JPH0375551B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to iron-
5, 10, 15, 20-tetra [α, α, α, α-o-
(Substituted amido)phenyl]porphyrin complex. Iron()porphyrin complexes present in hemoglobin and myoglobin reversibly adsorb and desorb oxygen molecules. Many studies have been reported on the synthesis of complexes with oxygen adsorption/desorption functions similar to those of natural porphyrin iron() complexes. Examples include J.P. Collman, Accounts of Chemical
Reseach, 10 , 265 (1977); F. Basolo, BM
Hoffman and JAIbers, ibid, 8 , 384 (1975)
etc. In particular, iron()-5,10,15,20-tetra[α,α,α,α-o-(pivalamide ) phenyl]porphyrin complex (JP Collman et al.,
Journal of the American Chemical Society
97, 1427 (1975)). However, if even a small amount of water coexists with this complex, it will be immediately oxidized and cannot produce an oxygen complex. For this reason, the present inventors have already developed a porphyrin iron () complex system that enables the formation of stable oxygen complexes even in aqueous solutions and can adsorb and desorb oxygen reversibly. ) and Japanese Patent Application No. 107461/1983 (Japanese Patent Application No. 10514/1983). However, the iron ()-5, 10, 15,
20-Tetra [α, α, α, α-o-(pivalamide)
Phenyl]porphyrin complexes are not water-soluble at all, and when considering the medical purpose of oxygen carriers, compounds that quickly become water-soluble in vivo are more desirable. Therefore, the purpose of the present invention is to promptly, after performing the necessary function as an oxygen carrier in the living body,
The object of the present invention is to provide an iron ()porphyrin complex as an intermediate for a substance that can be decomposed and water-solubilized by enzymes in the body. According to this invention, the above purpose is achieved by the general formula (Here, n is an integer from 0 to 10, R is hydrogen,
_{5,10,15,20 -} tetra [α, α, α, α-o
-(Substituted amido)phenyl]porphyrin iron ()
Alternatively, this can be achieved by providing an iron() complex (provided that in the case of an iron() complex, it has a counter ion (for example, a halogen ion such as chlorine, bromine, etc.)). To synthesize the complex of the present invention, first, JP
Collman et al., Journal of the American
Formula obtained according to Chemical Society, 97 , 1427 (1975) 5, 10, 15, 20-tetra [α, α,
α,α-o-aminophenyl]porphyrin (hereinafter abbreviated as H ₂ TamPP) and ω-alkoxycarbonylalkyl acid chloride are subjected to a condensation reaction in anhydrous tetrahydrofuran in the presence of pyridine,
5, 10, 15, 20-tetra [α, α, α, α-o-
(Substituted amido)phenyl]porphyrin is obtained.
Iron is introduced by heating this with iron bromide () in anhydrous tetrahydrofuran in the presence of a small amount of pyridine under reflux to give the desired iron ()-5,
A 10,15,20-tetra[α,α,α,α-o-(substituted amido)phenyl]porphyrin complex can be synthesized. In this case, the complex has a bromide ion as a counterion. The ω-alkoxycarbonylalkyl acid chloride used in the above reaction has the formula (Here, R and n are as described above. However, R is a group other than hydrogen) The corresponding carboxylic acid derivative represented by It can be easily obtained by reacting with Note that the carboxylic acid derivative represented by formula (3) and its acid chloride can be synthesized by the following method. (1) n=0 J.Bu¨chi et al., Helvetica Chemica Aeta, 35
75-82 (1952) using dimethylmalonic acid as a starting material, it can be synthesized, for example, according to the following reaction. (2) Using n=1 2,2-dimethylsuccinic acid as a starting material, it can be synthesized, for example, according to the following reaction. (3) n=2 Yu-Neng Kuo et al. Journal of the American
According to the method of Chemical Society 93 6321-6323 (1971), using methyl acrylate as a starting material, it can be synthesized, for example, according to the following reaction. (4) n=3～10 PLCreger, Journal of the American
Applying the synthesis method for 2,2-dimethylbutanoic acid described in Chemical Society 89 2500 (1967), the formula Br-(CH ₂ -) _o COOR (where R is as described above ( (excluding hydrogen), n is 3 to 10), for example, when n = 10, 11
-It can be synthesized using bromoundecanoic acid as a raw material and according to the following reaction. In addition, when R in formula (1) is hydrogen, Fe()-5,10,15,20-tetra[α,α,α,α-o-(substituted amido)phenyl obtained as above] ]Ester group (−
