JP2008094740A - Novel antimalaric - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a highly useful and safe antimalaric. <P>SOLUTION: The antimalaric contains as an active substance a bispyridinium salt compound represented by the general formula:(wherein two R<SB>1</SB>s, which may be same or different, each represents a 1-10C alkyl group or a 1-10C alkyl group with an aromatic ring, which may have a substituent, at the end, R<SB>2</SB>represents a phenylene group with a 1-20C linear alkylene or a 0-5C alkylene group at both ends, whose benzene ring may have a substituent, two R<SB>3</SB>s, which may be same or different, each represents a 1-20C alkyl group or a 1-10C alkyl group with an aromatic ring, which may have a substituent, at the end, and X<SP>-</SP>represents an anion). <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a novel antimalarial agent containing a bispyridinium salt compound as an active ingredient.

  Malaria caused by Plasmodium infection is the largest parasitic protozoan infection of mankind, and according to the latest statistics of the World Health Organization (WHO), 267 million people worldwide are infected with malaria every year, of which 200 Ten thousand people have died. Malaria-causing pathogens are protozoa belonging to the genus Plasmodium, such as P. falciparum distributed throughout the tropical regions of Africa, Asia and Latin America, and one of the tropical and temperate regions of the world. Protozoa such as Plasmodium falciparum (P. vivax) distributed in Japan, egg-shaped malaria parasite (P. ovale) mainly distributed in tropical West Africa, and P. malariae distributed in various parts of the world Infects humans through the mosquitoes. There is a tendency to increase as an imported infectious disease in Japan, and there are cases where death occurs due to delays in diagnosis and treatment.

  Quinine, chloroquine, pyrimethamine, artemisinin, mefloquine, etc. are used for the treatment and prevention of malaria. However, as represented by chloroquine retinopathy, conventional antimalarial agents have problems in terms of safety, and also have problems with drug resistance. In such a situation, WHO has set the development of new chemotherapeutic agents as an important goal. Therefore, development of a new type of antimalarial agent with low toxicity for treating malaria infection is desired. In addition, since malaria-infected areas include many developing countries, it is desired that antimalarial drugs can be synthesized and supplied at low cost.

  The inventors of the present invention have been searching for highly active antimalarial agents that can be synthesized in a low yield with a low number of steps using inexpensive raw materials, and have reported many results. In such a series of studies, the present inventors have disclosed an antimalarial agent containing a bisquaternary ammonium salt compound as an active ingredient in JP-A-2005-179212. Similarly, there is a report of Hamze et al. (J. Med. Chem., 2005, 48, 3639-3643) that a dimer-type compound, a bisthiazolium salt, has antimalarial activity.

  The present inventors have also reported N, N′-octamethylenebis (4-carbamoyl-1-hexylpyridinium bromide) having high antimalarial activity (Bioorg. Med. Chem. Lett., 2006, 16, 2758-2760). However, a bispyridinium salt compound having a structure in which the amide is arranged in the side chain and bonded at the pyridinium dication site instead of the amide bond which is the linker portion of this compound has not been studied so far.

  It is reported in JP-A-8-76314 that a compound containing a nitrogen-containing heterocycle is used as a silver halide photographic light-sensitive material, and the nitrogen-containing heterocycle may be bonded to form a bis-type structure. The effect is described. However, what is reported in JP-A-8-76314 is a silver halide photographic light-sensitive material, and this patent is not related to antimalarial activity.

JP-A-2005-179212 JP-A-8-76314 Hamze, A. et al., J. Med. Chem., 2005, 48, 3639-3643 Fujimoto, K. et al., Bioorg.Med.Chem.Lett., 2006, 16, 2758-2760

  Compounds having such an excellent antimalarial effect have been reported so far, but since malaria is an important disease, a novel antimalarial agent having an excellent effect and high safety is further added. It was sought after. Therefore, the object of the present invention is to develop studies on bis-quaternary ammonium salt compounds having a further structure and to study their biological activity. And thereby, it is providing the antimalarial agent useful for the prevention and / or treatment of the infectious disease by the malaria parasite, especially the novel antimalarial agent which has high usefulness and safety | security.

