JPH0597857A - Porphyrin derivative and its use - Google Patents

Abstract

PURPOSE:To provide a new (metallo)porphyrin useful as a sensitizer for photo- physico-chemical diagnosis and therapy (PDT) composed of single component, being excellent instability, water-solubility and accumulation to cancer tissue and quickly releasable from normal tissue to enable the reduction of phototoxicity. CONSTITUTION:The compound of formula I [R1 is CH(OR)CH3; R is alkyl; R2 is residue obtained by removing H from amino acid; M is 2H, Ga, Zn, etc.], its metal complex or a compound of formula II [R2 is OH, amino acid residue, etc.; R3 is CH=CH2, CH(OR)CH, etc.; R4 is CHO, CH(OH)OSO2Na, etc.] (including the position isomer obtained by exchanging the functional groups on the side chains of the ring A and the ring B), e.g. 2,4-bis(l-propoxyethyl)- deuteroporphinyl-diaspartic acid. The compound of formula I can be produced by introducing a metal to a porphyrin compound having R1 and bonding an amino acid residue R2 to the product. The compound has an absorption wavelength of <=600nm and >=670nm to enable the use of a titanium sapphire laser.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a porphyrin derivative and its use, and particularly to a photophysical chemical and diagnostic sensitizer and / or photophysical chemistry containing a novel porphyrin and metalloporphyrin derivative as an active ingredient. The present invention relates to a drug used for diagnosis and treatment of cancer.

[0002]

2. Description of the Related Art Photophysicochemical diagnostic treatment (PDT) is performed as a new treatment method for cancer. In this method, a porphyrin compound of a certain kind is administered by a method such as intravenous injection, and is retained in the cancer tissue, and then laser light is irradiated to selectively destroy only the cancer tissue. PDT utilizes two properties of porphyrin, that it is retained in cancer tissue for a longer time than in normal tissue and that it has a photosensitizing effect. Over the past 13 years, more than 3000 people have been treated with PDT for malignant tumors, and it is becoming established as one of the cancer treatment methods. Cancer types for which good treatment results have been reported by PDT include retinal cancer, skin cancer,
It is widespread, including esophageal cancer, superficial bladder cancer, and early stage lung cancer.

Drugs currently used in PDT are mainly hematoporphyrin derivatives (HPD) and their ether and / or ester dimers. HP
D is a mixture obtained by treating hematoporphyrin with sulfuric acid in acetic acid and further treating with 0.1N sodium hydroxide. The dimer mainly contains the highly hydrophobic component of HPD, is a complex mixture with HPD, and the active component is unknown. Further, the therapeutic effect is extremely unstable because the component ratio is not constant.

On the other hand, as a new porphyrin derivative for PDT, _{one in} which R ₁ is a lower alkyl group and R ₂ is a quaternary ammonium salt derivative is disclosed in JP-A-1-246286.
Nos. 63-145283, 62-205082, and 62-167873, the derivatives having an ether bond as R ₁ are disclosed in JP-A-62-2499.
No. 86, No. 62-246580, No. 62-24657
9 and 62-205081, and J. F.
Evensen et al. [Br. J. Cancer, 5
5, 483 (1987)]. Further, chlorin derivatives are disclosed in JP-A-1-250381 and Sho 63-2.
90881, Sho 62-5986, Sho 62-5985
No. 62-5924, 62-5912, 58
-981 and 57-185220 disclose porphyrin dimer derivatives in US Pat. No. 4,649,151 (19).
87), JP-A Nos. 62-63586 and 60-50.
No. 0132 discloses a porphyrin metal complex.
1382, Sho 63-104987 and Sho 57-3
1688. We also conducted various studies and investigated chlorin derivatives as disclosed in JP-A-61-7279 and JP-A-60-9.
No. 2287, a porphyrin metal complex was disclosed in JP-A-2-13.
8280, Hei2-76881, Sho 62-18266
3, Sho 62-174079 and Sho 61-8318.
No. 5, a bacteriochlorin derivative is disclosed in JP-A-63-19.
No. 6586. However, in order to use it as a sensitizer for PDT, the above-mentioned compounds are not suitable for synthesis, stability,
Practical application was difficult in terms of water solubility.

There is also a problem of tissue transparency of laser light used for PDT. HPD and its dimer have a maximum absorption wavelength of 630 nm and a low molar absorption coefficient of 3000.
With 630 nm light, the tissue permeability is poor, and the therapeutic effect of PDT is limited to superficial cancer of 5 to 10 mm.

On the other hand, the laser device also has a problem. The dye lasers that are most commonly used today have poor stability and are difficult to handle in operation. Using a titanium sapphire laser, which has recently been attracting attention, makes operation considerably easier. However, with this laser, 670 nm or more 60
It is limited to the absorption wavelength of 0 nm or less and cannot be applied to HPD or its dimer having the absorption wavelength near 630 nm.

Further, it is known that side effects of drugs cause temporary photosensitivity. Therefore, after administration of the drug, the patient must be kept in a dark place for a long time so that normal tissues such as skin are not destroyed by the photosensitizing effect. Since HPD and its dimer are slowly excreted from normal tissues, photosensitivity may remain for 6 weeks or longer at a long time. The drugs currently used have many of these problems, and development of new drugs to replace HPD and its dimer is strongly desired. Therefore, as a second-generation drug, a compound that is a single compound and has absorption in a longer wavelength region (650 to 800 nm) has been proposed to overcome the drawbacks of the above drugs. Various compounds such as azaporphyrins such as phthalocyanine, porphyrins such as chlorin and bacteriochlorin, ring-expanded porphyrins such as texaphyrin are currently available on HP.
It is being studied as an alternative drug for D and its dimer.

[0008]

DISCLOSURE OF THE INVENTION The present inventors have found that a single component, which is stable and has a good accumulation property to cancer tissues, has a fast elimination rate from normal tissues and reduces phototoxicity. Titanium sapphire laser (67
Various studies have been conducted for the purpose of providing a photosensitizer suitable for PDT by searching for a porphyrin derivative that can be used in a wavelength range of 0 nm to 600 nm).

[0009]

As a result, the derivative of the previous application [JP-A-2-138280, US application 375482 (1989), European application 89112955.3 (19)
89)], a certain metalloporphyrin derivative to which a specific side chain having a polyfunctional group is bound, and a certain alkoxyl group in chlorins synthetically derivatized from blood-derived protoporphyrin or By combining groups with various functional groups and specific side chains containing amino acid residues, a single component provides excellent accumulation in cancer tissue and better excretion than normal tissue, and metal porphyrin derivative Was found to have an absorption wavelength of 600 nm or less, and the synthetic chlorinated derivative had an absorption wavelength of 670 nm or more, and had a good PDT effect.

Further, the inventors of the present invention analyzed the ultraviolet absorption (UV) spectrum of a mixed solution of porphyrin and albumin during the course of this research and development, and found that the trend of the spectrum has a certain law and affinity to specific organs, especially cancer. I found that.

