1-tosyl-2-((3,3,5-trimethyl-3,4-dihydro-2H-pyrrol-2-yl)methyl)- 1H-Pyrrole (C-1f) In a round-bottom flask (RBF), a solution of compound C-1e (10.5 g, 22.3 mmol) in THF (105 mL) was added to HCl. The resulting suspension was treated with NH4 (28.1 g, 446 mmol) and zinc powder (28.3 g, 446 mmol). The reaction mixture was stirred vigorously at rt for 2 h. The reaction mixture was passed through a pad of silica (50 g) glass frit. The filter cake was eluted with EtOAc (500 mL). The filtrate was concentrated to give a fluffy A fluffy light brown solid was obtained. The residue was dissolved in CH2Cl2 and prepared as a silica cake. The cake was eluted on a 120 g SiO2 column with 75-80% hexane in EtOAc for 60 min. The single major product was combined, concentrated, and dried under high vacuum to give 6.29 g (67%) of 4-bromo- 1-Tosyl-2-((3,3,5-trimethyl-3,4-dihydro-2H-pyrrol-2-yl)methyl)-1H-pyrrole Compound (C-1f) was obtained as a pale brown solid. 1 H NMR (400 MHz, CDCl3) δ0.88(s, 3H), 1.07(s, 3H), 1.97(s, 3H), 2.28(AB , J = 16.8 Hz, 1H), 2.36(AB, J = 16.8 Hz, 1H), 2.41(s, 3H), 2.63(ABX, 2J = 16.1 Hz, 3J = 10.2 Hz, 1H), 2.92(ABX, 2J = 16.1 Hz, 3J = 3.8 Hz, 1H), 3.67 ~3.70(m, 1H), 6.25~6.28(m, 1H), 7.28~7.30(m, 3H), 7.68(AB, J = 8.2 Hz , 2H)

[0121] 4-Bromo-2-((3,3,5-trimethyl-3,4-dihydro-2H-pyrrol-2-yl)methyl)-1H-pyrrole Lu (C-1g) Compound C-1f (2.79 g, 6.59 mmol) was dissolved in tetra-n-butyl fluoride in a round-bottom flask (RBF). Treat with ammonium (TBAF, 19.8 mL, 1.0 M in THF, 19.8 mmol, 3 equiv.) and reflux the reaction. The mixture was stirred at room temperature (64-67°C) for 1 hour. To this was added a saturated solution of NaHCO3 (68 mL), followed by acetic acid. Ethyl acetate (68 mL) was added, and the mixture was extracted with ethyl acetate (134 mL). Dry (anhydrous Na2SO4), concentrate on a rotary evaporator to a dark brown oil, and simmer under high vacuum for 2 hours. The residue was dissolved in dichloromethane (DCM) and prepared as a silica cake. The cake was eluted on a 40 g SiO2 column with 25-33% EtOAc in hexane for 55 min. The main product fractions were combined, concentrated, and dried under high vacuum to give 1.56 g (88%) of compound 4-bromo-2-(4-methyl-2-propanol). Bromo-2-((3,3,5-trimethyl-3,4-dihydro-2H-pyrrol-2-yl)methyl)-1H-pyrrole (C -1g) was obtained as a light brown solid. 1 H NMR (400 MHz, CDCl3) δ0.92(s, 3H), 1.11(s, 3H), 2.03(s, 3H), 2.28(AB , J = 16.8 Hz, 1H), 2.38(AB, J = 16.8 Hz, 1H), 2.54(ABX, 2J = 14.9 Hz, 3J = 11.8 Hz, 1H), 2.69(ABX, 2J = 11.8 Hz, 3J = 2.5 Hz, 1H), 3.56-3.62(m, 1H), 5.85-5.94(m, 1H), 6.63-6.69(m, 1H), 9.72-10.01(br s, 1H)

