JP2012531443A - Photolabile compound - Google Patents

Abstract

In this invention, a photolabile compound and its usage are described. The photolabile compound has a light-emitting ligand, which can be biologically active and is light-emitted from the compound when exposed to light. In some embodiments, the photolabile compound comprises an optical antenna (a labeled molecule, an active derivative thereof, etc.). In one embodiment, the light is visible light that is not detrimental to the viability of the biological sample (cells, tissues, etc.), and the released organic molecules are bioactive and may have a therapeutic effect. In another embodiment, the light emitting ligand may be a label molecule such as a fluorescent molecule.
[Selection figure] None

Description

  This invention was made with the support of the US Government. The US government has certain rights in this invention.

  This patent disclosure includes material that is subject to copyright protection. The copyright holder has no objection to the facsimile reproduction of a patent document or patent disclosure by any person, as described in the US Patent and Trademark Office patent wrap or record, but all other copyrights are reserved. To do.

CROSS REFERENCE TO RELATED APPLICATIONS This application claims priority to US Provisional Patent Application No. 61 / 220,922, filed June 26, 2009, the entire disclosure of which is incorporated herein by reference. US Patent Application Publication No. 2008/0176940 is also fully incorporated herein by reference.

  The present invention generally relates to novel photolabile compounds and methods for uncaging organic molecules (such as bioactive molecules), which may have a variety of uses both in vitro and in vivo.

  Photolabile protecting groups (also called caging groups) are a class of protecting groups that are particularly useful in biochemistry. Since light can be accurately controlled both spatially and temporally, the bioactive molecule can be released or uncaged by cleavage of the protecting group from the bioactive molecule. Protecting groups typically mask or hide charged groups (eg, carboxylate, phosphate) or polar groups (eg, amine, hydroxyl, sulfhydryl) of the compound. In many cases, such functional groups increase the hydrophobicity and membrane permeability of the protected molecule. Prior to photolysis, photolabile compounds are typically chemically or biologically inert because at least one of the main functional groups of the compound is blocked. The pulse of light can induce the activity of the molecule and release the molecule from the light emitting compound. Thus, a photolabile protecting group can be removed from a protected compound by light irradiation, for example, to control the release of the compound at a desired timing and location in vivo or in vitro.

  Commercially available photolabile compounds typically require ultraviolet light (UV) to remove the compound from the cage. However, ultraviolet light is detrimental for in vivo use because it can damage organs, tissues and cells. For this reason, there is a need in the art to utilize novel photolabile compounds having ligands that can be released using light other than ultraviolet light, particularly for in vivo applications. The present invention provides novel photolabile compounds and methods of using the compounds, which are advantageous over current compounds that are photolabile only to ultraviolet light.

  One embodiment of the present invention is to provide a novel photolabile compound that protects an organic molecule (such as a physiologically active molecule). Upon exposure to light, the organic molecules are released and are useful in the methods described herein.

Ｉ
（式中、
ＭはＲｕであり、
各Ｌ¹は独立して
（ａ）環員の１つがＭと結合を形成する窒素原子である５員単環式芳香環、
（ｂ）環員の１つがＭと結合を形成する窒素原子である６員単環式芳香環、
（ｃ）２環式環の１つが芳香族であり且つＭと結合を形成する窒素原子員を有する８〜１０員２環式環、
（ｄ）窒素原子がＭと結合を形成する−ＮＨ₂基、又は
（ｅ）酸素原子の１つがＭと結合を形成する−ＣＯＯＨ基
を有する有機分子であり、
Ｌ²は（Ｒ⁹）₃Ｐ、（Ｒ⁹Ｏ）₃Ｐ又はＬ¹であり、ｍは２であり、あるいはＬ²は−ＣＮであり、ｍは１であり、あるいはＬ²は（Ｒ⁹）₂Ｐ若しくは（Ｒ⁹Ｏ）₂Ｐのリン原子を介してＭに結合される標識分子又はその活性誘導体であり、ｍは２であり、あるいはＬ²はＮＨＲ⁹、Ｎ（Ｒ⁹）₂、ピリジル、Ｃ（Ｒ⁹）＝ＮＨ、Ｃ（Ｒ⁹）＝ＮＲ⁹環状脂肪族アミン基若しくはニトリルの窒素原子を介してＭに結合される標識分子又はその活性誘導体であり、ｍは２であり、
各Ｒ⁹は独立して−Ｃ₁〜Ｃ₁₈アルキル、−Ｃ₃〜Ｃ₈シクロアルキル又はフェニルであり、
Ｒ¹〜Ｒ⁸は独立して−Ｈ、−Ｃ₁〜Ｃ₁₈アルキル、−ＮＨ₂、−ＣＯＯＨ、−（Ｃ₁〜Ｃ₁₈アルキル）−Ｏ−（Ｃ₁〜Ｃ₁₈アルキル）又は−ＯＣ（Ｏ）（Ｃ₁〜Ｃ₁₈アルキル）であり、
ＸはＣｌ^-、Ｆ^-、Ｂｒ^-、Ｉ^-、ＰＦ₆ ^-、ＣＦ₃ＳＯ₃ ^-、（Ｃ₁〜Ｃ₁₈アルキル）−ＣＯ₂ ^-又は（Ｃ₁〜Ｃ₁₈アルキル）−ＳＯ₃ ^-である）で示される化合物を提供する。 In one aspect, the invention provides a compound of formula I,

I
(Where
M is Ru,
Each L ¹ is independently (a) a 5-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with M;
(B) a 6-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with M;
(C) an 8- to 10-membered bicyclic ring having a nitrogen atom member in which one of the bicyclic rings is aromatic and forms a bond with M;
(D) an organic molecule having a —NH ₂ group in which a nitrogen atom forms a bond with M, or (e) a —COOH group in which one of the oxygen atoms forms a bond with M;
L ² is (R ⁹ ) ₃ P, (R ⁹ O) ₃ P or L ¹ , m is 2, or L ² is —CN, m is 1, or L ² is (R ⁹ ) A labeled molecule or an active derivative thereof bonded to M via a phosphorus atom of ₂ P or (R ⁹ O) ₂ P, m is 2, or L ² is NHR ⁹ , N (R ⁹ ) ₂ , pyridyl, C (R ⁹ ) = NH, C (R ⁹ ) = NR ⁹ is a labeled molecule or an active derivative thereof bonded to M via a nitrogen atom of a cyclic aliphatic amine group or a nitrile, and m is 2 And
Each R ⁹ is independently —C ₁ -C ₁₈ alkyl, —C ₃ -C ₈ cycloalkyl or phenyl;
R ^{1 to} R ⁸ are independently —H, —C _{1 to} C ₁₈ alkyl, —NH ₂ , —COOH, — (C _{1 to} C ₁₈ alkyl) —O— (C _{1 to} C ₁₈ alkyl) or —OC. an (O) (C ₁ ~C ₁₈ alkyl),
X is ^{Cl -, F -, Br -} , I -, PF 6 -, CF 3 SO 3 -, (C 1 ~C 18 alkyl) -CO ₂ ^- or (C ₁ -C ₁₈ alkyl) -SO ₃ ^- in A compound represented by (A).

In one embodiment of the compound of formula I:
M is Ru,
Each L ¹ is independently (a) a 5-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with M;
(B) a 6-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with M;
(C) an 8- to 10-membered bicyclic ring having a nitrogen atom member in which one of the bicyclic rings is aromatic and forms a bond with M;
(D) an organic molecule having a —NH ₂ group in which a nitrogen atom forms a bond with M, or (e) a —COOH group in which one of the oxygen atoms forms a bond with M;
L ² is (R ⁹ ) ₃ P, (R ⁹ O) ₃ P or L ¹ , m is 2, or L ² is —CN and m is 1.
R ^{1 to} R ⁸ are independently —H, —C _{1 to} C ₁₈ alkyl, —NH ₂ , —COOH, — (C _{1 to} C ₁₈ alkyl) —O— (C _{1 to} C ₁₈ alkyl) or —OC. an (O) (C ₁ ~C ₁₈ alkyl),
X is ^{Cl -, F -, Br -} , I -, PF 6 -, CF 3 SO 3 -, (C 1 ~C 18 alkyl) -CO ₂ ^- or (C ₁ -C ₁₈ alkyl) -SO ₃ ^- in is there.

In some embodiments of the compound of Formula I, when L ² is (R ⁹ ) ₃ P, each R ⁹ is different from each other.

In some embodiments of the compound of Formula I, when L ² is (R ⁹ ) ₃ P, the two R ⁹ are the same and different from the remaining R ⁹ .
In some embodiments of the compound of Formula I, when L ² is (Ph) ₃ P, each phenyl is not substituted with methyl.

ＩＩ
（式中、
各Ｌ¹は独立して
（ａ）窒素原子の１つがＲｕと結合を形成するテトラゾリル基、
（ｂ）ピリジル窒素原子がＲｕと結合を形成するニコチン若しくはカフェイン、
（ｃ）２環式環の１つが芳香族であり且つＲｕと結合を形成する窒素原子員を有する８〜１０員２環式環、
（ｄ）窒素原子がＲｕと結合を形成する−ＮＨ₂基、又は
（ｅ）酸素原子の１つがＲｕと結合を形成する−ＣＯＯＨ基
を有する有機分子であり、
Ｌ²は（Ｒ⁹）₃Ｐ、（Ｒ⁹Ｏ）₃Ｐ又はＬ¹であり、ｍは２であり、あるいはＬ²は−ＣＮであり、ｍは１であり、あるいはＬ²は（Ｒ⁹）₂Ｐ若しくは（Ｒ⁹Ｏ）₂Ｐのリン原子を介してＲｕに結合される標識分子又はその活性誘導体であり、ｍは２であり、あるいはＬ²はＮＨＲ⁹、Ｎ（Ｒ⁹）₂、ピリジル、Ｃ（Ｒ⁹）＝ＮＨ、Ｃ（Ｒ⁹）＝ＮＲ⁹環状脂肪族アミン基若しくはニトリルの窒素原子を介してＲｕに結合される標識分子又はその活性誘導体であり、ｍは２であり、
各Ｒ⁹は独立して−Ｃ₁〜Ｃ₁₈アルキル、−Ｃ₃〜Ｃ₈シクロアルキル又はフェニルであり、
Ｒ¹〜Ｒ⁸は独立して−Ｈ、−Ｃ₁〜Ｃ₁₈アルキル、−ＮＨ₂、−ＣＯＯＨ、−（Ｃ₁〜Ｃ₁₈アルキル）−Ｏ−（Ｃ₁〜Ｃ₁₈アルキル）又は−ＯＣ（Ｏ）（Ｃ₁〜Ｃ₁₈アルキル）であり、
ＸはＣｌ^-、Ｆ^-、Ｂｒ^-、Ｉ^-、ＰＦ₆ ^-、ＣＦ₃ＳＯ₃ ^-、（Ｃ₁〜Ｃ₁₈アルキル）−ＣＯ₂ ^-又は（Ｃ₁〜Ｃ₁₈アルキル）−ＳＯ₃ ^-である）の化合物を提供する。 In another embodiment, the present invention provides compounds of formula II,

II
(Where
Each L ¹ is independently (a) a tetrazolyl group in which one of the nitrogen atoms forms a bond with Ru;
(B) nicotine or caffeine in which the pyridyl nitrogen atom forms a bond with Ru;
(C) an 8 to 10 membered bicyclic ring having a nitrogen atom member in which one of the bicyclic rings is aromatic and forms a bond with Ru;
(D) an organic molecule having a —NH ₂ group in which a nitrogen atom forms a bond with Ru, or (e) a —COOH group in which one of the oxygen atoms forms a bond with Ru;
L ² is (R ⁹ ) ₃ P, (R ⁹ O) ₃ P or L ¹ , m is 2, or L ² is —CN, m is 1, or L ² is (R ⁹⁾ a ₂ P or (R ⁹ O) labeled molecule or an activated derivative thereof is coupled to Ru via the ₂ P phosphorus atoms, m is 2, or L ² is NHR ^9, N (R ⁹⁾ ₂ , pyridyl, C (R ⁹ ) ═NH, C (R ⁹ ) = NR ⁹ is a labeled molecule bonded to Ru via a cyclic aliphatic amine group or a nitrogen atom of a nitrile or an active derivative thereof, m is 2 And
Each R ⁹ is independently —C ₁ -C ₁₈ alkyl, —C ₃ -C ₈ cycloalkyl or phenyl;
R ^{1 to} R ⁸ are independently —H, —C _{1 to} C ₁₈ alkyl, —NH ₂ , —COOH, — (C _{1 to} C ₁₈ alkyl) —O— (C _{1 to} C ₁₈ alkyl) or —OC. an (O) (C ₁ ~C ₁₈ alkyl),
X is ^{Cl -, F -, Br -} , I -, PF 6 -, CF 3 SO 3 -, (C 1 ~C 18 alkyl) -CO ₂ ^- or (C ₁ -C ₁₈ alkyl) -SO ₃ ^- in A compound).

In one embodiment of the compound of formula II:
Each L ¹ is independently (a) a tetrazolyl group in which one of the nitrogen atoms forms a bond with Ru;
(B) nicotine or caffeine in which the pyridyl nitrogen atom forms a bond with Ru;
(C) an 8 to 10 membered bicyclic ring having a nitrogen atom member in which one of the bicyclic rings is aromatic and forms a bond with Ru;
(D) an organic molecule having a —NH ₂ group in which a nitrogen atom forms a bond with Ru, or (e) a —COOH group in which one of the oxygen atoms forms a bond with Ru;
L ² is (R ⁹ ) ₃ P, (R ⁹ O) ₃ P or L ¹ , m is 2, or L ² is —CN and m is 1.
R ^{1 to} R ⁸ are independently —H, —C _{1 to} C ₁₈ alkyl, —NH ₂ , —COOH, — (C _{1 to} C ₁₈ alkyl) —O— (C _{1 to} C ₁₈ alkyl) or —OC. an (O) (C ₁ ~C ₁₈ alkyl),
X is ^{Cl -, F -, Br -} , I -, PF 6 -, CF 3 SO 3 -, (C 1 ~C 18 alkyl) -CO ₂ ^- or (C ₁ -C ₁₈ alkyl) -SO ₃ ^- in is there.

In some embodiments of the compound of Formula II, when L ² is (R ⁹ ) ₃ P, each R ⁹ is different from each other.
In some embodiments of the compound of Formula II, when L ² is (R ⁹ ) ₃ P, the two R ⁹ are the same and different from the remaining R ⁹ .
In some embodiments of the compound of Formula II, when L ² is (Ph) ₃ P, each phenyl is not substituted with methyl.

ＩＩＩ
（式中、
Ｌ¹は、ピリジル窒素原子がＲｕと結合を形成する４−アミノピリジン（４−ＡＰ）であり、
Ｌ²は（Ｒ⁹）₃Ｐ、（Ｒ⁹Ｏ）₃Ｐ又はＬ¹であり、ｍは２であり、あるいはＬ²は−ＣＮであり、ｍは１であり、あるいはＬ²は（Ｒ⁹）₂Ｐ若しくは（Ｒ⁹Ｏ）₂Ｐのリン原子を介してＲｕに結合される標識分子又はその活性誘導体であり、ｍは２であり、あるいはＬ²はＮＨＲ⁹、Ｎ（Ｒ⁹）₂、ピリジル、Ｃ（Ｒ⁹）＝ＮＨ、Ｃ（Ｒ⁹）＝ＮＲ⁹環状脂肪族アミン基若しくはニトリルの窒素原子を介してＲｕに結合される標識分子又はその活性誘導体であり、ｍは２であり、
各Ｒ⁹は独立して−Ｃ₁〜Ｃ₁₈アルキル、−Ｃ₃〜Ｃ₈環状アルキル又はフェニルであり、
Ｒ¹〜Ｒ⁸は独立して−Ｈ、−Ｃ₁〜Ｃ₁₈アルキル、−ＮＨ₂、−ＣＯＯＨ、−（Ｃ₁〜Ｃ₁₈アルキル）−Ｏ−（Ｃ₁〜Ｃ₁₈アルキル）又は−ＯＣ（Ｏ）（Ｃ₁〜Ｃ₁₈アルキル）であり、
ＸはＣｌ^-、Ｆ^-、Ｂｒ^-、Ｉ^-、ＰＦ₆ ^-、ＣＦ₃ＳＯ₃ ^-、（Ｃ₁〜Ｃ₁₈アルキル）−ＣＯ₂ ^-又は（Ｃ₁〜Ｃ₁₈アルキル）−ＳＯ₃ ^-である）の化合物を提供する。 (0017)
In another embodiment, the present invention provides compounds of formula III,

III
(Where
L ¹ is 4-aminopyridine (4-AP) in which the pyridyl nitrogen atom forms a bond with Ru;
L ² is (R ⁹ ) ₃ P, (R ⁹ O) ₃ P or L ¹ , m is 2, or L ² is —CN, m is 1, or L ² is (R ⁹⁾ a ₂ P or (R ⁹ O) labeled molecule or an activated derivative thereof is coupled to Ru via the ₂ P phosphorus atoms, m is 2, or L ² is NHR ^9, N (R ⁹⁾ ₂ , pyridyl, C (R ⁹ ) ═NH, C (R ⁹ ) = NR ⁹ is a labeled molecule bonded to Ru via a cyclic aliphatic amine group or a nitrogen atom of a nitrile or an active derivative thereof, m is 2 And
Each R ⁹ is independently —C ₁ -C ₁₈ alkyl, —C ₃ -C ₈ cyclic alkyl or phenyl;
R ^{1 to} R ⁸ are independently —H, —C _{1 to} C ₁₈ alkyl, —NH ₂ , —COOH, — (C _{1 to} C ₁₈ alkyl) —O— (C _{1 to} C ₁₈ alkyl) or —OC. an (O) (C ₁ ~C ₁₈ alkyl),
X is ^{Cl -, F -, Br -} , I -, PF 6 -, CF 3 SO 3 -, (C 1 ~C 18 alkyl) -CO ₂ ^- or (C ₁ -C ₁₈ alkyl) -SO ₃ ^- in A compound).

(0018)
In one embodiment of the compound of formula III:
L ¹ is 4-aminopyridine (4-AP) in which the pyridyl nitrogen atom forms a bond with Ru;
L ² is (R ⁹ ) ₃ P, (R ⁹ O) ₃ P or L ¹ , m is 2, or L ² is —CN and m is 1.
R ^{1 to} R ⁸ are independently —H, —C _{1 to} C ₁₈ alkyl, —NH ₂ , —COOH, — (C _{1 to} C ₁₈ alkyl) —O— (C _{1 to} C ₁₈ alkyl) or —OC. an (O) (C ₁ ~C ₁₈ alkyl),
X is ^{Cl -, F -, Br -} , I -, PF 6 -, CF 3 SO 3 -, (C 1 ~C 18 alkyl) -CO ₂ ^- or (C ₁ -C ₁₈ alkyl) -SO ₃ ^- in is there.

(0019)
In some embodiments of compounds of Formula III, when L ² is (R ⁹ ) ₃ P, each R ⁹ is different from each other.
In some embodiments of the compound of Formula III, when L ² is (R ⁹ ) ₃ P, the two R ⁹ are the same and different from the remaining R ⁹ .
In some embodiments of the compound of formula III, when L ² is (Ph) ₃ P, each phenyl is not substituted with methyl.

ＩＶ
（式中、
Ｍ¹はＬｉ⁺、Ｎａ⁺又はＫ⁺であり、
Ｍ²はＲｕであり、
Ｌ¹は独立して
（ａ）環員の１つがＭ²と結合を形成する窒素原子である５員単環式芳香環、
（ｂ）環員の１つがＭ²と結合を形成する窒素原子である６員単環式芳香環、
（ｃ）２環式環の１つが芳香族であり且つＭ²と結合を形成する窒素原子員を有する８〜１０員２環式環、
（ｄ）窒素原子がＭ²と結合を形成する−ＮＨ₂基、又は
（ｅ）酸素原子の１つがＭ²と結合を形成する−ＣＯＯＨ基
を有する有機分子である）の化合物を提供する。 (0022)
In another embodiment, the present invention provides compounds of formula IV,

IV
(Where
M ¹ is Li ⁺ , Na ⁺ or K ⁺
M ² is Ru,
L ¹ is independently (a) a 5-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with M ² ;
(B) a 6-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with M ² ;
(C) an 8- to 10-membered bicyclic ring having a nitrogen atom member in which one of the bicyclic rings is aromatic and forms a bond with M ² ;
And (e) an organic molecule having a —NH ₂ group in which a nitrogen atom forms a bond with M ² or (e) a —COOH group in which one of the oxygen atoms forms a bond with M ² ).

Ｖ
（式中、
各Ｌ¹は独立して
（ａ）環員の１つがＲｕと結合を形成する窒素原子である５員単環式芳香環、
（ｂ）環員の１つがＲｕと結合を形成する窒素原子である６員単環式芳香環、
（ｃ）２環式環の１つが芳香族であり且つＲｕと結合を形成する窒素原子員を有する８〜１０員２環式環、
（ｄ）窒素原子がＲｕと結合を形成する−ＮＨ₂基、
（ｅ）酸素原子の１つがＲｕと結合を形成する−ＣＯＯＨ基、
（ｆ）リン原子がＲｕと結合を形成し、Ｒが独立して−Ｈ、−Ｃ₁〜Ｃ₁₈アルキル若しくはアリールである−ＰＲ₂基、又は
（ｇ）硫黄原子がＲｕと結合を形成し、Ｒが独立して−Ｈ、−Ｃ₁〜Ｃ₁₈アルキル若しくはアリールである−ＳＲ基
を有する有機分子であり、
Ｌ²は（Ｒ⁹）₃Ｐ、（Ｒ⁹Ｏ）₃Ｐ又はＬ¹であり、ｍは２であり、あるいはＬ²は−ＣＮであり、ｍは１であり、あるいはＬ²は（Ｒ⁹）₂Ｐ若しくは（Ｒ⁹Ｏ）₂Ｐのリン原子を介してＲｕに結合される標識分子又はその活性誘導体であり、ｍは２であり、あるいはＬ²はＮＨＲ⁹、Ｎ（Ｒ⁹）₂、ピリジル、Ｃ（Ｒ⁹）＝ＮＨ、Ｃ（Ｒ⁹）＝ＮＲ⁹環状脂肪族アミン基若しくはニトリルの窒素原子を介してＲｕに結合される標識分子又はその活性誘導体であり、ｍは２であり、
各Ｒ⁹は独立して−Ｃ₁〜Ｃ₁₈アルキル、−Ｃ₃〜Ｃ₈シクロアルキル又はフェニルであり、 (0023)
In another embodiment, the present invention provides compounds of formula V,

V
(Where
Each L ¹ is independently (a) a 5-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(B) a 6-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(C) an 8 to 10 membered bicyclic ring having a nitrogen atom member in which one of the bicyclic rings is aromatic and forms a bond with Ru;
(D) a —NH ₂ group in which the nitrogen atom forms a bond with Ru;
(E) a —COOH group in which one of the oxygen atoms forms a bond with Ru;
(F) phosphorus atoms form a bond and Ru, R is independently -H, -C ₁ ~C ₁₈ -PR ₂ groups are alkyl or aryl, or (g) a sulfur atom forms a bond with Ru , R is an organic molecule having a —SR group that is independently —H, —C ₁ -C ₁₈ alkyl or aryl,
L ² is (R ⁹ ) ₃ P, (R ⁹ O) ₃ P or L ¹ , m is 2, or L ² is —CN, m is 1, or L ² is (R ⁹⁾ a ₂ P or (R ⁹ O) labeled molecule or an activated derivative thereof is coupled to Ru via the ₂ P phosphorus atoms, m is 2, or L ² is NHR ^9, N (R ⁹⁾ ₂ , pyridyl, C (R ⁹ ) ═NH, C (R ⁹ ) = NR ⁹ is a labeled molecule bonded to Ru via a cyclic aliphatic amine group or a nitrogen atom of a nitrile or an active derivative thereof, m is 2 And
Each R ⁹ is independently —C ₁ -C ₁₈ alkyl, —C ₃ -C ₈ cycloalkyl or phenyl;

When L ² is a labeled molecule or an active derivative thereof linked to Ru via the phosphorus atom of P (phenyl) ₃ or P (phenyl) ₂ , each phenyl is independently —C ₁ -C ₁₈ alkyl, — ( C ₁ -C ₁₈ alkyl) -OH, aryl, - (C ₁ ~C ₁₈ alkyl) - oxy, - (C ₁ ~C ₁₈ alkyl) - amino, - (C ₁ ~C ₁₈ alkyl) - thio, -CO ₂ Y, —C (═O) Y, —C (═O) NY ₂ , —NH ₂ , —NO ₂ , —OH or —SH,
R ^{1 to} R ⁴ are independently —H, —C _{1 to} C ₁₈ alkyl, —NH ₂ , — (C _{1 to} C ₁₈ alkyl) —O— (C _{1 to} C ₁₈ alkyl), —OC (O). (C ₁ -C ₁₈ alkyl), - (C ₁ ~C ₁₈ alkyl) -OH, aryl, - (C ₁ ~C ₁₈ alkyl) - oxy, - (C ₁ ~C ₁₈ alkyl) - amino, - (C ₁ -C ₁₈ alkyl) - _{thio, -CO 2 Y, -C (=} O) Y, -C (= O) NY 2, an -NO ₂ or -SH, or R ¹ and and R ² and / Or R ³ and R ⁴ can combine to form a cyclic carbon substituted with one or more oxo groups,
When L ² is not P (phenyl) ₃ , R ^{1 to} R ⁴ are independently —H, — (C _{1 to} C ₁₈ alkyl) —OH, aryl, — (C _{1 to} C ₁₈ alkyl) -oxy, - (C ₁ -C ₁₈ alkyl) - amino, - (C ₁ ~C ₁₈ alkyl) - _{thio, -CO 2 Y, -C (=} O) Y, -C (= O) NY 2, -NO 2, -OH or -SH, or R ¹ and R ² and / or R ³ and R ⁴ may be combined to form a cyclic carbon substituted with one or more oxo groups, R ¹ to At least one of R ⁴ is not H,
X is ^{Cl -, F -, Br -} , I -, PF 6 -, CF 3 SO 3 -, (C 1 ~C 18 alkyl) -CO ₂ ^- or (C ₁ -C ₁₈ alkyl) -SO ₃ ^- in Yes,
Y is selected from the group consisting of —H, —C ₁ -C ₁₈ alkyl, aryl, — (C ₁ -C ₁₈ alkyl) -aryl, —C ₃ -C ₈ cycloalkyl, heteroaryl and heterocyclyl). The compound is provided.

