Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
  • C07D487/22—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains four or more hetero rings
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
  • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
  • A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
  • A61K47/6949—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
  • A61P7/06—Antianaemics
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B82—NANOTECHNOLOGY
  • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
  • B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery

Definitions

• the present invention relates to a novel porphyrin compounds such as a porphyrin metal complex, an albumin inclusion compound thereof, and an artificial oxygen carrier.
• Heme i.e., a porphyrin iron (II) complex, constituting the active center of hemoglobin or myoglobin that plays the role of carrying and storing oxygen within the living body, is capable of reversibly adsorbing and desorbing molecular oxygen in response to oxygen partial pressure.
• II porphyrin iron
• FeTpivPP complex 5,10,15,20-tetrakis( ⁇ , ⁇ , ⁇ , ⁇ -pivalamidophenylporphyrin iron (II) complex
• FeTpivPP complex is known as a porphyrin iron (II) complex that is reported to be capable of forming an oxygen complex that is stable under room temperature conditions (see J. P. Collman, et al., J. Am. Chem. Soc., 97, 1427 (1975).
• the FeTpivPP complex is capable of reversibly adsorbing and desorbing the oxygen molecule at room temperature within an organic solvent such as benzene, toluene, dichloromethane, tetrahydrofuran or N,N-dimethylformamide, if an excess amount of an axial base such as 1-alkylimidazole or 1-alkyl-2-methylimidazole is co-present.
• an organic solvent such as benzene, toluene, dichloromethane, tetrahydrofuran or N,N-dimethylformamide
• an axial base such as 1-alkylimidazole or 1-alkyl-2-methylimidazole
• the research group including the present inventors have took the position that a stable oxygen carrier can be provided without externally adding the axial base, if an imidazole is covalently bonded, as a side chain substituent, to the porphyrin iron (II) complex.
• a FeTpivPP analogue having a substituent in the 2-position of the porphyrin ring have been accurately synthesized.
• an inclusion compound having the FeTpivPP analogue included in the phospholipid vesicle or the human serum albumin has been prepared, providing an artificial oxygen carrier capable of reversibly adsorbing and desorbing oxygen (see Japanese Patent Disclosure (Kokai) No. 59-164791, Japanese Patent Disclosure No. 59-162924, Japanese Patent Disclosure No. 6-271577 and Japanese Patent Disclosure No. 8-301873).
• the group of the present inventors have also synthesized a derivative in which an alkylimidazole is covalently bonded to the protoporphyrin iron complex, and prepared an oxygenated complex in respect of the compound having the above-noted derivative included in albumin (see above mentioned Japanese Patent Disclosure No. 8-301873).
• the half-life period of its oxygenated complex is not longer than one hour under 25° C. It follows that the oxygenated complex leaves room for further improvement in terms of the stability in the case of using the oxygenated complex as an artificial oxygen carrier.
• the protoporphyrin iron complex derivative is advantageous also in the case where the administration into the living body is taken into account.
• Protoporphyrin iron (III) that has been no longer utilized in the living body is caught by heme oxidases so as to cleave the ⁇ -meso position of the porphyrin and thus is decomposed into biliberdin so as to be used in the metabolic process. Since the hydrogen atom in the meso position is substituted with the phenyl ring in the tetraphenylporphyrin iron complex, the tetraphenylporphyrin iron complex may not be decomposed in the metabolic mechanism.
• An object of the present invention is to provide a porphyrin compound capable of forming an oxygenated complex having a further improved stability, an albumin inclusion compound thereof, and an artificial oxygen carrier.
• the group of the present inventors have synthesized and reported various porphyrin metal complexes having the oxygen binding and dissociating function. Also, the group of the present inventors have provided the porphyrin metal complex-albumin inclusion compound having the porphyrin metal complex included in the serum albumin as an artificial oxygen carrier within water (see Japanese Patent Disclosure No. 8-301873 referred to previously).
• the group of the present inventors have clarified that, where an intramolecular axially coordinated base is constituted by a histidine derivative, the stability of the oxygenated complex in the porphyrin metal complex-albumin inclusion compound is improved, as compared to the case where the intramolecular axially coordinated base is constituted by an imidazolylalkyl (see T. Komatsu, et al., Bioconjugate Chem., 13, 397 (2002)).
• the present inventors have continued an extensive research based on the results of the research on the porphyrin metal complex capable of coupling oxygen and the research on the porphyrin metal complex inclusion compound having the porphyrin metal complex included therein as an active site, accomplishing the present invention.
