JP2002128781A - Substituted tetraphenylporphyrin compound having basic axial ligand - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a porphyrin compound which has oxygen affinity similar to erythrocyte, can form a stable oxygen complex, and effectively acts as an artificial oxygen carrier. SOLUTION: The substituted tetraphenylporphyrin compound is represented by the general formula (I) [R1 is a hydrocarbon group which may have a substituent; R2 is a 1 to 10C alkyl; R3 is a 1 to 18C alkyl; M is two hydrogen atoms or metal ion; X- is a halogen ion; (n) is 0 or the number of X-, and is an integer obtained by subtracting 2 from the valency of the metal ion].

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a tetraphenylporphyrin compound which can reversibly bind and desorb oxygen and can be used as a stable artificial oxygen carrier.

[0002]

BACKGROUND OF THE INVENTION Heme, which is present in hemoglobin and myoglobin, ie, a porphyrin iron (II) complex, can reversibly adsorb and desorb oxygen molecules. Synthetic porphyrin iron (I
I) Many studies to realize the complex have been reported (for example, JP Collman, Acc. Chem. Res., 10,
265 (1977), F. Basolo, BM Hoffman, JA Ibers,
ibid, 8, 384 (1975)). In particular, porphyrin iron (II) complexes reported to be capable of forming stable oxygen complexes under room temperature conditions include 5,10,15,20-tetrakis (α,
An α, α, α-o-pivalamidophenyl) porphyrin iron (II) complex (hereinafter referred to as FeTpivPP complex) is known (JP Collman, et al., J. Am. Chem. Soc., 197).
5, 97 , 1427).

[0003] When an axial base such as 1-alkylimidazole or 1-alkyl-2-methylimidazole coexists, the FeTpivPP complex is benzene, toluene, N, N-
Molecular oxygen can be reversibly bound at room temperature in an organic solvent such as dimethylformamide. This complex also
When embedded in a bilayer membrane vesicle composed of phospholipids, the same function is exhibited under physiological conditions (aqueous phase, pH 7.4, ≤40 ^° C) (E. Tsuchida, et al., J. Chem. . Soc., Dalton
Trans., 1984 , 1147). However, some imidazole derivatives required as axial bases for the FeTpivPP complex to reversibly bind and dissociate oxygen have pharmacological effects, and many of them have high internal toxicity. In addition, when phospholipid vesicles are used, there is also a problem that an excess imidazole derivative becomes a factor for destabilizing the form.

Therefore, as a method of minimizing the addition amount of the axial base, it is conceivable to introduce an imidazole derivative into the molecule through a covalent bond. The inventors of the present application believe that if an alkylimidazole derivative is introduced as a substituent into the porphyrin iron complex, a stable oxygen carrier can be supplied without externally adding an axial base. FeTpiv with substituent
A reversible oxygen adsorption / desorption reaction has been reported for a PP complex and a system in which this is bound to phospholipid endoplasmic reticulum or to human serum albumin (JP-A-59-164791, JP-A-59-164791). 162924, JP-A-8-301873).

When considering the use of aqueous solutions or dispersions of these porphyrin iron (II) complexes as substitutes for erythrocytes, oxygen affinity (50% of the total amount of heme It is important to adjust the oxygen partial pressure when oxygen binds to an appropriate value, that is, a value of oxygen affinity equivalent to that of red blood cells under physiological conditions. In general, when an N-substituted imidazole is bound in a molecule, it has a high oxygen affinity and cannot be used as a red blood cell substitute. Therefore, as a method of lowering the oxygen affinity, conventionally, a sterically hindered group such as a methyl group is introduced at the 2-position of the imidazole ring to loosen the bond between the imidazole nitrogen and the central iron (II).
Regulation of oxygen affinity has been widely practiced. However, on the other hand, there has been a tendency that the stability of the oxygen complex tends to decrease due to the presence of the methyl group. In other words, the fact is that the development of an intramolecularly basic porphyrin iron (II) complex having no sterically hindered group at the 2-position of the imidazole ring and exhibiting an appropriate value for oxygen affinity has been desired.

It is well known that the proximal base coordinated to the heme of hemoglobin is an imidazole of a histidine residue. However, when this is introduced into a synthetic porphyrin compound, particularly a tetraphenylporphyrin compound, In the past, N-substituted imidazole derivatives and pyridine derivatives have always been used (for example, Momentau et al.
l., Chem. Rev., 110, 7690 (1994), etc.). That is,
An example in which a histidine derivative is covalently bonded to a tetraphenylporphyrin compound has not been known so far.

Therefore, the invention of this application has been made in view of the circumstances described above, has solved the problems of the prior art, has an oxygen affinity close to the value of red blood cells, and has high stability. An object is to provide a porphyrin compound which can form an oxygen complex and effectively acts as an artificial oxygen carrier.

[0008]

Means for Solving the Problems The present inventors have conducted intensive studies on the molecular design and functional expression of a substituted porphyrin compound having a basic axial ligand capable of forming a more stable oxygen complex. , 10,15,20-Tetrakis (α, α, α, α-o
-Substituted amidophenyl) The porphyrin compound has a substituent necessary for effectively exhibiting oxygen adsorbing ability at two positions, that is, an imidazole derivative, particularly a histidine derivative, which is a basic axial ligand, is added in an amount of 1 mol per 1 mol of porphyrin. (A porphyrin having an axial base in the molecule), a new oxygen carrier that can form a more stable oxygen complex compared to the conventional system while maintaining the oxygen affinity at an appropriate value The present invention was completed, considering that it can be provided.

That is, the invention of the present application firstly has the general formula (I)

[0010]

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(Where R ₁ is a hydrocarbon group which may have a substituent, R ₂ is a C ₁ -C ₁₀ alkyl group, R ₃ is C ₁
Represents an alkyl group having -C _18, M is two hydrogen atoms X ^-
Is a halogen ion, and n is 0).