(0024)
In one embodiment of the compound of formula V:
Each L ¹ is independently (a) a 5-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(B) a 6-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(C) an 8 to 10 membered bicyclic ring having a nitrogen atom member in which one of the bicyclic rings is aromatic and forms a bond with Ru;
(D) a —NH ₂ group in which the nitrogen atom forms a bond with Ru;
(E) a —COOH group in which one of the oxygen atoms forms a bond with Ru;
(F) phosphorus atoms form a bond and Ru, R is independently -H, -C ₁ ~C ₁₈ -PR ₂ groups are alkyl or aryl, or (g) a sulfur atom forms a bond with Ru , R is an organic molecule having a —SR group that is independently —H, —C ₁ -C ₁₈ alkyl or aryl,
When L ² is P (phenyl) ₃ , each phenyl is independently -C ₁ -C ₁₈ alkyl,-(C ₁ -C ₁₈ alkyl) -OH, aryl,-(C ₁ -C ₁₈ alkyl) -oxy. , - (C ₁ ~C ₁₈ alkyl) - amino, - (C ₁ ~C ₁₈ alkyl) - _{thio, -CO 2 Y, -C (=} O) Y, -C (= O) NY 2, -NH 2 , -NO _2, substituted with -OH or -SH,
R ^{1 to} R ⁴ are independently —H, —C _{1 to} C ₁₈ alkyl, —NH ₂ , — (C _{1 to} C ₁₈ alkyl) —O— (C _{1 to} C ₁₈ alkyl), —OC (O). (C ₁ -C ₁₈ alkyl), - (C ₁ ~C ₁₈ alkyl) -OH, aryl, - (C ₁ ~C ₁₈ alkyl) - oxy, - (C ₁ ~C ₁₈ alkyl) - amino, - (C ₁ -C ₁₈ alkyl) - _{thio, -CO 2 Y, -C (=} O) Y, -C (= O) NY 2, an -NO ₂ or -SH, or R ¹ and and R ² and / Or R ³ and R ⁴ can combine to form a cyclic carbon substituted with one or more oxo groups,
When L ² is not P (phenyl) ₃ , R ^{1 to} R ⁴ are independently —H, — (C _{1 to} C ₁₈ alkyl) —OH, aryl, — (C _{1 to} C ₁₈ alkyl) -oxy, - (C ₁ -C ₁₈ alkyl) - amino, - (C ₁ ~C ₁₈ alkyl) - _{thio, -CO 2 Y, -C (=} O) Y, -C (= O) NY 2, -NO 2, -OH or -SH, or R ¹ and R ² and / or R ³ and R ⁴ may be combined to form a cyclic carbon substituted with one or more oxo groups, R ¹ to At least one of R ⁴ is not H,
X is ^{Cl -, F -, Br -} , I -, PF 6 -, CF 3 SO 3 -, (C 1 ~C 18 alkyl) -CO ₂ ^- or (C ₁ -C ₁₈ alkyl) -SO ₃ ^- in Yes,
Y is -H, ₁ -C ₁₈ alkyl, aryl, -C - (C ₁ ~C ₁₈ alkyl) - aryl, -C ₃ -C ₈ cycloalkyl, is selected from the group consisting of heteroaryl and heterocyclyl.

(0025)
In some embodiments of the compound of Formula V, when L ² is (R ⁹ ) ₃ P, each R ⁹ is different from each other.
In some embodiments of the compound of Formula V, when L ² is (R ⁹ ) ₃ P, the two R ⁹ are the same and different from the remaining R ⁹ .
In some embodiments of the compound of Formula V, when L ² is (Ph) ₃ P, each phenyl is not substituted with methyl.

ＶＩ
（式中、
各Ｌ¹は独立して
（ａ）環員の１つがＲｕと結合を形成する窒素原子である５員単環式芳香環、
（ｂ）環員の１つがＲｕと結合を形成する窒素原子である６員単環式芳香環、
（ｃ）２環式環の１つが芳香族であり且つＲｕと結合を形成する窒素原子員を有する８〜１０員２環式環、
（ｄ）窒素原子がＲｕと結合を形成する−ＮＨ₂基、
（ｅ）酸素原子の１つがＲｕと結合を形成する−ＣＯＯＨ基、
（ｆ）リン原子がＲｕと結合を形成し、Ｒが独立して−Ｈ、−Ｃ₁〜Ｃ₁₈アルキル若しくはアリールである−ＰＲ₂基、又は
（ｇ）硫黄原子がＲｕと結合を形成し、Ｒが独立して−Ｈ、−Ｃ₁〜Ｃ₁₈アルキル若しくはアリールである−ＳＲ基
を有する有機分子であり、
Ｌ²はＰ（フェニル）₃であり、各フェニルは独立して−Ｃ₁〜Ｃ₁₈アルキル、−（Ｃ₁〜Ｃ₁₈アルキル）−ＯＨ、アリール、−（Ｃ₁〜Ｃ₁₈アルキル）−オキシ、−（Ｃ₁〜Ｃ₁₈アルキル）−アミノ、−（Ｃ₁〜Ｃ₁₈アルキル）−チオ、−ＣＯ₂Ｙ、−Ｃ（＝Ｏ）Ｙ、−Ｃ（＝Ｏ）ＮＹ₂、−ＮＨ₂、−ＮＯ₂、−ＯＨ又は−ＳＨで置換され、あるいはＬ²は（Ｒ⁹）₂Ｐ若しくは（Ｒ⁹Ｏ）₂Ｐのリン原子を介してＲｕに結合される標識分子又はその活性誘導体であり、ｍは２であり、あるいはＬ²はＮＨＲ⁹、Ｎ（Ｒ⁹）₂、ピリジル、Ｃ（Ｒ⁹）＝ＮＨ、Ｃ（Ｒ⁹）＝ＮＲ⁹環状脂肪族アミン基若しくはニトリルの窒素原子を介してＲｕに結合される標識分子又はその活性誘導体であり、ｍは２であり、
各Ｒ⁹は独立して−Ｃ₁〜Ｃ₁₈アルキル、−Ｃ₃〜Ｃ₈シクロアルキル又はフェニルであり、 (0028)
In another embodiment, the present invention provides compounds of formula VI:

VI
(Where
Each L ¹ is independently (a) a 5-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(B) a 6-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(C) an 8 to 10 membered bicyclic ring having a nitrogen atom member in which one of the bicyclic rings is aromatic and forms a bond with Ru;
(D) a —NH ₂ group in which the nitrogen atom forms a bond with Ru;
(E) a —COOH group in which one of the oxygen atoms forms a bond with Ru;
(F) phosphorus atoms form a bond and Ru, R is independently -H, -C ₁ ~C ₁₈ -PR ₂ groups are alkyl or aryl, or (g) a sulfur atom forms a bond with Ru , R is an organic molecule having a —SR group that is independently —H, —C ₁ -C ₁₈ alkyl or aryl,
L ² is P (phenyl) ₃ and each phenyl is independently -C ₁ -C ₁₈ alkyl,-(C ₁ -C ₁₈ alkyl) -OH, aryl,-(C ₁ -C ₁₈ alkyl) -oxy. , - (C ₁ ~C ₁₈ alkyl) - amino, - (C ₁ ~C ₁₈ alkyl) - _{thio, -CO 2 Y, -C (=} O) Y, -C (= O) NY 2, -NH 2 , —NO ₂ , —OH or —SH, or L ² is a labeled molecule or an active derivative thereof bonded to Ru via the phosphorus atom of (R ⁹ ) ₂ P or (R ⁹ O) ₂ P Yes, m is 2, or L ² is NHR ⁹ , N (R ⁹ ) ₂ , pyridyl, C (R ⁹ ) ═NH, C (R ⁹ ) = NR ⁹ cyclic aliphatic amine group or nitrogen atom of nitrile A labeled molecule or an active derivative thereof bound to Ru via m, and m is 2.
Each R ⁹ is independently —C ₁ -C ₁₈ alkyl, —C ₃ -C ₈ cycloalkyl or phenyl;

R ^{1 to} R ⁴ are independently —H, —C _{1 to} C ₁₈ alkyl, —NH ₂ , — (C _{1 to} C ₁₈ alkyl) —O— (C _{1 to} C ₁₈ alkyl), —OC (O). (C ₁ -C ₁₈ alkyl), - (C ₁ ~C ₁₈ alkyl) -OH, aryl, - (C ₁ ~C ₁₈ alkyl) - oxy, - (C ₁ ~C ₁₈ alkyl) - amino, - (C ₁ -C ₁₈ alkyl) - _{thio, -CO 2 Y, -C (=} O) Y, -C (= O) NY 2, an -NO ₂ or -SH, or R ¹ and and R ² and / Or R ³ and R ⁴ can combine to form a cyclic carbon substituted with one or more oxo groups,
X is ^{Cl -, F -, Br -} , I -, PF 6 -, CF 3 SO 3 -, (C 1 ~C 18 alkyl) -CO ₂ ^- or (C ₁ -C ₁₈ alkyl) -SO ₃ ^- in Yes,
Y is selected from the group consisting of —H, —C ₁ -C ₁₈ alkyl, aryl, — (C ₁ -C ₁₈ alkyl) -aryl, —C ₃ -C ₈ cycloalkyl, heteroaryl and heterocyclyl). The compound is provided.

(0029)
In one embodiment of the compound of formula VI:
Each L ¹ is independently (a) a 5-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(B) a 6-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(C) an 8 to 10 membered bicyclic ring having a nitrogen atom member in which one of the bicyclic rings is aromatic and forms a bond with Ru;
(D) a —NH ₂ group in which the nitrogen atom forms a bond with Ru;
(E) a —COOH group in which one of the oxygen atoms forms a bond with Ru;
(F) phosphorus atoms form a bond and Ru, R is independently -H, -C ₁ ~C ₁₈ -PR ₂ groups are alkyl or aryl, or (g) a sulfur atom forms a bond with Ru , R is an organic molecule having a —SR group that is independently —H, —C ₁ -C ₁₈ alkyl or aryl,
L ² is P (phenyl) ₃ and each phenyl is independently -C ₁ -C ₁₈ alkyl,-(C ₁ -C ₁₈ alkyl) -OH, aryl,-(C ₁ -C ₁₈ alkyl) -oxy. , - (C ₁ ~C ₁₈ alkyl) - amino, - (C ₁ ~C ₁₈ alkyl) - _{thio, -CO 2 Y, -C (=} O) Y, -C (= O) NY 2, -NH 2 , -NO _2, substituted with -OH or -SH,
R ^{1 to} R ⁴ are independently —H, —C _{1 to} C ₁₈ alkyl, —NH ₂ , — (C _{1 to} C ₁₈ alkyl) —O— (C _{1 to} C ₁₈ alkyl), —OC (O). (C ₁ -C ₁₈ alkyl), - (C ₁ ~C ₁₈ alkyl) -OH, aryl, - (C ₁ ~C ₁₈ alkyl) - oxy, - (C ₁ ~C ₁₈ alkyl) - amino, - (C ₁ -C ₁₈ alkyl) - _{thio, -CO 2 Y, -C (=} O) Y, -C (= O) NY 2, an -NO ₂ or -SH, or R ¹ and and R ² and / Or R ³ and R ⁴ can combine to form a cyclic carbon substituted with one or more oxo groups,
X is ^{Cl -, F -, Br -} , I -, PF 6 -, CF 3 SO 3 -, (C 1 ~C 18 alkyl) -CO ₂ ^- or (C ₁ -C ₁₈ alkyl) -SO ₃ ^- in Yes,
Y is -H, ₁ -C ₁₈ alkyl, aryl, -C - (C ₁ ~C ₁₈ alkyl) - aryl, -C ₃ -C ₈ cycloalkyl, is selected from the group consisting of heteroaryl and heterocyclyl.

(0030)
In some embodiments of the compound of formula VI, when L ² is (R ⁹ ) ₃ P, each R ⁹ is different from each other.
In some embodiments of the compound of formula VI, when L ² is (R ⁹ ) ₃ P, the two R ⁹ are the same and different from the remaining R ⁹ .
In some embodiments of the compound of formula VI, when L ² is (Ph) ₃ P, each phenyl is not substituted with methyl.

ＶＩＩ
（式中、
各Ｌ¹は独立して
（ａ）環員の１つがＲｕと結合を形成する窒素原子である５員単環式芳香環、
（ｂ）環員の１つがＲｕと結合を形成する窒素原子である６員単環式芳香環、
（ｃ）２環式環の１つが芳香族であり且つＲｕと結合を形成する窒素原子員を有する８〜１０員２環式環、
（ｄ）窒素原子がＲｕと結合を形成する−ＮＨ₂基、
（ｅ）酸素原子の１つがＲｕと結合を形成する−ＣＯＯＨ基、
（ｆ）リン原子がＲｕと結合を形成し、Ｒが独立して−Ｈ、−Ｃ₁〜Ｃ₁₈アルキル若しくはアリールである−ＰＲ₂基、又は
（ｇ）硫黄原子がＲｕと結合を形成し、Ｒが独立して−Ｈ、−Ｃ₁〜Ｃ₁₈アルキル若しくはアリールである−ＳＲ基
を有する有機分子であり、
Ｌ²は（Ｒ⁹）₃Ｐ、（Ｒ⁹Ｏ）₃Ｐ又はＬ¹であり、各Ｒ⁵は独立して−Ｃ₁〜Ｃ₁₈アルキル、−Ｃ₃〜Ｃ₈シクロアルキル又はフェニルであり、ｍは２であり、Ｌ²はＰ（フェニル）₃ではなく、あるいはＬ²は−ＣＮであり、ｍは１であり、あるいはＬ²は（Ｒ⁹）₂Ｐ若しくは（Ｒ⁹Ｏ）₂Ｐのリン原子を介してＲｕに結合される標識分子又はその活性誘導体であり、ｍは２であり、あるいはＬ²はＮＨＲ⁹、Ｎ（Ｒ⁹）₂、ピリジル、Ｃ（Ｒ⁹）＝ＮＨ、Ｃ（Ｒ⁹）＝ＮＲ⁹環状脂肪族アミン基若しくはニトリルの窒素原子を介してＲｕに結合される標識分子又はその活性誘導体であり、ｍは２であり、
各Ｒ⁹は独立して−Ｃ₁〜Ｃ₁₈アルキル、−Ｃ₃〜Ｃ₈シクロアルキル又はフェニルであり、 (0033)
In another embodiment, the present invention provides compounds of formula VII,

VII
(Where
Each L ¹ is independently (a) a 5-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(B) a 6-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(C) an 8 to 10 membered bicyclic ring having a nitrogen atom member in which one of the bicyclic rings is aromatic and forms a bond with Ru;
(D) a —NH ₂ group in which the nitrogen atom forms a bond with Ru;
(E) a —COOH group in which one of the oxygen atoms forms a bond with Ru;
(F) phosphorus atoms form a bond and Ru, R is independently -H, -C ₁ ~C ₁₈ -PR ₂ groups are alkyl or aryl, or (g) a sulfur atom forms a bond with Ru , R is an organic molecule having a —SR group that is independently —H, —C ₁ -C ₁₈ alkyl or aryl,
L ² is (R ⁹ ) ₃ P, (R ⁹ O) ₃ P or L ¹ , and each R ⁵ is independently —C ₁ -C ₁₈ alkyl, —C ₃ -C ₈ cycloalkyl or phenyl. , M is 2, and L ² is not P (phenyl) ₃ , or L ² is —CN, m is 1, or L ² is (R ⁹ ) ₂ P or (R ⁹ O) _2. A labeled molecule or an active derivative thereof bonded to Ru via the phosphorus atom of P, m is 2, or L ² is NHR ⁹ , N (R ⁹ ) ₂ , pyridyl, C (R ⁹ ) ═NH , C (R ⁹ ) = NR ⁹ a labeled molecule bonded to Ru via a cyclic aliphatic amine group or a nitrogen atom of a nitrile, or an active derivative thereof, m is 2,
Each R ⁹ is independently —C ₁ -C ₁₈ alkyl, —C ₃ -C ₈ cycloalkyl or phenyl;

R ^{1 to} R ⁴ are independently -H,-(C ₁ -C ₁₈ alkyl) -OH, aryl,-(C ₁ -C ₁₈ alkyl) -oxy,-(C ₁ -C ₁₈ alkyl) -amino, - (C ₁ -C ₁₈ alkyl) - _{thio, -CO 2 Y, -C (=} O) Y, -C (= O) NY 2, -NO 2, -OH or -SH, or the R ¹ R ² and / or R ³ and R ⁴ can combine to form a cyclic carbon substituted with one or more oxo groups, at least one of R ^{1 to} R ⁴ is not H,
X is ^{Cl -, F -, Br -} , I -, PF 6 -, CF 3 SO 3 -, (C 1 ~C 18 alkyl) -CO ₂ ^- or (C ₁ -C ₁₈ alkyl) -SO ₃ ^- in Yes,
Y is selected from the group consisting of —H, —C ₁ -C ₁₈ alkyl, aryl, — (C ₁ -C ₁₈ alkyl) -aryl, —C ₃ -C ₈ cycloalkyl, heteroaryl and heterocyclyl). The compound is provided.

(0034)
In one embodiment of the compound of formula VII:
Each L ¹ is independently (a) a 5-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(B) a 6-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(C) an 8 to 10 membered bicyclic ring having a nitrogen atom member in which one of the bicyclic rings is aromatic and forms a bond with Ru;
(D) a —NH ₂ group in which the nitrogen atom forms a bond with Ru;
(E) a —COOH group in which one of the oxygen atoms forms a bond with Ru;
(F) phosphorus atoms form a bond and Ru, R is independently -H, -C ₁ ~C ₁₈ -PR ₂ groups are alkyl or aryl, or (g) a sulfur atom forms a bond with Ru , R is an organic molecule having a —SR group that is independently —H, —C ₁ -C ₁₈ alkyl or aryl,
L ² is (R ⁹ ) ₃ P, (R ⁹ O) ₃ P or L ¹ , and each R ⁵ is independently —C ₁ -C ₁₈ alkyl, —C ₃ -C ₈ cycloalkyl or phenyl. , m is 2, L ² is P (phenyl) rather than ₃ or L ^2, is -CN, m is 1,
R ^{1 to} R ⁴ are independently -H,-(C ₁ -C ₁₈ alkyl) -OH, aryl,-(C ₁ -C ₁₈ alkyl) -oxy,-(C ₁ -C ₁₈ alkyl) -amino, - (C ₁ -C ₁₈ alkyl) - _{thio, -CO 2 Y, -C (=} O) Y, -C (= O) NY 2, -NO 2, -OH or -SH, or the R ¹ R ² and / or R ³ and R ⁴ can combine to form a cyclic carbon substituted with one or more oxo groups, at least one of R ^{1 to} R ⁴ is not H,
X is Cl ⁻ , F ⁻ , Br ⁻ , I ⁻ , PF ₆ ⁻ , CF ₃ SO ₃ ⁻ , (C _{1 to} C ₁₈ alkyl) -CO ₂ ^— or (C _{1 to} C ₁₈ alkyl) —SO ₃ ^—. And
Y is -H, ₁ -C ₁₈ alkyl, aryl, -C - (C ₁ ~C ₁₈ alkyl) - aryl, -C ₃ -C ₈ cycloalkyl, is selected from the group consisting of heteroaryl and heterocyclyl.

(0035)
In some embodiments of the compound of formula VII, when L ² is (R ⁹ ) ₃ P, each R ⁹ is different from each other.
In some embodiments of the compound of formula VII, when L ² is (R ⁹ ) ₃ P, the two R ⁹ are the same and different from the remaining R ⁹ .
In some embodiments of the compound of formula VII, when L ² is (Ph) ₃ P, each phenyl is not substituted with methyl.