The present invention has been completed based on the above findings, and its gist is the general formula (I): ## STR1 ## wherein R ₁ is CH (OR) CH ₃ and R ₂ is hydrogen from an amino acid. R is alkyl, M is 2H, G
a, Zn, Pd, In, Sn), a porphyrin compound or a metal complex thereof. Formula (II) of 2 (wherein, _{R 3} is _{CH = CH 2, CH (OR} ) CH 3, C
HO, CH = NOH or CH ₂ OH, R ₄ is CH =
_{X, C (OH) OSO 2} Na, CH (SCH 2 -COO
H) ₂ , CH (OR) ₂ or Chemical formula 5, X is O, C (C
N) ₂ , N-W or C (Y) Z, W is OH, OCO
CH ₃ or NH-E, E is H, alkyl, COC ₅
H ₄ N, CONH ₂ , CSNH ₂ , COOCH ₃ , CO
CH ₂ NCl (CH ₃ ) ₂ or C (NH ₂ ) ═NH,
Y is H or alkyl, Z is NO ₂ or COF, or a cyclic structure represented by Chemical formula 6, F is methyl, phenyl,
A residue of aminophenyl or yonone, R ₂ is a residue obtained by removing hydrogen from OH, an amino sugar or an amino acid, and R is an alkyl). (However, in the formula, it also includes positional isomers in which the functional groups of the side chains of the A and B rings of the four tetrapyrrole rings are replaced with each other.)

[0012]

[Chemical 5]

[0013]

[Chemical 6]

The term "alkyl" used in the meaning of each symbol above has 20 or less carbon atoms, preferably 1 carbon atom.
~ 18 alkyl (eg methyl, ethyl, hexyl,
Octyl, decyl, undecyl, dodecyl, tetradecyl, octadecyl, etc.). The term "amino acid" means an essential amino acid.

The porphyrin compound of the present invention can be produced by a conventional method. In the porphyrin compound corresponding to the general formula (I), _one having R ₁ is constituted (step a), and then the process proceeds to the next step c as it is or a metal is introduced into this (step b) to obtain the compound. A residue of an amino acid is attached to R ₂ of the obtained porphyrin compound and their metal complex (step c). Further, it is not always necessary to sequentially react with the steps (a), (b) and (c), and for example, the steps (b), (a) and (c) or the steps (a), (c) and (b) are performed. The order of steps may be changed.

On the other hand, in the porphyrin compound corresponding to the general formula (II), one having R ₂ is first constituted (step d), and then this is derivatized into a compound having R _{4 and} especially an aldehyde group (step e), adding or condensing various compounds to the obtained chlorin derivative (step f),
Then, if necessary, amino sugar and amino acid residues are bound (step c).

The construction steps (a, b, d and e) are described in J.
E. Falk [Porphyrins and Me
talporphyrins] (Elsevier
Issued, 1975) and D.I. Dolphin [Th
e Porphyrins] (Academic Pr
This can be done by the conventional method described in Ess, 1978).

For example, the porphyrin compounds having R ₁ corresponding to (I) and their metal complexes are:
JP-A-61-2279, JP-A-61-83185,
Patent Publication No. 63-13997, Japanese Patent Laid-Open No. 2-76881
No. 2-138280 and Japanese Patent Application No. 2-168.
It may be prepared according to the method described in 4990. That is, in step (a), R may be introduced into the R ₁ side chain of the porphyrin compound (I), the Br derivative of (I) is prepared in advance, and the reaction proceeds between this and the hydroxyl group of the alcohol. Alternatively, a Lewis acid is preferably used for the reaction, but the reaction is not limited to this. For step (b), usually metal chlorides,
This is done using acetates, sulphates, nitrates and the like. As the kind of metal, Ga, Z which has an effect of extending the phosphorescence lifetime
Examples thereof include n, Pd, In, Sn and the like. Further, the metal introduction step (b) may be prepared before or after the step (a) if necessary. For example, first, the metal introduction step (b) is carried out, then a Br derivative of a metalloporphyrin compound is prepared and reacted with alcohols, or a Lewis acid is used to react with alcohols (step a). There is no problem with proceeding. Instead of artificially synthesizing
It may be harvested from natural sources such as plants and animals.

The porphyrin compound and the metal complex thereof constructed as described above are then subjected to the step (c) of binding amino acid residues. That is, a porphyrin compound in which R ₂ is a hydroxyl group or a metal complex (I) thereof is reacted with an amino acid to produce a porphyrin compound in which R ₂ is an amino acid or a metal complex (I) thereof. Izumiya et al. [Basics and experiments of peptide synthesis] (published by Maruzen, 1985)
This can be carried out by a conventional method described in, for example, JP-A 64-61481 and JP-A-2-768.
No. 81, JP-A-2-138280 and Japanese Patent Application No. 2-1.
It may be prepared according to the method described in 68499. Instead of artificially synthesizing it, it may be harvested from natural sources such as plants and animals.

[0020] In this case, since the short may be introduced residue of an amino acid in the side chain of a porphyrin compound, R ₂ of (I)
It is preferable to proceed the reaction between the carboxyl group of the side chain and the amino group of the amino acid, so that the former carboxyl group and / or the latter amino group is converted into a conventional reactive group or present in both. Appropriate protection of functional groups not desired to participate in the reaction may be considered. In any case, the use of a reaction accelerator such as a dehydrating agent or a deoxidizing agent or a condensing agent may be considered.

Hereinafter, the preparation of the porphyrin compound and the metal complex (I) thereof will be described more specifically with reference to representative examples. For example, a porphyrin compound in which R ₂ has a hydroxyl group and a complex thereof (Japanese Patent Application Laid-Open Nos. 61-7279 and 61-61).
No. 83185, Japanese Patent Publication No. 63-13997, Japanese Patent Application Laid-Open No. 2-76881, Japanese Patent Application Laid-Open No. 2-138280 or Japanese Patent Application No. 2-168499), a condensing agent such as dicyclohexylcarbodiimide (for example, dicyclohexylcarbodiimide ( DCC) and water-soluble carbodiimide (WS
C)] or the like to obtain a porphyrin compound having an amino compound bonded to the side chain of R ₂ or a metal complex (I) thereof. Specific examples thereof include the following.