式中、２つのＲ_１は同一もしくは異なり、それぞれ炭素数１から１０のアルキル基、又は置換基を有してもよい芳香環を末端に有する炭素数１から１０のアルキル基を表し、
Ｒ_２は、炭素数１から２０の直鎖のアルキレン基、又は炭素数０から５のアルキレン基を両端に有し、そのベンゼン環に置換基を有してもよいフェニレン基を表し、
２つのＲ_３は同一もしくは異なり、それぞれ炭素数１から２０のアルキル基、又は置換基を有してもよい芳香環を末端に有する炭素数１から１０のアルキル基を表し、
Ｘ^-はアニオンを表す。 Therefore, the present invention is an antimalarial agent characterized by containing a bispyridinium salt compound represented by the following general formula (1) as an active ingredient,

In the formula, two R _{1 s} are the same or different and each represents an alkyl group having 1 to 10 carbon atoms, or an alkyl group having 1 to 10 carbon atoms terminated with an aromatic ring which may have a substituent,
R ₂ represents a linear alkylene group having 1 to 20 carbon atoms or a phenylene group having an alkylene group having 0 to 5 carbon atoms at both ends and optionally having a substituent on the benzene ring;
Two R _{3 s} are the same or different and each represents an alkyl group having 1 to 20 carbon atoms, or an alkyl group having 1 to 10 carbon atoms having an aromatic ring which may have a substituent;
X ⁻ represents an anion.

式中、２つのＲ_１は同一もしくは異なり、それぞれ炭素数１から１０のアルキル基、又は置換基を有してもよい芳香環を末端に有する炭素数１から１０のアルキル基を表し、
Ｒ_２は、炭素数１から２０の直鎖のアルキレン基、又は炭素数０から５のアルキレン基を両端に有し、そのベンゼン環に置換基を有してもよいフェニレン基を表し、
２つのＲ_３は同一もしくは異なり、それぞれ炭素数１から２０のアルキル基、又は置換基を有してもよい芳香環を末端に有する炭素数１から１０のアルキル基を表し、
Ｘ^-はアニオンを表す。 Furthermore, the present invention is an antimalarial agent characterized by containing a bispyridinium salt compound represented by the following general formula (2) as an active ingredient,

In the formula, two R _{1 s} are the same or different and each represents an alkyl group having 1 to 10 carbon atoms, or an alkyl group having 1 to 10 carbon atoms terminated with an aromatic ring which may have a substituent,
R ₂ represents a linear alkylene group having 1 to 20 carbon atoms or a phenylene group having an alkylene group having 0 to 5 carbon atoms at both ends and optionally having a substituent on the benzene ring;
Two R _{3 s} are the same or different and each represents an alkyl group having 1 to 20 carbon atoms, or an alkyl group having 1 to 10 carbon atoms having an aromatic ring which may have a substituent;
X ⁻ represents an anion.

式中、Ｒ_１は炭素数１から２０の直鎖のアルキレン基、炭素数３から２０の分岐したアルキレン基、又は炭素数０から５のアルキレン基を両端に有し、そのベンゼン環に置換基を有してもよいフェニレン基を表し、
２つのＲ_２は同一もしくは異なり、それぞれ炭素数０から２０のアルキル基、又は置換基を有してもよい芳香環を末端に有する炭素数１から１５のアルキル基を表し、
Ｘ^-はアニオンを表す。 Furthermore, the present invention is an antimalarial agent characterized by containing a bispyridinium salt compound represented by the following general formula (3) as an active ingredient,

In the formula, R ₁ has a linear alkylene group having 1 to 20 carbon atoms, a branched alkylene group having 3 to 20 carbon atoms, or an alkylene group having 0 to 5 carbon atoms at both ends, and a substituent on the benzene ring. Represents a phenylene group which may have
Two R _{2 s} are the same or different and each represents an alkyl group having 0 to 20 carbon atoms, or an alkyl group having 1 to 15 carbon atoms having an aromatic ring which may have a substituent;
X ⁻ represents an anion.