ＶＩＩＩ
（式中、
各Ｌ¹は独立して（Ｒ⁹）₂Ｐ若しくは（Ｒ⁹Ｏ）₂Ｐのリン原子を介してＲｕに結合される標識分子又はその活性誘導体であり、ｍは２であり、あるいはＬ¹はＮＨＲ⁹、Ｎ（Ｒ⁹）₂、ピリジル、Ｃ（Ｒ⁹）＝ＮＨ、Ｃ（Ｒ⁹）＝ＮＲ⁹環状脂肪族アミン基若しくはニトリルの窒素原子を介してＲｕに結合される標識分子又はその活性誘導体であり、ｍは２であり、
各Ｒ⁹は独立して−Ｃ₁〜Ｃ₁₈アルキル、−Ｃ₃〜Ｃ₈シクロアルキル又はフェニルであり、
Ｌ²はＣｌ−、ホスフィン、ＯＨ₂又はピリジンであり、ｍは２であり、あるいはＬ²は−ＣＮであり、ｍは１であり、
Ｒ¹〜Ｒ⁴は独立して−Ｈ、−Ｃ₁〜Ｃ₁₈アルキル、−ＮＨ₂、−（Ｃ₁〜Ｃ₁₈アルキル）−Ｏ−（Ｃ₁〜Ｃ₁₈アルキル）、−ＯＣ（Ｏ）（Ｃ₁〜Ｃ₁₈アルキル）、−（Ｃ₁〜Ｃ₁₈アルキル）−ＯＨ、アリール、−（Ｃ₁〜Ｃ₁₈アルキル）−オキシ、−（Ｃ₁〜Ｃ₁₈アルキル）−アミノ、−（Ｃ₁〜Ｃ₁₈アルキル）−チオ、−ＣＯ₂Ｙ、−Ｃ（＝Ｏ）Ｙ、−Ｃ（＝Ｏ）ＮＹ₂、−ＮＯ₂又は−ＳＨであり、あるいはＲ¹とＲ²とは及び／又はＲ³とＲ⁴とは組み合わさって１つ以上のオキソ基で置換される環状炭素を形成し得て、
ＸはＣｌ^-、Ｆ^-、Ｂｒ^-、Ｉ^-、ＰＦ₆ ^-、ＣＦ₃ＳＯ₃ ^-、（Ｃ₁〜Ｃ₁₈アルキル）−ＣＯ₂ ^-又は（Ｃ₁〜Ｃ₁₈アルキル）−ＳＯ₃ ^-であり、
Ｙは、−Ｈ、−Ｃ₁〜Ｃ₁₈アルキル、アリール、−（Ｃ₁〜Ｃ₁₈アルキル）−アリール、−Ｃ₃〜Ｃ₈シクロアルキル、ヘテロアリール及びヘテロシクリルから成る群から選択される）で示される化合物を提供する。 (0038)
In another embodiment, the present invention provides compounds of formula VIII,

VIII
(Where
Each L ¹ is independently a labeled molecule or an active derivative thereof bonded to Ru via a phosphorus atom of (R ⁹ ) ₂ P or (R ⁹ O) ₂ P, and m is 2, or L ¹ NHR ⁹ , N (R ⁹ ) ₂ , pyridyl, C (R ⁹ ) ═NH, C (R ⁹ ) = NR ⁹ A labeled molecule bonded to Ru via a cyclic aliphatic amine group or a nitrogen atom of nitrile or An active derivative thereof, m is 2,
Each R ⁹ is independently —C ₁ -C ₁₈ alkyl, —C ₃ -C ₈ cycloalkyl or phenyl;
L ² is Cl-, phosphine, OH ₂ or pyridine, m is 2, or L ² is -CN, m is 1,
R ^{1 to} R ⁴ are independently —H, —C _{1 to} C ₁₈ alkyl, —NH ₂ , — (C _{1 to} C ₁₈ alkyl) —O— (C _{1 to} C ₁₈ alkyl), —OC (O). (C ₁ -C ₁₈ alkyl), - (C ₁ ~C ₁₈ alkyl) -OH, aryl, - (C ₁ ~C ₁₈ alkyl) - oxy, - (C ₁ ~C ₁₈ alkyl) - amino, - (C ₁ -C ₁₈ alkyl) - _{thio, -CO 2 Y, -C (=} O) Y, -C (= O) NY 2, an -NO ₂ or -SH, or R ¹ and and R ² and / Or R ³ and R ⁴ can combine to form a cyclic carbon substituted with one or more oxo groups,
X is ^{Cl -, F -, Br -} , I -, PF 6 -, CF 3 SO 3 -, (C 1 ~C 18 alkyl) -CO ₂ ^- or (C ₁ -C ₁₈ alkyl) -SO ₃ ^- in Yes,
Y is selected from the group consisting of —H, —C ₁ -C ₁₈ alkyl, aryl, — (C ₁ -C ₁₈ alkyl) -aryl, —C ₃ -C ₈ cycloalkyl, heteroaryl and heterocyclyl). The compounds shown are provided.

(0039)
In one embodiment of the compound of formula VIII:
Each L ¹ is independently a labeled molecule;
L ² is Cl-, phosphine, OH ₂ or pyridine, m is 2, or L ² is -CN, m is 1,
R ^{1 to} R ⁴ are independently —H, —C _{1 to} C ₁₈ alkyl, —NH ₂ , — (C _{1 to} C ₁₈ alkyl) —O— (C _{1 to} C ₁₈ alkyl), —OC (O). (C ₁ -C ₁₈ alkyl), - (C ₁ ~C ₁₈ alkyl) -OH, aryl, - (C ₁ ~C ₁₈ alkyl) - oxy, - (C ₁ ~C ₁₈ alkyl) - amino, - (C ₁ -C ₁₈ alkyl) - _{thio, -CO 2 Y, -C (=} O) Y, -C (= O) NY 2, an -NO ₂ or -SH, or R ¹ and and R ² and / Or R ³ and R ⁴ can combine to form a cyclic carbon substituted with one or more oxo groups,
X is ^{Cl -, F -, Br -} , I -, PF 6 -, CF 3 SO 3 -, (C 1 ~C 18 alkyl) -CO ₂ ^- or (C ₁ -C ₁₈ alkyl) -SO ₃ ^- in Yes,
Y is -H, ₁ -C ₁₈ alkyl, aryl, -C - (C ₁ ~C ₁₈ alkyl) - aryl, -C ₃ -C ₈ cycloalkyl, is selected from the group consisting of heteroaryl and heterocyclyl.

(0040)
In some embodiments of the compound of Formula VIII, when L ² is (R ⁹ ) ₃ P, each R ⁹ is different from each other.
In some embodiments of the compound of Formula VIII, when L ² is (R ⁹ ) ₃ P, the two R ⁹ are the same and different from the remaining R ⁹ .
In some embodiments of the compound of Formula VIII, when L ² is (Ph) ₃ P, each phenyl is not substituted with methyl.
In some embodiments of the compounds of Formulas I-III and V-VIII, L ² is rhodamine B-methylaminopropionitrile amide (RhodB-MAPN). In some embodiments of compounds of Formulas I-III and V-VIII, L ² is rhodamine G-methylaminopropionitrile amide (RhodG-MAPN).

（ＲｈｏｄＢ−ＭＡＭｅＰｙ）又はその塩化物塩（ＲｈｏｄＢ−ＭＡＭｅＰｙ−Ｃｌ）である。 (0044)
In one embodiment of compounds of formulas I-III and V-VIII, L ² is

(RhodB-MAMePy) or a chloride salt thereof (RhodB-MAMePy-Cl).

（Ｒｈｏｄ６Ｇ−ＭＡＭｅＰｙ）又はその塩化物塩（Ｒｈｏｄ６Ｇ−ＭＡＭｅＰｙ−Ｃｌ）である。 (0045)
In some embodiments of compounds of Formulas I-III and V-VIII, L ² is

(Rhod6G-MAMePy) or a chloride salt thereof (Rhod6G-MAMePy-Cl).

(0046)
In some embodiments of compounds of Formulas I-III and V-VIII, the labeling molecule is rhodamine B. In some embodiments of compounds of Formulas I-III and V-VIII, the labeling molecule is rhodamine 6G.
In some embodiments of compounds of Formulas I-III and V-VIII, the labeling molecule is body pie, dansyl, fluorescein, Texas red, cyanine dye, pyrene, coumarin, cascade blue TM, pacific blue, marina blue, oregon green. Selected from the group consisting of 4 ', 6-diamidino-2-phenylindole (DAPI), indopyra dye, lucifer yellow, propidium iodide, porphyrin and arginine. Compounds of Formulas I-VIII (“photolabile compounds”) release L ¹ when exposed to light.

(0048)
In another aspect, the present invention provides a composition comprising an effective amount of a photolabile compound and a physiologically acceptable carrier, vehicle, diluent or excipient.
In another aspect, the present invention provides a container containing a photolabile compound.
In yet another aspect thereof, the present invention provides a kit comprising a photolabile compound and instructions for use.
Another aspect of the invention provides a method of releasing an organic molecule or a labeled molecule from a photolabile compound, the method under conditions sufficient to release the organic molecule or the labeled molecule. Including exposure to light.

(0052)
In yet another aspect, the present invention provides a method of making an organic molecule bioavailable to a patient (patient) in need of the organic molecule, the method comprising administering a photolabile compound to the patient, Exposing the compound to light under conditions sufficient to release organic molecules from the compound.
In another aspect, the present invention provides a method of treating a patient's diseases and disorders comprising: (a) administering a photolabile compound to a patient; and (b) treating the photolabile compound with an organic molecule. Exposure to light under conditions sufficient to release the from the photolabile compound. In some embodiments, the diseases and disorders to be treated by the methods of the present invention include neurological, neurophysiological or neuromuscular diseases and conditions (such as epilepsy, multiple sclerosis), cancer, sweating and blood diseases. Is included.

(0054)
In another aspect, the invention provides a method for assaying an organic molecule, the method comprising (a) a photolabile compound and a biological sample under conditions sufficient to release the organic molecule from the photolabile compound. And (b) confirming the effect of this organic molecule on the biological sample.
In one embodiment, the present invention provides [Ru (bpy) 2 (RhodB-MAPN) Cl] Cl, [Ru (bpy) 2 (Rhod6G-MAPN) Cl] Cl, [Ru (bpy) 2 (RhodB-MAMePy) Cl. A compound selected from the group consisting of Cl.
In another embodiment, the present invention relates to [Ru (bpy) 2 (RhodB-MAPN) Cl] Z, [Ru (bpy) 2 (Rhod6G-MAPN) Cl] Z, [Ru (bpy) 2 (RhodB-MAMePy). Cl] Z, wherein Z is a compound selected from the group consisting of anions. In some embodiments, Z is Cl ⁻ , F ⁻ , Br ⁻ or I ⁻ .

(0057)
Additional aspects, features and advantages obtained by the present invention will become apparent from the following detailed description, figures and examples.

(0058-0077)
FIG. _{3 is} a partial ¹ H NMR spectrum of [Ru (bpy) ₂ (4AP) ₂ ] Cl ₂ described in the present specification (Example 1), showing a signal corresponding to 4-AP metahydrogen. ml: [Ru (bpy) ₂ (4AP) ₂ ] ²⁺ , m2: [Ru (bpy) ₂ (H ₂ O) (4AP) ₂ ] ²⁺ , m3: free ligand 4-AP. (Top) Action potentials (spikes) recorded in neurons of the medical hill for solutions of saline, [Ru (bpy) ₃ ] Cl ₂ , [Ru (bpy) ₂ (4AP) ₂ ] Cl ₂ ). (Bottom): Spike frequency. Arrows indicate light irradiation using a Xe flash lamp. (Intermediate): Composition of extracellular medium. ₂ shows a cyclic voltammetry (CV) profile of [Ru (bpy) ₂ (4AP) ₂ ] Cl ₂ in water. The supporting electrolyte was KNO ₃ (1M). DE / dt = 100 mV / sec in a glassy carbon electrode. The NMR spectrum of [Ru (bpy) ₂ (4AP) ₂ ] Cl ₂ in D ₂ O before light irradiation is shown. Bruker 500 MHz. The NMR spectrum of [Ru (bpy) ₂ (4AP) ₂ ] Cl ₂ in D ₂ O after light irradiation is shown. Bruker 500 MHz. The ultraviolet-visible light (UV-vis) spectrum of Ru (bpy) ₂ (4AP) ₂ before complete photolysis and after complete photolysis is shown. The photodegradation products after exposure to light were Ru (bpy) ₂ (4AP) (H ₂ O) and free 4AP. The complex did not undergo dark degradation beyond 20 hours. After 7 hours in the dark, the irradiated solution showed less than 4% 4AP recombination. The ultraviolet-visible spectrum of the filter used for the ganglion light irradiation experiment as described in Example 3 is shown. The action potentials and spike frequencies obtained from the study of the medulla of the medical hill (Hirudo medicalis) are shown. FIG. 6 shows a recording of Retzius neuron potential activity during perfusion of free 4AP on leech ganglia. [4AP] = 0, 10, 20 and 50 mM. Flow rate = 1 ml / min. Carrier: saline. As described in Example 3. The action potentials and spike frequencies obtained from the study of the medulla of the medical hill (Hirudo medicalis) are shown. FIG. 7 shows a record of Retzius neuron activity during exposure of green light through a filter of FIG. 6 to a 0.1 millisecond flash. Flow rate = 1 ml / min. Carrier: saline. As described in Example 3. Pulse energy: 0.5J. It relates to the spectral change of [Ru (bpy) ₂ (4AP) ₂ ] Cl ₂ during exposure to light. Change in spectrum of [Ru (bpy) ₂ (4AP) ₂ ] Cl ₂ during irradiation with 473 nm laser light. Output: 6.39 mW continuous. [Ru (bpy) ₂ (4AP) ₂ ] Cl ₂ starting concentration: 27.9 μM. A (473 nm) = 0.18. It relates to the spectral change of [Ru (bpy) ₂ (4AP) ₂ ] Cl ₂ during exposure to light. Fraction of [Ru (bpy) ₂ (4AP) (H ₂ O)] ²⁺ as a function of light irradiation time obtained from the spectrum of FIG. 8A. Fig. 4 is a graph of photoemitted 4AP versus pulse energy. The light source was a pulsed Xe lamp fitted with a bandpass filter. [Ru (bpy) ₂ (4AP) ₂ ] Cl ₂ = 44 μM. Volume = 3 mL. Data were obtained from ultraviolet and visible spectral analysis. FIG. ₄ is several two-photon fluorescence images of [Ru (bpy) ₂ (TzGly) (py)] Cl ₂ at different excitation wavelengths (magnification: about 20 ×). It is a graph of the excitation wavelength of a total two-photon fluorescence pair [Ru (bpy) ₂ (4AP) ₂ ] Cl ₂ . It is an ultraviolet and visible spectrum of TzGly before and after irradiation with light of 400 to 600 nm. The structure of TzGly is shown. FIG. 3 shows mouse cortical neuron spiking caused by adding TzGly (1 μM) to neurons by perfusion. The measurement results were obtained by the whole cell patch clamp method known and used in the art. [Ru (bpy) ₂ (TzGly) (py)] relates to experiments performed on neurons in contact with Cl ₂ . FIG. 2 is a fluorescence imaging micrograph of a neuron including an enlarged view of dendritic spines. [Ru (bpy) ₂ (TzGly) (py)] relates to experiments performed on neurons in contact with Cl ₂ . FIG. 2 is a fluorescence imaging micrograph of a neuron including an enlarged view of dendritic spines. [Ru (bpy) ₂ (TzGly) (py)] relates to experiments performed on neurons in contact with Cl ₂ . FIG. 2 is a fluorescence imaging micrograph of a neuron including an enlarged view of dendritic spines. [Ru (bpy) ₂ (TzGly) (py)] relates to experiments performed on neurons in contact with Cl ₂ . FIG. 6 shows the effect of laser irradiation (about 40 mW) on the spiking of one neuron in the presence of [Ru (bpy) ₂ (TzGly) ₂ ] Cl ₂ . [Ru (bpy) ₂ (TzGly) ₂ ] Cl ₂ concentration = 100 μM. Pulse width: 10 milliseconds. Output: 40 mW, wavelength: 720 nm. [Ru (bpy) ₂ (TzGly) (py)] relates to experiments performed on neurons in contact with Cl ₂ . 14A-D relates to the experiment performed as a control for the experiment of FIGS. It is an enlarged view of the dendritic spine of a neuron. [Ru (bpy) ₂ (TzGly) (py)] relates to experiments performed on neurons in contact with Cl ₂ . 14A-D relates to the experiment performed as a control for the experiment of FIGS. It is an enlarged view of the dendritic spine of a neuron. [Ru (bpy) ₂ (TzGly) (py)] relates to experiments performed on neurons in contact with Cl ₂ . . 14A-D relates to the experiment performed as a control for the experiment of FIGS. FIG. 6 is a plot showing the effect of laser irradiation on control neurons in the absence of [Ru (bpy) ₂ (TzGly) ₂ ] Cl ₂ . No activation of activity was observed. Pulse width: 10 milliseconds. Output: 40 mW. Wavelength: 720 nm. Figure ₅ shows the cyclic voltammetry (CV) profile of native [Ru (bpy) ₂ (PMe ₃ ) Glu] at 100 mV / sec with a Pt wire electrode in CH ₃ CN containing 100 mM TBAPF ₆ . (Top) Ultraviolet / visible spectrum of [Ru (bpy) ₂ (PMe ₃ ) Glu] (0.1 mM, pH = 7) during light irradiation for 360 seconds using 450 nm light. (Inset graph) The amount of glutamate released from the experimental data (circle). Fit to the theoretical equation (line). (Bottom) Absorbance change at 532 nm after flash photolysis (10 nanoseconds / pulse, average 256 pulses) of [Ru (bpy) ₂ (PMe ₃ ) Glu] (1 mM). The aromatic section of the ¹ H-NMR spectrum of [Ru (bpy) ₂ (PMe ₃ ) Glu] in D ₂ O before photolysis (top trace) and after photolysis (bottom trace) is shown, [Ru ( bpy) ₂ (PMe ₃ ) H ₂ O] ²⁺ aromatic signal. The aliphatic section of the ¹ H-NMR spectrum of [Ru (bpy) ₂ (PMe ₃ ) Glu] in D ₂ O before photolysis (upper trace) and after photolysis (lower trace). In the last case, the signals at 3.50, 2.30, 2.04 and 1.94 ppm correspond to free glutamate. ¹ H NMR aliphatic region spectrum of the complex [Ru (bpy) ₂ (RhodB-MAPN) Cl] ⁺ , (A) before light irradiation, (B) after 5 minutes light irradiation, (C) free It is a RhodB-MAPN spectrum. Note that in B, the signals of free ligands a, b and c are evident, indicating their light emission. Since the signals d and e corresponding to the ethyl group in RhodB-MAPN are away from the coordination center, they do not undergo a great change. A photolysis reaction diagram is added to make it easier to understand. It is an emission spectrum of 10 μM aqueous solution of the complex [Ru (bpy) ₂ (RhodB-MAPN) Cl] ⁺ during light irradiation of a cuvette using a 473 nm laser diode. The spectrum was measured every 10 seconds. The inset graph shows the light emission maximum during light irradiation.

(0078)
The present invention generally relates to a photolabile compound comprising an organic molecule or a labeled molecule and methods of using the photolabile compound. Organic molecules can be biologically active. In one embodiment of the invention, organic molecules (eg biologically active molecules) are protected and subsequently released by exposure to light, preferably visible light.
In contrast to known methods, visible light (eg, visible light pulses) can be used to release organic or labeled molecules from the photolabile compound. Thus, in this method, the sample (eg, organ, tissue, cell) or patient to which the photolabile compound is administered is exposed to ultraviolet light that has a detrimental effect on cellular components and ultimately cell growth and viability. , If any, is minimally exposed.

(0080)
In the present invention, although not intended to be bound by theory, since Ru-organic molecular bonds or Ru-labeled molecular bonds are usually weaker than covalent σ bonds, they can be cleaved by irradiation with low energy. Furthermore, although not intended to be bound by theory in the present invention, the energy required to release organic or labeled molecules upon exposure to light is relatively low. In the photolabile compound of the present invention, the organic molecule or the labeled molecule is light-emitted by light irradiation to the photolabile compound as described herein.
Also in the present invention, light emission can occur in vivo or in biological samples such as living cells and body fluids in the body, body samples (such as organ or tissue samples). Thus, photolabile compounds are particularly useful for in vivo biological applications, such as treating various diseases, conditions and disorders of the body. Through the use of a photolabile compound as described herein, the body, such as a living organ, tissue and tissue, while minimizing adverse effects on the biological sample or body or its organs, tissues and cellular components. The bioactive function or onset of bioactivity in the cell can be precisely controlled (ie within nanoseconds to milliseconds). In addition, exposure of the biological sample to light can be confined to the site where organic molecules are needed or desired. This is particularly beneficial for administration to patients, particularly human patients.
The photolabile compound is also suitable for use in non-biological systems such as solar cells, photovoltaic cells or optical memories (eg, three-dimensional optical memories).

Ｉ
（式中、Ｒ¹〜Ｒ⁸、Ｌ¹、Ｌ²、Ｘ、Ｍ及びｍは、式Ｉの化合物に関して上で定義した通りである）で示される化合物を含む。 (0083)
In one embodiment, the present invention provides compounds of formula I,

I
Wherein R ^{1 to} R ⁸ , L ¹ , L ² , X, M and m are as defined above for compounds of formula I.

ＩＩ
（式中、Ｒ¹〜Ｒ⁸、Ｌ¹、Ｌ²、Ｘ及びｍは、式ＩＩの化合物に関して上で定義した通りである）の化合物を含む。 (0084)
In another embodiment, the present invention provides compounds of formula II:

II
Wherein R ¹ -R ⁸ , L ¹ , L ² , X and m are as defined above for compounds of formula II.

ＩＩＩ
（式中、Ｒ¹〜Ｒ⁸、Ｌ¹、Ｌ²、Ｘ及びｍは、式ＩＩＩの化合物に関して上で定義した通りである）で示される化合物を含む。 (0085)
In another embodiment, the present invention provides compounds of formula III,

III
Wherein R ^{1 to} R ⁸ , L ¹ , L ² , X and m are as defined above for compounds of formula III.

ＩＶ
（式中、Ｍ¹、Ｍ²及びＬ¹は、式ＩＶで示される化合物に関して上で定義した通りである）で示される化合物を含む。 (0086)
In another embodiment, the present invention provides compounds of formula IV,

IV
Wherein M ¹ , M ² and L ¹ are as defined above with respect to the compound of formula IV.

Ｖ
（式中、Ｒ¹〜Ｒ⁴、Ｌ¹、Ｌ²、Ｘ及びｍは、式Ｖの化合物に関して上で定義した通りである）で示される化合物を含む。 (0087)
In one embodiment, the present invention provides compounds of formula V,

V
Wherein R ^{1 to} R ⁴ , L ¹ , L ² , X and m are as defined above for the compound of formula V.

ＶＩ
（式中、Ｒ¹〜Ｒ⁴、Ｌ¹、Ｌ²、Ｘ及びｍは、式ＶＩの化合物に関して上で定義した通りである）で示される化合物を含む。 (0088)
In another embodiment, the present invention provides compounds of formula VI,

VI
Wherein R ^{1 to} R ⁴ , L ¹ , L ² , X and m are as defined above for compounds of formula VI.

(0089)
In certain embodiments, the present invention includes compounds of Formula VI wherein L ² is P (phenyl) ₃ and each phenyl is independently substituted at the 3 or 4 position.
In another embodiment, the present invention includes compounds of Formula VI wherein L ² is P (phenyl) ₃ and at least one phenyl is substituted with — (C ₁ -C ₁₈ alkyl) —OH.
In another embodiment, the present invention includes compounds of Formula VI wherein L ² is P (phenyl) ₃ and each phenyl is substituted with — (C ₁ -C ₁₈ alkyl) —OH.
In another embodiment, the present invention includes compounds of Formula VI wherein L ² is P (phenyl) ₃ and at least one phenyl is replaced with —COOH.
In another embodiment, the present invention includes compounds of Formula II wherein L ² is P (phenyl) ₃ and each phenyl is substituted with —COOH.