Secondly, the invention of the present application relates to a compound of the above general formula (I) wherein R ₁ is a compound of the general formula (II)

[0013]

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(Wherein R ₄ is a C ₁ -C ₁₈ alkyl group) or a 1-methylcyclohexyl group.

Third, the invention of the present application is a compound represented by the general formula (I):

[0016]

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(Where R ₁ is a hydrocarbon group which may have a substituent, R ₂ is a C ₁ -C ₁₀ alkyl group, and R ₃ is C ₁
Represents an alkyl group having -C _18, M is a transition metal ion of the 4-5 period, X ^- is a halogen ion, n represents X ^- is an integer obtained by subtracting 2 from the valence of the metal ions which indicate the number of The present invention provides a substituted tetraphenylporphyrin metal complex represented by the formula:

The invention of this application is, fourthly, a substituted tetraphenylporphyrin metal complex according to the third invention wherein M is an ion of Fe or Co, and fifthly, the valence of Fe is +2 or The present invention provides a substituted tetraphenylporphyrin metal complex having a valence of +3, and sixthly, a substituted tetraphenylporphyrin metal complex having a valence of +2.

Seventh, the invention of this application also provides an artificial oxygen carrier containing at least one of the above substituted tetraphenylporphyrin metal complexes.

[0020]

BEST MODE FOR CARRYING OUT THE INVENTION The substituted tetraphenylporphyrin compound of the present invention has the following general formula (I)

[0021]

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(Where R ₁ is a hydrocarbon group which may have a substituent, R ₂ is a C ₁ -C ₁₀ alkyl group, R ₃ is C ₁
Represents an alkyl group of ＣC ₁₈ , M is two hydrogen atoms or transition metal ions of the 4th to 5th cycles, X ⁻ is a halogen ion, and n is the valence of the metal ion indicating the number of X ⁻ , minus 2. Which is an integer).

At this time, M is the fourth to fifth elements such as Co and Fe.
In the case of a periodic transition metal ion, the substituted tetraphenylporphyrin compound is a substituted tetraphenylporphyrin metal complex, in which an imidazole residue of histidine in the molecule is coordinated. Such a substituted tetraphenylporphyrin metal complex can exhibit oxygen binding ability only with the molecule. Further, in the substituted tetraphenylporphyrin metal complex of the invention of this application,
Since there is no steric hindrance group at the 2-position of the imidazole ring of the histidine derivative which is a proximal base, an oxygen coordination complex which is much more stable than the conventional system is generated while maintaining an appropriate oxygen affinity. Since such a histidine derivative is derived from a natural amino acid, it has excellent biocompatibility and is preferable.

As described above, the substituted tetraphenylporphyrin metal complex of the invention of the present application not only can reduce the axial base concentration to the maximum during use but also can eliminate the presence of free imidazole which is highly toxic to the body. And it works effectively as an artificial oxygen carrier.

The substituted tetraphenylporphyrin metal complex as described above further acts as a catalyst for a redox reaction, an oxygen oxidation reaction or an oxygen addition reaction. Therefore, the substituted tetraphenylporphyrin compound of the invention of this application can be used not only as an artificial oxygen carrier, but also as a gas adsorbent, a redox catalyst, an oxygen oxidation reaction catalyst, and an oxygenation reaction catalyst.

The substituted tetraphenylporphyrin compound as described above may be produced by any method. For example, the following general formula (III)

[0027]

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5,10,15,20-tetrakis (α,
α, α, α-o-aminophenyl) porphyrin can be synthesized as a starting material.

Specifically, Collman et al., J. Am. Che
m. Soc., 97, 1427 (1975), 5,10,15,20-tetrakis (α, α, α, α-o-aminophenyl) porphyrin synthesized according to the method described in For example, it is dissolved in dry tetrahydrofuran, and a base (eg, pyridine, triethylamine, 4-dimethylaminopyridine, etc.) and 2,2-dimethylalkanoic acid chloride, or 1-
Add methylcyclohexanoic acid chloride and stir in the dark, on ice or at room temperature. Thereafter, the solvent was removed under reduced pressure, and this was extracted with an organic solvent such as chloroform.
After washing with water and filtration, the product is fractionated and purified by silica gel column chromatography. Thus, 5,10,15,20-tetrakis
Of the four atropisomers of (o-aminophenyl) porphyrin, 5,10,15,20-tetrakis (α, α, α, α-o-
Only substituted amidophenyl) porphyrins are obtained.

Next, sufficiently degassed dimethylformamide is cooled with ice water, phosphorus oxychloride is added, and the mixture
Stir for hours to prepare Vilsmeier reagent. Here, 5,10,15,20 dissolved in dry dichloromethane or chloroform
-Tetrakis (α, α, α, α-o-substituted amidophenyl) porphyrin is added dropwise at room temperature. At this time, the reaction solution instantaneously changes from purple to green of the dication. This is 12-24
The mixture is refluxed at the boiling point for an hour, and after cooling, a saturated aqueous solution of sodium acetate is added to the reaction solution, and after confirming that the color changes from green to purple, stirring is further continued at 20 to 40 ° C for 20 to 120 minutes. This is extracted with an appropriate organic solvent, washed with water, dried, filtered, and then fractionated and purified by silica gel column chromatography. Thus, 2-formyl-5,10,15,20-tetrakis (α,
An α, α, α-o-substituted amidophenyl) porphyrin is obtained.