• R 1 denotes a C 1 -C 18 alkyloxy group, a C 1 -C 18 alkylamino group, or a peptide having 1-6 ⁇ -amino acids and having a hydroxyl group, a benzyl oxy group or a methoxy group at the C-terminal
• R 2 denotes a residual group after removal of an amino group and a carboxyl group from an ⁇ -amino acid
• R 3 denotes a C 1 -C 18 alkyloxy group, a C 1 -C 18 alkylamino group, or a peptide having 1-6 ⁇ -amino acids and having a hydroxyl group, a benzyloxy group or a methoxy group at the C-terminal
• each R 4 and each R 5 denote either a methyl group, or a hydrogen atom, a vinyl group, an ethyl group, a 1-methoxyethyl group, a 1-bromoe
• M in general formula (A) denotes Fe or Co
• the present invention provides a porphyrin metal complex-albumin inclusion compound having the porphyrin metal complex included in albumin.
• the present invention provides an artificial oxygen carrier containing, as an active component, the porphyrin metal complex-albumin inclusion compound of the present invention.
• porphyrin compound of the present invention is represented by general formula (A) given above.
• R 1 denotes a C 1 -C 18 alkyloxy group, a C 1 -C 18 alkylamino group, or a peptide having 1-6 ⁇ -amino acids and having a hydroxyl group, a benzyl oxy group or a methoxy group at the C-terminal.
• R 3 denotes a C 1 -C 18 alkyloxy group, a C 1 -C 18 alkylamino group, or a peptide having 1-6 ⁇ -amino acids and having a hydroxyl group, a benzyloxy group or a methoxy group at the C-terminal.
• Each R 4 and each R 5 denote either a methyl group, or a hydrogen atom, a vinyl group, an ethyl group, a 1-methoxy ethyl group, a 1-bromoethyl group or a formyl group, wherein, where each R 4 denotes a methyl group, each R 5 denotes a hydrogen atom, a vinyl group, an ethyl group, a 1-methoxyethyl group, 1-bromoethyl group or a formyl group, and where each R 4 denotes a hydrogen atom, a vinyl group, an ethyl group, a 1-methoxyethyl group, a 1-bromoethyl group or a formyl group, each R 5 denotes a methyl group.
• M denotes two hydrogen atoms bonded to two pyrrole nitrogen atoms or an ion of a transition metal belonging to the fourth to fifth periods in the Periodic Table such as chromium, manganese, iron, cobalt or ruthenium.
• a transition metal such as chromium, manganese, iron, cobalt or ruthenium.
• iron or cobalt is preferred.
• iron is particularly preferred.
• X ⁇ denotes a halogen ion that is present in the case where M denotes the transition metal ion, i.e., where the porphyrin compound of the present invention is a metal complex.
• n which denotes the number of X is the number obtained by subtracting 2 from the valency of the transition metal ion.
• the porphyrin compound of the present invention includes the compound represented by formula (A1) given below and the compound represented by formula (A2) given below:
• R 1 -R 3 , X ⁇ , and n are as defined above, and R 4 denotes a hydrogen atom, a vinyl group, an ethyl group, a 1-methoxyethyl group, a 1-bromoethyl group or a formyl group.
• R 1 -R 3 , X 31 , and n are as defined above, and R 5 denotes a hydrogen atom, a vinyl group, an ethyl group, a 1-methoxyethyl group, a 1-bromoethyl group or a formyl group.
• the porphyrin compound of the present invention can be synthesized by various methods.
• the compound can be synthesized by reacting a porphyrin represented by formula (1) given below with a histidine derivative represented by formula (2) given below:
• R 4 and R 5 are as defined above;
• R 2 and R 3 are as defined above.
• the porphyrin represented by formula (1) may be used in the form of, for example, a disodium salt.
• the histidine derivative represented by formula (2) may be used in the form of a hydrohalide salt.
• the porphyrin represented by formula (1) includes, for example, protoporphyrin IX, meso-porphyrin IX, and deuteroporphyrin IX.
• the histidine derivative represented by formula (2) can be prepared according to an ordinary method by subjecting histidine and the ⁇ -amino acid: H 2 N(R)CHCOOH noted above to amide forming reaction between the amino group of histidine and the carboxyl group of the ⁇ -amino acid, followed by modifying the carboxyl group of histidine.
• the histidine derivative can be obtained by the reaction between the histidine after reaction with ⁇ -amino acid and an alcohol: R 0 OH, where R 0 denotes a C 1 -C 18 alkyl group to esterify the histidine (where R 3 denotes a C 1 -C 18 alkyl group), or with an amine: R 0 NH 2 , where R 0 denotes a C 1 -C 18 alkyl group to amidate the histidine (where R 3 denotes a C 1 -C 18 alkylamino group), according to an ordinary method.