(0094)
In another embodiment, the present invention includes compounds of Formula VI wherein L ² is P (phenyl) ₃ and at least one phenyl is replaced with —OH.
In another embodiment, the present invention includes compounds of Formula VI wherein L ² is P (phenyl) ₃ and each phenyl is substituted with —OH.
In another embodiment, the present invention includes compounds of Formula VI wherein L ² is P (phenyl) ₃ and at least one phenyl is substituted with —NH ₂ .
In another embodiment, the present invention includes compounds of Formula VI wherein L ² is P (phenyl) ₃ and each phenyl is substituted with —NH ₂ .
In another embodiment, the present invention includes compounds of Formula VI wherein L ² is P (phenyl) ₃ and at least one phenyl is replaced with —NO ₂ .
In another embodiment, the present invention includes compounds of Formula VI wherein L ² is P (phenyl) ₃ and each phenyl is substituted with —NO ₂ .
In another embodiment, the invention includes a compound of formula VI, wherein L ¹ is an organic molecule comprising PMe _{2 in} which the phosphorus atom forms a bond with Ru.
In another embodiment, the invention comprises a compound of formula VI, wherein L ¹ is an organic molecule comprising P (phenyl) _{2 in} which the phosphorus atom forms a bond with Ru.

ＶＩＩ
（式中、Ｒ¹〜Ｒ⁴、Ｌ¹、Ｌ²、Ｘ及びｍは、式ＶＩＩの化合物に関して上で定義した通りである）で示される化合物を含む。 (0102)
In another embodiment, the present invention provides compounds of formula VII,

VII
Wherein R ^{1 to} R ⁴ , L ¹ , L ² , X and m are as defined above for compounds of formula VII.

(0103)
In another embodiment, the present invention includes compounds of Formula VII where L ² is P (methyl) (phenyl) ₂ .
In another embodiment, the present invention includes compounds of Formula VII where L ² is P (methyl) ₂ (phenyl).
In another embodiment, the invention comprises a compound of formula VII, wherein L ¹ is an organic molecule comprising PMe _{2 in} which the phosphorus atom forms a bond with Ru.
In another embodiment, the invention comprises a compound of formula VII, wherein L ¹ is an organic molecule comprising P (phenyl) _{2 in} which the phosphorus atom forms a bond with Ru.

ＶＩＩＩ
（式中、Ｒ¹〜Ｒ⁴、Ｌ¹、Ｌ²、Ｘ及びｍは、式ＶＩＩＩの化合物に関して上で定義した通りである）で示される化合物を含む。 (0107)
In another embodiment, the present invention provides compounds of formula VIII,

VIII
Wherein R ^{1 to} R ⁴ , L ¹ , L ² , X and m are as defined above for compounds of formula VIII.

(0108)
The photolabile compounds of formulas I-VIII can exist in cis or trans structures. Thus, Formulas I-VIII include both cis and trans forms of photolabile compounds.
In the compounds of the present invention, the term “— (C ₁ -C ₁₈ ) alkyl” refers to a saturated straight chain or branched acyclic hydrocarbon having 1 to 18 carbon atoms. Representative saturated straight chain-(C ₁ -C ₁₈ ) alkyl includes -methyl, -ethyl, -n-propyl, -n-butyl, -n-pentyl, -n-hexyl, -n-heptyl,- n-octyl, -n-nonyl, -n-decyl, -n-undecyl, -n-dodecyl, -n-tridecyl, -n-tetradecyl, -n-pentadecyl, -n-hexadecyl, -n-heptadecyl and- n-octadecyl is included. Representative saturated branched-(C ₁ -C ₁₈ ) alkyl includes -isopropyl, -sec-butyl, -isobutyl, -tert-butyl, -isopentyl, -2-methylbutyl, -3-methylbutyl, -2,2 -Dimethylbutyl, -2,3-dimethylbutyl, -2-methylpentyl, -3-methylpentyl, -4-methylpentyl, -2-methylhexyl, -3-methylhexyl, -4-methylhexyl, -5 -Methylhexyl, -2,3-dimethylbutyl, -2,3-dimethylpentyl, -2,4-dimethylpentyl, -2,2-dimethylhexyl, -2,3-dimethylhexyl, -2,4-dimethyl Hexyl, -2,5-dimethylhexyl, -2,2-dimethylpentyl, -3,3-dimethylpentyl, -3,3-dimethylhexyl, -4,4-di Methylhexyl, -2-ethylpentyl, -3-ethylpentyl, -2-ethylhexyl, -3-ethylhexyl, -4-ethylhexyl, -2-methyl-2-ethylpentyl, -2-methyl-3-ethylpentyl, -2-methyl-4-ethylpentyl, -2-methyl-2-ethylhexyl, -2-methyl-3-ethylhexyl, -2-methyl-4-ethylhexyl, -2,2-diethylpentyl, -3,3- Diethylhexyl, -2,2-diethylhexyl, -3,3-diethylhexyl and the like are included.

(0110)
In the compounds of the present invention, the term “— (C ₃ -C ₈ ) cycloalkyl” refers to a saturated cyclic hydrocarbon having 3 to 8 carbon atoms. The (C ₃ ~C ₈₎ cycloalkyl, - - typical cyclopropyl, - cyclobutyl, - cyclopentyl, - cyclohexyl, - cyclooctyl hydrocarbon - cycloheptyl and.
In the compounds of the present invention, the term “aryl” refers to an aromatic group containing 1 to 3 aromatic rings fused or bonded.

(0112)
In the compounds of the present invention, the term “heterocyclic group” or “heterocyclic” or “heterocyclyl” or “heterocyclo” as used herein refers to at least one heteroatom in a ring containing at least one carbon atom. Having a fully saturated or partially or fully unsaturated aromatic (ie, “heteroaryl”) cyclic group (eg, 4-7 membered monocyclic, 7-11 membered bicyclic) Formula, a 10-16 membered tricyclic ring system). Each ring of the heterocyclic group having a heteroatom may have 1, 2, 3 or 4 heteroatoms selected from a nitrogen atom, an oxygen atom and / or a sulfur atom, and the nitrogen and sulfur heteroatom is It can optionally be oxidized and the nitrogen heteroatom can optionally be quaternized. The heterocyclic group may be attached to the remainder of the molecule at any heteroatom or carbon atom position in the ring or ring system. Exemplary heterocyclic groups include azepanyl, azetidinyl, aziridinyl, dioxolanyl, furanyl, furazanyl, homopiperazinyl, imidazolidinyl, imidazolinyl, isothiazolyl, isoxazolyl, morpholinyl, oxadiazolyl, oxazolidinyl, oxazolidinyl, pyrimidinyl, pyrimidinyl, pyrimidinyl, pyrimidinyl, pyrimidinyl, pyrimidinyl Lolinyl, piperazinyl, piperidinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazolyl, pyridoimidazolyl, pyridothiazolyl, pyridinyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, quinuclidinyl, tetrahydrofuranyl, thiadiazinyl, thiadiazinyl, thiadiazyl Thienothiazolyl, Thienooxazolyl, Thienoimida Lil, thiomorpholinyl, thiophenyl, but triazinyl and triazolyl, and the like. Exemplary bicyclic heterocyclic groups include indolyl, isoindolyl, benzothiazolyl, benzoxazolyl, benzooxadiazolyl, benzothienyl, quinuclidinyl, quinolinyl, tetrahydroisoquinolinyl, isoquinolinyl, benzimidazolyl, benzopyranyl, indolizinyl, Benzofuryl, benzofurazanyl, chromonyl, coumarinyl, benzopyranyl, cinnolinyl, quinoxalinyl, indazolyl, pyrrolopyridyl, furopyridinyl (furo [2,3-c] pyridinyl, furo [3,2-b] pyridinyl], furo [2,3-b] pyridinyl Etc.), dihydroisoindolyl, dihydroquinazolinyl (3,4-dihydro-4-oxo-quinazolinyl and the like), triazinylazepinyl, tetrahydroquinolinyl and the like. Exemplary tricyclic heterocyclic groups include carbazolyl, benzidolyl, phenanthrolinyl, acridinyl, phenanthridinyl, xanthenyl and the like.

(0113)
As used herein, the term “cycloaliphatic amine” or “cycloaliphatic amine group” refers to a non-aromatic secondary amine or cyclic tertiary amine group. Examples of cycloaliphatic amines include, but are not limited to, aziridine and piperidine.
As used herein, the term “active derivative” or “active derivative of a labeling molecule” refers to a chemical derivative of a labeled molecule, and the labeling function of the labeled molecule is (For example, its fluorescence characteristics). In some embodiments, such active derivatives provide better properties for binding to Ru via a phosphine group, aliphatic amine group, imine group, pyridyl group or nitrile group. A specific example of an active derivative of rhodamine (labeled molecule) is B-methylaminopropionitrileamide (RhodB-MAPN). One skilled in the art will create the derivative by modifying the labeled molecule, and then test the activity of the resulting derivative in the same manner as it tests the activity of the labeled molecule to determine whether the derivative is active. Can be determined.

(0115)
An amino acid group (α-amino acid or the like) is an organic molecule having an amino group (—NH ₂ ) and a carboxylic acid group. Amino acids include 20 common α-amino acids (Gly, Ala, Val, Leu, Ile, Ser, Thr, Asp, Asn, Lys, Glu, Gln, Arg, His, Phe, Cys, Trp, Tyr, Met. , Pro) or one of the other natural amino acids such as norleucine, ethylglycine, ornithine, γ-aminobutyric acid and phenylglycine.
Examples of 6-membered monocyclic aromatic rings in which one of the ring members is a nitrogen atom include pyridyl, pyrimidinyl, pyridazinyl and pyrazinyl rings.
Examples of the 5-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom include pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, oxadiazolyl, thiazolyl and thiadiazolyl rings.

(0118)
Examples of 8- to 10-membered bicyclic aromatic rings where one of the rings is aromatic and has a nitrogen atom member include indolizinyl, isoindolyl, 3H-indolyl, indolyl, indazolyl, purinyl, quinolyl, isoquinolyl, phthalazinyl, naphthyridinyl, Quinoxalinyl, 1,3,7-trimethyl-2,6-dioxopurinyl, quinazolinyl, cinnolinyl, pteridinyl, 6-amino-1H-purinyl and 2-aminohypoxanthinyl bicyclic aromatic rings are included.
In general, examples of organic molecules useful in the present photolabile compounds include a variety of substances, including, but not limited to, pharmaceuticals, small molecules, drugs, neurochemicals, peptides, proteins, and chemotherapy It is an agent.

(0120)
Exemplary organic molecules further include luciferin, enzyme inhibitors, fatty acids (eg, arachidonic acid), protein kinase C activators (eg, dioctanoyl glycerol), tubulin association promoters (eg, paclitaxel), antibiotics Substance (eg, penicillin, A23187), neurotransmitter (eg, L-glutamic acid, aspartic acid, carbamylcholine, dopamine, epinephrine, GABA, glutamic acid, glycine, haloperidol, isoproterenol, kainic acid, NMDA, NMDA receptor Antagonists MK-801, norepinephrine, phenylephrine, propranolol), 4-aminopyridine (4AP), serotonin (5-hydroxytryptamine, 5HT), (RS)-(tetrazol-5-yl) glycine (Tz) ly), tetrazolyl-α-amino-3-hydroxy-5-methyl-4-isoxazoleproprionic acid ((tetrazol-5-yl) AMPA), nicotine, nicotinic acid, isoxazole, fluorescent dyes (eg, fluorescein, HPTS, rhodamine, succinimidyl ester of carboxy-Q-rhodamine and sulfosuccinimidyl ester or rhodamine green), nucleotides (eg ATP, ADP, cAMP, GDP, GTP, cGMP, GTP-γ-S, GDP-β , CAMP or cGMP 8-substituted derivatives such as 8-bromo-cAMP, 8-bromo-cGMP, 8-chloro-cAMP, 8-chloro-cGMP, 8-parachlorophenylthio (cCPT) cAMP or cGMP, phosphate (eg , Phosphate, Scan phosphate esters), phenyl phosphate (PPh _3), Py, nucleoside, nucleoside derivative, nucleotide derivative (e.g., CADP- ribose, 8-amino -cADP ribose, 8-bromo -cADP- ribose), cyclitols (e.g., inositol) Cyclitol phosphate (e.g., myo-inositol phosphate, myo-inositol-1,4,5-triphosphate, myo-inositol-1,3,4,5-tetrakisphosphate, myo-inositol-3,4,5,6-tetrakisphosphate ), NO (eg, derived from degradable compounds of HON = N (O) (Net ₂ )), chelants (eg, EDTA, EGTA) and ionophores (eg, nigericin). Organic molecules are described, for example, by Furuta et al., Biochem. Biophys. Res. Commun. 228: 193-198 (1996).

(0121)
Other useful examples of organic molecules include adenosine 5′-diphosphate ADP, adenosine 5′-triphosphate ATP, adenosine 5′-monophosphate AMP, aminobutyric acid, L-glutamic acid, cyclic adenosine 5′-diphosphate ribose. , Adenosine 3 ′, 5′-cyclic monophosphate, fluorescein, methyl-D-aspartate, tyramine, tryptophan, 4-aminopyridine (4AP), epinephrine, norepinephrine, dopamine, serotonin (5 hydroxytryptamine, 5HT), (RS )-(Tetrazol-5-yl) glycine (TzGly) (a potent N-methyl-D-aspartate receptor (NMDA) agonist), tetrazolyl-α-amino-3-hydroxy-5-methyl-4-isoxa Zolproprionic acid (Tetrazol-5-yl) AMPA), include caffeine and nicotine.

(0122)
In the present invention, it has been demonstrated that organic molecular ligands glutamate, γ-aminobutyric acid (GABA), alaninate, glycinate and the like are light-released from a photolabile compound after exposure to visible light. Such photolabile compounds are stable in solution in addition to water. Organic molecules having —NH ₂ groups or —COOH groups can be released into solvents other than water, such as alcohols (eg, methanol, ethanol), acetone, and the like.
The organic molecules of the present invention, the phosphorus atom contains phosphorus derivatives having -PR ₂ groups form a bond with Ru, R is independently -H, -C ₁ -C ₁₈ alkyl or aryl. Exemplary phosphorus derivatives include dimethylphosphinyl, diethylphosphinyl and diphenylphosphinyl.
The organic molecule of the present invention includes a sulfur derivative having a —SR group in which a sulfur atom forms a bond with Ru, and R is independently —H, —C ₁ -C ₁₈ alkyl or aryl. Exemplary sulfur derivatives include isopropyl β-D-1-thiogalactopyranoside (IPTG), thioether, thiolate, methylthio, ethylthio and phenylthio.

(0125)
In general, examples of labeling molecules useful in the present photolabile compounds include a variety of materials, such as, but not limited to, fluorescent groups, bioluminescent groups, chemiluminescent groups, colorimetric groups, or radioactive groups. Is a molecule.
Fluorescent molecules include body pie, dansyl, fluorescein, rhodamine, Texas red, cyanine dye, pyrene, coumarin, cascade blue (registered trademark), pacific blue, marina blue, oregon green, 4 ', 6-diamidino-2-phenylindole ( DAPI), indopyra dyes, lucifer yellow, propidium iodide, porphyrins, arginine and variations and derivatives thereof. See, for example, Richard P. Haughland's Handbook of Fluorescent Probes and Research Chemicals (6th edition, Molecular Probes, 1996) for more information on fluorescent labeling moieties and fluorescent techniques. This document is incorporated herein by reference in its entirety.
The photolabile compounds of Formulas I-III and V-VIII where L ₂ is other than L ₁ are about molar equivalents of Ru (bdt) ₂ Cl ₂ (bdt is as defined in Formulas I-III. R _{1 to} R ₈ groups, or bipyridine or phenanthroline substituted with R _{1 to} R ₄ groups as defined in Formulas V to VIII) with about a molar equivalent of organic or labeled molecule and refluxed water , Ethanol, methanol, isopropyl alcohol, ethylene glycol, acetone, methylene chloride or a mixture thereof under nitrogen. After about 4 to about 8 hours, the resulting solution is cooled and at least about an equivalent amount of L ₂ is added thereto. The resulting mixture is heated at reflux for about 4 to about 20 hours. After cooling to room temperature, the resulting solution is diluted with water and an excess of NH ₄ PF ₆ is added thereto. The resulting precipitate is filtered, purified by silica gel chromatography, dried and dissolved in acetone. (N-Bu) ₄ NH ₄ ⁺ X (X is defined by Formulas I-III and V-VIII) is added to the acetone solution and the resulting Formulas I-III where L ₂ is other than L _1. And the photolabile compound shown by V-VIII is filtered.

(0128)
The photolabile compounds of Formulas I-III and V-VIII where L ₂ is L ₁ are about molar equivalents of Ru (bdt) ₂ Cl ₂ (bdt is R as defined in Formulas I-IIII). _{1 to} R ₈ groups, or bipyridine or phenanthroline substituted with R _{1 to} R ₄ groups as defined in formulas V to VIII) with an excess of organic or labeled molecules and refluxed water, ethanol , Methanol, isopropyl alcohol, ethylene glycol, acetone, methylene chloride or a mixture thereof under nitrogen. After about 4 to about 8 hours, the resulting solution is cooled to room temperature. The resulting mixture is diluted with water and an excess of NH ₄ PF ₆ is added thereto. The resulting precipitate is filtered, purified by silica gel chromatography, dried and dissolved in acetone. (N-Bu) ₄ NH ₄ ⁺ X (X is defined by Formulas I-III and V-VIII) is added to the acetone solution and the resulting Formulas I-III and L ₂ are L ₁ and The photolabile compound of V to VIII is filtered.

(0129)
The photolabile compound of formula IV, where M ² is Ru, is about 1 molar equivalent of (M ¹ ) ₃ [M ² (CN) ₅ NH ₃ ] 2H ₂ O (M ¹ is defined by formula IV). , And dissolved in about 15 mL of argon deoxygenated ethanol / water (1: 1) containing about 10 molar equivalents of organic molecules. The resulting mixture is maintained under argon at about room temperature for about 1 hour and concentrated under vacuum to a volume of about 1 mL at about room temperature. When a cold saturated solution of M ¹ I in ethanol is added to the resulting concentrate, precipitation of the photolabile compound of formula IV with M ² Ru occurs, which is washed with ethanol and diethyl ether.
In the context of the present invention, light emission is generally rapid following exposure to visible light of the appropriate wavelength, for example from about a few nanoseconds to about 500 milliseconds (Salierno et al., J Inorg Biochem. 2010 104 (4): (See 418-22) Can happen later. Suitable wavelengths of light for effectively emitting the organic molecule or label molecule from the photolabile compound range from about 300 to about 500 nm, from about 300 to about 360 nm, or from about 450 to about 500 nm (eg, 473 nm). Can be extended to up to 700 nm. Suitable light sources include those capable of emitting light of the appropriate wavelength, such as commercially available tungsten lamps (Cole-Parmer), arc lamps, xenon continuous lamps, lasers (eg blue or green lasers) or Although it is a photooptic light source, it is not limited to these. Such light sources are commercially available (CrystaLaser, Reno, Nevada, Lasever, Koto, Ningbo, China). Other forms of light (sunlight, infrared light, pulsed infrared light, ultraviolet light, etc.) can be used in the present invention as needed or desirable.

(0131)
Devices and systems suitable for exposing photolabile compounds to light (especially visible or infrared light) further include imaging probes, imaging catheters and fiber optic probes, particularly US Pat. Utzinger et al. Biomed. Optics, 8 (1): 121-147 (2003) and described in Fujimoto et al. Photonic Materials, Devices and Systems-Laser Medicine and Medical Imaging Group, Rle Progress Report 27-27. Includes those with index or graded index (GRIN) lenses (Sp3 plus, UK). Light suitable for light emission of organic or labeled molecules upon exposure to a photolabile compound includes a wavelength of about 300 to about 500 nm, about 300 to about 360 nm, or about 450 to about 500 nm. Suitable light includes visible light or infrared light.

(0132)
Furthermore, in the present invention, the organic molecule or the labeled molecule can be released from the photolabile compound by one-photon or two-photon photolysis. Optical memories that utilize two-photon excitation are described, for example, in Stickler and Webb, Optics Letters, 16: 1780-1782 (1991). A feature of two-photon excitation is the elimination of out-of-focus background (see, for example, W. Denk et al., 1990, Science, 248: 73-76). Thus, by two-photon uncaging, organic or labeled molecules can be released only at the focal plane (eg, W. Denk et al., 1994, Proc. Natl. Acad. Sci. USA, 91: 6629-6633). checking).
In certain embodiments, the present invention includes compounds of Formula I wherein the organic molecule is 4-AP. In another embodiment, the present invention includes compounds of Formula I wherein the organic molecule is TzGly. In another embodiment, the present invention includes compounds of Formula I wherein the organic molecule is (tetrazol-5-yl) AMPA. In another embodiment, the invention includes a compound of formula I, wherein the organic molecule is nicotine or caffeine. In another embodiment, the invention includes a compound of formula I, wherein the organic molecule is serotonin, epinephrine, norepinephrine, or dopamine. In another embodiment, the invention provides that the organic molecule is adenosine 5′-diphosphate ADP, adenosine 5′-triphosphate ATP, adenosine 5′-monophosphate AMP, cyclic adenosine 5′-diphosphate ribose or adenosine 3 ′, Including compounds of formula I that are 5'-cyclic monophosphates. In another embodiment, the invention includes a compound of formula I, wherein the organic molecule is aminobutyric acid or L-glutamic acid or methyl-D-aspartic acid.

(0134)
In certain embodiments, the present invention includes compounds of Formula II or Formula III wherein the organic molecule is 4-AP. In another embodiment, the invention includes a compound of formula II, wherein the organic molecule is TzGly. In another embodiment, the invention includes a compound of formula II, wherein the organic molecule is (tetrazol-5-yl) AMPA. In another embodiment, the invention includes a compound of formula II, wherein the organic molecule is nicotine or caffeine. In another embodiment, the invention includes a compound of formula II, wherein the organic molecule is serotonin, epinephrine, norepinephrine, or dopamine. In another embodiment, the invention provides that the organic molecule is adenosine 5′-diphosphate ADP, adenosine 5′-triphosphate ATP, adenosine 5′-monophosphate AMP, cyclic adenosine 5′-diphosphate ribose or adenosine 3 ′, Including compounds of formula II that are 5′-cyclic monophosphates. In another embodiment, the invention includes a compound of formula II, wherein the organic molecule is aminobutyric acid, L-glutamic acid, or methyl-D-aspartic acid.

(0135)
In certain embodiments, the present invention includes compounds of Formula IV wherein the organic molecule is 4-AP. In another embodiment, the invention includes a compound of formula IV, wherein the organic molecule is TzGly. In another embodiment, the invention includes a compound of formula IV, wherein the organic molecule is (tetrazol-5-yl) AMPA. In another embodiment, the invention includes a compound of formula IV, wherein the organic molecule is nicotine or caffeine. In another embodiment, the invention includes a compound of formula IV, wherein the organic molecule is serotonin, epinephrine, norepinephrine, or dopamine. In another embodiment, the invention provides that the organic molecule is adenosine 5′-diphosphate ADP, adenosine 5′-triphosphate ATP, adenosine 5′-monophosphate AMP, cyclic adenosine 5′-diphosphate ribose or adenosine 3 ′, Compounds of formula IV that are 5′-cyclic monophosphates are included. In another embodiment, the invention includes a compound of formula IV, wherein the organic molecule is aminobutyric acid, L-glutamic acid, or methyl-D-aspartic acid.

(0136)
In certain embodiments, the present invention includes compounds of Formula V wherein the organic molecule is 4-AP. In another embodiment, the invention includes a compound of formula V, wherein the organic molecule is TzGly. In another embodiment, the invention includes a compound of formula V, wherein the organic molecule is (tetrazol-5-yl) AMPA. In another embodiment, the invention includes a compound of formula V, wherein the organic molecule is nicotine or caffeine. In another embodiment, the invention includes a compound of formula V, wherein the organic molecule is serotonin, epinephrine, norepinephrine, or dopamine. In another embodiment, the invention provides that the organic molecule is adenosine 5′-diphosphate ADP, adenosine 5′-triphosphate ATP, adenosine 5′-monophosphate AMP, cyclic adenosine 5′-diphosphate ribose or adenosine 3 ′, Including compounds of formula V that are 5'-cyclic monophosphates. In another embodiment, the invention includes a compound of formula V, wherein the organic molecule is aminobutyric acid, L-glutamic acid, or methyl-D-aspartic acid.