This 2-formyl-5,10,15,20-tetrakis
(α, α, α, α-o-Substituted amidophenyl) porphyrin is dissolved in a suitable organic solvent (eg, dichloromethane, chloroform, benzene, etc.), methanol is added, and the mixture is sufficiently degassed with nitrogen. After adding sodium borohydride under ice-cooling and stirring for 5 to 15 minutes, water is added to stop the reaction. This is extracted with an organic solvent such as chloroform, washed with water, dried, filtered and fractionated and purified by silica gel column chromatography. Thus 2-hydroxymethyl-
5,10,15,20-Tetrakis (α, α, α, α-o-substituted amidophenyl) porphyrin is obtained.

The thus obtained 2-hydroxymethyl-
Introduction of a histidine derivative into the 2-hydroxyethyl group of 5,10,15,20-tetrakis (α, α, α, α-o-substituted amidophenyl) porphyrin can be achieved, for example, by the following method.

2-Hydroxymethyl-5,10,15,20-tetrakis (α, α, α, α-o-substituted amidophenyl) porphyrin is dissolved in a suitable dry organic solvent (eg, tetrahydrofuran,
Dissolved in dichloromethane, benzene, etc.) and cyclic anhydride of dicarboxylic acid (eg, glutaric anhydride), 4-
Add dimethylaminopyridine, and under nitrogen atmosphere, 10 ~
Allow to boil for 24 hours. After the solvent is removed under reduced pressure, the residue is extracted with an organic solvent such as chloroform, washed with water, dried, filtered and fractionated and purified by silica gel column chromatography. Thus, 2-carboxyalkanoyloxymethyl-5,10,15,20-tetrakis (α, α, α, α-o-substituted amidophenyl) porphyrin is obtained.

The 2-carboxyalkanoyloxymethyl-5,10,15,20-tetrakis (α, α, α, α-o-substituted amidophenyl) porphyrin and triethylamine are dissolved in anhydrous dimethylformamide, and the solution is dissolved for 10 to 30 minutes. Nitrogen bubbles. BOP as a coupling reagent is added, and the mixture is further stirred under a nitrogen atmosphere for 10 to 30 minutes. Thereafter, L-histidinealkyl dihydrochloride is added, and the mixture is stirred at room temperature for 10 to 18 hours in a nitrogen atmosphere while shielding from light. After confirming the progress of the reaction by TLC, this is dropped into pure water to precipitate porphyrin. The precipitate is filtered off with a G4 glass filter, and the filtrate is dissolved in chloroform and collected. After drying over anhydrous sodium sulfate, the solvent is removed under reduced pressure, and the residue is purified by silica gel column chromatography. Thus, the target product 2- (L-
(Alkyl histidine amide alkanoyloxymethyl)-
5,10,15,20-Tetrakis (α, α, α, α-o-substituted amidophenyl) porphyrin is obtained.

Since the imidazole ring is decomposed by singlet oxygen in this product, the purification operation is preferably performed in a dark place.

The introduction of the central metal into the porphyrin thus obtained is described, for example, in D. Dolphin, edited by The Porphyrin, 197
Eight years, achieved by the general method described in Academic Press Co., etc., and obtained as a considerable porphyrin compound. In general, porphyrin iron (III) complex in the case of iron complex, and porphyrin cobalt (I
I) A complex is obtained.

When the above substituted tetraphenylporphyrin metal complex is in the form of an iron (III) complex, the central metal can be prepared by a conventional method using an appropriate reducing agent (eg, sodium dithionite, ascorbic acid). Is reduced from trivalent to divalent, oxygen binding activity can be imparted.

The iron (II) complex of the substituted tetraphenylporphyrin compound of the invention of the present application can be embedded in a bilayer vesicle composed of phospholipid molecules, encapsulated in a phospholipid-coated fat emulsion, and added to human serum albumin. Inclusion, inclusion in recombinant human serum albumin, or inclusion in albumin multimers may be used. The substituted tetraphenylporphyrin iron (II) complex can also quickly form a stable oxygen complex upon contact with oxygen in these embedded materials, inclusions, inclusions, and the like.

Further, the substituted tetraphenylporphyrin metal complex of the invention of this application is capable of adsorbing and desorbing oxygen depending on the oxygen partial pressure. That is, such oxygen bond dissociation can be performed repeatedly and reversibly.
The substituted tetraphenylporphyrin metal complex of the invention of this application effectively functions as an oxygen adsorbing / desorbing agent or an oxygen carrier.

The substituted tetraphenylporphyrin metal complex as described above is not limited to oxygen as long as it is a gas capable of coordinating with a metal. For example, carbon monoxide, nitrogen monoxide,
Nitrogen dioxide or the like can also be combined. Therefore,
The substituted tetraphenylporphyrin metal complex of the invention of this application can be applied as a catalyst for a redox reaction or a gas adsorbent in a homogeneous or heterogeneous system.

Hereinafter, the present invention will be described in detail with reference to examples. It goes without saying that the present invention is not limited to the embodiment.

[0042]

[Example] <Reference Example 1> 5,10,15,20-tetrakis (α,
α, α, α-o-aminophenyl) porphyrin 0.4 g (0.59
mmol) in 55 mL of tetrahydrofuran,
1.7 mL (20.7 mmol) and 2.11 g (14.8 mmol) of 1-methylcyclohexanoic acid chloride were added, and the mixture was stirred at room temperature in a dark place for 2 hours. Thereafter, the solvent is removed under reduced pressure, extracted with chloroform, washed with 5% aqueous ammonia, and then washed several times with pure water. The chloroform layer is dried over anhydrous sodium sulfate, filtered, and the solvent is removed under reduced pressure. The residue was dissolved in a small amount of chloroform, fractionated and purified on a silica gel column (chloroform / ethyl acetate: 10/1 (volume / volume)), and the desired product was collected and dried in vacuo. Thus, purple 5,
0.56 g of 10,15,20-tetrakis (α, α, α, α-o- (1-methylcyclohexylamido) phenyl) porphyrin
(82% yield).