COOR) with an alkali such as sodium hydroxide or an acid such as trifluoroacetic acid. The complex of this invention described above has a central iron of 3
By reducing the valent to divalent and coordinating an appropriate basic ligand to the axial position, it is oxygenated in a suitable organic solvent such as benzene at room temperature, and reversible adsorption and desorption of oxygen is possible. Can be used as Furthermore, when this complex is included in a liposome, it becomes possible to adsorb and desorb oxygen in an aqueous medium (eg, water, physiological saline, phosphate buffered water, etc.) under physiological conditions at room temperature. Iron-5,10,15,20-tetra[α,α,α,α-o-(substituted amido)phenyl] represented by formula (1)
As is well known, the carboxylic acid ester group contained in the substituted amide moiety of the porphyrin complex is easily decomposed in vivo by oxygen from esterases, lipases, etc., and becomes water-soluble. It has been confirmed that all of them become soluble in water when hydrolyzed to carboxylate salts, which is extremely advantageous when applied to living organisms. In addition, the basic ligand used is the formula {Here, R ¹ is hydrogen or C ₁ - C ₃ alkyl group,
Each of R ² , R ³ and R ⁴ is preferably an imidazole ligand represented by hydrogen, an alkyl group, or a hydrophobic substituent (eg, trityl group, phenyl group, phenyl alkyl group, alkoxycarbonyl alkyl group). In this case, when R ¹ is hydrogen, two imidazoles are coordinated to the two axial positions, while R ¹ is
In the case of a _C1 - _C3 alkyl group, only one imidazole is coordinated to one axis position. In addition, iron ()-5, 10, 15, 20-tetra [α,
In order to reduce the α,α,α-o-(substituted amido)phenyl]porphyrin complex to the corresponding iron()porphyrin complex, the iron()porphyrin complex and an excess amount of the imidazole of formula (2) are treated with a suitable organic Dissolve in a solvent such as benzene, add a small amount of 10% palladium/carbon as a catalyst, stir for several minutes under a hydrogen gas atmosphere, and then remove the catalyst with a filter under hydrogen gas. In this way, a benzene solution of an iron()porphyrin-imidazole complex in which the central iron is reduced and imidazole is coordinated is obtained. When oxygen gas is passed through this, a stable oxygen complex is formed, and oxygen can be adsorbed and desorbed repeatedly by performing an appropriate deoxidation operation (by blowing nitrogen gas or freezing and degassing). Furthermore, the iron()-5,10,15,20-tetra[α,α,α,α-o-(substituted amido)phenyl]
When a porphyrin-imidazole complex is included in a phospholipid liposome, it forms an oxygen complex that is stable in water at room temperature. In order to obtain such a liposome, the iron()porphyrin complex, an excess amount of the imidazole of formula (2), and a sufficient amount of phospholipid to solubilize them are dissolved in an appropriate solvent, and the above-mentioned hydrogen After the reduction operation with a gas, the solvent is distilled off from the resulting solution, and then an aqueous medium (e.g., water, phosphate buffer aqueous solution,
Physiological saline) is added and shaken to obtain an emulsified liposome containing an iron()porphyrin-substituted imidazole complex. This is further subjected to ultrasonication under an inert gas atmosphere to obtain a transparent liposome aqueous solution containing the iron()porphyrin complex. By blowing air or oxygen into this aqueous solution, an oxygen complex is formed, and reversible adsorption and desorption can be repeated. Note that the iron ()-tetra [α,
Liposomes containing α,α,α-o-(substituted amido)phenyl]porphyrin complexes have been disclosed in patent applications filed in 1983-
It can also be prepared by the reduction method using sodium dithionite described in No. 89312 or the reduction method using an enzyme system described in Japanese Patent Application No. 107462/1989. The iron-5,10,15,20-tetra[α,α,α,α-o-(substituted amido)phenyl]porphyrin complex of the present invention described above provides such a bioapplicable oxygen adsorbent. It is useful as an intermediate for Examples of this invention will be described below. Example 1 Methyl 2,2-dimethylmalonic acid chloride was prepared in the literature (J. Bu¨chi, G. Ene´zian, H. Eichenberger and
R. Lieberherr, Helvetica Chemica Acta, 35 ,
75-82 (1952)). Boiling point 58-61
℃/13mmHg. To a solution of 0.50 g of H ₂ TamPP and 2 ml of pyridine in anhydrous tetrahydrofuran (50 ml), 1.5 g of methyl 2,2-dimethylmalonic acid chloride (9.12 times molar equivalent) was added.