(0137)
In certain embodiments, the present invention includes compounds of Formula VI wherein the organic molecule is 4-AP. In another embodiment, the present invention includes compounds of Formula VI wherein the organic molecule is TzGly. In another embodiment, the present invention includes compounds of Formula VI wherein the organic molecule is (tetrazol-5-yl) AMPA. In another embodiment, the invention includes a compound of formula VI, wherein the organic molecule is nicotine or caffeine. In another embodiment, the invention includes a compound of formula VI, wherein the organic molecule is serotonin, epinephrine, norepinephrine, or dopamine. In another embodiment, the invention provides that the organic molecule is adenosine 5′-diphosphate ADP, adenosine 5′-triphosphate ATP, adenosine 5′-monophosphate AMP, cyclic adenosine 5′-diphosphate ribose or adenosine 3 ′, Compounds of formula VI that are 5′-cyclic monophosphates are included. In another embodiment, the invention includes a compound of formula VI, wherein the organic molecule is aminobutyric acid, L-glutamic acid, or methyl-D-aspartic acid.

(0138)
In certain embodiments, the present invention includes compounds of Formula VII where the organic molecule is 4-AP. In another embodiment, this invention includes a compound of formula VII, wherein the organic molecule is TzGly. In another embodiment, the present invention includes compounds of Formula VII wherein the organic molecule is (tetrazol-5-yl) AMPA. In another embodiment, the invention includes a compound of formula VII, wherein the organic molecule is nicotine or caffeine. In another embodiment, the invention includes a compound of formula VII, wherein the organic molecule is serotonin, epinephrine, norepinephrine, or dopamine. In another embodiment, the invention provides that the organic molecule is adenosine 5′-diphosphate ADP, adenosine 5′-triphosphate ATP, adenosine 5′-monophosphate AMP, cyclic adenosine 5′-diphosphate ribose or adenosine 3 ′, Including compounds of formula VII that are 5'-cyclic monophosphates. In another embodiment, the invention includes a compound of formula VII, wherein the organic molecule is aminobutyric acid, L-glutamic acid, or methyl-D-aspartic acid.

(0139)
In certain embodiments, the present invention includes compounds of Formulas I-III and V-VIII, wherein the labeling molecule is rhodamine. In another embodiment, the present invention includes compounds of Formulas I-III and V-VIII, wherein the labeling molecule is fluorescein. In another embodiment, the present invention includes compounds of Formulas I-III and V-VIII, wherein the labeling molecule is iodoeosin. In another embodiment, the invention includes compounds of Formulas I-III and V-VIII, wherein the labeling molecule is a fluorescent molecule having a -CN group in which the nitrogen atom forms a bond with Ru. In another embodiment, the invention includes compounds of formulas I-III and V-VIII, wherein the labeling molecule is a fluorescent molecule having a pyridyl group in which the nitrogen atom forms a bond with Ru. In another embodiment, the invention includes compounds of formulas I-III and V-VIII, wherein the labeling molecule is a fluorescent molecule having an amino group in which the nitrogen atom forms a bond with Ru. In another embodiment, the invention includes compounds of formulas I-III and V-VIII, wherein the labeling molecule is a fluorescent molecule having a phosphine group in which the nitrogen atom forms a bond with Ru.

(0140)
In another embodiment, the invention includes compounds of Formulas I-III and V-VIII, wherein the labeling molecule is rhodamine having a -CN where the nitrogen atom forms a bond with Ru. In another embodiment, the invention includes compounds of Formulas I-III and V-VIII, wherein the labeling molecule is a rhodamine having a pyridyl group in which the nitrogen atom forms a bond with Ru. In another embodiment, the invention includes compounds of Formulas I-III and V-VIII, wherein the labeling molecule is rhodamine having an amino group in which the nitrogen atom forms a bond with Ru. In another embodiment, the invention includes compounds of Formulas I-III and V-VIII, wherein the labeling molecule is rhodamine having a phosphine group in which the nitrogen atom forms a bond with Ru.

（ＲｈｏｄＢ−ＭＡＭｅＰｙ）又はその塩化物塩（ＲｈｏｄＢ−ＭＡＭｅＰｙ−Ｃｌ）である。 (0141)
In some embodiments of compounds of Formulas I-III and V-VIII, L ² is

(RhodB-MAMePy) or a chloride salt thereof (RhodB-MAMePy-Cl).

（Ｒｈｏｄ６Ｇ−ＭＡＭｅＰｙ）又はその塩化物塩（Ｒｈｏｄ６Ｇ−ＭＡＭｅＰｙ−Ｃｌ）である。 (0142)
In some embodiments of compounds of Formulas I-III and V-VIII, L ² is

(Rhod6G-MAMePy) or a chloride salt thereof (Rhod6G-MAMePy-Cl).

(0143)
In some embodiments, the labeling molecule or an active derivative thereof, other coordinating group -NH _2, is coupled to the Ru to Ru via the pyridyl or phosphine group.
In some embodiments of the photolabile compound, L ² comprises a labeled molecule or an active derivative thereof, serves as an optical antenna, and transfers energy to the site of the compound that results in light emission of the moiety L ^1. Can do. In this way, light collection is decoupled from the chemical release process, allowing orthogonal chemical strategies for both processes. Thus, in some embodiments, the photolabile compound has two functions, the first being an optical antenna and the second being a light emitting entity.
In one aspect, photolabile compounds can be used in a combined chemical strategy. For example, a series of systematic derivatives can be generated in the light emitting part of the molecule and used to perform large-scale screening. Alternatively or in addition, the light emitting part of the molecule can be fixed while derivatizing the optical antenna part.
In another embodiment, the magnetic and / or radiopaque properties of the photolabile compound are used as a means of its identification or manipulation in living cells. In one such embodiment, the photolabile compound is used as a contrast agent for MRI (magnetic resonance imaging).

(0147)
When L ² contains a labeled molecule such as a fluorophore or a derivative thereof, L ² serves as an antenna and can extend the light irradiation spectrum to lower energy with very high absorption and efficiency. In certain embodiments, the fluorescent dye is coupled through derivatization with a non-releasing ligand (ie, phosphine) (: P (R) ₂ -fluorophore). In these embodiments, the light is collected by a fluorescent moiety that functions as an antenna, moved to the Ru center, and populates the MLCT or dd excited state. Once the excited state is reached, the other ligand (ie L ^{1, which} can be a biomolecule or drug) is released.
In this case, the fact that the active wavelength is determined not by the Ru center but by the high absorption of the fluorescent dye (approximately 100000 M ⁻¹ cm ⁻¹ ) is due to the different wavelengths (400-600 nm) of different complexes with different photouncaging components. ) Can be adjusted, and orthogonality can be obtained.

(0149)
Coordination of the appropriate organic dye to ruthenium adjacent to the coordinating fluorescent dye (ie, the labeling molecule) allows selective quenching of fluorescence due to its proximity. In this case, absorption of light through the Ru-bpyMLCT band (no antenna effect; fluorescence is quenched by an adjacent non-fluorescent dye) causes the fluorescent dye to emit light, dramatically increasing fluorescence, and the fluorophore Can be “uncaged”. Alternatively, non-fluorescent quenchers can be light emitted, with somewhat similar results (however, depending on conditions, the presence of a metal center can quench the fluorophore).
Furthermore, in the present invention, the compounds of formulas V to VIII on which R ¹ and R ² and / or R ³ and R ⁴ combine to form a cyclic carbon substituted with one or more oxo groups It can bind and is useful for delivery of compounds to surfaces. For example, such compounds can be bound to nanoparticles or nanopowder and used to deliver biologically active organic molecules or labeled molecules. Nanoscale delivery systems have numerous applications in controlled targeted delivery of drugs. Drugs can be selectively delivered to target cells by nanoparticles with specific surface functionalization. Nanoparticles can penetrate cell membranes and overcome biological physiological barriers, and nanoparticles can also improve drug solubility and bioavailability.

(0151)
In another embodiment, the present invention provides a method for improving the solubility of an organic molecule, the method comprising producing a photolabile compound by complexing an organic molecule with a photolabile caging group. Organic molecules are released from the compound upon exposure to light of the compound under sufficient conditions. In certain embodiments, the organic molecule is
(I) a 5-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(Ii) a 6-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(Iii) an 8 to 10 membered bicyclic ring having a nitrogen atom member in which one of the bicyclic rings is aromatic and forms a bond with Ru;
(Iv) a —NH ₂ group in which the nitrogen atom forms a bond with Ru;
(V) a —COOH group in which one of the oxygen atoms forms a bond with Ru;
(Vi) the phosphorus atom form a bond and Ru, R is independently -H, -C ₁ ~C ₁₈ -PR ₂ groups are alkyl or aryl, or (vii) a sulfur atom forms a bond with Ru , R has an —SR group which is independently —H, —C ₁ -C ₁₈ alkyl or aryl.

(0152)
In certain embodiments, the invention includes a composition comprising an effective amount of a photolabile compound and a physiologically acceptable carrier, vehicle, diluent or excipient. Suitable carriers, vehicles, diluents or excipients are known to those skilled in the art and include, but are not limited to, physiologically sterile saline and the like as described herein.

(0153)
In another embodiment, the present invention provides a container containing a photolabile compound. The container may further store a biological sample, for example, hair, organ specimen, tissue or cell (eg nerve tissue, nerve cell), tumor, cancer or neoplastic tissue or cell, patient or subject of interest. A tissue or cell removed from a subject. For example, a tissue specimen sectioned with a microtome is an example of a suitable biological sample.
Any type of container that is capable of transmitting light of the wavelength used to release the organic molecule or labeling molecule constituting the photolabile compound and is inert to the solvent in which the photolabile compound is suspended. Suitable for use. For example, the container can be formed of glass, plastic, acrylic, quartz, noble metal, or the like. In addition, if the container is constructed of metal (eg, aluminum, titanium, stainless steel) or covered with metal, exposure to light is from a “window” or other light that opens from the top of the container or into the container. Is carried out through an opening that can be passed through. In the case of a solid-like material, for example, an acrylic resin or an acrylamide / bisacrylamide gel can be used as a medium for containing a photolabile compound. For example, an acrylic resin coating prepared using an acrylic CHCl ₃ solution and a Ru (bpy) complex changed its spectrum after light irradiation to enable light emission in the solid state. For such solid state embodiments of the invention, the temperature can be maintained at 4K.

(0155)
Suitable solvents that can be exposed to light with a photolabile compound include aqueous solvents, water, acetonitrile, alcohols (eg, methanol, ethanol), acetone, chlorinated solvents (CH ₂ Cl ₂ , CHCl ₃ etc.) or Dimethyl sulfoxide is included.
Suitable temperatures for exposing the photolabile compound to light generally range from about 0 ° C to about 100-150 ° C.
In another embodiment, the invention includes a method of releasing an organic molecule or a labeled molecule from a photolabile compound. The method includes exposing the photolabile compound to light under conditions sufficient to cause the organic molecule or label molecule to be released from the compound. In this method, the light comprises a wavelength of about 300 to about 500 nm, about 300 to about 360 nm, or about 450 to about 500 nm. Further, the exposure can occur at a temperature from about 0 to about 150 ° C. In certain embodiments, a method of the invention comprises a photolabile compound (eg, a compound of Formulas I-VIII), light at a wavelength of about 300 to about 500 nm, L ² that is L ² and about 0 to about 150 ° C. Includes temperature. In another embodiment, the method comprises a photolabile compound, light at a wavelength of about 300 to about 360 nm, L ¹ that is L ² and a temperature that is about 0 to about 150 ° C. In another embodiment, the method comprises photolabile compound, the temperature of about 450 to about 500nm wavelengths of light, L ¹ and about 0 to about 0.99 ° C. is L ^2. In another embodiment, the method comprises photolabile compounds, visible or infrared light, the L ¹ and about 0 temperature of about 0.99 ° C. is L ^2.

(0158)
In certain embodiments, the invention includes a method of assaying an organic molecule, wherein the method exposes the photolabile compound and biological sample to light under conditions sufficient to release the organic molecule from the photolabile compound. And (b) confirming the action of this organic molecule on the biological sample. The sample can be a biological sample, for example a sample excised, removed or otherwise collected from the patient's body. The patient biological sample can be, for example, a hair sample, an organ or tissue sample (eg, obtained by biopsy, autopsy) or a cell sample. In addition, the biological sample can be a body fluid sample. Body fluid samples include, but are not limited to, blood, serum, plasma, lymph, saliva, sputum, tears, semen or urine. Biological samples further include brain tissue, brain cells, muscle tissue, muscle cells, muscle fibers, fibroblasts, tissue sections or fine tissue specimens obtained from any organ of the body, sarcoplasmic reticulum, skin tissue, membrane specimens Or a fragment etc. may be included, but it is not limited to these.

(0159)
The light for exposing the compound according to the method of the invention can be sunlight, photooptic light or laser light. Advantageously, in the method of the invention, the light for exposure of the compound is other than ultraviolet light. Thus, for example, the light can be visible light or infrared light, including one-photon light and two-photon light. Light can be emitted from a variety of light sources, including but not limited to laser light sources, tungsten light sources, photooptic light sources, and the like. Another advantage of visible light for exposure or light irradiation of the compounds of the present invention is the convenience and observation of the sample into which the compound is introduced, for example, and the ligand that is light-released from the compound after exposure to visible light. It relates to the fact that a visible light microscope can be used for visualization or monitoring with a microscope. Since many microscopes do not transmit ultraviolet light, it is advantageous to be able to use a non-quartz microscope in the present invention. Yet another advantage of using visible light is that visible light is not harmful to living cells and tissues and is beneficial for use in a patient's body. In addition, when used on a patient, light can be specifically directed to the area where the photolabile compound is introduced or administered by the use of laser technology, fibers, probes, tubes, and the like. Such probes, fibers or tubes can be inserted, for example, directly into a body cavity or opening or transcutaneously to expose the photolabile compound to light.

(0160)
In another embodiment thereof, the present invention includes a method of making an organic molecule bioavailable to a patient. Organic molecules can be made bioavailable systemically in a local body region or area of the patient or throughout the body. Local bioavailability of the photolabile compound is achieved, for example, via delivery devices and methods that allow direct administration of the photolabile compound (eg, inserted into a body cavity or opening or transcutaneously). . The method of this embodiment involves administering a photolabile compound to a patient and exposing the organic molecule to the patient and / or patient by exposing the compound to light under conditions sufficient to release the organic molecule from the compound. With bioavailability in the body part or area of the body. Exposure to light can include the use of probes, fibers, tubes, etc., which can direct light to specific areas on or within the body. Alternatively, the photolabile compound can be administered to a patient that remains in the dark. To light-emit organic molecules, the patient can be moved to a bright place where light exposure and light emission occurs. In an embodiment of the method, the organic molecule is
(A) a 5-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(B) a 6-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(C) an 8 to 10 membered bicyclic ring having a nitrogen atom member in which one of the bicyclic rings is aromatic and forms a bond with Ru;
(D) a —NH ₂ group in which the nitrogen atom forms a bond with Ru;
(E) a —COOH group in which one of the oxygen atoms forms a bond with Ru;
(F) phosphorus atoms form a bond and Ru, R is independently -H, -C ₁ ~C ₁₈ -PR ₂ groups are alkyl or aryl, or (g) a sulfur atom forms a bond with Ru , R has an —SR group which is independently —H, —C ₁ -C ₁₈ alkyl or aryl.

(0161)
In another embodiment, the organic molecule is
(A) a tetrazolyl group in which one of the nitrogen atoms forms a bond with Ru;
(B) nicotine or caffeine in which the pyridyl nitrogen atom forms a bond with Ru;
(C) an 8 to 10 membered bicyclic ring having a nitrogen atom member in which one of the bicyclic rings is aromatic and forms a bond with Ru;
(D) a —NH ₂ group in which the nitrogen atom forms a bond with Ru;
(E) one of the oxygen atoms forms a bond with Ru —COOH group (f) a phosphorus atom forms a bond with Ru, and R is independently —H, —C ₁ -C ₁₈ alkyl or aryl— PR ₂ group, or (g) a sulfur atom forms a bond with Ru, with R is independently -H, an -SR group is -C ₁ -C ₁₈ alkyl or aryl.

(0162)
In related embodiments, the photolabile compound is useful for the release of organic molecules (drugs, pharmaceuticals, biologically active small molecules, etc.) as described above. Release of the organic molecule from the photolabile compound renders the organic molecule bioavailable to a subject or patient suffering from a disease, disorder, condition or condition. Photolabile compounds and organic molecules are useful in animal and human drugs. Diseases, disorders, conditions or conditions that treat, recover, reduce, eliminate, ameliorate or prevent organic molecules by making them bioavailable are further described below, but include, as non-limiting examples, peripheral and central nervous system disorders Neuropathy and related disorders, neurodegenerative disorders and related disorders, epilepsy, seizures, migraines, headache, stroke, anxiety, depression, limited brain function, addiction disorders, neurosis, psychosis, pruritus , Parkinson's disease, Huntington's disease, cognitive impairment, lack of memory, Alzheimer's disease, dementia, dyskinesia, muscle spasticity, retinopathy, vomiting, cancer, neoplasm, tumor, vascular disease and cardiovascular disease.

(0163)
As yet another non-limiting example, the organic molecule is a neurochemical that blocks potassium channels, for example, for use in the treatment of neurodegeneration or neurological diseases or disorders. In certain embodiments, the organic molecule is 4-AP, which is a calcium channel inhibitor. In another embodiment, the organic molecule is TzGly, which is a more potent NMDA receptor agonist than NMDA. In one embodiment, in order to make the organic molecule of the present invention bioavailable to patients in need thereof, exposure of the photolabile compound to light is performed at the site of the disease, disorder, condition or condition (tumor, new It is possible to release the organic molecules locally and more accurately at the necessary positions by causing them to occur in living organisms or cancer lesions or growth sites. In another embodiment, the photolabile compound can be exposed to light at the site of a blood disorder in order to make the organic molecule of the invention bioavailable to a patient in need thereof.

(0164)
In other related embodiments, the present invention provides an effective amount of a photolabile compound or a physiologically acceptable composition comprising a photolabile compound to a patient to make the organic molecule bioavailable to the patient. Provides methods of treatment, therapy and prevention. A photolabile compound can be substantially purified (eg, substantially free of substances that limit its effect or cause undesirable side effects). Advantageously, in a method of administering a photolabile compound to a patient, light (e.g., infrared light, laser light, visible light) for photoemissioning an organic molecule to make it bioavailable to the patient, Through the use of optical devices, probes and fibers (such as those known in the art and those described above), they can be directed to the intended internal site or region. One of ordinary skill in the art can utilize, manipulate, and direct the instrument to expose the photolabile compound to light after administering the photolabile compound to the patient.

(0165)
In the methods of the invention comprising treatment of a patient and / or disease, disorder, condition or condition, therapy or prophylaxis, the patient is preferably an animal and is a mammal such as a human, non-human primate, cow, pig, horse. , Goats, sheep, rabbits, chickens, cats, dogs, guinea pigs, rats, mice and the like. The methods of the present invention particularly include human therapy.
Various delivery systems are known and can be used to administer photolabile compounds, such as the use of sterile solutions, liposomes, microparticles, encapsulation in microcapsules or receptor-dependent endocytosis ( See, for example, Wu and Wu, 1987, J. Biol. Chem., 262: 4429-4432). Intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, topical, transdermal, parenteral, intrathecal, intravaginal, rectal, colorectal, oral , Intracranial, retro-orbital, intrasternal route, or combinations thereof.

(0167)
The photolabile compound or composition can be administered by any convenient route or mode, for example, continuous infusion, non-continuous infusion, bolus administration, absorption through epithelium or mucocutaneous skin (eg, oral mucosa, epidermis, rectal mucosa, Intestinal mucosa. It can also be administered with another biologically active and / or therapeutic agent. Administration can be systemic or local. In addition, it is also desirable to introduce the photolabile compound or composition into the central nervous system by any suitable route, including intraventricular and intrathecal administration. Intraventricular administration can be facilitated by an intraventricular catheter attached to a reservoir, such as, for example, an Ommaya reservoir. It can also be administered from the lung, for example, by use of an inhaler or nebulizer and formulation with an aerosolizing agent.
In certain embodiments, it may be desirable to administer the photolabile compound or composition locally to the area in need of treatment. This includes, for example, topical injection during surgery, topical application (eg, with a post-surgical wound dressing), injection, catheter, suppository or implant (which includes a membrane or fiber such as a sialastic membrane, Porous, non-porous or gelatin-like materials).

(0169)
In another embodiment involving topical administration, a transdermal patch can be used. In this embodiment, the photolabile compound remains unexposed to light until the patient or healthcare provider exposes all or part of the patch containing the photolabile compound to light. Thus, by opening the patch, for example, by moving the patient from a dark room to a bright room or from a dark area to a bright area, or by exposing the entire patch or part thereof directly to a suitable light source Alternatively, the patient can expose or “activate” the bioactive molecule from the compound after exposure to light by exposing the entire patch or a portion thereof to sunlight. Various types of transdermal patches are known and used by those skilled in the art. Alternatively, for topical administration, the photolabile compound can be prepared into a photosensitive composition that is stored in a dark shading container and applied topically in the dark to the area of interest (eg, Applied or rubbed into the patient's skin). Following topical application in the dark, the organic molecules of the photolabile compound are released by exposing the area of interest to light or a suitable light source, or moving the patient to a bright place.

(0170)
In another embodiment, the photolabile compound or composition can be delivered in vesicles, particularly liposomes (eg, Langer, 1990, Science, 249: 1527-1533, Treat et al.,). Liposomes in the Therapeutic of Infectious Disease and Cancer, Lopez-Berstein and Fiddler (eds.), Liss, New York, pp. 353-365 (1989), Lopez-Ber, 27. See the same book). In yet another embodiment, the photolabile compound or composition can be delivered in a controlled release system. For example, a pump can be used (Langer, Sefton, 1987, CRC Crit. Ref. Biomed. Eng. 14: 201, Buchwald et al., 1980, Surgary, 88: 507, Saudek et al., 1989, NEJM. , Med. 321: 574 (1989)). Alternatively, polymeric materials can be used (e.g., Medical Applications of Controlled Release, Langer and Wise (eds.), CRC Press, Boca Raton, Fla. (1974), Controlled DrugDrivable Drug Bioavailability. Smolen and Ball (eds.), Wiley, New York (1984), Ranger and Peppas, 1983, J. Macromol. Sci. Rev. Macromol. Chem., 23:61, Levy et al., 1985, 228: 190, During et al., 1989. Neurol, 25:.. 351, Howard et al, 1989, J.Neurosurg, 71:. 105 see). Furthermore, since a controlled release system can be placed in the vicinity of a therapeutic target (eg, the brain), only a fraction of the total body mass is required (eg, Goodson, 1984, Medical Applications of Controlled Release, Vol. 2, pp. 115-138). For further guidance, other controlled release systems are described in Langer, 1990, Science, 249: 1527-1533.