5,10,15,20-tetrakis (α, α, α, α-o
The analysis results of-(1-methylcyclohexylamido) phenyl) porphyrin were as follows.

Thin layer chromatography (chloroform /
Ethyl acetate: 10/1 (vol / vol): Rf: 0.50 (monospot)).

Infrared absorption spectrum (cm ^-1 ): 1691 (ν
_{C = O} (amide)), 3428 (ν _NH ).

UV-visible absorption spectrum (CHCl ₃ , λ _max :
423, 516, 549, 590, 647 nm). FAB-MS spectrum (m /
z): 1172 [M] ^<+> .

¹ H-NMR spectrum (CDCl ₃ , TMS standard, δ (p
pm)): -2.3 (s, 2H, innerH), 0.2 (s, 12H, 1-methy
l), 0.3-1.2 (m, 40H, cyclohexyl), 7.5 (t, 4H, ph
enyl-4), 7.6 (s, 4H, amide-H), 7.8 (t, 4H, pheny
l-5), 7.9 (d, 4H, phenyl-3), 8.8 (d, 4H, phenyl-
6), 8.9 (s, 8H, pyrrole-βH). Reference Example 2 5,10,15,20-tetrakis (α, α, α, α-o
Dissolve 0.54 g (0.461 mmol) of-(1-methylcyclohexylamido) phenyl) porphyrin in 10 mL of chloroform,
177 mg (1.15 mmol) of copper chloride dihydrate and 233 mg (2.3
(1 mmol) in methanol (2 mL) was added and refluxed at 65 ° C. for 1 hour. In the visible absorption spectrum of the reaction solution,
After confirming that copper ions were inserted into the porphyrin, the solvent was removed under reduced pressure, and this was extracted with chloroform,
It was washed several times with pure water. The chloroform layer was dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The residue was dissolved in a small amount of chloroform and fractionated and purified on a silica gel column (chloroform / ethyl acetate: 5/1 (vol / vol)). did. Collect the desired product and dry it in vacuum to obtain a purple 5,10,15,2
0.45-g yield of 0-tetrakis (α, α, α, α-o- (1-methylcyclohexylamido) phenyl) porphyrin copper (II)
(79% yield).

The 5,10,15,20-tetrakis (α, α, α,
The analysis results of α-o- (1-methylcyclohexylamido) phenyl) porphyrin copper (II) were as follows.

Thin layer chromatography (chloroform /
Ethyl acetate: 10/1 (vol / vol): Rf: 0.74 (monospot)).

Infrared absorption spectrum (cm ^-1 ): 1687 (ν
_{C = O} (amide)), 3434 (ν _NH ).

UV-visible absorption spectrum (CHCl ₃ , λ _max :
423, 541, 576 nm).

FAB-MS spectrum (m / z): 1233 [M] ⁺ . <Example 1> 6.3 mL of distilled dimethylformamide sufficiently degassed with nitrogen was cooled with ice water, 7.4 mL of phosphorus oxychloride was added, and the mixture was stirred at room temperature for 1 hour to give a red-orange Vilsmeier.
Reagent was obtained. There dissolved in 14 mL of dichloromethane
5,10,15,20-tetrakis (α, α, α, α-o- (1-methylcyclohexylamido) phenyl) porphyrin copper (II) 0.45
g (0.36 mmol) was added dropwise at room temperature. As the reaction proceeded, the color of the solution changed from purple to dark green, and the formation of iminium salt was confirmed. After refluxing this for 18 hours,
After allowing to cool and neutralizing by adding 300 ml of a saturated aqueous sodium acetate solution, the color of the reaction solution changed from green to purple. Then, stirring was further continued at 40 ° C. for 3 hours. This was extracted with chloroform, washed several times with pure water, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. Silica gel column (chloroform / ethyl acetate: 5
/ 1 (volume / volume)), and the target product was collected and dried under vacuum. Thus, purple 2-formyl-5,10,15,20-tetrakis (α, α, α, α-o- (1-methylcyclohexylamido) phenyl) porphyrin copper (II) yield 0.22 g
(50% yield).

This 2-formyl-5,10,15,20-tetrakis
The analysis results of (α, α, α, α-o- (1-methylcyclohexylamido) phenyl) porphyrin copper (II) were as follows.

Thin layer chromatography (chloroform /
Ethyl acetate: 5/1 (volume / volume): Rf: 0.53 (monospot)).

Infrared absorption spectrum (cm ^-1 ): 1672 (νC =
O (formyl)), 1692 (ν _{C = O} (amide)), 3428 (ν _NH ).

UV-visible absorption spectrum (CHCl ₃ , λ _max :
430, 552, 593 nm).

FAB-MS spectrum (m / z): 1262 [M] ⁺ . <Example 2> 2-formyl-5,10,15,20-tetrakis (α,
0.22 g (0.175 mmol) of α, α, α-o- (1-methylcyclohexylamido) phenyl) porphyrin copper (II) was dissolved in 8 mL of dichloromethane, and 2 mL of concentrated sulfuric acid was added thereto.
Upon vigorous stirring at room temperature for 10 minutes, the color of the solution turned green. This solution was dropped into a two-layer solution of chloroform and ice water and neutralized with sodium carbonate, and the chloroform layer returned to purple. The chloroform layer was washed several times with pure water, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The residue was dissolved in a small amount of chloroform, and the residue was dissolved in a silica gel column (chloroform / ethyl acetate: 5.
/ 2 (volume / volume)), and the target product was collected and dried in vacuo. Thus, purple 2-formyl-5,10,15,20-
0.18 g of tetrakis (α, α, α, α-o- (1-methylcyclohexylamido) phenyl) porphyrin was obtained (yield 84
%).

The above-mentioned 2-formyl-5,10,15,20-tetrakis (α, α, α, α-o- (1-methylcyclohexylamide))
The analysis results of (phenyl) porphyrin were as follows.