was added dropwise over 10 minutes and allowed to react at room temperature for 4 hours. After adding 100 ml of 4% aqueous sodium hydrogen carbonate solution to this, the mixture was extracted with 200 ml of chloroform. This extract was added to a 4% aqueous sodium bicarbonate solution (2 x 200 ml).
After washing and separating, the chloroform solution was dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated to dryness under reduced pressure. 50g of silica gel was added to the resulting oily residue,
It was purified by column chromatography using chloroform as a solvent. Collect the first elution fraction,
It was concentrated under reduced pressure. This was recrystallized from a chloroform-ether mixed solvent system, and the crystals were collected, washed with ether, and then dried in vacuum. Thus, 5,
10, 15, 20-tetra [α, α, α, α-o-(2,
2-dimethyl-2-methoxycarbonylacetamido)phenyl]porphyrin was obtained as purple needle-like crystals in an amount of 0.74 g and a yield of 84%. FDMS spectrum (M+H) ⁺ :1187 (Molecular formula C ₆₈ H ₆₆ N ₈ O ₁₂ = 1186) IR spectrum (KBr) ν3380, 1750, 1695,
1590, 1525, 1450, 1160, 1140, 970, 810,
765cm ^-1 Visible spectrum (CHCl ₃ ) λmax 418, 512, 544,
586, 645nm PMR spectrum ( _CDCl3 ) δ (ppm) −2.54 (2H,
s), 0.61 (24H, s), 2.04 (12H, s), 7.4~
8.7 (16H, m), 8.80 (8H, s). CMR spectrum ( _CDCl3 ) No. δ (ppm) No. δ (ppm) α 147.0 7 173.08 β 131.39 8 50.14 m 114.89 9 169.32 1 131.39 10 51.43 2 138.09 11 22.72 3 121.53 4 129.81 5 1 23.35 6 134.92 Elemental analysis value (%) C ₆₈ H ₆₆ C: 68.74 (68.79) for N ₈ O ₁₂ ; H: 5.75 (5.60); N: 9.17
(9.44). However, the values in parentheses are calculated values. Iron introduction is 5, 10, 15, 20-tetra [α, α,
After bubbling nitrogen gas into a solution of 0.38 g of α,α-o-(2,2-dimethyl-2-methoxycarbonylacetamido)phenyl]porphyrin and 0.5 ml of pyridine in anhydrous tetrahydrofuran (40 ml) for 2 hours, ferrous bromide was added. - The reaction was carried out by adding 1.2 g (large excess) of dihydrate and refluxing for 4 hours.
The reaction product was extracted with 200 ml of chloroform, washed with water (2 x 100 ml), separated, and the chloroform solution was dried over anhydrous sodium sulfate and dried under reduced pressure to obtain a black solid. Alumina column chromatography was performed using chloroform as a solvent, and the first eluted portion was collected, shaken with 3 ml of 48% hydrogen bromide, dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure. Recrystallization is performed from a mixed solvent of chloroform-ether-petroleum ether, and the crystals are collected and dried under vacuum to obtain the desired bromo{5,10,15,20-tetra[α,α,α,α-o-(2, 2-dimethyl-2-
methoxycarbonylacetamide) phenyl]porphyrinat}iron () (hereinafter referred to as Fe(TacePP _OCH3 )
It is abbreviated as Br. ) was obtained in a yield of 0.22 g and 52%. FDMS spectrum (M+H) ⁺

Claims (1)

[Claims] 1. General formula (Here, n is an integer from 0 to 10, R is hydrogen,
_{5,10,15,20 -} tetra [α, α, α, α-o
-(Substituted amido)phenyl]porphyrin iron ()
or an iron() complex (provided that it has a counter ion in the case of an iron() complex).