(0171)
The photolabile compound is also provided in an effective amount in a pharmaceutical composition comprising a pharmaceutically acceptable carrier, diluent, excipient or vehicle, eg for use as a therapeutic agent. In one embodiment, the term “pharmacologically acceptable” is approved by a federal or state government regulatory authority, or is used by the United States Pharmacopeia or other general public for use in animals, particularly humans. It means that it is described in a recognized pharmacopoeia. The term “vehicle, carrier or excipient” refers to a diluent or adjuvant that includes or is administered with a therapeutic agent. Such pharmaceutical carriers, vehicles or excipients are sterile liquids (water, oil etc., including those of petroleum, animal, plant or synthetic origin, eg peanut oil, soybean oil, mineral oil, sesame oil etc.) obtain. When the pharmaceutical composition is administered intravenously and is water soluble, the preferred carrier is water. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid carriers, particularly for injectables. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dry skim milk, glycerol, propylene , Glycol, water, ethanol and the like. Where necessary or desirable, the compositions of the present invention may also contain minor amounts of wetting agents, emulsifying agents or pH buffering agents.

(0172)
The photolabile compound and composition of the present invention can be prepared into solutions, suspensions, emulsions, tablets, pills, capsules, powders, sustained-release preparations and the like. The composition can be formulated as a suppository, with traditional binders and carriers such as triglycerides. Oral formulations can include standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate and the like. Examples of suitable pharmaceutical carriers and the like are described in E.I. W. It is described in the latest edition of Martin's Remington's Pharmaceutical Sciences. Such compositions usually contain a therapeutically effective amount, preferably a purified form of the photolabile compound, together with a suitable amount of carrier to obtain a dosage form for proper administration to the patient. The formulation must be compatible with the method of administration.

(0173)
In another embodiment, the photolabile compound of the present invention is formulated according to conventional methods as a pharmaceutical composition adapted for intravenous administration to humans. Typically, compositions for intravenous administration are solutions in sterile isotonic aqueous buffer. If desired, the composition may also include a solubilizing agent and a local anesthetic such as lidocaine to ease pain at the site of the injection. In general, the ingredients are supplied separately, for example as a lyophilized powder or an anhydrous concentrate (such as an ampoule or sachet indicating the amount of active agent) in a sealed container, or mixed in a unit dosage form. Where the composition is to be administered by infusion, it can be dispensed with an infusion bottle containing sterile pharmaceutical grade water or saline. Where the composition is administered by injection, an ampoule of sterile water for injection or saline can be provided so that the ingredients may be mixed prior to administration. Sterility of the composition when administered for therapeutic purposes is readily achieved by filtration through a sterile filtration membrane (eg, a 0.2 micron membrane). The therapeutic agent is typically placed in a container with a sterile access port (eg, a drug bag or vial for intravenous infusion with a stopper that can be pierced with a hypodermic needle). The therapeutic agent is typically stored as an aqueous solution or lyophilized formulation for reconstitution in a single or multiple dose container (eg, a sealed ampoule or vial). If desired, ampoules or vials are essentially light impermeable. As an example of a lyophilized therapeutic formulation, a 10 ml vial is filled with 5 ml of sterile filtered 1% (w / v) aqueous therapeutic agent solution and the resulting mixture is lyophilized. An infusion solution is prepared by reconstitution of this lyophilized therapeutic with bacteriostatic water for injection.

(0174)
The amount of photolabile compound effective in the treatment, recovery, reduction, elimination, inhibition or prevention of a particular disease, condition, condition or disorder associated with the use and biological activity of organic molecules is determined by standard clinical techniques. be able to. “Effective amount” or “pharmacologically effective amount” of the photolabile compound of the present invention means treatment, recovery, or recovery of a disease, condition, pathological condition or disorder to which the compound is used. An amount that is effective for relief, remission, elimination, or prevention. In certain embodiments, an effective amount is an amount effective to make the organic molecules of the invention bioavailable to the patient. When another therapeutic agent is used with the photolabile compound, an effective amount of the therapeutic agent is an amount that is effective to obtain the therapeutic effect of the therapeutic agent. The exact dosage used in the formulation will depend on the route of administration and individual patient circumstances (patient age, health and vital statistics, severity of the disease, condition or disorder, etc.) . Medication should be determined according to the physician's judgment based on patient assessment, consideration of the patient's physiological status and medical history. In addition, optional in vitro assays can be performed to help determine optimal dosage ranges. Effective doses can be extrapolated from dose-response curves derived from in vitro or in vivo animal model test systems.

(0175)
In general, the total effective amount of a photolabile compound administered parenterally is in the range of about 1 μg / kg / day to 10 mg / kg / day relative to the patient's body weight. However, as described above, this amount is determined based on the patient's condition and the above variables. The therapeutic amount can be at least 0.01 mg / kg / day or even about 0.01-1 mg / kg / day, particularly for human administration. When administered continuously, the therapeutic agent is typically administered by one to four injections per day at a dosage rate of about 1 μg / kg / hour to about 50 μg / kg / hour or by continuous subcutaneous infusion using, for example, a minipump. Be administered. Intravenous infusion bag solutions may also be used. The duration of treatment required to observe the change and the interval between responses and subsequent response can vary depending on the effect desired. In some embodiments, a suitable effective dosage is about 10 μg to about 2500 mg every about 4 hours. However, the amount is typically about 100 mg or less. In one embodiment, the effective dose of the photolabile compound is about 0.01 mg to about 100 mg every about 4 hours. In another embodiment, an effective dose of a compound of the invention is about 0.020 mg to about 50 mg every about 4 hours, and in another embodiment about 0.025 mg to about 20 mg every about 4 hours. . An effective dosage is the total amount administered. Therefore, when administering two or more photolabile compounds, the effective dosage corresponds to the total amount administered.

(0176)
In another embodiment, when the photolabile compound is contacted with a biological sample in vitro, the effective amount is typically about 0 of a solution or suspension of a pharmaceutically acceptable carrier, diluent or excipient. 0.01 μg / L to about 5 mg / L, in another embodiment from about 0.01 μg / L to about 2.5 mg / L, and in another embodiment from about 0.01 μg / L to about 0. 5 mg / L, and in yet another embodiment from about 0.01 μg / L to about 0.25 mg / L. In certain embodiments, the volume of the solution or suspension is from about 1 μL to about 1 mL, and in another embodiment, the volume of the solution or suspension is about 200 μL.

(0177)
Examples of neoplastic or hyperproliferative diseases, disorders, conditions or conditions that can be treated, recovered, reduced, ameliorated, eliminated, inhibited, prevented and / or diagnosed using photolabile compounds and light-released organic molecules Is the colon, abdomen, bone, breast, digestive system, esophagus, liver, pancreas, peritoneum, endocrine glands (adrenal, parathyroid, pituitary, testis, ovary, cervix, thymus, thyroid), eyes, head and neck, Includes, but is not limited to, neoplasms (cancer or tumor) located in the nerves (central nerve, peripheral nerve), lymphatic system, pelvis, skin, soft tissue, spleen, thoracic region, bladder and genitourinary system. Cancers that can be treated using photolabile compounds include follicular lymphomas, p53-mutated cell tumors and hormone-dependent tumors, colon cancer, heart tumors, pancreatic cancer, melanoma, retina Blastoma, glioblastoma, lung cancer, intestinal cancer, testicular cancer, gastric cancer, neuroblastoma, myxoma, myoma, lymphoma, endothelial tumor, osteoblastoma, giant cell tumor, chondrosarcoma, adenoma Breast cancer, prostate cancer, Kaposi's sarcoma, ovarian cancer or metastases thereof, but is not limited thereto. Autoimmune diseases, disorders or conditions can be treated with photolabile compounds and can also be treated with multiple sclerosis, Sjogren's syndrome, Hashimoto's thyroiditis, biliary cirrhosis, Behcet's disease, Crohn's disease, polymyositis, systemic lupus erythematosus, immune Glomerulonephritis, rheumatoid arthritis, ischemic injury (eg, caused by myocardial infarction, stroke and reperfusion injury), liver injury (eg, liver injury related to hepatitis, ischemia / reperfusion injury, cholestasis) This includes cholestosis (bile duct injury, liver cancer), toxin-induced liver disease (eg caused by alcohol), septic shock, cachexia and anorexia. Viral infections (herpesviruses, poxviruses, adenoviruses, etc.), inflammation, graft versus host disease (GVH), acute graft rejection and chronic graft rejection can also be treated with photolabile compounds.

(0178)
Additional diseases or conditions associated with abnormal and elevated cell survival that can be treated, recovered, reduced, ameliorated, eliminated, inhibited, prevented and / or diagnosed using photolabile compounds include malignant tumor progression and / or Metastasis and related disorders such as leukemia (eg acute leukemia (eg acute lymphoblastic leukemia, acute myeloid leukemia. Myeloblastic leukemia, promyelocytic leukemia, myelomonocytic leukemia, monocytic leukemia, erythroleukemia) Including), chronic leukemia (eg, including chronic myelogenous (granulocyte) leukemia, chronic lymphocytic leukemia), polycythemia vera, lymphoma (eg, Hodgkin's disease, non-Hodgkin's disease), multiple myeloma, Walden Solid tumors including, but not limited to, stromal macroglobulinemia, heavy chain disease, sarcomas and carcinomas such as fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteosarcoma, chordoma, blood vessels Tumor, endothelial sarcoma, lymphangiosarcoma, lymphatic endothelial sarcoma, synovial tumor, mesothelioma, Ewing tumor, leiomyosarcoma, rhabdomyosarcoma, colon cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer , Squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinoma, cystadenocarcinoma, medullary carcinoma, bronchogenic lung cancer, renal cell carcinoma, hepatocellular carcinoma, cholangiocarcinoma, Choriocarcinoma, seminoma, fetal cancer, Wilms tumor, cervical cancer, testicular tumor, lung cancer, small cell lung cancer, bladder cancer, epithelial cancer, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pine These include, but are not limited to, keratinoma, hemangioblastoma, acoustic neuroma, oligodendroma, menangioma, melanoma, neuroblastoma and retinoblastoma.

(0179)
In another embodiment, the photolabile compound is needed as a therapeutic agent to stimulate epithelial cell proliferation and basal keratinocytes, and to stimulate hair follicle formation and skin wound healing for wound healing. Can be done. The photolabile compounds of the present invention are clinically useful in stimulating wound healing, which includes surgical wounds, excision wounds, deep wounds with dermal and epidermal damage, ocular tissue wounds, dental tissues Wounds, oral wounds, diabetic ulcers, skin ulcers, elbow ulcers, arterial ulcers, venous stasis ulcers, heat exposure or chemical burns and other abnormal wound healing conditions such as uremia, malnutrition, vitamins Includes deficiencies and complications related to systemic treatment with steroids, radiation therapy, antineoplastic drugs and antimetabolites.

(0180)
Other diseases, disorders or conditions that can be treated, recovered, reduced, ameliorated, eliminated, inhibited, prevented and / or diagnosed with a photolabile compound include AIDS, neurodegenerative diseases, disorders and / or conditions (Alzheimer's disease, Parkinson Disease, amyotrophic lateral sclerosis (ALS), multiple sclerosis, retinitis pigmentosa (RP), cerebellar degeneration, brain tumor or prior related diseases, etc.).
In one embodiment, diseases and conditions that can be treated using calcium channel inhibitors (eg, 4AP) include heart disease, hypertension, angina pectoris, chest pain, cardiovascular disease such as coronary artery disease, cardiomyopathy, heart Valvular disease, kidney disease, Peyronie's disease, neurological, neurophysiological or neuromuscular disease and condition, including but not limited to amyotrophic lateral sclerosis (ALS), multiple sclerosis, epilepsy .
In another embodiment, diseases treatable using NMDA receptor agonists or antagonists (eg, TzGly) include neurological, neurodegenerative or neurophysiological diseases, disorders and conditions such as Alzheimer's disease, Parkinson's disease. , Include, but are not limited to, Huntington's disease and dyskinesia. In another embodiment, neurological, neurodegenerative and neurophysiological diseases such as Parkinson's disease, Alzheimer's disease and the like can be treated using TzAMPA.

(0183)
In another embodiment, the present invention relates to a kit comprising a photolabile compound and instructions for use. The kit can be used for diagnostic, screening or test assays. A kit can also be a pharmaceutical pack, particularly for use in the treatment or prevention of a disease, disorder, condition or condition. Kits for pharmaceutical use are typically sterile and contain an amount of a photolabile compound effective for the treatment or prevention of a disease, disorder, condition or condition and a pharmaceutically acceptable carrier, diluent or Containing excipients. The kit or pharmaceutical pack comprises one or more containers or containers (eg, unit dosage forms) filled with an effective amount of one or more photolabile compounds or compositions of the invention, and a pharmaceutically acceptable carrier. , Diluents or excipients. The kit or pharmaceutical pack can further include a label. In addition, the kit or pharmaceutical pack may also include a unit dosage form of another therapeutic agent, such as a container containing an effective amount of the other therapeutic agent. The kit or drug pack may further optionally be accompanied by a notice in the form prescribed by the government agency that regulates the manufacture, use or sale of the drug or biologic. Represents approval by a government agency for use or sale. The kit or pharmaceutical pack can also include devices useful for administration of unit dosage forms. Examples of such devices include, but are not limited to, syringes, drip bags, patches, inhalers, and enema bags or containers.

(0184)
The examples described below are intended to illustrate the present invention and are not intended to limit the present invention.
Example 1
Synthesis of [Ru (bpy) ₂ (4AP) ₂ ] Cl ₂ 159 mg of Ru (bpy) ₂ Cl ₂ (bpy = 2,2′-bipyridine) was suspended in 7 mL of water at 85 ° C. under N ₂ . . After dissolution, 66 mg 4-aminopyridine (4AP) was added and the resulting solution was heated above about 50-80 ° C. for about 20 minutes. A molar excess of NH ₄ PF ₆ was added and the resulting red solid was washed with water and dried. This red solid was dissolved in a minimum amount of acetone, and tetraethylammonium chloride was added to the acetone solution to precipitate [Ru (bpy) ₂ (4AP) ₂ ] Cl ₂ (yield 79%).

(0186)
Example 2
_{[Ru (bpy) 2 (TzGly} ) 2] Synthesis of Cl _{2 [Ru (bpy) 2 (TzGly} ) 2] and Cl _2, described in Example _{1 [Ru (bpy) 2 (} 4AP) 2] of Cl ₂ Produced according to the procedure used in the production. However, (RS)-(tetrazol-5-yl) glycine (TzGly) was used instead of 4AP.

(0187)
Example 3
_{[Ru (bpy) 2 (5HT} ) 2] Synthesis of Cl _{2 [Ru (bpy) 2 (5HT} ) 2] and Cl _2, described in Example _{1 [Ru (bpy) 2 (} 4AP) 2] of Cl ₂ Generate according to the procedure used for generation. However, serotonin (5HT) is used instead of 4AP.

(0188)
Example 4
Synthesis of [Ru (bpy) ₂ (4AP) (PPh ₃ )] Cl ₂ Ru (bpy) ₂ Cl ₂ (bpy = 2,2′-bipyridine) was added to water at a concentration of 10 mg / mL at 85 ° C. under N _2. And suspended. After dissolution, 1 equivalent of PPh ₃ was added and the resulting solution was heated above about 50-80 ° C. for about 60 minutes. 1.1 equivalents of 4AP was subsequently added and heating continued for another 30 minutes. A molar excess of NH ₄ PF ₆ was added and the resulting orange solid was washed with water and dried. The orange solid was dissolved in a minimum amount of acetone and tetraethylammonium chloride was added to the acetone solution to precipitate [Ru (bpy) ₂ (4AP) (PPh ₃ )] Cl ₂ .
Example 5
_{[Ru (bpy) 2 (TzGly} ) (PPh 3)] Synthesis of _{Cl 2 [Ru (bpy) 2} (TzGly) (PPh 3)] to Cl _2, described in Example _{4 [Ru (bpy) 2 (} 4AP ) (PPh ₃ )] Cl ₂ was produced according to the procedure used for the production of Cl ₂ . However, TzGly was used instead of 4AP.
Example 6
_{[Ru (bpy) 2 (5HT} ) (PPh 3)] Synthesis of _{Cl 2 [Ru (bpy) 2} (5HT) (PPh 3)] to Cl _2, described in Example _{4 [Ru (bpy) 2 (} 4AP ) (PPh ₃ )] Cl ₂ is produced according to the procedure used for the production of Cl ₂ . However, serotonin is used instead of 4AP.

(0191)
Example 7
[Ru (bpy) ₂ (nicotine) (PPh _3)] Synthesis of Cl ₂ [Ru (bpy) ₂ (nicotine) (PPh _3)] to Cl _2, described in Example 4 [Ru (bpy) ₂ (4AP ) (PPh ₃ )] Cl ₂ is produced according to the procedure used for the production of Cl ₂ . However, nicotine is used instead of 4AP.

(0192)
Example 8
_{[Ru (bpy) 2 (TzGly} ) (py)] Synthesis of _{Cl 2 [Ru (bpy) 2} (TzGly) (py)] Cl 2 the (py = pyridine), as described in Example 4 [Ru (bpy) ₂ (4AP) (PPh ₃ )] Prepared according to the procedure used for the generation of Cl ₂ . However, TzGly was used instead of 4AP, and pyridine was used instead of PPh ₃ .

(0193)
Example 9
_{[Ru (bpy) 2 (4AP} ) (py)] Synthesis of _{Cl 2 [Ru (bpy) 2} (4AP) (py)] Cl 2 the (py = pyridine), as described in Example 4 [Ru (bpy) ₂ (4AP) (PPh ₃ )] Produced according to the procedure used for the production of Cl ₂ . However, pyridine is used instead of PPh ₃ .

(0194)
Example 10
_{[Ru (bpy) 2 (5HT} ) (py)] Synthesis of _{Cl 2 [Ru (bpy) 2} (5HT) (py)] Cl 2 the (py = pyridine), as described in Example 4 [Ru (bpy) ₂ (4AP) (PPh ₃ )] Produced according to the procedure used for the production of Cl ₂ . However, 5HT is used instead of 4AP, and pyridine is used instead of PPh ₃ .

(0195)
Example 11
[Ru (bpy) ₂ (nicotine) (py)] Synthesis of Cl ₂ [Ru (bpy) ₂ (nicotine) (py)] Cl ₂ the (py = pyridine), as described in Example 4 [Ru (bpy) ₂ (4AP) (PPh ₃ )] Produced according to the procedure used for the production of Cl ₂ . However, nicotine is used instead of 4AP, and pyridine is used instead of PPh ₃ .
Example 12
Synthesis of Co (DMG) ₂ (5HT) (Cl) CoCl ₂ was dissolved in a 1: 1 v / v mixture of water / ethanol at a final concentration of about 0.2M. Two equivalents of dimethylglyoxime (DMG) were added and the resulting mixture was stirred under N ₂ until dissolved. One equivalent of 5HT was added and air was bubbled through the resulting mixture for 6 hours to precipitate Co (DMG) ₂ (5HT) (Cl). The precipitated product was filtered and washed.

(0197)
Example 13
Photoemission of 4AP from [Ru (bpy) ₂ (4AP) ₂ ] Cl ₂ UV and visible spectra in water were obtained using a HP 8453 diode array spectrophotometer. RMN ¹ H spectra were obtained using a Bruker 500 MHz instrument. CV measurements were made using a PAR 273A potentiostat. Light irradiation was performed using a pulsed Xe lamp (pulse energy: about 0.5 J) equipped with a 480 nm low-pass filter. Similar results were obtained with light irradiation using a 473 nm DPSS laser.

(0198)
[Ru (bpy) ₂ (4AP) ₂ ] Cl ₂ is very well soluble in water and stable in the dark, while its metal-ligand charge transfer (MLCT) centered at 489 nm when irradiated with visible light. Resolved in the band (in spite of characterization in previous studies to be 450 nm, in CH ₃ CN solution, the absorption band shifts red to 492 nm, which is consistent with the low polarity of the solvent. Light exposure of [Ru (bpy) ₂ (4AP) ₂ ] Cl ₂ in CH ₃ CN produced a yellow compound with an absorption maximum at 450 nm, which was previously misinterpreted for this compound. especially-obtained codes (D.Chun-Ying et al, 1999 , J.Coord.Chem, 46:.. 301-312), photoproduct complex [Ru (bpy) ₂ (4 P) is considered to be CH ₃ CN] ^2+). Some ruthenium polypyridyl complexes exhibit this behavior (DV Pinnick et al., 1984, Inorg. Chem., 23: 1440-1445).

(0199)
At pH 7, the spectrum of the irradiated complex is very similar to the spectrum of the first complex, but the shoulder decay at 470 nm is evident. In order to know the nature of the photochemical reaction, NMR spectra were taken before and after irradiation with visible light. FIG. 1 shows the signal assigned to metahydrogen [Ru (bpy) ₂ (4AP) ₂ ] Cl ₂ (m1). After irradiation, this signal decreased and two new signals appeared in the figure below. One signal corresponds to the free ligand (m3) and the other corresponds to the aco-4AP complex (m2), which indicates 4AP light emission. These latter two signals are integrated into the initial signal 0.30 and 0.27, corresponding to 60% photochemical reaction.

（４７３ｎｍ）が得られる。 (0200)
The redox potential of the couple Ru ^III / Ru ^II for [Ru (bpy) ₂ (4AP) ₂ ] Cl ₂ measured in water is E = 0.76 V versus Ag / AgCl, which is compared to pyridine Consistent with the higher basicity of 4AP. Therefore, the redox and photochemistry of [Ru (bpy) ₂ (4AP) ₂ ] Cl ₂ is completely consistent with the results obtained corresponding to the Ru (bpy) ₂ XY system (X, Y are monodentate coordinations) (See, eg, ES Dodsworth et al., 1986, Chem. Phys. Lett., 124: 152-158). The photoactivity of these compounds has been described in a reaction pathway involving a transition between the MLCT state and the low energy dd state, which promotes ligand release. There is a direct correspondence between the energy of the MLCT transition and the quantum yield of the photochemical reaction. For [Ru (bpy) ₂ (4AP) CH ₃ CN] ²⁺ , the photochemical reaction yield is about Φ _PR = 0.4. Since [Ru (bpy) ₂ (4AP) ₂ ] Cl ₂ exhibits a red-shifted band, a lower photochemical reaction yield is expected. From estimates based on past experiments,

(473 nm) is obtained.

(0201)
Example 14
Neurophysiological activity of 4AP photo-emitted from [Ru (bpy) ₂ (4AP) ₂ ] Cl ₂ Using standard equipment for measuring intracellular voltage and using medical medical hills (Hirudo medicalis) Thus, the neurophysiological activity of light-released 4AP was demonstrated. The medical hill (Hirudo medicalis) has a central nerve cord with several ganglia, each ganglion containing about 400 neurons arranged in a known pattern (W.-R.Schule et al.). al., 1980, J. Exp. Biol., 82: 23-34). The entire ganglion was placed on the dish. The membrane potential of a single cell (neuron) in the ganglion was inserted into the neuron with a glass micropipette with a micrometer-sized end filled with a saturated aqueous KCl solution that served as a Lugin bridge for the Ag / AgCl electrode. Recorded. Another Ag / AgCl electrode was used as the reference electrode. The signal was captured using an AM-System 1600 amplifier and the entire device was covered with a Faraday cage. Data was digitized using an ad hoc program written in Quick Basic using a 12-bit A / D collection card.

(0202)
Perfuse dish with low Ca ²⁺ / high Mg ²⁺ saline (NaCl: 102 mM, KCl: 4 mM, CaCl ₂ : 1 mM, MgCl ₂ : 10 mM, Tris base, pH 5.4 adjusted to 7.4) It was. [Ru (bpy) ₂ (4AP) ₂ ] Cl ₂ and free ligand 4AP were injected into the mainstream at a controlled timing. The solution was irradiated with light using a pulsed Xe lamp located under the dish. Ultraviolet light was removed using a 500 nm bandpass filter. FIG. 2 shows the behavior of the membrane potential recorded with one of the Retzius cells (Rz) in the ganglion. The upper graph in FIG. 2 is raw data, showing a resting potential period and a very fast spike (action potential), which are caused by changes in membrane ion permeability. The lower graph shows the instantaneous spike frequency of each time.