Thin layer chromatography (chloroform /
Ethyl acetate: 5/2 (volume / volume): Rf: 0.60 (monospot)).

Infrared absorption spectrum (cm ^-1 ): 1672 (ν
_{C = O} (formyl)), 1691 (ν _{C = O} (amide)), 3429
(Ν _NH ).

UV-visible absorption spectrum (CHCl ₃ , λ _max :
432, 527, 555, 597, 662 nm).

FAB-MS spectrum (m / z): 1201 [M] ⁺ .

¹ H-NMR spectrum (CDCl ₃ , TMS standard, δ (p
pm)): -2.3 (s, 2H, innerH), 0.2 (d, 12H, 1-methy
l), 0.3-1.0 (m, 40H, cyclohexyl), 7.4-7.6 (m, 8
H, amide-H, phenyl-4), 7.8-7.9 (m, 8H, phenyl-3,
5), 8.7-8.9 (m, 10H, pyrrole-βH, phenyl-6), 9.4
(s, 1H, pyrrole-H), 9.6 (s, 1H, formyl). Example 3 2-Formyl-5,10,15,20-tetrakis (α,
177 mg (0.147 mmol) of α, α, α-o- (1-methylcyclohexylamido) phenyl) porphyrin was dissolved in 2.5 mL of dichloromethane, 7 mL of methanol was added, and the mixture was sufficiently degassed with nitrogen. Under ice cooling, sodium borohydride 53.6 mg
(1.47 mmol) was added and stirred for 10 minutes, and water was added to stop the reaction. This was extracted with chloroform, washed several times with pure water, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The residue was dissolved in a small amount of chloroform, and the residue was dissolved in a silica gel column (chloroform / methanol: 30
/ 1 (volume / volume)), and the target product was collected and dried under vacuum. Thus, purple 2-hydroxymethyl-5,1
The yield of 0,15,20-tetrakis (α, α, α, α-o- (1-methylcyclohexylamido) phenyl) porphyrin is 89.3 m
g (yield 50%).

The above-mentioned 2-hydroxymethyl-5,10,15,20-
The analysis results of tetrakis (α, α, α, α-o- (1-methylcyclohexylamido) phenyl) porphyrin were as follows.

Thin layer chromatography (chloroform /
Methanol: 30/1 (volume / volume): Rf: 0.29 (monospot)).

Infrared absorption spectrum (cm ^-1 ): 1690 (ν
_{C = O} (amide)), 3427 (ν _NH ).

UV-visible absorption spectrum (CHCl ₃ , λ _max :
423, 517, 548, 591, 645 nm).

FAB-MS spectrum (m / z): 1219 [M] ⁺ .

¹ H-NMR spectrum (CDCl ₃ , TMS standard, δ (p
pm)): -2.6 (s, 2H, innerH), 0.1 (m, 12H, 1-methy
l), 0.3-1.0 (m, 40H, cyclohexyl), 4.9 (q, 2H, -C
H ₂ OH), 7.4-7.6 (m, 8H, amide-H, phenyl-4), 7.7-
7.8 (m, 8H, phenyl-3,5), 8.6-8.8 (m, 10H, pyrrole
-βH, phenyl-6), 9.0 (s, 1H, pyrrole-H). <Example 4> 2-hydroxymethyl-5,10,15,20-tetrakis (α, α, α, α-o- (1-methylcyclohexylamide)
Glutaric anhydride (155 m) was added to a distilled tetrahydrofuran solution (15 mL) of 0.54 g (0.45 mmol) of phenyl) porphyrin.
g, 0.67 mmol), 4-dimethylaminopyridine (32.7 mg,
(0.27 mmol), and reacted at 60 ° C. for 18 hours under a nitrogen atmosphere. After removing the solvent under reduced pressure, it is extracted with chloroform, washed several times with pure water, dried over anhydrous sodium sulfate, filtered, and the solvent is removed under reduced pressure. The residue was dissolved in a small amount of chloroform, fractionated and purified on a silica gel column (chloroform / methanol: 10/1 (volume / volume)), and the target product was collected and dried in vacuo. Thus, purple 2
-(3-carboxypentanoyloxy) methyl-5,10,15,20
-Tetrakis (α, α, α, α-o- (1-methylcyclohexylamido) phenyl) porphyrin was obtained in an amount of 0.45 g (yield 79
%).

The above-mentioned 2- (3-carboxypentanoyloxy) methyl-5,10,15,20-tetrakis (α, α, α, α-o
The analysis results of-(1-methylcyclohexylamido) phenyl) porphyrin were as follows.

Thin layer chromatography (chloroform /
Methanol: 5/1 (volume / volume): Rf: 0.49 (monospot)).

Infrared absorption spectrum (cm ^-1 ): 1688 (ν
_{C = O} (amide)), 1734 (ν _{C = O} (ester)), 3427
(Ν _NH ).

UV-visible absorption spectrum (CHCl ₃ , λ _max :
423, 517, 547, 591, 646 nm).

FAB-MS spectrum (m / z): 1333 [M] ⁺ .