(1) 2,4-bis (1-propoxyethyl) -deuteroporfinyl diaspartic acid (hereinafter referred to as C ₃ -DP-diAsp) (2) 2,4-bis (1-butoxyethyl) - Deuteroxyl Pol nyl di-aspartate (hereinafter _C 4 -DP-
(3) 2,4-bis (1-pentyloxyethyl) -deuteroporfinyl diaspartic acid (hereinafter C ₅ -D)
(P-diAsp) (4) 2,4-bis (1-hexyloxyethyl) -deuteroporfinyl diaspartic acid (hereinafter C ₆ -D
(P-diAsp) (5) 2,4-bis (1-octyloxyethyl) -deuteroporfinyl diaspartic acid (hereinafter C ₈ -D)
P-diAsp) (6) 2,4-bis (1-decyloxyethyl) -deuteroporfinyl diaspartic acid (hereinafter referred to as C ₁₀ -D)
P-diAsp) (7) 2,4-bis (1-butoxyethyl) -Ga-deuteroporfinyl diaspartic acid (hereinafter C ₄ -G)
a-DP-diAsp) (8) 2,4-bis (1-pentyloxyethyl) -G
a- Dew terrorism Pol alkylsulfinyl Jiserin (below _{C 5} -
Ga-DP-diSer) (9) 2,4-bis (1-pentyloxyethyl) -G
a-deuteroporfinyl diaspartic acid (hereinafter referred to as C ₅ -Ga-DP-diAsp) (10) 2,4-bis (1-hexyloxyethyl)-
Ga-deuteroporfinyl diaspartic acid (hereinafter referred to as C ₆ -Ga-DP-diAsp) (11) 2,4-bis (1-heptyloxyethyl)-
Ga-deuteroporfinyl diaspartic acid (hereinafter referred to as C ₇ -Ga-DP-diAsp) (12) 2,4-bis (1-octyloxyethyl)-
Ga-deuteroporfinyl diglycine (hereinafter C ₈
-Ga-DP-diGly) (13) 2,4-bis (1-octyloxyethyl)-
Ga- Dew terrorism Pol alkylsulfinyl Jiserin (below _{C 8} -
Ga-DP-diSer) (14) 2,4-bis (1-octyloxyethyl)-
Ga- Deuteroxyl Pol nyl di-aspartate (hereinafter referred to as _{C 8 -Ga-DP-diAsp)} (15) 2,4- bis (1-octyloxy-ethyl) -
Ga- Deuteroxyl Pol nyl di glutamic acid (hereinafter referred to as _{C 8 -Ga-DP-diGlu)} (16) 2,4- bis (1-octyloxy-ethyl) -
Ga-deuteroporfinil dityrosine (hereinafter C ₈
-Ga-DP-diTyr) (17) 2,4-bis (1-nonyloxyethyl) -G
a-deuteroporfinyl diaspartic acid (hereinafter referred to as C ₉ -Ga-DP-diAsp) (18) 2,4-bis (1-decyloxyethyl) -G
a-Deuteroporfinyl diaspartic acid (hereinafter referred to as C ₁₀ -Ga-DP-diAsp) (19) 2,4-bis (1-dodecyloxyethyl)-
Ga-deuteroporfinyl diaspartic acid (hereinafter referred to as C ₁₂ -Ga-DP-diAsp) (20) 2,4-bis (1-hexyloxyethyl)-
Zn-deuteroporfinyl diaspartic acid (hereinafter referred to as C ₆ -Zn-DP-diAsp) (21) 2,4-bis (1-hexyloxyethyl)-
Pd-deuteroporfinyl diaspartic acid (hereinafter referred to as C ₆ -Pd-DP-diAsp) (22) 2,4-bis (1-hexyloxyethyl)-
In-deuteroporphinyl diaspartic acid (hereinafter referred to as C ₆ -In-DP-diAsp) (23) 2,4-bis (1-hexyloxyethyl)-
Sn-deuteroporfinyl diaspartic acid (hereinafter referred to as C ₆ -Sn-DP-diAsp) (24) 2,4-bis (1-decyloxyethyl) -Z
n-deuteroporfinyl diaspartic acid (hereinafter referred to as C ₁₀ -Zn-DP-diAsp) (25) 2,4-bis (1-decyloxyethyl) -P
d-deuteroporfinyl diaspartic acid (hereinafter referred to as C ₁₀ -Pd-DP-diAsp) (26) 2,4-bis (1-decyloxyethyl) -I
n-deuteroporfinyl diaspartic acid (hereinafter referred to as C ₁₀ -In-DP-diAsp) (27) 2,4-bis (1-decyloxyethyl) -S
n-deuteroporfinyl diaspartic acid (hereinafter referred to as C ₁₀ -Sn-DP-diAsp)

On the other hand, the porphyrin compound having R of R ₃ corresponding to (II) may be prepared according to the method described in the above step a. See also R ₃
Aldehyde group of the compound described below, 1-hydroxy-2-formylethylidene-protoporphyrin dimethyl ester (hereinafter referred to as photoprotoporphyrin dimethyl ester) by photochemical reaction treatment of protoporphyrin dimethyl ester (hereinafter referred to as PP-Me). (Step e) is prepared, and then this is reduced to give 2-formyl-4-vinyl-deuteroporphyrin dimethyl ester (hereinafter referred to as spirography porphyrin dimethyl ester) with an oxidizing agent such as periodic acid (hereinafter referred to as "spirography porphyrin dimethyl ester"). Step d). (However, the functional groups of the side chains of the A and B rings of the four tetrapyrrole rings were replaced with each other.
-Including formyl-2-vinyl form. ) It is preferable to use a photochemical reaction for the chlorination step (e),
A chlorin compound (photoprotoporphyrin) in which R ₄ has an aldehyde group is obtained. Instead of artificially synthesizing it, it may be harvested from natural sources such as plants and animals.

Next, the chlorin compound constituted as described above is subjected to the addition and / or condensation step (f). That is, in the porphyrin compound (II) in which R ₄ is an aldehyde group, sodium bisulfite, mercaptal,
An alcohol porphyrin compound is reacted to produce an adduct porphyrin compound, and a hydroxylamine, hydrazine, malonic acid derivative or the like is reacted to produce a condensed porphyrin compound. This can be carried out by a conventional method described in the general organic chemistry experimental manual [addition or condensation reaction of alcohol, hydroxylamine, hydrazone, semicarbazone, aminothiophenol, malonic acid ester and the like with an aldehyde compound]. it can. In any case, the use of a reaction accelerator such as a dehydrating agent or a deoxidizing agent or a condensing agent may be considered.

The preparation of the porphyrin compound (II) will be described in more detail below with reference to representative examples. For example, chlorin compound (photoprotoporphyrin derivative) in which R ₄ has an aldehyde group is reacted with sodium hydrogen sulfite or the like in a solvent to obtain a porphyrin compound (II) to which sodium hydrogen sulfite is added. On the other hand, compounds having an amino group in the photoprotoporphyrin derivative (for example, hydroxylamine, hydrazine derivative, semicarbazide, aminothiophenol, etc.) and compounds having an active hydrogen group (for example, malonnitrile, barbituric acid, acetone, acetophenone, aminoacetophenone, Yonone or the like) is reacted in a solvent with a condensing agent (eg, pyridine, piperidine, acid, alkali, etc.) to obtain a porphyrin compound (II) in which the side chain of R ₄ is condensed with these compounds. Specific examples thereof include the following.

(28) 2-vinyl-3-hydroxy-4
- formyl ethylidene - protoporphyrin dimethyl ester (photo protoporphyrin dimethyl ester) sodium bisulfite adduct (hereinafter NaHSO ₃ -
(29) Photoprotoporphyrin dimethyl ester acetic acid mercaptal (hereinafter referred to as HOAcS-P-Me) (30) Photoprotoporphyrin diethyl acetal (hereinafter referred to as EtO-P) (31) 2- (1- Hexyloxyethyl) -3-hydroxy-4-formylethylidene-protoporphyrin (C ₆ -photoprotoporphyrin) (hereinafter referred to as C ₆ -P) (32) C ₆ -photoprotoporphyrin diaspartic acid (hereinafter referred to as C _6-) P-diAsp say) (33) 2- (1-decyl-oxy-ethyl) -3-hydroxy-4-formyl-ethylidene - protoporphyrin _{(C 10} - photo protoporphyrin) (hereinafter _{C 10} -
(34) C ₁₀ -photoprotoporphyrin diaspartic acid (hereinafter referred to as C ₁₀ -P-diAsp) (35) Photoprotoporphyrin oxime (hereinafter referred to as N)
OH-P) (36) 2-formyl-3-hydroxy-4-formylethylidene-protoporphyrin (spirographis photoprotoporphyrin) (hereinafter SP) (37) photoprotoporphyrin trimethylacetohydrazone chloride (hereinafter G) 'THZ-P' (38) Photoprotoporphyrin hydrooxamic acid (hereinafter referred to as N2OH-P) (39) Spirographies photoprotoporphyrin
Dioxime (hereinafter referred to as 2NOH-SP) (40) Photoprotoporphyrin acetate oxime (hereinafter referred to as NOAc-P) (41) Photoprotoporphyrin ^t -butylhydrazone (hereinafter referred to as BuHZ-P) (42) Photoprotoporphyrin Guanylhydrazone (hereinafter referred to as GHZ-P) (43) Photoprotoporphyrin hydrazone nicotinic acid (hereinafter referred to as NHZ-P) (44) Photoprotoporphyrin nicotinic acid hydrazone diaspartic acid (hereinafter NHZ-P-diAsp)
(45) Photoprotoporphyrin carbonate hydrazone (hereinafter referred to as CHZ-P) (46) Photoprotoporphylidene barbituric acid (hereinafter referred to as BA-P) (47) Photoprotoporphyrin semicarbazone (hereinafter referred to as NH ₂ CONHN- (48) Photoprotoporphyrin thiosemicarbazone (hereinafter referred to as NH ₂ CSNHN-P) (49) Photoprotoporphyrin benzothiazole (hereinafter referred to as BT-P) (50) Photoprotoporphyrin malonnitrile (hereinafter referred to as “P”) MCN-P say) (51) referred to as photo protoporphyrinogen den nitromethylene (hereinafter _NO 2 -P) (52) referred to as photo protoporphyrinogen Den nitroethylene (hereinafter MeNO ₂ -P) (53) photo protoporphyrinogen Den Nitoropuropire (Hereinafter referred to as Etno ₂ -P) (54) (hereinafter referred to as _Me 2 CO-P) Photo protoporphyrinogen Den acetone (55) Photo protoporphyrinogen Den acetonedicarboxylic aspartic acid (hereinafter referred to as _Me 2 _CO-P-diAsp) (56) Photoprotoporphylidene ionone diaspartic acid (hereinafter referred to as Io-P-diAsp) (57) Photoprotoporphylidene ionone oxime diaspartic acid (hereinafter NOH-Io-P-diAs)
(58) Photoprotoporphylidene acetophenone (hereinafter PhCO-P) (59) Photoprotoporphylidene aminoacetophenone (hereinafter NH ₂ PhCO-P) (60) Photoprotoporphyrinyl oxime aminoglucose (Hereinafter referred to as NOH-P-NHglu)