[0122] Scheme 5 (Figure 5) shows the synthesis of the eastern half of chlorin and its reaction with the western half. Shows cyclization reaction. 5-(4-(methoxycarbonyl)phenyl)dipyrromethane (C-1h) In a 500 mL round-bottom flask (RBF), pyrrole (183.9 g, 2741 mmol, 190 mL, 45 equiv.) and methyl- A mixture of 4-formylbenzoate (10.0 g, 60.9 mmol) was degassed under argon for 30 minutes. Trifluoroacetic acid (TFA, 1.18 mL, 0.25 equivalents) was added to the mixture, and the mixture was further stirred under an argon atmosphere. The mixture was stirred for 30 minutes. An aliquot of the reaction mixture was subjected to thin film filtration using CH2Cl2 / hexane (2:1). TLC showed complete conversion of methyl-4-formyl benzoate to C-1h. The reaction mixture was diluted with 400 mL of CH2Cl2 and washed with 200 mL of 0.1N NaOH. The organic layer was washed with brine (200 mL), dried (20 g of anhydrous NaSO), filtered, and evaporated on a rotary evaporator. The crude solid was recrystallized from 200 mL of methanol / water (10:1) and Filtration by vacuum filtration gave a light brown solid. After recrystallization, 13.5 g (79%) of 5-(4-(methoxyphenyl)-2-methyl-2-propanol was obtained by filtration through a 150 mL glass frit. Alternatively, this residue was dissolved in CH2Cl2 to give The silica cake was dissolved and prepared as a silica cake. The cake was diluted with 75-100% CHCl in hexane gradient. The product was eluted over 70 min on a 120 g SiO column containing 2. The main product fractions were combined, concentrated, and purified. Drying under air gave a light brown solid. 1-Formyl-5-(4-(methoxycarbonyl)phenyl)dipyrromethane (C-1i) The Vilsmeier reagent was prepared according to the procedure reported by Laha et al., J. Org. hem. 2006, 71, 4092-4102. A sample of anhydrous DMF (8.0 mL) was heated under argon. POCl3 (1.90 mL, 1.2 equiv., 20.3 mmol) in an oven-dried 50 mL round-bottom flask. The mixture was stirred at RT for 20 min at 0° C. The resulting mixture was added (via cannula) to DMF (35 mL) at 0° C. Solution of 5-(4-(methoxycarbonyl)phenyl)dipyrromethane (C-1h, 5.43, 19.4 mmol) in A second round-bottom flask (250 mL) was vacuum evacuated and argon-flushed with oven-dried HCl. After 1.5 hours, saturated NaHCO3 solution (87 mL) was added slowly to the mixture. The mixture was stirred overnight and extracted with ethyl acetate. The organic layers were combined, washed (brine), and It was dried over Na2SO4, filtered, and concentrated. The residue was dissolved in CH2Cl2 and a silica cake (10 g The cake was purified by elution on an 80 g SiO2 column containing 0-40% EtOAc in hexane. The main product fractions were combined, concentrated, and dried under high vacuum to give 3.37 g (5 6%) of 1-formyl-5-(4-(methoxycarbonyl)phenyl)dipyrromethane (C-1i) in a light brown solution. Obtained as a solid.

[0123] 8,9-Dibromo-1-formyl-5-(4-(methoxycarbonyl)phenyl)dipyrromethane (C-1j) In an oven-dried round-bottom flask (250 mL), C-1i (1.40 g, 4.54 mL) in THF (45 mL) was dissolved. (1.62 g, 9.08 mmol) was treated with recrystallized N-bromosuccinimide (NBS) (1.62 g, 9.08 mmol) at -78 °C. After 1 h, the cooling bath was removed and the reaction mixture was allowed to warm to -20 °C and then cooled to 100 °C. The resulting mixture was diluted with ethyl acetate, washed with brine, and dried. The residue was dissolved in CH2Cl2 and formed into a silica cake or slurry (approximately 6.0 g). This cake was diluted with 80 g of S using a 0-20% EtOAc in hexane gradient. The iO2 column was eluted for 86 minutes. The main product fractions were combined, concentrated, and dried under high vacuum to give 2.00 g (94%) of 8,9-dibromo-1-formyl-5-(4-(methoxycarbonyl)fluor)-2,4-dibromo-1,4-dibromo ... (phenyl)dipyrromethane (C-1j) was obtained. 3,13-Dibromo-10-(4-(methoxycarbonyl)-18,18-dimethylchlorin Zn(II) (C-1k) 3,4,5,6-tetrahydro-1,3,3-trimethyldipyrrin (C-1g, 0.63 g) in CHCl (35 mL) , 2.35 mmol) and 8,9-dibromo-1-formyl-5-(4-(methoxycarbonyl)phenyl)dipyrromethyl A suspension of toluene (C-1j, 1.09 g, 2.35 mmol) was added to p-toluenesulfonyl ether (C-1j, 1.09 g, 2.35 mmol) in methanol (12 mL). The sample was treated with phosphonic acid monohydrate (2.23 g, 11.7 mmol) and stirred at room temperature for 40 min. The resulting mixture was analyzed using a 2,2,6,6-tetramethylisothiazolinone (2,2,6,6-tetramethylisothiazolinone) spectrophotometer. The reaction mixture was treated with dimethylpiperidine (7.92 mL, 46.9 mmol) for 5-10 minutes. The mixture was concentrated in a centrifuge, and the resulting brown solid was suspended in acetonitrile (235 mL). Zinc acetate (6.47 g, 35.2 mmol), 2,2,6,6-tetramethylpiperidine (15.9 mL, 93.9 mmol) The resulting suspension was treated with silver trifluoromethanesulfonate (1.82 g, 7.04 mmol). The reaction mixture was refluxed for 8-24 hours. The reaction mixture was cooled and concentrated to dissolve the residue. was dissolved in CH2Cl2 and filtered through a silica bed on a glass frit until the eluate was clear. This residue was prepared as a silica cake and dissolved in 120 g of SiO2 cake with 50% CH2Cl2 in hexane. The gradient was eluted with 70% CH2 in hexane for 3 minutes and held at this gradient for an additional 15 minutes. The gradient was then changed to Cl2 in 3 minutes and held at that gradient for an additional 5 minutes. The gradient was changed to 0% CH2Cl2 for 5 min and held for 20 min. Thin Layer Chromatography (TLC) All green fractions were then combined to obtain 3,13-dibromo-10-(4-(methoxycarbonyl)-18,18- Dimethylchlorin Zn(II) (C-1k) was obtained as a green solid (318 mg; 20% yield). 3,13-Dibromo-10-(4-(methoxycarbonyl)-18,18-dimethylchlorin (C-1l) A round-bottom flask was charged with C-1k (49 mg, 0.070 mmol). 5.0 mL of a mixture of CH2Cl2 / TFA (4.90 / 0. 10 ml) was added to the flask and the resulting mixture was stirred for 2 hours. The reaction was quenched with aqueous CO3 (20 mL), washed with brine, dried (Na2SO4), filtered, and concentrated. The concentrate was recrystallized from hexane / CH2Cl2 (20 mL) to give the compound 3,13-dibromo-10-(4-( Methoxycarbonyl)-18,18-dimethylchlorin (C-11) was obtained as a dark powder (34 mg, 75%).