(0203)
After inserting the electrode into the cell, reproducibility was ensured by conducting numerous experiments on the cell. After 5000 seconds, the cells showed low activity, as seen on the left of the graph. At t = 5200 seconds, about 100 μM Ru (bpy) ₃ Cl ₂ was added to the saline, but there was no significant change in activity. At 5500 seconds after 300 seconds, the ganglion was flashed. The rapid increase in the frequency of the action potential was mainly due to the temperature pulse, but the activity decreased to the basal level after a short time. After washing by perfusion, further pulse irradiation (t = 6000 seconds) showed a very similar pattern. At t = 6250 seconds, approximately 100 mM of [Ru (bpy) ₂ (4AP) ₂ ] Cl ₂ was added to the saline, but the activity remained unchanged. However, rapid activity was recorded after a new flash (t = 6400 seconds) and remained high after 300 seconds. A second light pulse at 6750 seconds promoted higher activity, which decreased only after cleansing perfusion with pure saline.

(0204)
A similar increase in frequency also occurred when free 4AP was perfused onto the cell nodes, demonstrating that the release of 4AP caused this sustained increase in frequency. Calibration of cell activity using a solution of 4AP showed that 10-15 μM 4AP was released from [Ru (bpy) ₂ (4AP) ₂ ] Cl ₂ during the previous experiment with each light irradiation. During the experiment, no neuronal toxicity or adverse effects were observed. These results indicate that [Ru (bpy) ₂ (4AP) ₂ ] Cl ₂ (an exemplary photolabile compound) that photoreleases organic molecules with neurophysiological activity can be used to stimulate neuronal responses. .
Example 15
Procedure of _{[Ru (bpy) 2 (TzGly} ) (py)] Light emission TzGly from Cl _{2 [Ru (bpy) 2 (TzGly} ) (py)] of TzGly from Cl ₂ light emission, in Example 13 It is the same as that used for the light emission of 4AP from the above-mentioned [Ru (bpy) ₂ (4AP) ₂ ] Cl ₂ . However, the irradiation spot was very localized (diameter <1 micron). TzGly was photo-emitted by irradiation of [Ru (bpy) ₂ (TzGly) (py)] Cl ₂ at 470 nm.

(0206)
Example 16
[Ru (bpy) ₂ (TzGly) (py)] Neurophysiological activity of TzGly photo-released from Cl _{2 The} neurophysiological activity of photo-released TzGly was tested in the same manner as described above in Example 14. Evaluated by doing. Therefore, a standard device for measuring intracellular voltage was used to demonstrate the neurophysiological activity in the radii of TzGly leeches radiated using medical hills (Hirudo medicalis).
Example 17
Synthesis of [Ru (bpy) ₂ (PMe ₃ ) Cl] PF ₆ 520 mg of [Ru (bpy) ₂ Cl ₂ ] was dissolved in 20 mL methanol / water mixture (1: 1) and refluxed under N ₂ . . 1.2 mL of trimethylphosphine 1M in THF (Aldrich, 324108) was added via syringe. The reaction was followed by UV / visible spectroscopy. In some cases, more phosphine solution was added. Once the UV-visible spectrum was stable, methanol and excess phosphine were distilled using a rotary evaporator under vacuum. The resulting aqueous solution was filtered to remove solids and precipitated using an excess of KPF ₆ on ice. The dark orange solid was washed 3 times with cold water and dried.

(0208)
Example 18
Synthesis of [Ru (bpy) ₂ (PMe ₃ ) GlutH ₂ ] (PF ₆ ) ₂ 110 mg of [Ru (bpy) ₂ (PMe ₃ ) Cl] PF ₆ was dissolved in 2 mL of acetone. 200 mg of a chloride-containing anion exchange resin DOWEX 2 × 8 suspension was added and stirred for 10 minutes. The obtained [Ru (bpy) ₂ (PMe ₃ ) Cl] Cl solution was filtered. 500 mg monosodium glutamate and 4.4 mL NaOH 1M were added and heated for 3 hours. 1 mL of saturated KPF ₆ was added and the resulting precipitate was discarded. The solution was cooled to 0 ° C. and acidified to pH 2 with HCl 5M. When an excess amount of KPF ₆ was added, [Ru (bpy) ₂ (PMe ₃ ) GlutH ₂ ] (PF ₆ ) ₂ precipitated. The yellowish orange solid was washed 3 times with cold water and dried.
Example 19
Photoemission of glutamate from [Ru (bpy) ₂ (PMe ₃ ) GlutH ₂ ] (PF ₆ ) ₂ [Ru (bpy) ₂ (PMe ₃ ) GlutH ₂ ] (PF ₆ ) ₂ exhibits a bright orange color, pH > 6 showing high solubility in water (as deprotonated species [Ru (bpy) ₂ (PMe3) Glu]). The aqueous solution exhibits a strong metal-to-ligand charge transfer band (MLCT band) at 450 nm, characteristic of this Ru polypyridine system. Cyclic voltammetry of the compound dissolved in acetonitrile (FIG. 15) shows three redox steps at 0.98, 1.46 and 1.56 V (vs. NHE), each of which is the first complex. Corresponding to the Ru (III) / Ru (II) couple of the complex and the Ru (III) / Ru (II) couple of the complex with the oxidation product of glutamate.

(0210)
FIG. 16 (upper) is an ultraviolet / visible spectrum of the complex at pH = 7 during light irradiation with a 450 ± 20 nm LED. As the photochemical reaction proceeded, the formation of an aco-complex was recorded and was complete after about 4 minutes. Factor analysis for the presence and spectrum of the two isosbestic points shows that exactly two colored species are present in the solution, according to what would be expected with a single photoacation process. The spectrum of the photoproduct was found to be the same as that of the complex [Ru (bpy) ₂ (PMe ₃ ) (H ₂ O)] ₂ ⁺ , which can be synthesized by refluxing the chloro complex in water. it can. Similar behavior was observed at pH = 11. In this case, however, the product was a hydroxo complex [Ru (bpy) ₂ (PMe ₃ ) (OH)] ⁺ .
The inset graph in FIG. 16 (top) shows the yield of free glutamate generated by photolysis obtained by spectral analysis. The irradiance of the light source was calibrated using the known efficiency of photolysis of [Ru (bpy) (Py) ₂ ] ²⁺ . The amount of glutamate released was fitted using a two parameter single exponential function: y = a [1-exp (−bx)]. The quantum yield (φPC) was obtained as a · b, and a value of 0.13 at pH = 7 and a value of 0.10 at pH = 11 were adopted.

(0212)
To estimate the uncaging time, a flash photolysis experiment was carried out using the second harmonic (532 nm, 10 nanosecond pulse) of Nd-YAG laser in a basic aqueous solution of [Ru (bpy) ₂ (PMe ₃ ) Glu]. I went there. The results are shown in FIG. 16 (bottom). Photocleavage occurs within the first 50 nanoseconds and is the fastest caged compound known to have been reported. The fact that several tens of nanoseconds are required to complete the glutamate uncaging reaction means that the lifetime of the excited state measured for similar compounds containing pyridine instead of glutamate is 10-100 nanoseconds. This is consistent with the observation that
The photochemical reaction was also followed by NMR spectroscopy. In the aromatic region of the 1H-NMR spectrum of the complex (FIG. 17), 16 signals corresponding to the bpy protons could be distinguished and the signal was found to be doubled, which was obtained This is because the material is an approximately 1: 1 mixture of diastereomers resulting from the coordination of L-glutamate with a racemic mixture of Λ and ΔRu-bpy enantiomers. No attempt was made to separate diastereomers, and both chemical and biological tests suggest similar behavior.

(0214)
The aliphatic region corresponding to the glutamate moiety is depicted in the upper trace of FIG. 18, with the —NH ₂ protons appearing at 4.50, 4.12, 4.00, and 3.57 ppm. The presence of these signals in D ₂ O solution (providing the absence of isotope exchange in the complex) is that glutamate coordination is carried out via the amine nitrogen, preventing further protonation. Is clearly shown. This behavior is different from that of free glutamate, which quickly exchanges D + with H + under the same conditions. After 2 months in D ₂ O at room temperature, no exchange is seen, demonstrating that the compound is truly inert.
After photolysis, many new signals appear in the aromatic region corresponding to the acocomplex. At the same time, analysis of the aliphatic region (Figure 18, bottom trace) clearly demonstrates the appearance of free glutamate signal. The coordinated —NH ₂ signal is attenuated by light irradiation, and the characteristic signals of methylene protons of free glutamate appear at 3.50, 2.30, 2.04 and 1.94 ppm.
Further addition of glutamate does not generate a new signal, only the intensity of the existing signal increases, demonstrating that the intrinsic aliphatic photoproduct is glutamate.

(0217)
A biocompatibility test was performed to evaluate the potential toxicity of the complex. 350 μM [Ru (bpy) ₂ (PMe ₃ ) Glu] (concentration used in the two-photon uncaging experiment) was incubated with living neocortical pyramidal neurons in mouse brain slices, and the morphology and electrophysiology of the neurons The physical characteristics were carefully monitored by two-photon microscopy and whole cell patch recording. After 1 hour incubation, no detectable effect was observed and the neurons remained normal. This means that this complex does not change the integrity of the membrane or has a detrimental effect on living neurons.
All patent applications, published patent applications, patents granted and granted patents, documents and references cited herein are incorporated herein by reference in their entirety, and the state of the art to which this invention pertains Is described in more detail.
Since various changes may be made to the above methods and compositions without departing from the scope and spirit of the invention as described, they are included in the above description, depicted in the accompanying drawings, or accompanying It is intended that all subject matter defined in the following claims be interpreted as illustrative and not limiting.

(0220)
Example 20
Ruthenium-Rhodamine Complex as an Activable Fluorescent Probe This example describes a caged fluorescent probe based on a ruthenium-bipyridine core and having a modified rhodamine as an active derivative of a labeled molecule. This complex behaves as a caged fluorescent probe, increasing its fluorescence approximately 6-fold when excited by visible light. Therefore, in this example, a fluorescent molecule of a photolabile compound containing a labeling molecule or an active derivative thereof or an active derivative thereof (rhodamine B, rhodamine 6G, RhodB-MAPN, Rhod-6G-MAPN, RhodB-MAMePy, Rhod- 6G-MAMePy etc.) demonstrates the advantage in increasing fluorescence.
All reagents were purchased from Sigma-Aldrich and used as they were. Ru (bpy) ₂ Cl ₂ was synthesized as described using water as the solvent as in the literature (Viala C., Coudret C, Inorg. Chim. Acta 2006, 359: 984-989). UV and visible spectra were obtained using a HP8453 diode array spectrophotometer. NMR spectra were obtained using a 500 MHz Bruker AM-500. Fluorescence emission measurements were performed on a PTI Quantama spectrofluorometer and corrected for instrument response function. Rhodamine B was used as the fluorescence standard for luminescence quantum yield (φ = 0.31 in aqueous solution). The quantum yield of the complex was obtained directly from the ratio of spectral areas (showing the same shape).

(0222)
The measurement of photouncaging quantum yield was performed at a constant output of 6.3 mW using an Nd: YAG diode pumped solid state laser doubled to 473 nm. The light was collimated and sent with stirring through a 1 cm light path into a fluorescent glass cuvette. Total irradiation energy was measured using a Coherent Fieldmaster FM light meter with a visible light diode model SR45.
Photoelectrophysiology, comprising a micromanipulator used to insert a capillary with a tip diameter of about 1 μm into a neuron body and a CCD web cam as a recording device for observing the ganglia of Hill (Hirudomedicinalis). It was done with custom-made equipment. Isolated leeched ganglia of segments 7-14 were fixed in a 35 mm Petri dish coated with Sirgard. Retzius cells were identified from their location, size and firing behavior. Intracellular recordings were obtained using a sharp capillary microelectrode of approximately 20 MΩ, a Neuroprobe 1600 (A-M Systems) amplifier and A / D signal input board (1 kHz sampling rate) and custom software. For dye iontophoretic injection, microelectrodes were filled with a recording solution supplemented with 1 mM caged fluorescent dye (chloride form). A 1 nA, 2 Hz square biphasic pulse was applied for less than 5 minutes.

(0224)
Photographs of the spectrophotometric thin layer flow cell and FIA capillaries were taken using a Nikon TS-100 fitted with a fluorescent adapter. A consumer electronics type compact digital camera (Casio, Exilim EX-FC100) focused with a normal eyepiece through a custom adapter was used for video shooting with ISO 800 sensitivity. Video and photo analysis was performed using published ImageJ software. The fluorescence was activated at 405 nm and 6 mW, and the laser diode was controlled by a pulse generator using a fast reed relay to obtain a 20 millisecond light pulse. The laser beam was focused on the cell surface and monitored with an inverted microscope (Nikon, TS100, with fluorescent adapter).
Synthetic rhodamine B-methylaminopropionitrile amide (RhodB-MAPN)
550 mg rhodamine B.I. HCl was dissolved in 10 ml anhydrous 1,2-dichloroethane. The system was purged with N ₂ and 300 μL of phosphorus oxychloride was added. The mixture was refluxed for 5 hours. The solvent was distilled under vacuum and the solid was quickly dissolved in anhydrous acetonitrile. 500 μL of triethylamine and 108 μL of N-methylamine propionitrile were added and the mixture was refluxed for 12 hours. The solvent was removed under vacuum, redissolved in water, filtered to remove solids, and precipitation by addition of excess KPF ₆ was performed. The dark red solid was washed several times with distilled water and dried over silica gel. The exchange of Cl with PF ₆ to obtain a water soluble ligand is described in RhodB-MAPN. This was done by stirring PF ₆ in acetone / water solution (1: 1) with Dowex 22 anion resin overnight and lyophilizing the resulting solution. RhodB-MAPN chloride salt is slightly hygroscopic and must be stored in a damp environment.

(0226)
Total yield: 55%, 1H NMR (acetone-d6): 1Hδ 1.37 (t, 12H), 2.37 (t, 2H), 3.16 (s, 3H), 3.53 (t, 2H), 3.80 (m, 8H), 6.98 (d, 2H), 7.20 (dd, 2H), 7.37 (dd, 2H), 7.62 (m, 1H), 7.77 (m 1H), 7.82 (t, 2H)

(0227, 0228)
[Ru (bpy) 2 (L) Cl] PF ₆
L = rhodamine B-methylaminopropionitrileamide 20 mg Ru (bpy) ₂ Cl ₂ is suspended in 2 mL EtOH / water (2: 1) mixture and the suspension is completely dissolved up to 80 ° C. Until heated. The formation of [Ru (bpy) ₂ (H ₂ O) Cl] ⁺ complex was confirmed by its absorption at 490 nm (in water). After formation of the chloro-aquo complex, 2 equivalents of RhodB-MAPN chloride salt was added and the reaction followed by TLC using a silica plate and a water / EtOH / nBuOH (1: 1: 1) mixture as eluent. It was. The solution was heated at 80 ° C. for about 2 hours until no TLC change was observed. All subsequent procedures were performed in the dark. The solution was filtered to remove insoluble particles and the solvent was removed under vacuum. The resulting oil was redissolved in 4 mL acetone / methanol mixture (3: 1) and precipitated by the addition of excess THF.

(0229)
The THF fraction containing unreacted ligand was discarded and the precipitate was washed several times with THF, re-dissolved in water and precipitated with saturated KPF ₆ . Yield: 26%, NMR (acetone-d6): 1Hδ 1.35 (m, 12H), 2.98 (t, 2H), 3.10 (s, 3H), 3.40 to 3.55 (dm, 2H), 3.80 (m, 8H), 6.97 (d, 2H), 7.20-7.40 (m, 6H), 7.60 (d, 1H), 7.67 (t, 1H) ), 7.70-7.85 (m, 4H), 7.88-7.98 (m, 4H), 8.17 (t, 1H), 8.30 (t, 1H), 8.56 ( dd, 2H), 8.69 (d, 1H), 8.72 (d, 1H), 9.55 (d, 1H), 10.08 (d, 1H).

(0230)
L = vinylacetonitrile (VACN) 20 mg Ru (bpy) ₂ Cl ₂ was suspended in 2 mL 96% EtOH and the suspension was heated at 80 ° C. until completely dissolved. The formation of [Ru (bpy) ₂ (H ₂ O) Cl] ⁺ complex was confirmed by its band at 490 nm (in water). 1.5 equivalents of VACN diluted with 4 mL of EtOH was added and the mixture was maintained at 80 ° C. for 2 hours. The solvent was removed by distillation under vacuum, the residue was redissolved in 2 mL water, the solid was removed by centrifugation and precipitation with saturated KPF ₆ was performed. The yield was 66%.

(0231)
NMR (acetone-d6): 1Hδ 3.65 (d, 2H), 5.13 (d, 2H), 5.75 (m, 1H), 7.29 (t, 1H), 7.32 (t, 1H) ), 7.70 (d, 1H), 7.81 (t, 1H), 7.92 (3t, 3H), 7.97 (d, 1H), 8.20 (t, 1H), 8.31 (T, 1H), 8.56 (d, 1H), 8.60 (d, 1H), 8.69 (d, 1H), 8.74 (d, 1H), 9.53 (d, 1H) 10.08 (d, 1H).

(0232)
Rhodamine is a useful fluorophore due to its close to 1 emission quantum yield and high resistance to photobleaching. Rhodamine is composed of a xanthene moiety that provides fluorescence and benzoic acid that adjusts its spectral properties. Scheme 1A shows the structure of rhodamine B selected as the starting point for this example.

(0233)
Ruthenium-bipyridyl complexes can be easily coordinated to donor nitrogen (aliphatic amine, pyridine, imine, and even nitrile nitrogen, etc.), but the aniline nitrogen is coordinated even if it is primary. do not do. Rhodamine B diethylaniline does not form a stable complex with the Ru center. Carboxylate can coordinate to the Ru-bpy core, but such complexes are easily hydrolyzed at room temperature in aqueous solution.
In view of these properties, the chemical structure of rhodamine B was modified by the addition of a “sticky tail” to coordinate the fluorescent molecule to the metal center. In the first attempt to synthesize a modified rhodamine having a coordinating group, the carboxylate was treated with the aminopropionitrile by the Adamczyk method (Adamczyk, MJ Bioorg. Med. Chem. Lett. 2000, 10: 1539-1541. ) Was used for amidation. The resulting compound is RhodB-APN shown in Scheme 1B. However, although this ligand can coordinate to a Ru-bpy complex via a nitrile, it causes isomerization to cyclic spirolactam at physiological pH (Scheme 1C) and becomes non-fluorescent. Rhodamine 6G can be modified to Rhodamine B as described above.

(0235)
Since secondary amides cannot isomerize into cyclic forms, RhodB-MAPN (Scheme 1D) obtained through amidation of rhodamine B acid chloride with aminopropionitrile exhibits very high yellow fluorescence, while The terminal nitrile allows coordination to the Ru-bpy center.
The complex cis- [Ru (bpy) ₂ (L) Cl] PF ₆ , L) RhodB-MAPN is obtained as a dark purple solid, which is slightly soluble in water and very soluble in acetone. it was high. Due to its extremely high molar extinction coefficient, its solubility in water was high enough for most experimental situations. For FIA imaging experiments, the complex was used in this form. A much more soluble chloride salt (more suitable for biological experiments) was prepared by batch ion exchange using Dowex 22 chloride resin in acetone / water (1: 1). The salt was used in biological experiments. The solution of the complex exhibits a weak yellowish fluorescence. Preliminary investigations have shown that this complex is photosensitive and increases its fluorescence when irradiated in aqueous ethanol or acetone.

(0237)
FIG. 19 (top) shows the aliphatic region of the ¹ HNMR spectrum of a Ru complex with RhodB-MAPN as a ligand, showing the number of signals and integrals expected for the coordinated RhodB-MAPN. . Two multiplets (denoted a ′) at 3.40 to 3.55 ppm correspond to the methylene group closest to the coordination point. The 2.98 ppm triplet (b ′) and 3.10 ppm singlet (c ′) correspond to the other methylene and N-methyl groups, respectively. Similar signals were obtained as long as the aminonitrile tail was coordinated to the Ru-bpy core. After 5 minutes of light irradiation in the NMR tube using a 450 nm LED (middle trace), new signal appears at 2.37, 3.16 and 3.53 ppm due to the release of RhodB-MAPN ligand. These three signals perfectly match the signal of the free ligand (labeled a, c, b, respectively, bottom trace). The four ethyl groups of RhodB-MAPN are not completely equivalent and appear as multiplets (—CH ₂ —) or broad triplets (—CH ₃ ) and show little change after light irradiation, although these groups This is because it is far from the coordination center.

によって求められ、式中、δｔは照射時間であり、Ｉは１秒あたりの光子のモル数での光強度であり、φ_PUは、光アンケージングの量子収率であり、Ａｂｓ_Rは、４７３ｎｍでの光活性種［Ｒｕ（ｂｐｙ）₂（ＲｈｏｄＢ−ＭＡＰＮ）Ｃｌ］⁺の吸光度であり、Ａｂｓ_Tは照射された溶液の総吸光度である。前出の方程式の有限要素法計算によって、光アンケージング量子収率φ_PU＝０．１２（２５℃）が計算された。この値は、同様の錯体に関して報告されている量子収率と十分に一致する。 (0238)
In the aromatic region, the expected number of signals and integrals were obtained for an asymmetric cis complex with RhodB-MAPN. After photolysis, the signal from the free RhodB-MAPN appears at 6.98, 7.20, 7.37, 7.62, 7.77, and 7.82 as the aco-complex [Ru (bpy) ₂ (H ₂ O) Appears with the same chemical shift as from Cl] ⁺ .
A diluted aqueous solution of [Ru (bpy) ₂ (RhodB-MAPN) Cl] ⁺ exhibits weak fluorescence with a maximum at 592 nm (excitation: 518 nm).
FIG. 20 is an emission spectrum of the complex during light irradiation. The maximum and shape of the initial emission spectrum is the same as that of the free ligand RhodB-MAPN, but the emission quantum yield is much lower, about φt = 0.04. The quantum yield is easily calculated by integrating the spectral area. After light irradiation with a 473 nm laser, RhodB-MAPN is released and the solution increases its fluorescence by 6 times to φt = 0.24. The inset graph shows the maximum of the spectrum as a function of irradiation time.
The quantum yield of rhodamine light emission was calculated after analysis of fluorescence emission data during photouncaging at 473 nm. A complete spectrum was obtained every second. Absorption of light by the complex [Ru (bpy) ₂ (RhodB-MAPN) Cl] ⁺ results in the acocomplex [Ru (bpy) ₂ (H ₂ O) Cl] ⁺ and free RhodB-MAPN. During each light irradiation period, the amount of aco-complex and free RhodB-MAPN produced is given by the following formula:

Where δt is the irradiation time, I is the light intensity in moles of photons per second, φ _PU is the quantum yield of photouncaging, and Abs _R is 473 nm Is the absorbance of the photoactive species [Ru (bpy) ₂ (RhodB-MAPN) Cl] ⁺ , and Abs _T is the total absorbance of the irradiated solution. The optical uncaging quantum yield φ _PU = 0.12 (25 ° C.) was calculated by the finite element method calculation of the above equation. This value is in good agreement with the reported quantum yields for similar complexes.