The pharmaceutical preparation of the porphyrin derivative according to the present invention is produced by a method known per se, and the derivative according to the present invention may be dissolved in a suitable buffer solution. Suitable additives are, for example, pharmaceutically acceptable solubilizers (eg organic solvents), pH adjusters (eg acids, bases, buffers),
Stabilizers (eg ascorbic acid), excipients (eg glucose), tonicity agents (eg sodium chloride) and the like may be added.

The drug according to the present invention has necessary and sufficient properties as a drug for PDT, ie, long phosphorescence lifetime, affinity for albumin, specific accumulation property for specific organs, especially cancer, photokilling effect, absorption wavelength, water solubility, purity, etc. Is fully satisfied. The good water solubility of the drug according to the invention is
It allows the production of highly concentrated solutions (50 mg / ml), and the drug according to the invention shows a high stability not only in vitro but also in vivo. Generally, it is desirable to administer the drug of the present invention in an amount of 1 mg to 5 mg / kg body weight for application as a drug for PDT.

[0029]

The porphyrin compound according to the present invention has a chemical structure in that the side chain of the porphyrin skeleton has an alkoxy residue / amino acid residue, or an aldehyde residue and its adduct / condensate, or a metal complex in the porphyrin skeleton. And thus exerts various physiological or pharmacological properties.

These porphyrin derivatives selectively accumulate in cancer cells and are slowly excreted from cancer cells. Since it is rapidly excreted from normal organs and cells, it does not damage them. Originally, most of the porphyrin derivatives have a strong effect on light, but by introducing a polyfunctional compound residue into the side chain of the porphyrin derivative according to the present invention, it is possible to enhance excretion from normal tissues. , A derivative designed to suppress the expression of phototoxicity as much as possible has become possible. In addition, the porphyrin was derivatized with chlorin, and the wavelength was red-shifted, so that the therapeutic effect could be deepened. Based on these characteristics (cancer affinity, photocell killing effect, absorption wavelength, water solubility), the porphyrin derivative of the present invention is useful as a PDT drug for specific organs, particularly cancer and malignant tumors.

[0031]

【Example】

Example 1 Synthesis of Porphyrin Compound (I) JP-A-2-138280 and Japanese Patent Application No. 2-16849
It was synthesized by the method described in No. 9. As a starting material, 5 g of protoporphyrin dimethyl ester (hereinafter referred to as PP-Me) was used, and 10% hydrobromic acid / acetic acid 30
ml was added and stirred overnight. Then, the reaction solution is concentrated under reduced pressure, and alcohols (for example, hexyl alcohol C) are added to the residue.
₆ H ₁₃ OH) (18 ml) was added, and the mixture was reacted with stirring at room temperature for 24 hours. (Introduction of alcohol residue into porphyrin derivative) After the reaction, the residue was hydrolyzed by adding 60 ml of 2N potassium hydroxide / ethanol solution. (Saponification of ester) After neutralization with 1N hydrochloric acid, the mixture was separated with chloroform, and the chloroform layer was concentrated under reduced pressure. Silica gel column chromatography of the concentrate (eluent; n-hexane-ethyl acetate)
Stepwise method), and 2,4-bis (1-
Hexyloxyethyl) -deuteroporphyrin was obtained. (3.3 g) The total amount of the obtained alkoxy compound was converted into a DCHA salt with dicyclohexylamine (DCHA) by a conventional method. This DCHA
The total amount of salt is 30 ml of chloroform and 1 of acetonitrile.
5 ml was added and dissolved. Next, 3 g of aspartic acid dimethyl ester [Asp (OMe) ₂ ] hydrochloride was added, and 1.8 g of WSC was gradually added with stirring to react for 2 hours. After the reaction, the reaction solution was washed with water, separated, and the chloroform layer was concentrated under reduced pressure. The obtained concentrate was recrystallized from methanol-ethyl acetate-n-hexane to obtain 2,4-bis (1-hexyloxyethyl) -deuteroporfinyl diasparagic acid tetramethyl ester.
The total amount of the obtained amide compound was hydrolyzed with a 2N potassium hydroxide / ethanol solution by a conventional method to obtain a crude crystal of a target substance. Then, the crude crystals were purified by octyl silica gel (C ₈ ) medium pressure high speed preparative liquid chromatography [eluent; methanol-water (9: 1)] to carry out 2.
4-bis (1-hexyloxyethyl) -deuteroporfinyl diaspartic acid [C ₆ -DP-diAs
p (4)] was obtained. (3.43 g, total yield 40.6%)

Example 2 Synthesis of metalloporphyrin compound (I) JP-A-2-138280 and Japanese Patent Application No. 2-16849
It was synthesized by improving the method described in No. 9. After using 30 g of PP-Me as a starting material and forming a gallium complex,
300 ml of 0% hydrobromic acid / acetic acid was added and stirred overnight. Then, the reaction solution was concentrated under reduced pressure, and the residue was mixed with methanol
00 ml was added, and the reaction was carried out at room temperature with stirring overnight. (Introduction of a methoxy group into a porphyrin derivative) Water was added to form a reddish purple precipitate, which was collected by filtration. (30 g) Alcohols (eg decyl alcohol C ₁₀ ) in the precipitate
H ₂₃ OH) (150 ml) was added, and the mixture was reacted with heating and stirring under a Lewis acid (for example, stannic chloride) catalyst. (Exchange Reaction of Alcohol Residue to Methoxyporphyrin Derivative) After the reaction, the reaction solution was extracted and concentrated under reduced pressure, and the residue was hydrolyzed by adding 60 ml of 2N potassium hydroxide / ethanol solution. (Saponification of Ester) After neutralization with 1N hydrochloric acid, the mixture was allowed to stand in the refrigerator for a whole day and night to form a reddish purple precipitate. The precipitate was collected by filtration and purified by silica gel column chromatography (eluent: n-hexane-ethyl acetate stepwise method) to give 2,4-bis (1
A gallium complex of -decyloxyethyl) -deuteroporphyrin was obtained. (14.4 g) The total amount of the obtained complex was subjected to amidation, hydrolysis and purification operations in the same manner as in Example 1 to give 2,4-bis (1-decyloxyethyl) -Ga-deuteroporfinyl diasparagine. to obtain an acid _{[C 10 -Ga-DP-diAsp} (18)]. (15.7 g, total yield 25.5%)