[0124] Next, compound C-11 can be used to prepare intermediate compounds C-1m and C-2m, as shown in Scheme 6 (Figure 6). Compound C1 was prepared via C-1n. Suzuki Coupling Precursor (C-1m) Compound C-11 (63.2 mg, 100 μmol), coupling partner CP-1 (102 mg, 220 μmol), P A mixture of d(PPh3)4 (69.4 mg, 60.0 μmol) and Cs2CO3 (196 mg, 60.0 μmol) was placed in a 25 mL round-bottom flask. The flask was placed under high vacuum for 1 hour, and then subjected to three evacuation-refill cycles. To this was added toluene / DMF (2:1, 10 mL in total) using a syringe, and the solution was The solution was heated at 90°C for 18 hours. After cooling to room temperature, the reaction mixture was diluted with ethyl acetate and washed with aqueous NaHCO3 solution. The resulting mixture was concentrated and purified by chromatography (silica 40 g, hexahydrate). Compound C-1m (94.9 mg, 83%) was obtained as a green solid by subjecting the solution to a 200 mL aqueous solution of ethanol / ethyl acetate (0-75%). Suzuki coupling precursor (C-1n) with a t-butyl-protected β-alanine linker Compound C-1m (38.1 mg, 33.3 μmol) in THF (13.3 mL) and MeOH (6.7 mL) was dissolved in 1.0 M NaOH. The reaction mixture was stirred in the dark at room temperature under argon for 3.5 hours. After this time, the reaction mixture was diluted with EtOAc, washed with 1.0% aqueous HCl, dried over Na2SO4, and concentrated. did. The crude residue was treated with O-(N-succinimidyl)-N,N,N',N'-tetramethyluronate. A mixture of 1000mg of tetrafluoroborate (TSTU, 40.1mg, 133µmol), CH2Cl2 (8.3mL) and TEA (18.5mL) was used. The reaction mixture was stirred in the dark at room temperature under argon for 1 hour. Afterwards, the reaction mixture was diluted with EtOAc, washed with brine, dried over Na2SO4 and concentrated. The resulting residue was treated with β-alanine t-butyl ester hydrochloride (45.3 mg, 333 μmol) and CsC O3 (109 mg, 333 μmol) was added. The system was evacuated and flushed with argon, and CH2C l2 (8.3 mL) was added. The reaction mixture was stirred in the dark at room temperature under argon. After 3 hours The reaction mixture was diluted with EtOAc, washed with brine, dried over Na2SO4, and concentrated. Compound C-1 was obtained by chromatography [silica 12 g, hexane / ethyl acetate (0-100%)]. n (40.2 mg, 97%) was obtained as a green solid. MS: obsd 1257.04, calcd 1256.68 [(M + H) + , M = C 72 H 89 N9O 11 ].