式中、Ｒ_１は炭素数１から２０の直鎖のアルキレン基、炭素数３から２０の分岐したアルキレン基、又は炭素数０から５のアルキレン基を両端に有し、そのベンゼン環に置換基を有してもよいフェニレン基を表し、
２つのＲ_２は同一もしくは異なり、それぞれ炭素数０から２０のアルキル基、又は置換基を有してもよい芳香環を末端に有する炭素数１から１５のアルキル基を表し、
Ｘ^-はアニオンを表す。 Furthermore, the present invention is an antimalarial agent characterized by containing a bispyridinium salt compound represented by the following general formula (4) as an active ingredient,

In the formula, R ₁ has a linear alkylene group having 1 to 20 carbon atoms, a branched alkylene group having 3 to 20 carbon atoms, or an alkylene group having 0 to 5 carbon atoms at both ends, and a substituent on the benzene ring. Represents a phenylene group which may have
Two R _{2 s} are the same or different and each represents an alkyl group having 0 to 20 carbon atoms, or an alkyl group having 1 to 15 carbon atoms having an aromatic ring which may have a substituent;
X ⁻ represents an anion.

式中、Ｒ_１は炭素数１から２０の直鎖のアルキレン基、炭素数３から２０の分岐したアルキレン基、又は炭素数０から５のアルキレン基を両端に有し、そのベンゼン環に置換基を有してもよいフェニレン基を表し、
４つのＲ_２は同一もしくは異なり、それぞれ炭素数０から２０のアルキル基、又は置換基を有してもよい芳香環を末端に有する炭素数１から１５のアルキル基を表し、
Ｘ^-はアニオンを表す。 Furthermore, the present invention is an antimalarial agent characterized by containing a bispyridinium salt compound represented by the following general formula (5) as an active ingredient,
General formula (5)

In the formula, R ₁ has a linear alkylene group having 1 to 20 carbon atoms, a branched alkylene group having 3 to 20 carbon atoms, or an alkylene group having 0 to 5 carbon atoms at both ends, and a substituent on the benzene ring. Represents a phenylene group which may have
Four R _{2 s} are the same or different and each represents an alkyl group having 0 to 20 carbon atoms or an alkyl group having 1 to 15 carbon atoms having an aromatic ring which may have a substituent;
X ⁻ represents an anion.

  Since the bispyridinium salt compound of the present invention has high safety and exhibits antimalarial activity at a low concentration, an antimalarial agent containing the compound is useful for the prevention and / or treatment of infectious diseases caused by malaria parasites. It is.

  Hereinafter, the antimalarial agent of the present invention will be described in more detail.

一般式（２）

で示されるビスピリジニウム塩化合物を有効成分として含有することを特徴とする。 The antimalarial agent of the present invention has the general formula (1) or the general formula (2).
General formula (1)

General formula (2)

It contains the bispyridinium salt compound shown by these as an active ingredient.

In the formula, two R _{1 s} may be the same or different, and each represents an alkyl group having 1 to 10 carbon atoms, or an alkyl group having 1 to 10 carbon atoms terminated with an aromatic ring which may have a substituent. To express. Preferred examples of the aromatic ring include benzene ring, naphthalene ring, pyridine ring, pyrimidine ring, quinoline ring, isoquinoline ring, quinazoline ring, furan ring, pyrrole ring, thiophene ring, benzofuran ring, indole ring, benzothiophene ring, A coumarin ring, an isocoumarin ring, etc. can be mentioned. The alkyl group may be linear or branched, for example, methyl, ethyl, propyl, iso-propyl, butyl, iso-butyl, sec-butyl, tert-butyl, pentyl Group, iso-pentyl group, sec-pentyl group, hexyl group, heptyl group, octyl group, 2-ethylhexyl group, nonyl group, decyl group, isodecyl group, dodecyl group, tetradecyl group, hexadecyl group, octadecyl group, etc. It is done. Examples of the substituent that the aromatic ring may have include, for example, amino group, nitro group, cyano group, carboxyl group, trifluoro group, methyl group, ethyl group, n-propyl group, isopropyl group, and n-butyl group. , Tert-butyl group, halogen and the like.