Example 3 Synthesis of Photoprotoporphyrin R. K. The method of Dinello et al. [The Porph
yrins, Academic Press, Vo
l. 1.303 (1978)]. PP
-Me 100g was melt | dissolved in chloroform 10l, and it was made to react under light irradiation for 1 week. (Porphyrin to chlorin derivatization) After the reaction, it was concentrated under reduced pressure to obtain a residue. The obtained residue was purified by silica gel column chromatography (eluent: n-hexane-chloroform) to give 1 (3) -hydroxy-2 (4) -formylethylidene-protoporphyrin dimethyl ester (photoprotoporphyrin dimethyl ester). , Hereinafter referred to as P-Me) was obtained. (5
0.0 g) Then, this was hydrolyzed in a pyridine / methanol mixture to obtain dark green crystalline photoprotoporphyrin (hereinafter referred to as P). (43.0 g, total yield 47.
4%)

Example 4 300 mg of P-Me obtained in Example 3 was added to pyridine 15
It was dissolved in 0 ml, and 30 ml of 30% aqueous sodium hydrogen sulfite solution was gradually added with stirring to carry out a reaction for 3 hours. The reaction solution was neutralized with a 20% citric acid solution, extracted with chloroform, washed with water, and concentrated under reduced pressure. The obtained concentrate was crystallized from methanol-ethyl acetate-n-hexane to give NaHS.
O ₃ was obtained -P-Me (28). (50 mg, 14.3
%)

Example 5 The procedure of Example 1 was repeated except that 3% of 10% hydrobromic acid / acetic acid was used in place of 2- (1-hexyloxyethyl) as a synthetic intermediate. ) -4-Vinyl-deuteroporphyrin from 2- (1-decyloxyethyl) -4 from decyl alcohol.
4 g each of vinyl-deuteroporphyrin were obtained. All the obtained monoalkoxy compounds were subjected to a light irradiation reaction and post-treatment in the same manner as in Example 3 to obtain C ₆ -P (31) and C ₁₀ -P (33). (0.88g, 21.0
% And 1.1 g, 6.5%) These photooxidation reaction products [31 (0.8 g), 33 (1.0 g)] were separately operated in the same manner as in Example 1 to carry out amidation and hydrolysis treatments. And C ₆ -P-diAsp (32) and C ₁₀
-P-diAsp (34) was obtained separately. (450m
g, 42.3% and 160 mg, 12.3%)

Example 6 300 mg of P obtained in Example 3 was added to 100 ml of pyridine.
The resulting solution was dissolved in, and a 30% hydroxylamine hydrochloride solution was added with stirring at room temperature to react for 30 minutes. After the reaction, chloroform was added to the reaction solution, the solution was washed with water and separated, and the chloroform layer was concentrated under reduced pressure. The obtained concentrate was reprecipitated with methanol-ethyl acetate-n-hexane to obtain NOH-P (35).
Got (250 mg, 81.3%)

Example 7 P-Melg was dissolved in 300 ml of chloroform, 20 ml of a 5% sodium borohydride methanol solution was added dropwise with stirring at room temperature, and the mixture was reacted for 30 minutes. After the reaction
A 10% aqueous citric acid solution was added to the reaction solution, the solution was washed with water and separated, and the chloroform layer was concentrated under reduced pressure. The obtained concentrate was dissolved in 100 ml of dioxane, 10 ml of 10% aqueous sodium periodate solution and 40 ml of 3% hydrochloric acid were added, and the mixture was stirred at room temperature for 1 hour.
It was left for 8 hours. The purple crystals recrystallized in the reaction solution were collected by filtration, washed with water, dried, dissolved in 120 ml of chloroform, and reacted under light irradiation for 48 hours. After the reaction, the mixture was concentrated under reduced pressure to obtain a residue. The obtained residue was purified by silica gel chromatography (eluent: chloroform-acetone), and SP
150 mg of dimethyl ester was obtained. The total amount of the obtained SP dimethyl ester was dissolved in 30 ml of pyridine and 10
Hydrolyze by adding 5 ml of 5% sodium hydroxide aqueous solution,
After neutralizing with 20% aqueous citric acid solution, add chloroform,
After liquid separation by washing with water, the chloroform layer was concentrated under reduced pressure. The obtained concentrate is reprecipitated with ethyl acetate-n-hexane, and S
P (36) was obtained. (120 mg, 12.5%)

Example 8 200 mg of P was dissolved in 40 ml of methanol, 400 mg of Girard's reagent T was added with stirring at room temperature, and the mixture was reacted for 1 hour. After the reaction, a 20% aqueous citric acid solution and chloroform were added to the reaction solution, the solution was washed with water and separated, and the chloroform layer was concentrated under reduced pressure. The obtained concentrate was reprecipitated with methanol-ethyl acetate-n-hexane to obtain G'THZ-P (37). (200 mg, 79.9%)

Example 9 300 mg of P was dissolved in methanol, 300 mg of benzenesulfohydroxamic acid and 4 ml of 1N sodium hydroxide were added with stirring at room temperature, and the mixture was reacted for 18 hours. After the reaction, a 20% aqueous citric acid solution and chloroform were added to the reaction solution, the solution was washed with water and separated, and the chloroform layer was concentrated under reduced pressure. The obtained concentrate was reprecipitated with methanol-ethyl acetate-n-hexane to obtain N2OH-P (38). (60m
g, 19.0%)

Example 10 500 mg of SP dimethyl ester obtained as an intermediate of Example 7 was dissolved in 100 ml of pyridine, and 10 ml of 30% hydroxylamine hydrochloride aqueous solution was added with stirring at room temperature to react for 30 minutes. 20% in the reaction solution after the reaction
Aqueous citric acid solution and chloroform were added, the solution was washed with water and separated, and the chloroform layer was concentrated under reduced pressure. The obtained concentrate was reprecipitated with ethyl acetate-n-hexane to obtain 450 mg of NOH-SP dimethyl ester. NOH-S obtained
The P dimethyl ester was hydrolyzed by a conventional method.
After neutralization with 1N hydrochloric acid, the mixture was separated with chloroform, and the chloroform layer was concentrated under reduced pressure. The obtained concentrate is methanol-
Re-precipitate with ethyl acetate-n-hexane, 2NOH
-SP (39) was obtained. (180 mg, 35.9%)

Example 11 100 mg of NOH-P obtained in Example 6 was added to 20 m of pyridine.
1 ml of acetic anhydride was added with stirring at room temperature and reacted for 6 hours. After the reaction, chloroform was added to the reaction solution, the solution was washed with water and separated, and the chloroform layer was concentrated under reduced pressure. The obtained concentrate was reprecipitated with ethyl acetate-n-hexane to obtain NOAc-P (40). (60 mg, 19.
0%)

Example 12 P (200 mg) was dissolved in pyridine (50 ml), and t-butylhydrazine (400 mg) and 10% aminoguanidine hydrochloride aqueous solution (4 ml) were separately added and reacted for 18 hours while stirring at room temperature. After the reaction, chloroform was added to each reaction solution, the solution was washed with water and separated, and the chloroform layer was concentrated under reduced pressure. Each of the obtained concentrates was added to ethyl acetate-n-
Re-precipitation was performed with hexane to obtain ^t BuHZ-P (41) and GHZ-P (42). (80 mg, 35.7
% And 190 mg, 87.2%)