• R 3 denotes a peptide
• the histidine after reaction with the ⁇ -amino acid is successively subjected to the reaction with the ⁇ -amino acid: H 2 N(R)CHCOOH according to an ordinary method, and the C-terminal is converted into a benzyl ester or an amino ester according to an ordinary method, as required.
• the histidine derivative represented by formula (2) is commercially available.
• the porphyrin represented by formula (1) is dissolved in a suitable organic solvent such as pyridine or dimethylformamide (DMF), followed by adding an activating agent for activating the carboxyl group of the porphyrin represented by formula (1), for example, (benzotriazole-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphoric acid (BOP), to the resultant solution, and the mixture thus obtained is stirred for, e.g., 10 minutes to one hour.
• a suitable organic solvent such as pyridine or dimethylformamide (DMF)
• an activating agent for activating the carboxyl group of the porphyrin represented by formula (1) for example, (benzotriazole-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphoric acid (BOP)
• the activating agent such as BOP can be used in an amount of at least 2 moles relative to one mole of porphyrin represented by formula (1) in the case where the histidine derivative represented by formula (2) is reacted with one of the two carboxyl groups of porphyrin, and the other carboxyl group is modified as described herein later.
• activating agent such as BOP can be used in an amount of 1.0-2.0 moles relative to one mole of the porphyrin represented by formula (1) in the case where the histidine derivative represented by formula (2) is reacted with one of the two carboxyl groups of the porphyrin and the other carboxyl group is not modified.
• the histidine derivative represented by formula (2) which is dissolved in a suitable organic solvent such as DMF or pyridine, is slowly added by using, for example, a dropping funnel to the reaction mixture after the stirring noted above.
• the histidine derivative can be used in an amount of 0.5 to 1.0 mole relative to one mole of the porphyrin represented by formula (1).
• the reaction mixture is stirred at room temperature for, e.g., 1 to 12 hours under the condition that the light is shielded so as to carry out the desired reaction.
• a porphyrin compound represented by formula (3) given below can be obtained by this reaction:
• the porphyrin compound represented by formula (3) is reacted with an alcohol: R 0 OH, where R 0 denotes a C 1 -C 18 alkyl group, to esterify the carboxyl group (where R 1 is a C 1 -C 18 alkyl oxy group), or with an amine: R 0 NH 2 , where R 0 denotes a C 1 -C 18 alkyl group, to amidate the carboxyl group (where R 1 is a C 1 -C 18 alkyl amino group).
• R 1 denotes the peptide
• the histidine after the reaction with the ⁇ -amino acid is successively reacted with the ⁇ -amino acid: H 2 N(R)CHCOOH so as to convert the C-terminal into a benzyl ester or an amino ester by an ordinary method, as required.
• a porphyrin compound represented by formula (4) given below is obtained in which R 1 denotes an atomic group other than the hydroxyl group:
• the iron (III) complex can be reduced into the iron (II) complex by using a suitable reducing agent such as sodium dithionite or ascorbic acid according to an ordinary method so as to develop an oxygen binding activity.
• a suitable reducing agent such as sodium dithionite or ascorbic acid
• the porphyrin compound of the present invention comprises a metal free porphyrin compound in which M denotes two hydrogen atoms, and a porphyrin metal complex in which M denotes a transition metal.
• M denotes two hydrogen atoms
• M denotes a transition metal.
• the central metal M is in +2 in the latter metal complex
• the imidazole group of the histidine derivative covalently bonded to the propionic acid residue bonded to the porphyrin ring is coordinated with the central metal M as a proximal base.
• the porphyrin metal complex of the present invention the +2 central metal M can exhibit an oxygen binding capability by itself so as to make it unnecessary to add from the outside an imidazole derivative as an axial base.
• the porphyrin metal complex comprises iron or cobalt as the central metal M
• the porphyrin metal complex can be encapsulated in a phospholipid bi-layer vesicle by an ordinary method (see Japanese Patent Disclosure No. 8-301873 referred to previously), or can be enclosed in a phospholipid-encapsulated fat emulsion (see Japanese Patent Disclosure No. 6-184156).
• the porphyrin metal complex can also be included in albumins such as a bovine serum albumin, a human serum albumin, a recombinant human serum albumin, and a polymer albumin (see T. Komatsu, et al., Bioconjugate Chem., 13, 387 (2002)).