[0125] Compound C-1 Compound C-1n (12.6 mg, 10.0 μmol) in a 10 mL round-bottom flask (RBF) was dissolved in dioxane (2.5 m The reaction mixture was stirred in the dark at room temperature under argon. After 1 hour, the reaction mixture was placed under high vacuum for 16 hours. The resulting residue was treated with CH3(OC2H4) 12 CONHS (mPEG 12 -NHS, 54.9 mg, 80.0 μmol) and Cs2CO3( 52.2 mg, 160 μmol) was added, and the flask was evacuated and flushed with inert gas. 5 mL) was added and the reaction mixture was stirred at room temperature. After 1 hour, the reaction mixture was chromatographed. After filtration [C18 gold, 15.5 g, H2O / CH3CN (0-40%)], compound C-1 was obtained as a green solid ( 4.0mg, 13%). Then, the synthesis of compound C-1 was started from compound C-1n (155.3 mg, 124 μmol) and the reaction The reaction was scaled up to obtain 252 mg of compound C-1 (yield 66%). MS: Observed 1541.94, Calculated 1541.86 [M + 2H] 2+ ; Observed value 1553.24, calculated value 1552.85 [( M + H + Na] 2+ ; Observed 1563.89, Calculated 1563.84 [(M + 2Na] 2+ ; λ abs 417, 651 nm(P BS); λ em 656 nm (PBS). Quantum yield: 26% (PBS); FWHM: 20 nm (PBS). The solubility of chlorins is assessed by measuring the absorbance of a series of dilutions of the chlorin stock solution. Jiang et al. (2014) Organic & Biomolecular Chemistry 12:86-103. See, The solubility of compound C-1 was >10 mg / mL (PBS).

[0126] Example 3 Synthesis of compound 4 Sonogashira Coupling NIRvana680 Precursor (2) Compound 4, also referred to herein as NIRvana 680, is shown in Scheme 7 (Figure 7). It was synthesized as follows: 5-ethynyl-1,3-benzenedicarboxylic acid (500 mg, 2.63 mmol), t-butyl N- (2-aminoethyl)carbamate (2.08 mL, 13.2 mmol), 1-ethyl-3-(3-dimethylaminopropyl) pyridinium carbonate (EDCI, 2.00 g, 10.4 mmol) and 4-dimethylaminopyridine (DMAP, 1. Compound C237 was prepared from a mixture of 48 g of methyl methylpropional (48 g, 13.2 mmol) and dissolved in DMF (3.3 mL). The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was directly loaded onto silica gel and chromatographed. Chromatography [silica, CH2Cl2 / MeOH (0-10%)] gave a white solid. , 1 H NMR revealed that it contained DMAP, so it was redissolved in ethyl acetate and added 1.0% HCl The mixture was washed with aqueous solution, dried over Na2SO4, and concentrated to give compound C237 as a white solid (938 mg, 75%). And got it. Compound 1 (60.0 mg, 0.095 mmol), coupling partner C237 (225 mg, 0.474 mmol, 5.0 A mixture of PdCl2(PPh3)2 (17 mg, 0.024 mmol, 0.25 equiv.) and PdCl2(PPh3)2 (17 mg, 0.024 mmol, 0.25 equiv.) was added to an oven-dried 100 ml The flask was placed under high vacuum for 0.5 hours and then evacuated three times. The mixture was further degassed by refilling cycles. To this was added toluene / Et3N (2:1, 9 mL total) via syringe. The solution was heated at 85-90°C for 24 hours. After cooling to room temperature, the reaction mixture was diluted with vinegar. The residue was diluted with ethyl acetate, washed with aqueous NaHCO3, brine, dried over Na2SO4, and concentrated. The material was dissolved in a minimum amount of CH2Cl2 and prepared as a silica cake. Elution on a 40 g silica column containing H (0-1%) gave compound 2 (132 mg, 98%) as a dark solid. I got it. MS: [M + H] + calc. 1419.68; obs. 1419.86.