Example 13 P (200 mg) was dissolved in pyridine (20 ml), nicotinic acid hydrazide (730 mg) and carbazic acid methyl ester were added separately, and the mixture was reacted under heating at 50 ° C. for 3 hours. After the reaction, a 20% aqueous citric acid solution and chloroform were added to each reaction solution, the solution was washed with water and separated, and the chloroform layer was concentrated under reduced pressure. Each of the obtained concentrates was reprecipitated with methanol-ethyl acetate-n-hexane,
NHZ-P (43) and CHZ-P (45) were obtained.
(33 mg, 13.8% and 130 mg, 58.0
%)

Example 14 930 mg of NHZ-P obtained in Example 13 was subjected to D by a conventional method.
CHA salt. The total amount of this DCHA salt is chloroform 50
ml and 25 ml of acetonitrile were added and dissolved.
Then aspartic acid dimethyl ester hydrochloride 930
mg was added and 1.5 g of WSC was gradually added to react with stirring. After the reaction, the reaction solution was washed with water and separated, and the chloroform layer was concentrated under reduced pressure. The obtained concentrate was dissolved in pyridine and hydrolyzed with a 10% aqueous sodium hydroxide solution by a conventional method. After neutralization with a 20% aqueous citric acid solution, chloroform was added, the solution was washed with water and separated, and the chloroform layer was concentrated under reduced pressure. The obtained concentrate was reprecipitated with methanol-ethyl acetate-n-hexane, and NHZ-P-diAsp (4
4) was obtained. (870 mg, 71.0%)

Example 15 150 mg of P was dissolved in 20 ml of pyridine, and 150 mg of barbituric acid and 20 ml of malonnitrile were stirred at room temperature.
ml was added separately and reacted for 30 minutes. After the reaction, a 20% aqueous citric acid solution and chloroform were added to each reaction solution, the solution was washed with water and separated, and the chloroform layer was concentrated under reduced pressure. Each of the obtained concentrates was reprecipitated with ethyl acetate-n-hexane to obtain BA-P (46) and MC.
NP (50) was obtained. (20 mg, 11.2% and 50 mg, 30.9%)

Example 16 200 mg of P was dissolved in 20 ml of pyridine, and 200 mg of semicarbazide hydrochloride and 400 mg of thiosemicarbazide hydrochloride were added separately with stirring at room temperature and reacted for 50 minutes. After the reaction, a 20% aqueous citric acid solution and chloroform were added to each reaction solution, the solution was washed with water and separated, and the chloroform layer was concentrated under reduced pressure. Each of the obtained concentrates was reprecipitated with pyridine-chloroform-ethyl acetate-n-hexane, and NH ₂ CONHN-P (47) and NH
₂ CSNHN-P (48) was obtained. (160 mg, 7
3.0% and 70 mg, 31.2%)

Example 17 300 mg of P was dissolved in 50 ml of pyridine, 10 ml of o-aminobenzenethiol was added, and the mixture was heated and stirred at 60 ° C. 1
Reacted for 8 hours. After the reaction, a 20% aqueous citric acid solution and chloroform were added to the reaction solution, the solution was washed with water and separated, and the chloroform layer was concentrated under reduced pressure. The obtained concentrate is ethyl acetate-n-
Reprecipitation was performed with hexane to obtain BT-P (49).
(350 mg, 98.9%)

Example 18 P (100 mg) was dissolved in viridine (8 ml), and nitromethane, nitroethane and nitropropane (8 ml) were separately added under room temperature stirring, and sodium ethylate (600 mg) was further added.
The reaction was allowed to proceed for 24 minutes. After the reaction, a 20% aqueous citric acid solution and chloroform were added to each reaction solution, the solution was washed with water and separated, and the chloroform layer was concentrated under reduced pressure. The resulting respective concentrate chloroform - reprecipitation with ethyl acetate -n- _{hexane, NO 2 -P (51),} MeNO 2 -
P (52) and Etno ₂ was obtained -P (53). (6
0 mg, 56.0%, 40 mg, 36.5% and 70
mg, 62.5%)

Example 19 P-Me 3 g Acetone 1.1 l and methanol 300 ml
It was dissolved in the mixed solution of, and 365 ml of a 10% sodium hydroxide aqueous solution was added with stirring at room temperature. Then, the mixture was neutralized with a 20% aqueous citric acid solution, chloroform was added thereto, the mixture was washed with water and separated, and the chloroform layer was concentrated under reduced pressure. The obtained concentrate was reprecipitated with chloroform-ethyl acetate-n-hexane to give Me.
₂ CO-P (54) was obtained. (2.6g, 85.0%)

Example 20 2.6 g of Me ₂ CO-P obtained in Example 19 was converted into a DCHA salt by a conventional method. The total amount of this DCHA salt is chloroform 5
0 ml and 25 ml of acetonitrile were added and dissolved. Then aspartic acid dimethyl ester hydrochloride 3
05 g was added, and 3.0 g of WSC was gradually added to react with stirring. After the reaction, the reaction solution was washed with water and separated, and the chloroform layer was concentrated under reduced pressure. The obtained concentrate was dissolved in pyridine and hydrolyzed with a 10% aqueous sodium hydroxide solution by a conventional method. After neutralization with a 20% aqueous citric acid solution, chloroform was added, the solution was washed with water and separated, and the chloroform layer was concentrated under reduced pressure. The obtained concentrate was reprecipitated with methanol-ethyl acetate-n-hexane to give Me ₂ CO-P-diAsp.
(55) was obtained. (870 mg, 24.6%)

Example 21 P-Me (100 mg) was dissolved in pyridine (8 ml), β-ionone (8 ml) was added with stirring at room temperature, 10% aqueous sodium hydroxide solution (6 ml) was added, and the mixture was reacted for 20 minutes.
After the reaction, a 20% aqueous citric acid solution and chloroform were added to the reaction solution, the solution was washed with water and separated, and the chloroform layer was concentrated. The obtained residue is subjected to silica gel chromatography (eluent; n-
It was purified with hexane-ethyl acetate) to obtain 80 mg of Io-P dimethyl ester. The obtained Io-P dimethyl ester was dissolved in pyridine and hydrolyzed with a 10% aqueous sodium hydroxide solution by a conventional method. After neutralization with a 20% aqueous citric acid solution, chloroform was added, the solution was washed with water and separated, and the chloroform layer was concentrated under reduced pressure. The obtained concentrate was reprecipitated with methanol-ethyl acetate-n-hexane to give I
70 mg of o-P was obtained. The obtained Io-P was converted into a DCHA salt by a conventional method. The total amount of this DCHA salt is chloroform 1
0 ml and 5 ml of acetonitrile were added and dissolved.
Then aspartic acid dimethyl ester hydrochloride 70m
g was added, and 50 mg of WSC was gradually added to react with stirring. After the reaction, the reaction solution was washed with water and separated, and the chloroform layer was concentrated under reduced pressure. The obtained concentrate was dissolved in pyridine and hydrolyzed with a 10% aqueous sodium hydroxide solution by a conventional method. After neutralization with a 20% aqueous citric acid solution, chloroform was added, the mixture was washed with water and separated, and the chloroform layer was concentrated. The obtained concentrate was reprecipitated with methanol-ethyl acetate-n-hexane to obtain Io-P-diAsp (56). (56 mg, 35.0%)