• the porphyrin iron or cobalt complex of the present invention is capable of promptly forming a stable oxygenated complex upon contact with oxygen even in an aqueous system.
• the porphyrin metal complex of the present invention is capable of adsorbing and desorbing oxygen in accordance with the oxygen partial pressure. The binding and dissociating of oxygen can be reversibly effected repeatedly.
• a derivative of histidine, which is one of the natural amino acids is used as a proximal base, and a porphyrin having the meso-position not modified is used as the porphyrin. It follows that the porphyrin metal complex of the present invention can provide a useful artificial oxygen carrier that is highly excellent in the adaptability to the living body.
• porphyrin metal complex of the present invention is utilized as an artificial oxygen carrier in, for example, a substitute for a transfusing blood, a blood diluent before the surgical operation, a replenishing solution in an external circulating circuit such as an artificial lung, a perfusion solution of a transplanted internal organs, an oxygen supply solution into the ischemia portion, a treating agent of a chronic amenia, a perfusion solution for a liquid ventilation, a sensitizing agent for curing cancer, and a culture solution for a regenerated tissue cell. It is also expected for the porphyrin metal complex of the present invention to be utilized in a rare blood type patient, a patient who denies blood transfusion for religious reasons, and an animal therapy.
• the porphyrin compound of the present invention is in the form of a complex of an ion of a metal belonging to, for example, the fourth period of the Periodic Table
• the complex can be utilized as a catalyst in an oxygen reducing reaction, an oxygen oxidizing reaction or an oxygen addition reaction.
• the porphyrin metal complex of the present invention can be used as, particularly, an adsorbing-desorbing agent of an oxygen gas, as a redox catalyst, as a catalyst for an oxygen oxidizing reaction, and as a catalyst for an oxygen addition reaction in addition to the use as an artificial oxygen carrier.
• a gas capable of coordination with the central metal M such as carbon monoxide, nitrogen monoxide or nitrogen dioxide can form a coordinated complex with the porphyrin metal complex of the present invention. It follows that the porphyrin metal complex of the present invention can be used as an adsorbent of such a gas.
• Protoporphyrin IX 400 mg (0.71 mmol) was dissolved in pyridine (40 mL), and the resultant solution was stirred at room temperature for 10 minutes. Then, BOP (840 mg (1.9 mmol)) was added to the solution, and the solution was further stirred for 10 minutes. Further, a solution of glycyl-L-histidine methyl ester dihydrochloride (129 mg (0.57 mmol)) in DMF (15 mL) was slowly added dropwise into the resultant solution by using a dropping funnel.
• the reaction mixture was stirred for 2.5 hours at room temperature under a light-shielded condition, followed by adding ethanol (4.2 mL (71 mmol)) to the reaction mixture, and the mixture was stirred for 18 hours.
• Deuteroporphyrin IX dihydrochloride 400 mg (0.69 mmol) was dissolved in pyridine (50 mL), and the resultant solution was stirred at room temperature for 10 minutes. Then, BOP (816 mg (1.8 mmol)) was added to the resultant solution, followed by further stirring the solution for 10 minutes. After a solution of valyl-L-histidinyl leucyl-leucine methyl ester dihydrochloride (3741 mg (0.55 mmol)) in pyridine (15 mL) was slowly added dropwise into the resultant solution by using a dropping funnel, the solution was stirred at room temperature for 6 hours under a light-shielded condition.
• porphyrin iron (II) complex 20 ⁇ M; serum albumin: 20 ⁇ M
• a dispersion of the porphyrin iron complex-albumin inclusion compound in a phosphate buffer (porphyrin iron (II) complex: 20 ⁇ M; serum albumin: 20 ⁇ M) was put in a cell made of quartz for a spectral measurement, and the cell was hermetically sealed under a nitrogen gas atmosphere.
• the visible light absorption spectrum of the dispersion showed ⁇ max at 423; 539; and 569 nm, supporting that the included porphyrin iron (II) complex forms an Fe(II) high spin 5-coordinated complex having a single intramolecular axial base coordinated therein.
• the present invention provides a porphyrin compound capable of forming an oxygenated complex having a further improved stability.
• the albumin inclusion body of the porphyrin compound can function as an artificial oxygen carrier.

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• 2003
  • 2003-03-14 JP JP2003069760A patent/JP3945428B2/ja not_active Expired - Fee Related
• 2004
  • 2004-03-11 DE DE602004002975T patent/DE602004002975T2/de not_active Expired - Lifetime
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  • 2004-03-11 EP EP04005777A patent/EP1466915B1/de not_active Expired - Lifetime
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