¹ H-NMR spectrum (CDCl ₃ , TMS standard, δ (p
pm)): -2.6 (s, 2H, innerH), 0.1 (m, 12H, 1-methy
l), 0.3-1.0 (m, 40H, cyclohexyl), 2.0 (t, 2H, -C
H ₂ COOH), 2.4 (m, 4H, -CH ₂ CH ₂ C (= O) O-), 5.3 (q, 2H,
-CH ₂ OC (= O)-), 7.4-7.5 (m, 8H, amide-H, phenyl-
4), 7.7-7.8 (m, 8H, phenyl-3,5), 8.6 (s, 1H, pyr
role-βH), 8.7-8.8 (m, 10H, pyrrole-βH, phenyl-
6). Example 5 2- (3-carboxypentanoyloxy) methyl-5,10,15,20-tetrakis (α, α, α, α-o- (1-methylcyclohexylamido) phenyl) porphyrin (46 m
g, 35 μmol) and TEA (37.7 μL, 0.31 mmol) in distilled DMF
(4.6 mL) was degassed by bubbling nitrogen through for 10 minutes, and then BOP (1
51 mg, 0.35 mmol) and further stirred at room temperature for 10 minutes under a nitrogen atmosphere. Then, histidine methyl ester 2
Hydrochloride 25.1 mg (0.11 mmol) was added and under a nitrogen atmosphere,
The mixture was stirred for 12 hours while shielding the light at room temperature. TL for the progress of the reaction
After confirmation by C, the reaction solution was dropped into pure water to precipitate porphyrin. The precipitate was separated by filtration with a G4 glass filter, and the precipitate was dissolved in chloroform and collected. The chloroform layer was dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The residue was dissolved in a small amount of chloroform, and the residue was dissolved in a silica gel column (chloroform / methanol: 30
/ 1 (volume / volume)), and the target product was collected and dried under vacuum. Thus, purple 2- (3-methylhistidinamidopentanoyloxy) methyl-5,10,15,20-tetrakis (α, α, α, α-o- (1-methylcyclohexylamide)
(Phenyl) porphyrin was obtained in a yield of 19.1 mg (37% yield).

2- (3-methylhistidinamidopentanoyloxy) methyl-5,10,15,20-tetrakis (α, α, α,
The analysis results of α-o- (1-methylcyclohexylamido) phenyl) porphyrin were as follows.

Thin layer chromatography (chloroform /
Methanol: 20/1 (volume / volume): Rf: 0.16 (monospot)).

Infrared absorption spectrum (cm ^-1 ): 1684 (ν
_{C = O} (amide)), 1740 (ν _{C = O} (ester)), 3428
(ν _NH ).

Ultraviolet-visible absorption spectrum (CHCl ₃ , λ _max :
423, 517, 549, 591, 646 nm).

FAB-MS spectrum (m / z): 1469 [M] ⁺ .

[0081]¹H-NMR spectrum (CDCl_Three, TMS standard, δ (p
pm)): -2.6 (s, 2H, innerH), 0.0-0.1 (m, 12H, 1-me
thyl), 0.3-1.0 (m, 40H, cyclohexyl), 1.9 (t, 2H,
 -CH _TwoCONH-), 2.3-2.5 (m, 4H, -CH _TwoCH _TwoC (= O) O-), 3.2
(s, 2H, Im-CH_Two-), 3.7 (m, 3H, His-OCH_Three), 4.9 (m, 1
H, His-CH_TwoCH-), 5.2-5.4 (q, 2H, -CH_TwoOC (= O)-), 7.2
(s, 1H, Im), 7.4-7.5 (m, 8H, amide-H, phenyl-4),
 7.7 (s, 1H, Im), 7.8 (m, 8H, phenyl-3,5), 8.6-8.
8 (m, 11H, pyrrole-βH, phenyl-6). <Example 6> The reaction of introducing iron into porphyrin is, for example,
D. Dolphin, The Porphyrin, 1978, Academic
-It can be achieved by the general method described in the press company.

After sufficiently degassing and deoxygenating 1.3 mL of a 47% aqueous solution of hydrobromic acid, 87 mg of electrolytic iron (1.56 m
mol), and the mixture was heated to 80 ° C. and stirred for 1 hour. The color of the solution changed from colorless and transparent to light green. When the electrolytic iron was completely dissolved, the temperature was raised to 130 ° C. to evaporate hydrobromic acid and water. When water disappears, ferrous bromide as a white solid is obtained. When the water was completely removed, the reaction vessel was allowed to cool. 2- (3-methylhistidinamidopentanoyloxy) methyl-5,10,15,20-tetrakis (α, α, α,
19 mg (13 μmol) of α-o- (1-methylcyclohexylamido) phenyl) porphyrin was dissolved in a tetrahydrafuran solution, and 17.2 μL (0.16 mmol) of 2,6-lutidine was added.
Degassed enough. This is added dropwise to the prepared ferrous bromide in a nitrogen atmosphere, and the mixture is refluxed at the boiling point for 2 hours. After removing the solvent under reduced pressure,
This was extracted with chloroform, washed with water, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. Silica gel column (chloroform / methanol (10/1)
(Volume / volume)), fractionation and purification, and the target product was collected and vacuum dried. Thus, 2- (3-methylhistidineamidopentanoyloxy) methyl-5,10,15,20-tetrakis (α, α,
α, α-o- (1-Methylcyclohexylamido) phenyl) porphyrin iron (III) bromide yield 9.1 mg (46% yield)
I got it.

2- (3-methylhistidineamidopentanoyloxy) methyl-5,10,15,20-tetrakis (α, α, α,
The analysis results of α-o- (1-methylcyclohexylamido) phenyl) porphyrin iron (III) bromide were as follows.

Thin layer chromatography (chloroform /
Methanol: 10/1 (volume / volume): Rf: 0.44 (monospot)).

Infrared absorption spectrum (cm ^-1 ): 1693 (ν
_{C = O} (amide)), 1740 (ν _{C = O} (ester)), 3420
(ν _NH ).

UV-visible absorption spectrum (CHCl ₃ , λ _max :
420, 505, 581 nm).

FAB-MS spectrum (m / z): 1523 [M-Br] ⁺ . <Example 7> In Example 1, pivaloyl chloride was used in place of 1-methylcyclohexanoic acid chloride. In Example 5, histidine octadecyl ester · 2 was used instead of histidine methyl ester · 2 hydrochloride.
Except that the hydrochloride was used, 2- (3-octadecylhistidinamidopentanoyloxy) methyl-5,10,15,20-tetrakis (α, α,
α, α-o-Pivalamidophenyl) porphyriniron (III)
Bromide was synthesized quantitatively.