Example 22 180 mg of Io-P obtained as an intermediate of Example 21
Was dissolved in 60 ml of pyridine, 1.2 g of hydroxylamine hydrochloride was added under stirring with heating at 50 ° C., and the mixture was reacted for 40 minutes. After the reaction, a 20% aqueous citric acid solution and chloroform were added to the reaction solution, the solution was washed with water and separated, and the chloroform layer was concentrated under reduced pressure. The obtained residue is purified by silica gel chromatography (eluent: ethyl acetate-methanol), and NOH-
110 mg of Io-P was obtained. The obtained NOH-Io-P
Was converted into DCHA salt by a conventional method. The whole amount of this DCHA salt was dissolved by adding 20 ml of chloroform and 10 ml of acetonitrile, and then 110 mg of aspartic acid dimethyl ester hydrochloride was added, and with stirring, 10 mg of WSCl was added.
Was gradually added to react. After the reaction, the reaction solution was washed with water and separated, and the chloroform layer was concentrated under reduced pressure. The obtained concentrate was dissolved in ethanol and hydrolyzed with a 2N aqueous potassium hydroxide solution by a conventional method. After neutralization with a 20% aqueous citric acid solution, chloroform was added, the mixture was washed with water and separated, and the chloroform layer was concentrated. The obtained concentrate is methanol-ethyl acetate-
Re-precipitation was carried out with n-hexane, NOH-Io-P-d
iAsp (57) was obtained. (80 mg, 33.8%)

Example 23 P-Me (300 mg) was dissolved in pyridine (50 ml), and acetophenone (5 ml) and p-aminoacetophenone were added separately with stirring at room temperature. 6 ml of 10% aqueous sodium hydroxide solution was added to each, and the reaction was allowed to proceed for 3 hours. After the reaction, 15 ml of a 10% aqueous sodium hydroxide solution was added to each reaction solution, neutralized with a 20% aqueous citric acid solution, extracted with chloroform, washed with water and separated, and the chloroform layer was concentrated under reduced pressure. Each of the obtained concentrates was reprecipitated with ethyl acetate-n-hexane, and PhCO-P (5
8) and _NH 2 to give PhCO-P and (59). (22
0 mg, 65.4% and 180 mg, 52.6%)

Example 24 NOH-P (150 mg) was dissolved in tetrahydrofuran (5 ml), and DCHA (90 mg) was dissolved in diethyl ether (1.5 ml).
The solution dissolved in was added to give a DCHA salt by a conventional method. Book DC
The total amount of HA salt was dissolved in 18 ml of dimethylformamide,
Add 4 ml of 10% D-glucosamine hydrochloride aqueous solution, and add 50
6m of 2.5% DCC solution in chloroform with stirring at ℃
1 was gradually added and reacted for 2.5 hours. After the reaction, a 20% aqueous citric acid solution and chloroform were added to the reaction solution, the solution was washed with water and separated, and the chloroform layer was concentrated under reduced pressure. The obtained concentrate was reprecipitated with methanol-chloroform-ethyl acetate-n-hexane to obtain NOH-P-NHglu (60).
Got (40 mg, 20.6%)

Example 25 Mass Spectrometry The mass of this derivative was measured by FAB mass spectrometry. Table 1 shows the main results of the measurement. As typical examples thereof, C ₆ -Pd-DP-diAsp (21) and NO
FAB mass spectrometric spectra of HP (35) dimethyl ester are shown in FIGS. 1 and 2.

[0056]

[Table 1]

Example 26 Nuclear Magnetic Resonance Analysis This derivative was measured by the nuclear magnetic resonance ( ¹ H-NMR) method. As a typical example thereof, C ₆ -DP-diAsp (4)
And ¹ H of C ₁₀ -In-DP-diAsp (26)
-NMR spectra are shown in Figures 3 and 4.

Example 27 Ultraviolet absorption spectrum analysis (albumin test) It is known that a porphyrin compound forms a monomer or a dimer in an albumin solution. This property can be seen from the fact that the maximum absorption value shifts or the absorption coefficient changes when the analysis is performed while changing the albumin concentration variously. Therefore, it is a simple screening test for examining the affinity with cancer cells. Albumin 54 mg 3
Dissolve it in ml of physiological saline to make a concentration of 1.8%. Then, this was diluted 10-fold to 0.18% and diluted with a common ratio of 3 to obtain albumin concentrations (1.8, 0.18, 0.0
6, 0.02, 0.0066, 0.0022%) was prepared. On the other hand, 1 mg of the porphyrin derivative was dissolved in 1 ml of a phosphate buffer (pH 8.0), and the solution was diluted with physiological saline to 10 mg.
It was set to 0 ml. Then, 2 ml of the albumin diluent and 2 ml of the porphyrin solution were mixed to give albumin concentrations of 0.9, 0.09, 0.03, 0.01, 0.003.
3, 0.0011% and ultraviolet absorption spectrum measurement (3
50-900 nm). Also, instead of the albumin diluting solution, the same measurement was performed in physiological saline and methanol solution. The results of these measurements are shown in Table 2. As a typical example thereof, the ultraviolet absorption spectrum of C ₁₀ -Ga-DP-diAsp (18) is shown in FIGS. 5 and 6.

[0059]

[Table 2]

Example 28 Determination of photocell killing effect by light irradiation (in vitro) A test was carried out by the method described in JP-A-2-138280. 1 × 10 ⁴ HGC-27 cells (1 ml) were placed in a Petri dish (3.5 cm) and cultured for 2 days. Each drug was adjusted to various concentrations, added to the Petri dish, incubated for 30 minutes, and washed with phosphate buffer. After leaving for 2-3 minutes col
d spot PICL-SX (halogen la
Light was irradiated for 5 minutes (5.8 mw / cm ² ) (mp 150 W). The light irradiation was performed by cutting the wavelength of 600 nm or less. Then, the cells were cultured for 48 hours and the number of surviving cells was counted. On the other hand, as a control, a group was provided in which light was blocked by aluminum foil during light irradiation. Photocell destruction results in ID ₅₀ (50%
Cell growth inhibition rate). In FIG. 7, C ₆ -DP-d
iAsp (4) and C ₁₀ -Zn-DP-di in FIG.
The graph of the cell growth inhibition rate by Asp (24) is shown.

Example 29 C ₆ -DP-diAsp (4), C ₁₀ -Ga-DP-
diAsp (18) and NOH-P (35) the preparation of injectables _{C 6 -DP-diAsp (4)} , C 10 -Ga-DP-
DiAsp (18) and 5 g of NOH-P (35) were separately dissolved by adding 90 ml of a phosphate buffer (pH 8.0), and then 10 ml of 0.1N sodium hydroxide was added for pH adjustment to bring the total amount to each. 100 ml (5
0 mg / ml, pH 7.3). Then, 5 ml each was dispensed into a sterile vial while performing aseptic filtration to obtain a drug for PDT. Furthermore, after dispensing as needed, the samples were frozen and stored until use.

Example 30 TLC and H of C ₆ -DP-diAsp (4) Injection
Purity in PLC An appropriate amount of the C ₆ -DP-diAsp (4) injection solution obtained in Example 29 was added to an octyl silica gel (C ₈ ) thin plate (RP).
-8, manufactured by Merck), methanol-water mixture (9: 1)
It was developed using. As a result, spots were observed only near Rf 0.5. Furthermore, HPLC analysis [column; Wakosil 5C ₈ 4.0 × 150 mm, eluent; methanol: water: acetic acid (80: 15: 5), flow rate; 1.0 ml /
min, detection wavelength: 395 nm], Rt
One peak was observed at 6.3 minutes and the purity was 90% or more.