[0127] Sonogashira Coupling NIRvana680 Precursor, t-Butyl-Protected β-Alanine Linker (3) Compound 2 (45.8 mg, 32.3 μmol) in THF (6.0 mL) and MeOH (3.0 mL) was dissolved in 1.0 M aqueous NaOH. (3.0 mL). The reaction mixture was stirred at room temperature in the dark. After 4.5 hours, the reaction mixture The mixture was quenched with 1.0 M aqueous HCl (4.5 mL), diluted with EtOAc (25 mL), and washed with brine (25 mL). The crude residue was dried over NaSO (4.0 g), filtered, concentrated, and dried under high vacuum. TSTU (19.4 mg, 64.4 μmol, 2.0 equiv.), CHCl (10.0 mL, 3.2 mM), and triethylamine (9 The reaction mixture was stirred for approximately 1 hour at room temperature under argon in the dark. The resulting residue was stirred for 1 hour. β-Alanine t-butyl ester hydrochloride (17.6 mg, 96.8 1 μmol, 3.0 equiv.), triethylamine (27.0 μL, 64.4 μmol, 2.0 equiv.), and the reaction was heated to room temperature. The mixture was stirred at room temperature overnight. To this was further added TEA (54 μL, 12 equivalents) and β-alanine t-butyl ester. C. Hydrochloride (52.8 mg, 9.0 equiv.) was added and the reaction was stirred overnight. The reaction mixture was heated to reflux, diluted with CH2Cl2 (15 mL), washed with brine, and the organic layer was separated. The extract was separated, dried over anhydrous Na2SO4, filtered, and concentrated. Compound 3 (38.2 mg, 77%) was obtained as a dark solid by elution with CH2Cl2 / MeOH (0-1%). Got it as a body. MS: [M + H] + calc. 1532.76; obs. 1533.11. NIRvana 680 (compound 4) Compound 3 (38.2 mg, 24.9 μmol) was evacuated and flushed with argon. A 4.0 M solution of ethanol in 100 ml of ethanol (2.5 mL) was added. The reaction mixture was stirred in the dark at room temperature under argon. After 5 hours, the reaction mixture was placed under a high flow of argon through an outlet needle to remove the solvent. After this was complete, the vial was dried under vacuum overnight. The resulting residue was treated with CH3O(OC2H4 ) 12 CH2CH2COONHS ester (109.1 mg, 149.5 μmol, 6.0 equiv.) and Cs2CO3 (81.2 mg, 249.2 The flask was evacuated and flushed with inert gas. DMF (3.0 mL) was added. The reaction mixture was then protected from light and stirred under argon at room temperature. After 1.5 hours, CH3O(OC2H4) 12 CH2C H2COONHS ester (109.1 mg, 149.5 μmol, 6.0 equiv.) and Cs2CO3 (81.2 mg, 249.2 μmol, 1 0 equiv.), the vial was evacuated and flushed with argon for 10 minutes, and the argon atmosphere was Stirring was continued under reduced pressure. After 3.5 hours, the reaction was concentrated. The sample was diluted with water (2.0 mL). The solution was dissolved in 30% ACN in water and subjected to reversed-phase preparative LC using a 30-80% ACN in water gradient over 45 min. The main product peak was combined, concentrated, transferred to a storage vial of ACN, concentrated, and purified under high vacuum. Drying at rt gave 17.5 mg (20%) of compound 4 as a green residue. MS: [M+2H] 2+ calc. 1767.45; obs. 1767.54.

[0128] Example 4 Flow cytometry device The sample was measured using seven lasers (355, 405, 488, 532, 561, 594, and 633 nm). A 19-parameter LSR-II SORP flow cytometer (BD Biosciences, San Jose, California) was used. fornia, USA) or LSRFortes equipped with five lasers (355, 405, 488, 561, and 640 nm) sa (BD Biosciences, San Jose, California, USA) and FACSDiva 8.0 acquisition. The data for compound C-1 were analyzed using 630 nm long-pass (LP) filter. Channel A of a 100 mW 405 nm laser equipped with a 660 / 20 nm bandpass (BP) filter. Data for chlorin H2C12-PEG6-NHS were collected using 635 nm LP and 655 / 40 nm BP filters. Used and collected. antibody bioconjugates : As described above for the preparation of the intermediate NHS ester in the synthesis of compound C-1n, The NHS ester of compound C-1 was prepared by TSTU (i.e., O-(N-succinimidyl)-N,N,N') in CH2Cl2. , N'-tetramethyluronium tetrafluoroborate) and TEA, and HP Liquid chromatography mass spectrometry (LC / MS) showed a single peak (MS: obsd. 1591 [M+2H] 2 + , 1602 [M+H+Na] 2+ 9.4 mg / mL (1.0 mg) of anti-human CD8 mouse monoclonal antibody (clon e UCHT-4, Leinco Technologies, Inc., St. Louis, Missouri, USA) 106 μL, 1 M bicarbonate Multiple PEGylated dye NHS esters (5–20 molar equivalents) were prepared from 15 μL of salt (pH 8.4) and 44 μL of PEGylated dye NHS esters (5–20 molar equivalents) in PBS. The solution was prepared in a microcentrifuge tube. The tube was shielded from light and left to stand at room temperature for 1-2 hours. The mixture was incubated at room temperature for an additional hour by adding 15 μL of 200 μM Tris. The reaction was stopped and the bioconjugate was purified by elution with Sephadex G50M, G75M or G1 The product was purified using a size exclusion chromatography column of either 0.05 or 0.1M. Antibody bioconjugates prepared from dyes with 12 or more units were incubated in G75M or G100M media. The column fractions were purified by measuring the absorbance at 280 nm (protein) and the absorbance maximum of the red or NIR dye. The pooled fractions were characterized by fluorophore-to-protein (F / P) yield. The labeling ratio was determined from these two maxima by correcting for the dye absorbance at 280 nm.