The obtained 2- (3-octadecylhistidinamidopentanoyloxy) methyl-5,10,15,20-tetrakis (α, α, α, α-o-pivalamidophenyl) porphyrin iron (III) The bromide analysis results were as follows.

Thin layer chromatography (chloroform /
Methanol: 10/1 (volume / volume): Rf: 0.50 (monospot)).

Infrared absorption spectrum (cm ^-1 ): 1693 (ν
_{C = O} (amide)), 1741 (ν _{C = O} (ester)), 3420
(ν _NH ).

UV-visible absorption spectrum (CHCl ₃ , λ _max :
420, 506, 582 nm).

FAB-MS spectrum (m / z): 1599 [M-Br] ⁺ . <Example 8> The procedure of Example 1 was repeated except that 2,2-dimethyloctadecanoic acid chloride was used instead of 1-methylcyclohexanoic acid chloride, and that succinic anhydride was used instead of glutaric anhydride in Example 4. Example 1
5- (3-methylhistidinamidopropanoyloxy) methyl-5,10,15,20-tetrakis (α, α, α, α-o-2,2-dimethyloctadecanamidophenyl) ) Porphyrin was synthesized quantitatively.

Thin layer chromatography (chloroform /
Methanol: 20/1 (volume / volume): Rf: 0.11 (monospot)).

Infrared absorption spectrum (cm ^-1 ): 1684 (ν
_{C = O} (amide)), 1739 (ν _{C = O} (ester)), 3428
(ν _NH ).

UV-visible absorption spectrum (CHCl ₃ , λ _max :
423, 516, 548, 592, 646 nm).

¹ H-NMR spectrum (CDCl ₃ , TMS standard, δ (p
pm)): -2.6 (s, 2H, innerH), -1.5-0.5 (m, 24H, 2,2
-dimethyl), 0.5-1.5 (m, 52H,-(CH ₂ ) ₅ CH ₃ ), 1.9
(t, 2H, -CH ₂ CONH-), 2.3 (m, 2H, -C H ₂ C (= O) O-), 3.2
(s, 2H, Im-CH _2- ), 3.7 (m, 3H, His-OCH ₃ ), 4.9 (m, 1
H, His-CH ₂ C H- ), 5.2-5.4 (q, 2H, -CH ₂ OC (= O)-), 7.2
(s, 1H, Im), 7.4-7.5 (m, 8H, amide-H, phenyl-4),
7.7 (s, 1H, Im), 7.8 (m, 8H, phenyl-3,5), 8.6-8.
8 (m, 11H, pyrrole-βH, phenyl-6).

FAB-MS spectrum (m / z): 1573 [M] ⁺ . Example 9 2- (3-Methylhistidinamidopropanoyloxy) methyl-5,10,15,20-tetrakis (α, α, α, α-o-2,2-dimethyl) synthesized in Example 8 (Octadecanamidophenyl) porphyrin is reacted with cobalt chloride in dry tetrahydrofuran containing 2,6-lutidine, and a cobalt complex, 2- (3-methylhistidineamidopropanoyloxy) methyl-5,10,15,20- Tetrakis (α, α,
α, α-o-2,2-Dimethyloctadecanamidophenyl) porphyrin cobalt (II) was synthesized quantitatively.

The obtained 2- (3-methylhistidinamidopropanoyloxy) methyl-5,10,15,20-tetrakis (α,
The analysis results of α, α, α-o-2,2-dimethyloctadecanamidophenyl) porphyrin cobalt (II) were as follows.

Thin layer chromatography (chloroform /
Methanol: 10/1 (volume / volume): Rf: 0.61 (monospot)).

Infrared absorption spectrum (cm ^-1 ): 1693 (ν
_{C = O} (amide)), 1740 (ν _{C = O} (ester)), 3420
(Ν _NH ).

UV-visible absorption spectrum (CHCl ₃ , λ _max :
403, 521, 553 nm).

FAB-MS spectrum (m / z): 1630 [M] ⁺ . <Application Example 1> 2- (3-methylhistidinamidopentanoyloxy) methyl-5,10,15,20-tetrakis (α, α, α, α-o- (1-methylcyclohexyl) synthesized in Example 6 Amido) phenyl) porphyrin iron (III) bromide 48.1 μg (
0.03 μmol) was converted to 10 mL of anhydrous toluene solution, and after replacement with nitrogen, mixed with an aqueous solution of dithionite in a heterogeneous system for about 2 hours with stirring to reduce the solution to iron (II).

Under a nitrogen atmosphere, only the toluene layer was extracted, dehydrated and dried with anhydrous sodium sulfate, filtered, and the obtained toluene solution was transferred to a measurement cell and sealed. Thus, 2- (3-
(Methylhistidinamidopentanoyloxy) methyl-5,
A toluene solution of 10,15,20-tetrakis (α, α, α, α-o- (1-methylcyclohexylamido) phenyl) porphyrin iron (II) complex was obtained.

The visible absorption spectrum of this solution was λ _max : 4
At 42, 545 and 565 nm, the complex corresponded to a 5-coordinate deoxy form with one imidazole coordination.

As soon as oxygen gas was blown into this solution, the spectrum changed, and a spectrum having λ _{max of} 431 and 553 nm was obtained. This confirmed that the complex was an oxygenated complex.

By blowing nitrogen gas into this oxygenated complex solution for 1 minute, the visible absorption spectrum was reversibly changed from the oxygenated spectrum to the deoxy spectrum, and it was confirmed that the absorption and desorption of oxygen occurred reversibly. did. Furthermore, when the operation of blowing oxygen and then blowing nitrogen was repeated, it was confirmed that oxygen absorption and desorption could be performed continuously.