Example 31 TL of C ₁₀ -Ga-DP-diAsp (18) Injection
Purity in C and HPLC C ₁₀ -Ga-DP-diAsp obtained in Example 29
(18) An appropriate amount of the injection solution was developed on an octyl silica gel (C ₈ ) thin layer plate (RP-8, manufactured by Merck Ltd.) using a methanol-water mixture (95: 5). As a result, spots were observed only near Rf 0.5. Furthermore, HPLC analysis [column; Wakosil 5C ₈ 4.0 × 150 mm, eluent; methanol: water: acetic acid (85: 10: 5), flow rate;
1.0 ml / min, detection wavelength; 405 nm], one peak was observed at Rt 5.9 minutes and the purity was 9
It was 5.0% or more.

Example 32 Purity of NOH-P (35) Injection by TLC and HPLC An appropriate amount of NOH-P (35) Injection obtained in Example 29 was added to octadecyl silica gel (C ₁₈ ) thin layer plate (RP-). 1
8, manufactured by Merck & Co., Inc.) and developed using a methanol-water mixture (4: 1). As a result, spots were observed only near Rf 0.5. Furthermore, HPLC analysis [column; Wakosil 5C ₁₈ 4.0 × 250 mm, eluent: methanol: water: acetic acid (80: 15: 5), flow rate; 0.5 ml /
min, detection wavelength; 405 nm], Rt
One peak was observed at 7.8 minutes, and the purity was 96.0% or more.

Example 33 C ₆ -DP-diAsp (4), C ₁₀ -Ga-DP-
diAsp (18) and NOH-P (35) in a in vitro of injectables stability _{C 6 -DP-diAsp (4)} , C 10 -Ga-DP-
The in vitro stability of this drug was evaluated by TLC analysis and HPLC analysis of the purity of the diAsp (18) and NOH-P (35) injections, respectively, over time. C ₆ -DP- prepared according to Example 29
_{diAsp (4), C 10 -Ga} -DP-diAsp
(18) and NOH-P (35) injections were each diluted 5-fold with physiological saline to a concentration of 10 mg / ml, and allowed to stand at room temperature and in the dark, at the time of preparation of this drug, for 1 day, 1 week, and After one month, TLC and HPLC purity of each drug were measured according to Example 32. As a result, the chemical purity of each drug at each measurement time was 1 spot except 4 in TLC analysis, about 95.0% and 95.0% by HPLC analysis, and each drug was stable for at least 1 month. It turned out. In addition, 4 was stable for at least 1 week.

Example 34 C ₆ -DP-diAsp (4), C ₁₀ -Ga-DP-
diAsp (18) and NOH-P (35) Injection of fresh frozen plasma (in vivo) in at Stability _{C 6 -DP-diAsp (4)} , C 10 -Ga-DP-
The plasma purity of diAsp (18) and NOH-P (35) injections were analyzed by TLC analysis and H, respectively, over time.
Pseudo in vivo of each drug by PLC analysis
The stability inside was evaluated. C ₆ -DP-diAsp (4) and C ₁₀ -Ga-DP- prepared in the same manner as in Example 29.
DiAsp (18) and NOH-P (35) injections were diluted 2.5 times with physiological saline to obtain 20 mg / ml.
And add the same amount of fresh frozen plasma (10
mg / ml, pH 7.1), body temperature (36.5 ° C.), leave it in the dark, and prepare each drug for 1 day, 1 week, 2 weeks, 3
Weekly and 1 month later, according to Example 32, T
LC and HPLC purity was measured. As a result, the chemical purity of each drug at each measurement time point was 1 spot for TLC analysis, and about 90.0% and 95.0% for HPLC analysis.
% And 95.0%, and each drug was found to be stable for at least 1 month.

[0067]

INDUSTRIAL APPLICABILITY The porphyrin derivative of the present invention has a property of accumulating in cancer cells, a reactivity to external energy and a destructive action of cancer cells, and does not exhibit toxicity to normal cells. It is useful as a drug or a cancer diagnostic agent.

[Brief description of drawings]

FIG. 1 is a diagram showing a mass spectrometry spectrum (FAB-NBA) of C ₆ -Pd-DP-diAsp (21).

FIG. 2 is a diagram showing a mass spectrometry spectrum (FAB-NBA) of NOH-P (35) dimethyl ester.

FIG. 3 is a diagram showing a nuclear magnetic resonance spectrum ( ¹ H-NMR) of C ₆ -DP-diAsp (4).

FIG. 4 is a diagram showing a nuclear magnetic resonance spectrum ( ¹ H-NMR) of C ₁₀ -In-DP-diAsp (26).

FIG. 5 is a diagram showing an ultraviolet absorption spectrum of C ₁₀ -Ga-DP-diAsp (18).

FIG. 6 is a diagram showing an ultraviolet absorption spectrum of C ₁₀ -Ga-DP-diAsp (18).

FIG. 7 is a diagram showing a cell growth inhibition rate after C ₆ -DP-diAsp (4) administration.

FIG. 8 is a diagram showing the cell growth inhibition rate after administration of C ₁₀ -Zn-DP-diAsp (24).

[Explanation of symbols]

 1 mixed solution of porphyrin solution and physiological saline (albumin concentration 0%) 2 mixed solution of porphyrin solution and albumin solution (albumin concentration 0.0011%) 3 mixed solution of porphyrin solution and albumin solution (albumin concentration 0.0033%) 4 porphyrin solution Mixed solution of albumin and albumin solution (0.01% albumin concentration) 5 Mixed solution of porphyrin solution and albumin solution (albumin concentration 0.03%) 6 Mixed solution of porphyrin solution and albumin solution (albumin concentration 0.09%) 7 Porphyrin solution and albumin Mixture of solutions (albumin concentration 0.9%) 8 Mixture of porphyrin solution and methanol 9 With light irradiation 10 Without light irradiation

Claims (4)

[Claims] 1. A compound represented by the following general formula (I): wherein R ₁ is CH (OR) CH ₃ , R ₂ is a residue obtained by removing hydrogen from an amino acid, R is alkyl, M is 2H, G
a, Zn, Pd, In, Sn), a porphyrin compound or a metal complex thereof. [Chemical 1] 2. A compound represented by the general formula (II): embedded image wherein R ₃ is CH═CH ₂ , CH (OR) CH, or C.
HO, CH = NOH or CH ₂ OH, R ₄ is CH =
_{X, C (OH) OSO 2} Na, CH (SCH 2 -COO
H) ₂ , CH (OR) ₂ or Chemical formula 3, X is O, C (C
N) ₂ , N-W or C (Y) Z, W is OH, OCO
CH ₃ or NH-E, E is H, alkyl, COC ₅
H ₄ N, CONH ₂ , CSNH ₂ , COOCH ₃ , CO
CH ₂ NCl (CH ₃ ) ₂ or C (NH ₂ ) ═NH,
Y is H or alkyl, Z is NO ₂ or COF,
Alternatively, a cyclic structure represented by Chemical formula 4, F is a residue of methyl, phenyl, aminophenyl or ionone, R ₂ is O
H, amino sugar or a residue obtained by removing hydrogen from an amino acid,
A porphyrin compound represented by R is alkyl. (However, in the formula, positional isomers in which the functional groups of the side chains of the A and B rings of the four tetrapyrrole rings are replaced are also included.) [Chemical 3] [Chemical 4] 3. A photophysicochemical diagnostic and / or therapeutic sensitizer comprising the porphyrin and the metalloporphyrin compound according to claim 1. 4. The sensitizer for photophysical chemistry according to claim 3, which is used for diagnosis and / or treatment of cancer.