[0129] cell stainingCryopreserved human peripheral blood mononuclear cells (PBMCs) were purchased from ZenBio, Inc. (Research Tria (United Park, North Carolina, USA; Product SER-PBMC-F), thawed, and then used in this The cells were prepared for staining according to the manufacturer's guidelines. The cells were separated into 2 sections and centrifuged at 400 x g (2000 rpm) for 5 minutes. The cells were washed with washing buffer (0.5% BSA). The cells were washed three times with PBS containing trypsin and resuspended in 0.5 mL of wash buffer. Dilute 1:2 with Lew and measure cell number and viability by counting a 4 nL square in a hemocytometer. The viability was usually >96%. The cells were diluted to 1x10 6 Dilute to 50µL / mL L (500,000 cells) was dispensed into a microcentrifuge tube. The maximum labeled antibody concentration was 4.74 μg / 5x10 5 The cells (designated 3.16X) were diluted in half-logarithmic dilutions. This dilution was used for all antibodies except the control antibody. For all assays, 15 μL was added to the cell aliquot. The tubes were incubated at room temperature for 30 minutes with stirring. Each tube was washed twice with 1 mL of washing buffer. Afterwards, the cells were resuspended in 0.5 mL of wash buffer containing 1% formaldehyde. Prior to cytometric characterization, samples were filtered through a nylon filter cloth to Placed in a flow cytometry tube.

[0130] If desired, a positive control bioconjugate was prepared using CD8 (UCHT-4)-fluorescein isothiocyanate. Fluorescence Intensity Change (FITC) (Leinco Technologies, Inc., St. Louis, Missouri, USA; catalog no. C119), CD4(RPA-T4)-BUV737 antibody (BD Biosciences, San Jose, California, USA; Catalog Log number 564306) and / or CD8(UCHT-4)-DY650 antibody (Leinco Technologies, Inc., St Louis, Missouri, USA; Catalog No. C2064) and peripheral blood samples were collected using the same general procedure. The staining index (SI) was calculated using the Cytome method described by Maecker et al. (2004). The mean fluorescence intensity (MFI) was calculated as follows according to the method of Example 1. SI = (mean: positive - mean: background) / (2 × SD background) Table 1 below shows the staining index data for titration of anti-CD8 bioconjugates of compound C-1 and 10-phenylalanine. The previously published chlorin, H2C12-PEG6-NHS, which has three PEG6s attached via hydroxyl groups, was used as an anti- Staining index data for titration of CD8 bioconjugates are shown. Liu et al ., Molecules, 2018, 23, 13 For comparison, FITC (Leinco Technologies, Inc., St. Louis, Missouri) was used. Data for an anti-CD8 bioconjugate prepared from Sigma-Aldrich (Santa Cruz, CA; Cat. No. C119) are also provided. [Table 1] These results demonstrate the significant improvement in performance of the PEGylated formulation of compound C-1. do.

[0131] References All references listed herein include all patents, patent applications and publications. and the methods employed therein, including but not limited to scientific journal articles. to the extent that it supplements, explains, provides background to, or teaches the theory, techniques, and / or compositions of matter. is incorporated herein by reference in its entirety. Various details of the invention disclosed herein may be changed without departing from the scope of the invention. It should be understood that it is possible. Furthermore, the foregoing description is for illustrative purposes only. and is not intended to limit the invention.