R ₂ represents a linear alkylene group having 1 to 20 carbon atoms or a phenylene group having an alkylene group having 0 to 5 carbon atoms at both ends and optionally having a substituent on the benzene ring. Note that when the number of carbon atoms of the alkylene group R ₂ is zero R ₂ is a phenylene group which may have a substituent. The alkylene group - _{(CH 2) n} - are preferred those represented by. The phenylene group may be any of 1,4-, 1,3-, or 1,2-phenylene groups. Examples of the substituent that the benzene ring may have include, for example, an amino group, a nitro group, a cyano group, a carboxyl group, a trifluoro group, a methyl group, an ethyl group, an n-propyl group, an isopropyl group, and an n-butyl group. , Tert-butyl group, halogen and the like.

Two R _{3 s} may be the same or different and each represents an alkyl group having 1 to 20 carbon atoms or an alkyl group having 1 to 10 carbon atoms having an aromatic ring which may have a substituent. Preferred examples of the aromatic ring include benzene ring, naphthalene ring, pyridine ring, pyrimidine ring, quinoline ring, isoquinoline ring, quinazoline ring, furan ring, pyrrole ring, thiophene ring, benzofuran ring, indole ring, benzothiophene ring, A coumarin ring, an isocoumarin ring, etc. can be mentioned. The alkyl group may be linear or branched, for example, methyl group, ethyl group, propyl group, iso-propyl group, butyl group, iso-butyl group, sec-butyl group, tert-butyl group, pentyl Group, iso-pentyl group, sec-pentyl group, hexyl group, heptyl group, octyl group, 2-ethylhexyl group, nonyl group, decyl group, isodecyl group, dodecyl group, tetradecyl group, hexadecyl group, octadecyl group, etc. It is done. Examples of the substituent that the aromatic ring may have include, for example, amino group, nitro group, cyano group, carboxyl group, trifluoro group, methyl group, ethyl group, n-propyl group, isopropyl group, and n-butyl group. , Tert-butyl group, halogen and the like.

X ⁻ represents an anion. The anions include inorganic anions such as bromine ions and chlorine ions, organic anions such as acetate anions, propionate anions, oxalate anions and succinate anions.

一般式（４）

一般式（５）

で示されるビスピリジニウム塩化合物を有効成分として含有することを特徴とする。 Furthermore, the antimalarial agent of the present invention is represented by general formula (3), general formula (4), or general formula (5).
General formula (3)

General formula (4)

General formula (5)

It contains the bispyridinium salt compound shown by these as an active ingredient.

In the formula, R ₁ has a linear alkylene group having 1 to 20 carbon atoms, a branched alkylene group having 3 to 20 carbon atoms, or an alkylene group having 0 to 5 carbon atoms at both ends, and a substituent on the benzene ring. Represents a phenylene group which may have Note that when the number of carbon atoms of the alkylene group R ₁ is 0 R ₁ is a phenylene group which may have a substituent. The alkylene group - _{(CH 2) n} - are preferred those represented by. The phenylene group may be any of 1,4-, 1,3-, or 1,2-phenylene groups. Further, the substituents that the benzene ring may have include amino group, nitro group, cyano group, carboxyl group, trifluoro group, methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, tert- A butyl group, halogen, etc. can be mentioned.

Two or four R _{2 s} may be the same or different, and each represents an alkyl group having 0 to 20 carbon atoms, or an alkyl group having 1 to 15 carbon atoms terminated with an aromatic ring which may have a substituent. To express. Note that when the number of carbon atoms of the alkyl group R ₂ is zero R ₂ is hydrogen. Preferred examples of the aromatic ring include benzene ring, naphthalene ring, pyridine ring, pyrimidine ring, quinoline ring, isoquinoline ring, quinazoline ring, furan ring, pyrrole ring, thiophene ring, benzofuran ring, indole ring, benzothiophene ring, A coumarin ring, an isocoumarin ring, etc. can be mentioned. The alkyl group may be linear or branched, for example, methyl group, ethyl group, propyl group, iso-propyl group, butyl group, iso-butyl group, sec-butyl group, tert-butyl group, pentyl Group, iso-pentyl group, sec-pentyl group, hexyl group, heptyl group, octyl group, 2-ethylhexyl group, nonyl group, decyl group, isodecyl group, dodecyl group, tetradecyl group, hexadecyl group, octadecyl group, etc. It is done. Examples of the substituent that the aromatic ring may have include, for example, amino group, nitro group, cyano group, carboxyl group, trifluoro group, methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, Examples thereof include a tert-butyl group and halogen.