(0242)
The cause of this quenching can be attributed to the presence of an adjacent ruthenium-bipyridine moiety, which shows an MLCT transition that overlaps that of rhodamine emission. However, direct examination of the absorption band in this wavelength range of this complex is not possible because the molar extinction coefficient of the fluorescent ligand is very high (approximately 105 M ^-1 cm ^-1 ) and the absorption is weakened by the metal center. Is possible.
To measure this absorption, a similar complex was synthesized using vinylacetonitrile (VACN) instead of RhodB-MAPN. It can be seen that the energy of the MLCT band of the Ru-bpy complex depends largely on the nature and basicity of the ligand close to the Ru center, but is almost independent of the farther fragment. The complex [Ru (bpy) ₂ (VACN) Cl] ⁺ shows a typical MLCT band, spreads over 100 nm in the low energy region centering around 460 nm, and overlaps with the emission spectrum of the rhodamine ligand. This absorption is sufficient to explain the observed quenching. This similar complex emits VACN with a higher quantum yield φ _PU = 0.21.

(0244)
This example provides an activatable fluorescent probe that exhibits surprising sensitivity in the visible light region. This probe increases its intrinsic fluorescence up to 6 times after blue light irradiation. This probe is physiologically innocuous, and even excitable cells such as neurons can be injected into living cells and showed no signs of acute toxicity in short-term experiments (about 2 hours). This probe was used to image laminar flow inside the spectrophotometric laminar flow cell and visualize parabolic flow in FIA capillaries. This probe reveals not only the plume spread, but also the memory effect due to the accumulation of analyte near the wall where the flow velocity approaches zero. Simply put, the complex [Ru (bpy) ₂ (RhodB-MAPN) Cl] Cl is a surprisingly effective tool for imaging all types of systems that require fluorescence manipulation.
[Ru (bpy) ₂ (Rhod6G-MAPN) Cl] Cl and [Ru (bpy) ₂ (RhodB-MAMePy) Cl] are generated using the procedures described herein and [Ru (bpy) ₂ (RhodB-MAPN) Cl] showed similar results to those described herein for the photodissociation of formulas I, II, III, V, VI, VII and VIII described herein. Exemplifies the applicability of the optical antenna concept to sex compounds (L ² includes a labeled molecule or an active derivative thereof).

(0246)
The same surprising properties demonstrated in this example for [Ru (bpy) ₂ (RhodB-MAPN) Cl] Cl are the light of formulas I, II, III, V, VI, VII and VIII described herein. It is expected that this also applies to dissociable compounds (L ² includes a labeled molecule or an active derivative thereof).

Claims (69)

V
(Where
Each L ¹ is independently (a) a 5-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(B) a 6-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(C) an 8 to 10 membered bicyclic ring having a nitrogen atom member in which one of the bicyclic rings is aromatic and forms a bond with Ru;
(D) a —NH ₂ group in which the nitrogen atom forms a bond with Ru;
(E) a —COOH group in which one of the oxygen atoms forms a bond with Ru;
(F) phosphorus atoms form a bond and Ru, R is -H independently, -C ₁ ~C ₁₈ -PR ₂ groups are alkyl or aryl, or (g) a sulfur atom, a bond with Ru A —SR group, wherein R is independently —H, —C ₁ -C ₁₈ alkyl or aryl,
An organic molecule having
L ² is (R ⁹ ) ₃ P, (R ⁹ O) ₃ P or L ¹ , m is 2, or L ² is —CN, m is 1, or L ² is (R ⁹ ) A labeled molecule or an active derivative thereof bonded to Ru via a phosphorus atom of ₂ P or (R ⁹ O) ₂ P, m is 2, or L ² is NHR ⁹ , N (R ⁹ ) ₂ , pyridyl, C (R ⁹ ) ═NH, C (R ⁹ ) ═NR ⁹ is a labeled molecule or an active derivative thereof bonded to Ru via a cyclic aliphatic amine group or a nitrogen atom of a nitrile, 2
Each R ⁹ is independently —C ₁ -C ₁₈ alkyl, —C ₃ -C ₈ cycloalkyl or phenyl;
When L ² is a labeled molecule or an active derivative thereof linked to Ru via the phosphorus atom of P (phenyl) ₃ or P (phenyl) ₂ , each phenyl is independently —C ₁ -C ₁₈ alkyl, — (C ₁ -C ₁₈ alkyl) -OH, aryl, - (C ₁ -C ₁₈ alkyl) - oxy, - (C ₁ -C ₁₈ alkyl) - amino, - (C ₁ -C ₁₈ alkyl) - thio, - CO ₂ Y, —C (═O) Y, —C (═O) NY ₂ , —NH ₂ , —NO ₂ , —OH or —SH,
R ^{1 to} R ⁴ are independently —H, —C _{1 to} C ₁₈ alkyl, —NH ₂ , — (C _{1 to} C ₁₈ alkyl) —O— (C _{1 to} C ₁₈ alkyl), —OC (O). (C ₁ -C ₁₈ alkyl), - (C ₁ ~C ₁₈ alkyl) -OH, aryl, - (C ₁ ~C ₁₈ alkyl) - oxy, - (C ₁ ~C ₁₈ alkyl) - amino, - (C ₁ -C ₁₈ alkyl) - _{thio, -CO 2 Y, -C (=} O) Y, -C (= O) NY 2, an -NO ₂ or -SH, or R ¹ and and R ² and / Or R ³ and R ⁴ can combine to form a cyclic carbon substituted with one or more oxo groups,
When L ² is not P (phenyl) ₃ , R ^{1 to} R ⁴ are independently —H, — (C _{1 to} C ₁₈ alkyl) —OH, aryl, — (C _{1 to} C ₁₈ alkyl) -oxy, - (C ₁ -C ₁₈ alkyl) - amino, - (C ₁ ~C ₁₈ alkyl) - _{thio, -CO 2 Y, -C (=} O) Y, -C (= O) NY 2, -NO 2, -OH or -SH, or R ¹ and R ² and / or R ³ and R ⁴ may be combined to form a cyclic carbon substituted with one or more oxo groups, R ¹ to At least one of R ⁴ is not H,
X is Cl ⁻ , F ⁻ , Br ⁻ , I ⁻ , PF ₆ ⁻ , CF ₃ SO ₃ ⁻ , (C _{1 to} C ₁₈ alkyl) -CO ₂ ^— or (C _{1 to} C ₁₈ alkyl) —SO ₃ ^—. And
Y is selected from the group consisting of —H, —C ₁ -C ₁₈ alkyl, aryl, — (C ₁ -C ₁₈ alkyl) -aryl, —C ₃ -C ₈ cycloalkyl, heteroaryl and heterocyclyl. ) The compound according to claim 1, wherein the organic molecule L ¹ is 4-aminopyridine.   The compound of claim 1, wherein the organic molecule is (RS)-(tetrazol-5-yl) glycine.   The compound of claim 1, wherein the organic molecule is (tetrazol-5-yl) AMPA.   The compound according to claim 1, wherein the organic molecule is nicotine or caffeine.   The compound according to claim 1, wherein the organic molecule is serotonin, epinephrine, norepinephrine or dopamine.   The organic molecule is adenosine 5′-diphosphate ADP, adenosine 5′-triphosphate ATP, adenosine 5′-monophosphate AMP, cyclic adenosine 5′-diphosphate ribose or adenosine 3 ′, 5′-cyclic monophosphate. The compound according to claim 1.   The compound according to claim 1, wherein the organic molecule is aminobutyric acid, L-glutamic acid or methyl-D-aspartic acid. A method of releasing organic molecules from a photolabile compound,
A method comprising exposing the compound of claim 1 to light under conditions sufficient to release organic molecules.   The method of claim 9, wherein the light comprises a wavelength of about 300 to about 500 nm.   The method of claim 9, wherein the light comprises a wavelength of about 300 to about 360 nm.   The method of claim 9, wherein the light comprises a wavelength of about 450 to about 500 nm. The method of claim 9, wherein L ² is L ¹ .   The method of claim 9, wherein the light comprises visible light or infrared light.   The method of claim 9, wherein the exposure occurs at a temperature of about 0 ° C. to about 150 ° C. A method of making organic molecules bioavailable to patients,
(A) administering the compound of claim 1 to a patient;
(B) exposing the compound to light under conditions sufficient to release organic molecules from the compound;
The organic molecule is
(I) a 5-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(Ii) a 6-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(Iii) an 8- to 10-membered bicyclic ring having one nitrogen atom member that is aromatic and forms a bond with Ru, one of the bicyclic rings;
(Iv) a —NH ₂ group in which the nitrogen atom forms a bond with Ru;
(V) a —COOH group in which one of the oxygen atoms forms a bond with Ru;
(Vi) the phosphorus atom forms a bond with Ru, R is independently -H, -C ₁ ~C ₁₈ -PR ₂ groups are alkyl or aryl, or (vii) a sulfur atom, a bond with Ru A —SR group, wherein R is independently —H, —C ₁ -C ₁₈ alkyl or aryl,
A method characterized by comprising:   The method according to claim 16, wherein the light is sunlight, photooptic light, or laser light.   The method of claim 16, wherein the light is visible light or infrared light.   17. The method of claim 16, wherein the exposure occurs at the site of a tumor, cancer or neoplasm.   The method of claim 16, wherein the exposure occurs at the site of a blood disorder.   The administration is intravenous, topical, intradermal, intramuscular, transdermal, subcutaneous, intranasal, parenteral, intrathecal, intravaginal, rectal, colorectal, oral, intracranial, retroorbital, sternum 17. The method of claim 16, wherein the method is performed by an internal route or by injection.   17. The method of claim 16, wherein the administration is performed via a transdermal patch.   A composition comprising the compound of claim 1 and a physiologically acceptable carrier, vehicle, diluent or excipient.   A container storing the compound according to claim 1.   25. A container according to claim 24, further storing a biological sample.   26. A container according to claim 25, wherein the biological sample is an organ, tissue, cell or hair sample.   27. A container according to claim 26, wherein the tissue is neural tissue.   The container according to claim 26, wherein the cell is a nerve cell.   27. The container of claim 26, wherein the tissue or cell is a tumor, cancer or neoplastic tissue or cell.   26. A container according to claim 25, wherein the biological sample is a body fluid sample.   The container according to claim 30, wherein the body fluid sample is blood, serum, plasma, lymph, saliva, sputum, tears, semen or urine.   A kit comprising the compound of claim 1 and instructions for use of the compound. A method of treating a disorder and disease in a patient comprising
(A) administering to a patient a therapeutically effective amount of the compound of claim 1;
(B) exposing the compound to light under conditions sufficient to release organic molecules from the compound;
The organic molecule is
(I) a 5-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(Ii) a 6-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(Iii) an 8- to 10-membered bicyclic ring having one nitrogen atom member that is aromatic and forms a bond with Ru, one of the bicyclic rings;
(Iv) a —NH ₂ group in which the nitrogen atom forms a bond with Ru;
(V) -COOH group in which one of the oxygen atoms forms a bond with Ru;
(Vi) the phosphorus atom forms a bond with Ru, R is independently -H, -C ₁ ~C ₁₈ -PR ₂ groups are alkyl or aryl, or (vii) a sulfur atom, a bond with Ru A —SR group, wherein R is independently —H, —C ₁ -C ₁₈ alkyl or aryl,
A method characterized by comprising:   34. The method of claim 33, wherein the disorder and disease are neurological, neurophysiological or neuromuscular disease and condition.   35. The method of claim 34, wherein the neurological, neurophysiological or neuromuscular disease and condition is epilepsy.   35. The method of claim 34, wherein the neurological, neurophysiological or neuromuscular disease and condition is multiple sclerosis.   35. The method of claim 34, wherein the disorder is sweating.   35. The method of claim 34, wherein the disease is cancer. A method for improving the solubility of organic molecules,
Generating the compound of claim 1 by complexing an organic molecule with a photolabile caging group,
When the compound is exposed to light under sufficient conditions, the organic molecule is released from the compound, and the organic molecule is
(I) a 5-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(Ii) a 6-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(Iii) an 8- to 10-membered bicyclic ring having one nitrogen atom member that is aromatic and forms a bond with Ru, one of the bicyclic rings;
(Iv) a —NH ₂ group in which the nitrogen atom forms a bond with Ru;
(V) -COOH group in which one of the oxygen atoms forms a bond with Ru;
(Vi) the phosphorus atom forms a bond with Ru, R is independently -H, -C ₁ ~C ₁₈ -PR ₂ groups are alkyl or aryl, or (vii) a sulfur atom, a bond with Ru A method of forming and having a —SR group wherein R is independently —H, —C ₁ -C ₁₈ alkyl or aryl.

VI
(Where
Each L ¹ is independently (a) a 5-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(B) a 6-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(C) an 8- to 10-membered bicyclic ring having one nitrogen atom member that is aromatic and forms a bond with Ru, one of the bicyclic rings;
(D) a —NH ₂ group in which the nitrogen atom forms a bond with Ru;
(E) -COOH group in which one of the oxygen atoms forms a bond with Ru;
(F) phosphorus atoms, to form a bond with Ru, R is, independently -H, -C ₁ ~C ₁₈ -PR ₂ groups are alkyl or aryl, or (g) a sulfur atom, bound to Ru And R is independently an organic molecule having a —SR group that is —H, —C ₁ -C ₁₈ alkyl or aryl,
L ² is P (phenyl) ₃ , and each phenyl is independently -C ₁ -C ₁₈ alkyl,-(C ₁ -C ₁₈ alkyl) -OH, aryl,-(C ₁ -C ₁₈ alkyl)- oxy, - (C ₁ ~C ₁₈ alkyl) - amino, - (C ₁ ~C ₁₈ alkyl) - _{thio, -CO 2 Y, -C (=} O) Y, -C (= O) NY 2, -NH ₂ , —NO ₂ , —OH or —SH, or L ² is a labeled molecule or an active derivative thereof bonded to Ru via the phosphorus atom of (R ⁹ ) ₂ P or (R ⁹ O) ₂ P And m is 2, or L ² is NHR ⁹ , N (R ⁹ ) ₂ , pyridyl, C (R ⁹ ) ═NH, C (R ⁹ ) = NR ⁹ cyclic aliphatic amine group or nitrile A labeled molecule or an active derivative thereof bonded to Ru via a nitrogen atom, m is 2,
Each R ⁹ is independently —C ₁ -C ₁₈ alkyl, —C ₃ -C ₈ cycloalkyl or phenyl;
R ^{1 to} R ⁴ are independently —H, —C _{1 to} C ₁₈ alkyl, —NH ₂ , — (C _{1 to} C ₁₈ alkyl) —O— (C _{1 to} C ₁₈ alkyl), —OC (O ) (C ₁ -C ₁₈ alkyl), - (C ₁ -C ₁₈ alkyl) -OH, aryl, - (C ₁ -C ₁₈ alkyl) - oxy, - (C ₁ -C ₁₈ alkyl) - amino, - ( C ₁ -C ₁₈ alkyl) -thio, —CO ₂ Y, —C (═O) Y, —C (═O) NY ₂ , —NO ₂ or —SH, or R ¹ and R ² are And / or R ³ and R ⁴ can be combined to form a cyclic carbon substituted with one or more oxo groups;
X is Cl ⁻ , F ⁻ , Br ⁻ , I ⁻ , PF ₆ ⁻ , CF ₃ SO ₃ ⁻ , (C _{1 to} C ₁₈ alkyl) -CO ₂ ^— or (C _{1 to} C ₁₈ alkyl) —SO ₃ ^—. And
Y is selected from the group consisting of —H, —C ₁ -C ₁₈ alkyl, aryl, — (C ₁ -C ₁₈ alkyl) -aryl, —C ₃ -C ₈ cycloalkyl, heteroaryl and heterocyclyl. ) L ² is a P (phenyl) _3, each phenyl is substituted at the 3-position or 4-position, independently, a compound according to claim 40. L ² is a P (phenyl) _3, at least one phenyl - is substituted with (C ₁ -C ₁₈ alkyl) -OH, A compound according to claim 40. L ² is P (phenyl) _3, each phenyl, - is substituted by (C ₁ -C ₁₈ alkyl) -OH, A compound according to claim 40 L ² is P (phenyl) _3, at least one phenyl is substituted by -COOH, the compound according to claim 40. L ² is a P (phenyl) _3, each phenyl is substituted with -COOH, the compound according to claim 40. L ² is a P (phenyl) _3, at least one phenyl is substituted by -OH, A compound according to claim 40. L ² is a P (phenyl) _3, each phenyl is substituted with -OH, A compound according to claim 40. L ² is a P (phenyl) _3, at least one phenyl is substituted with -NH _2, A compound according to claim 40. L ² is a P (phenyl) _3, each phenyl is substituted with -NH _2, A compound according to claim 40. L ² is P (phenyl) _3, at least one phenyl is substituted by -NO _2, A compound according to claim 40. L ² is P (phenyl) _3, each phenyl is substituted with -NO _2, A compound according to claim 40.

VII
(Where
Each L ¹ is independently (a) a 5-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(B) a 6-membered monocyclic aromatic ring in which one of the ring members is a nitrogen atom that forms a bond with Ru;
(C) an 8- to 10-membered bicyclic ring having one nitrogen atom member that is aromatic and forms a bond with Ru, one of the bicyclic rings;
(D) a —NH ₂ group in which the nitrogen atom forms a bond with Ru;
(E) a —COOH group in which one of the oxygen atoms forms a bond with Ru;
(F) phosphorus atoms, to form a bond with Ru, R is, independently -H, -C ₁ ~C ₁₈ -PR ₂ groups are alkyl or aryl, or (g) a sulfur atom, bound to Ru And R is independently an organic molecule having a —SR group that is —H, —C ₁ -C ₁₈ alkyl or aryl,
L ² is (R ⁹ ) ₃ P, (R ⁹ O) ₃ P or L ¹ , and each R ⁵ is independently —C ₁ -C ₁₈ alkyl, —C ₃ -C ₈ cycloalkyl or phenyl. And m is 2, L ² is not P (phenyl) ₃ , or L ² is —CN, m is 1, or L ² is (R ⁹ ) ₂ P Or a labeled molecule or an active derivative thereof bound to Ru via a phosphorus atom of (R ⁹ O) ₂ P, m is 2, or L ² is NHR ⁹ , N (R ⁹ ) ₂ , Pyridyl, C (R ⁹ ) = NH, C (R ⁹ ) = NR ⁹ is a labeled molecule bonded to Ru via a cyclic aliphatic amine group or a nitrogen atom of a nitrile or an active derivative thereof, and m is 2 Yes,
Each R ⁹ is independently —C ₁ -C ₁₈ alkyl, —C ₃ -C ₈ cycloalkyl or phenyl;
R ^{1 to} R ⁴ are independently -H,-(C ₁ -C ₁₈ alkyl) -OH, aryl,-(C ₁ -C ₁₈ alkyl) -oxy,-(C ₁ -C ₁₈ alkyl) -amino. , — (C ₁ -C ₁₈ alkyl) -thio, —CO ₂ Y, —C (═O) Y, —C (═O) NY ₂ , —NO ₂ , —OH or —SH, or R ^1. And R ² and / or R ³ and R ⁴ may combine to form a cyclic carbon substituted with one or more oxo groups, wherein at least one of R ^{1 to} R ⁴ is not H;
X is Cl ⁻ , F ⁻ , Br ⁻ , I ⁻ , PF ₆ ⁻ , CF ₃ SO ₃ ⁻ , (C _{1 to} C ₁₈ alkyl) -CO ₂ ^— or (C _{1 to} C ₁₈ alkyl) —SO ₃ ^—. And
Y is selected from the group consisting of —H, —C ₁ -C ₁₈ alkyl, aryl, — (C ₁ -C ₁₈ alkyl) -aryl, —C ₃ -C ₈ cycloalkyl, heteroaryl and heterocyclyl. ) L ² is a ₂ P (methyl) (phenyl) The compound according to claim 52. L ² is a P (methyl) ₂ (phenyl) The compound according to claim 52.

VIII
(Where
Each L ¹ is independently a labeled molecule or an active derivative thereof bound to Ru via a phosphorus atom of (R ⁹ ) ₂ P or (R ⁹ O) ₂ P, m is 2, or L ¹ is bonded to Ru via NHR ⁹ , N (R ⁹ ) ₂ , pyridyl, C (R ⁹ ) ═NH, C (R ⁹ ) = NR ⁹ cyclic aliphatic amine group or nitrile nitrogen atom. A labeling molecule or an active derivative thereof, m is 2,
Each R ⁹ is independently —C ₁ -C ₁₈ alkyl, —C ₃ -C ₈ cycloalkyl or phenyl;
L ² is Cl—, phosphine, OH ₂ or pyridine, m is 2, or L ² is —CN, m is 1.
R ^{1 to} R ⁴ are independently —H, —C _{1 to} C ₁₈ alkyl, —NH ₂ , — (C _{1 to} C ₁₈ alkyl) —O— (C _{1 to} C ₁₈ alkyl), —OC (O ) (C ₁ -C ₁₈ alkyl), - (C ₁ -C ₁₈ alkyl) -OH, aryl, - (C ₁ -C ₁₈ alkyl) - oxy, - (C ₁ -C ₁₈ alkyl) - amino, - ( C ₁ -C ₁₈ alkyl) -thio, —CO ₂ Y, —C (═O) Y, —C (═O) NY ₂ , —NO ₂ or —SH, or R ¹ and R ² are And / or R ³ and R ⁴ can be combined to form a cyclic carbon substituted with one or more oxo groups;
X is Cl ⁻ , F ⁻ , Br ⁻ , I ⁻ , PF ₆ ⁻ , CF ₃ SO ₃ ⁻ , (C _{1 to} C ₁₈ alkyl) -CO ₂ ^— or (C _{1 to} C ₁₈ alkyl) —SO ₃ ^—. And
Y is selected from the group consisting of —H, —C ₁ -C ₁₈ alkyl, aryl, — (C ₁ -C ₁₈ alkyl) -aryl, —C ₃ -C ₈ cycloalkyl, heteroaryl and heterocyclyl. )   56. The compound of claim 55, wherein the fluorescent molecule contains a -CN group in which a nitrogen atom forms a bond with Ru.   56. The compound of claim 55, wherein the fluorescent molecule contains rhodamine.   56. The compound of claim 55, wherein the fluorescent molecule contains fluorescein.   56. The compound of claim 55, wherein the fluorescent molecule contains iodoeosin. A method of releasing fluorescent molecules from a photolabile compound,
56. A method comprising exposing the compound of claim 55 to light under conditions sufficient to release a fluorescent molecule.   61. The method of claim 60, wherein the light comprises a wavelength of about 300 to about 500 nm.   61. The method of claim 60, wherein the light comprises a wavelength of about 300 to about 360 nm.   61. The method of claim 60, wherein the light comprises a wavelength of about 450 to about 500 nm.   61. The method of claim 60, wherein the light comprises visible light or infrared light.   61. The method of claim 60, wherein the exposure occurs at a temperature from about 0C to about 150C. L ² is a labeling molecule or a derivative thereof, The method of any of claims 1,40,52 or 55. L ² is rhodamine B- methyl aminopropionitrile amide (RhodB-MAPN), rhodamine 6G- methyl aminopropionitrile amide (Rhod6G-MAPN), a RhodB-MAMePy, Rhod6G-MAMePy or salts thereof, according to claim 66. The method according to 66. [Ru (bpy) ₂ (RhodB-MAPN) Cl] Cl, [Ru (bpy) ₂ (Rhod6G-MAPN) Cl] Cl, [Ru (bpy) ₂ (RhodB-MAMePy) Cl] Cl Compound. Z is an anion [Ru (bpy) ₂ (RhodB-MAPN) Cl] Z, [Ru (bpy) ₂ (Rhod6G-MAPN) Cl] Z, [Ru (bpy) ₂ (RhodB-MAMePy) Cl] Z A compound selected from the group consisting of:

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