Claims

1. A compound represented by the following formula (I): 【Chemistry 1】 wherein M is a metal or two hydrogen atoms; R ５ , R １０ and R １５ are each independently a hydrogen atom, an alkoxy group, or a group represented by the following formula: a linker group selected from the group consisting of: -L １ -(X １ -L ２ ) ｐ -G where p is 0 or 1, and L １ is alkylidene, and X １ is —C(═O)NH— or -NHC(=O)-, and L ２ Ha-(CH ２ CH ２ O) ｑ -alkylene- (wherein q is 1 to 24), alkylene, or substituted alkylene (optionally, the substituted alkylene The alkylene may be bonded to one or more groups containing a polyoxyethylene chain and / or an amide group. G is a bioconjugable group; and R ２ , R ３ , R １２ and R １３ are each independently a hydrogen atom, cyano, halo, or perhydrogen atom. alkyl, sulfonate, sulfonamide, ester, carboxylic acid, formyl, acetyl Lu, formula-L １ -(X １ -L ２ ) ｐ selected from linker groups and solubilizing groups represented by G; wherein the solubilizing group is -aryl-(R ｓ ) ｗ and -alkynyl-aryl-(R ｓ ) ｗ where w is an integer from 0 to 5, and R ｓ is a group represented by the following formula: 。 -X ２ -(L ３ ) ｚ -R １７ 、 where z is 0 or 1, and X ２ Ha-CH ２ NHC(=O)-, -C(=O)NH-alkyl L is phenyl-NH- or triazolyl; ３ is -C(=O)-alkylene-C(=O)-NH - and R １７ Ha-(C ２ H ４ O) ｍ -R １８ , -C(=O)C ２ H ４ -(OC ２ H ４ ) ｍ OR １８ and -(C ２ H ４ O) ｎ -C ２ H ４ -C(=O)NH-C(R １９ ) ３ Selected from where m is an integer of 4 or greater (optionally, m is an integer of 8 or greater), and n is a number from 1 to 5. is an integer, and R １８ is lower alkyl (optionally methyl), and R １９ Ha-CH ２ O-C ２ H ４ -C(=O)NH-(C ２ H ４ O) ｍ R １８ It is.) ) ; However, R ２ , R ３ , R １２ and R １３ At least one of -aryl-(R ｓ ) ｗ or -alkynyl-aryl-(R ｓ ) ｗ It is.)

2. R ５ , R １０ and R １５ is a hydrogen atom, methoxy and the formula -L １ -(X １ -L ２ ) ｐ -Represented by G 2. The compound of claim 1, wherein the linker group is selected from the group consisting of:

3. R １０ But, formula-L １ -(X １ -L ２ ) ｐ 3. The method according to claim 1 or 2, wherein the linker group is represented by -G. The compound described.

4. R １０ But, formula-L １ -(X １ -L ２ ) ｐ -G (in the formula, L １ is phenylene, p is 1, , X １ is —C(═O)NH—, and L ２ is alkylene, and G is a carboxylic acid or an active ene 4. The compound of claim 3, wherein the linker group is selected from the group consisting of aryl, aryl, aryl esters ...

5. R １０ The compound according to claim 4, wherein: 【Chemistry 15】

6. R ３ and R １３ are each -aryl-(R ｓ ) ｗ (Optionally, R ３ and R １３ Gaso -phenyl-(R ｓ ) ２ The compound according to any one of claims 1 to 5,

7. Each R ｓ The compound according to claim 6, wherein is a group represented by the following formula: -X ２ -(L ３ ) ｚ -R １７ (wherein z is 0 and X ２ Ha-CH ２ NHC(=O)-, and R １７ Ha-(C ２ H ４ O) ｍ -R １８ (wherein m is an integer from 12 to 24.)

8. R ３ and R １３ The compound according to claim 7, wherein each of 【Chemistry 16】

9. The compound according to any one of claims 1 to 8, wherein the compound is represented by the following formula: 【Chemistry 17】 or [Chemistry 18]

10. A composition comprising a covalently bonded conjugate formed between: (a) R ２ , R ３ , R ５ , R １０ , R １２ , R １３ and R １５ At least one of the A compound represented by formula (I) according to claim 1, wherein (b) small molecules, antigens, microparticles, nanoparticles, polymers, peptides, proteins, antibodies or one or more of the group consisting of antibody fragments, nucleic acids, hormones and growth factors 。

11. A compound of claim 1 or a conjugate of claim 10, and a pharmaceutically acceptable carrier. A pharmaceutical composition comprising:

12. A method for detecting a target, wherein the target is a compound, a cell or a particle, the method comprising the steps of:

11. A method comprising labeling a target with the conjugate of claim 10.

13. The method of claim 12, wherein the method comprises the use of flow cytometry.

14. 10. A method for imaging a cell, tissue or organism, comprising the step of: A method comprising the use of a conjugate as described above.