The oxygen coordination complex was gradually oxidized and degraded by a trace amount of water contained in toluene, but had a half life of about 12 hours (25 ° C.). In addition, oxygen affinity (P
_1/2 ) is 1.7 Torr (25 ° C), the value of red blood cells (8 Torr)
Close to. <Application Example 2> 2- (3-methylhistidinamidopentanoyloxy) methyl-5,10,15,20-tetrakis (α, α, α, α-o- (1-methylcyclohexyl) synthesized in Example 6 Amido) phenyl) porphyrin iron (III) bromide 48.1 μg (0.
03 μmol) and 2.2 mg (3.0 μmol) of dipalmitoylphosphatidylcholine were converted into 15 μL of a methanol solution. After purging with nitrogen, carbon monoxide was passed through the mixture, and an aqueous solution of ascorbic acid was added to reduce the mixture to form a carbon monoxide complex. This 3
By pouring into mL of water (70 ° C.), a vesicle dispersion of the carbon monoxide complex was obtained (λ _max : 427, 543 n
m).

Further, when CO was desorbed by light irradiation in a nitrogen atmosphere, the visible absorption spectrum was λ _max : 440,
Shifted to 542 and 563 nm and coordinated one of imidazole 5
It was confirmed to be a coordinated deoxy type. Electron microscope observation of this dispersion confirmed formation of uniform bilayer vesicles having a particle size of 100 to 200 nm.

The oxygen gas was blown into this solution.
The spectrum changes immediately,_{ma x}A: 426, 551 nm
The spectrum was obtained. This confirms the formation of oxygenated complex
Was done. Nitrogen gas is blown into this oxygenated complex solution for 1 minute
The visible absorption spectrum is oxygenated
From reversible to deoxy-type spectra
It was confirmed that oxygen adsorption and desorption occurred reversibly.
Was.

When the operation of blowing oxygen and then blowing nitrogen was repeated, it was confirmed that oxygen absorption / desorption could be performed continuously. <Application Example 3> 2- (3-Methylhistidinamidopentanoyloxy) methyl-5,10,15,20-synthesized according to Example 6
Tetrakis (α, α, α, α-o- (1-methylcyclohexylamido) phenyl) porphyrin iron (III) bromide (20μM)
Was encapsulated in human serum albumin (2.5 μM) according to the method described in JP-A-8-301873, and an albumin-heme complex (porphyrin / albumin: 8 (mol / mol)) in a phosphate buffered aqueous solution (pH 7) was prepared. .4, 1/30 mM) 3 mL was transferred to a quartz spectrometry cell and sealed under a nitrogen atmosphere. Its visible absorption spectrum has a _λmax of 443, 540, and 567 nm, and the included porphyrin iron (II) complex forms a Fe (II) high-spin five-coordinate complex in which one axial base is coordinated. It was confirmed that.

When oxygen was passed through the dispersion, the λ _{max of the} visible absorption spectrum shifted to 425,551 nm, indicating the formation of an oxygenated complex. By injecting nitrogen gas into this oxygenated complex solution for 1 minute, the visible absorption spectrum was reversibly changed from an oxygenated spectrum to a deoxy-type spectrum, indicating that oxygen absorption and desorption occurred reversibly. Was done.

When the operation of blowing oxygen and then blowing nitrogen was repeated, it was confirmed that oxygen absorption / desorption could be performed continuously.

The oxygen coordination complex has a half-life of about 2
4 hours (25 ° C.). This was longer than the conventional system.

[0114]

According to the invention of this application, a substituted tetraphenylporphyrin compound having an oxygen affinity close to that of erythrocytes and having high stability of an oxygen complex is provided. Since this compound has a histidine derivative that functions as a basic axis ligand at the 2-position, it exhibits excellent oxygen binding without adding free imidazole, and in addition to an artificial oxygen carrier,
It also functions effectively as a gas adsorbent, an oxygen adsorbing / desorbing agent, a redox catalyst, an oxygen oxidation reaction catalyst and the like.

 ──────────────────────────────────────────────────続 き Continued on the front page F term (reference) 4C050 PA06 4C086 AA01 AA03 BC38 CB04 MA01 MA04 MA16 NA14 ZA52 4H050 AB99 WB14 WB21

Claims (7)

[Claims] 1. A compound of the general formula (I) (Where R ₁ represents an optionally substituted hydrocarbon group, R ₂ represents a C ₁ -C ₁₀ alkyl group, R ₃ represents a C ₁ -C ₁₈ alkyl group, and M represents two A hydrogen atom, X ⁻ is a halogen ion, and n is 0). 2. R ₁ is a group represented by the general formula (II): (Where R ₄ is a C ₁ -C ₁₈ alkyl group) or
The substituted tetraphenylporphyrin compound according to claim 1, which is a 1-methylcyclohexyl group. 3. A compound of the general formula (I) (Where R ₁ represents a hydrocarbon group which may have a substituent, R ₂ represents a C ₁ -C ₁₀ alkyl group, R ₃ represents a C ₁ -C ₁₈ alkyl group, and M represents a transition metal ion of 5 cycles, X ^- is a halogen ion, n represents X ^- porphyrin metal complex represented number integer a is) obtained by subtracting 2 from the valence of the metal ions which indicate the. 4. The substituted tetraphenylporphyrin metal complex according to claim 3, wherein M is an ion of Fe or Co. 5. The substituted tetraphenylporphyrin metal complex according to claim 4, wherein the valence of Fe is +2 or +3. 6. The substituted tetraphenylporphyrin metal complex according to claim 4, wherein the valence of Co is +2. 7. An artificial oxygen carrier containing at least the substituted tetraphenylporphyrin metal complex according to any one of claims 3 to 6.