15. 1. A method of treating a disease in a patient in need thereof, the method comprising: administering to a patient a compound of claim 1, a conjugate of claim 10, or a pharmaceutical composition of claim 11. and - irradiating at least a portion of the patient with light (Optionally, the disease is a hyperproliferative disease, and further optionally, the disease is cancer.)

16. having a solubility of greater than about 1 mg / ml in aqueous solution (optionally greater than about 2.5 mg / ml in aqueous solution); / ml or more, and optionally have a solubility of about 10 mg / ml or more in aqueous solution. (having greater solubility than this), water-soluble chlorin dyes.

17. A method for producing a synthetic intermediate of a compound represented by the following formula (I): 【Chemistry 1】 wherein M is a metal or two hydrogen atoms; R ５ , R １０ and R １５ are each independently a hydrogen atom, an alkoxy group, or a group represented by the following formula: a linker group selected from the group consisting of: -L １ -(X １ -L ２ ) ｐ -G where p is 0 or 1, and L １ is alkylidene, and X １ is —C(═O)NH— or -NHC(=O)-, and L ２ Ha-(CH ２ CH ２ O) ｑ -alkylene- (wherein q is 1 to 24), alkylene, or substituted alkylene (optionally, the substituted alkylene The alkylene may be bonded to one or more groups containing a polyoxyethylene chain and / or an amide group. G is a bioconjugable group; and R ２ , R ３ , R １２ and R １３ are each independently a hydrogen atom, halo, cyano, or perhydrogen atom. alkyl, sulfonate, sulfonamide, ester, carboxylic acid, formyl, acetyl Lu, formula-L １ -(X １ -L ２ ) ｐ selected from linker groups and solubilizing groups represented by G; wherein the solubilizing group is -aryl-(R ｓ ) ｗ and -alkynyl-aryl-(R ｓ ) ｗ where w is an integer from 0 to 5, and R ｓ is a group represented by the following formula: 。 -X ２ -(L ３ )z-R １７ 、 where z is 0 or 1, and X ２ Ha-CH ２ NHC(=O)-, -C(=O)NH-alkyl L is phenyl-NH- or triazolyl; ３ is -C(=O)-alkylene-C(=O)-NH - and R １７ Ha-(C ２ H ４ O) ｍ -R １８ , -C(=O)C ２ H ４ -(OC ２ H ４ ) ｍ OR １８ and -(C ２ H ４ O) ｎ -C ２ H ４ -C(=O)NH-C(R １９ ) ３ Selected from where m is an integer of 4 or greater (optionally, m is an integer of 8 or greater), and n is a number from 1 to 5. is an integer, and R １８ is lower alkyl (optionally methyl), and R １９ Ha-CH ２ O-C ２ H ４ -C(=O)NH-(C ２ H ４ O) ｍ R １８ It is.) ) ; However, R ２ , R ３ , R １２ and R １３ At least one of -aryl-(R ｓ ) ｗ or -alkynyl-aryl-(R ｓ ) ｗ It is.) A method comprising the following steps: (a) providing a compound represented by formula (I'): 【Transformation 5】 wherein M is a metal or two hydrogen atoms; R ５ ', R １０ ' and R １５ ' are each independently a hydrogen atom, an alkoxy, 【Transformation 6】 Selected from: R ２ ', R ３ ', R １２ ' and R １３ ' are each independently a hydrogen atom, an ester, a carboxylic acid, or a carboxylic acid, formyl, acetyl, 【Transformation 7】 is selected from However, R ２ ', R ３ ', R １２ ' and R １３ At least one of the ' 【Transformation 8】 It is.) (b) The compound provided in step (a) is dissolved in a solution containing 4 molar (M) HCl in dioxane. with a solution to provide a compound of formula (I"): 【Chemistry 9】 wherein M is a metal or two hydrogen atoms; R ５ '', R １０ '' and R １５ '' are each independently a hydrogen atom, an alkoxy, 【Chemistry 10】 Selected from: R ２ '', R ３ '', R １２ '' and R １３ '' each independently represent a hydrogen atom, an ester, Carboxylic acid, formyl, acetyl, 【Chemistry 11】 is selected from However, R ２ '', R ３ '', R １２ '' and R １３ At least one of the 【Chemistry 12】 It is.)

18. A compound represented by one of the following formulae: 【Chemistry 17】 and [Chemistry 18] or a conjugate thereof, wherein the compound is a small molecule, an antigen, a microparticle, a nanoparticle, a polymer, -, peptides, proteins, antibodies or antibody fragments, nucleic acids, hormones and growth factors A compound conjugated to one or more of the groups consisting of:

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