  In addition, the following compounds can be mentioned as specific examples of the compound of the present invention that can be used as an active ingredient having antimalarial activity. These compounds are novel substances.

1,1 ′-(1,6-hexamethylene) bis [4-{(n-butylamino) carbonyl} pyridinium bromide].
1,1 '-(1,8-octamethylene) bis [4-{(n-butylamino) carbonyl} pyridinium bromide].
1,1 ′-(1,10-Decamethylene) bis [4-{(n-butylamino) carbonyl} pyridinium bromide].
1,1 ′-(1,12-dodecamethylene) bis [4-{(n-butylamino) carbonyl} pyridinium bromide].
1,1 ′-(1,6-hexamethylene) bis [4-{(n-hexylamino) carbonyl} pyridinium bromide].
1,1 ′-(1,8-octamethylene) bis [4-{(n-hexylamino) carbonyl} pyridinium bromide].
1,1 ′-(1,10-Decamethylene) bis [4-{(n-hexylamino) carbonyl} pyridinium bromide].
1,1 ′-(1,12-dodecamethylene) bis [4-{(n-hexylamino) carbonyl} pyridinium bromide].
1,1 ′-(1,6-hexamethylene) bis [4-{(n-octylamino) carbonyl} pyridinium bromide].
1,1 ′-(1,8-octamethylene) bis [4-{(n-octylamino) carbonyl} pyridinium bromide].
1,1 ′-(1,10-Decamethylene) bis [4-{(n-octylamino) carbonyl} pyridinium bromide].
1,1 '-(1,12-dodecamethylene) bis [4-{(n-octylamino) carbonyl} pyridinium bromide].
N, N′-dimethyl-N, N′-hexamethylenebis (4-carbamoyl-1-n-hexylpyridinium bromide).
N, N′-di-n-butyl-N, N′-hexamethylenebis (4-carbamoyl-1-n-hexylpyridinium bromide).
N, N′-dibenzyl-N, N′-hexamethylenebis (4-carbamoyl-1-n-hexylpyridinium bromide).
N, N′-dimethyl-N, N′-hexamethylenebis (4-carbamoyl-1-n-octylpyridinium bromide).
N, N′-di-n-butyl-N, N′-hexamethylenebis (4-carbamoyl-1-n-octylpyridinium bromide).
N, N′-dibenzyl-N, N′-hexamethylenebis (4-carbamoyl-1-n-octylpyridinium bromide).
N, N′-dimethyl-N, N′-octamethylenebis (4-carbamoyl-1-n-hexylpyridinium bromide).
N, N′-di-n-butyl-N, N′-octamethylenebis (4-carbamoyl-1-n-hexylpyridinium bromide).
N, N′-dibenzyl-N, N′-octamethylenebis (4-carbamoyl-1-n-hexylpyridinium bromide).
N, N′-dimethyl-N, N′-octamethylenebis (4-carbamoyl-1-n-octylpyridinium bromide).
N, N′-di-n-butyl-N, N′-octamethylenebis (4-carbamoyl-1-n-octylpyridinium bromide).
N, N′-dibenzyl-N, N′-octamethylenebis (4-carbamoyl-1-n-octylpyridinium bromide).

The compounds of the present invention can be synthesized with reference to the synthesis methods described in the following Examples and the academic papers listed below. Compounds other than those described in the following examples can also be synthesized by appropriately modifying the synthesis examples described in the present specification and the knowledge of the following academic papers as necessary. This can be done by a contractor.
1) Okazaki, K. et al., Chem. Pharm. Bull., 1997, 45, 1970-1974.
2) Maeda, T. et al., Biol. Pharm. Bull., 1998, 21, 1057-1061.
2) Maeda, T. et al., Chem. Pharm. Bull., 1999, 47, 1020-1023.

  As is apparent from the following examples, the bispyridinium salt compound of the present invention has a malaria parasite growth inhibitory action as well as chloroquine and quinine, which are existing antimalarial agents currently used against malaria parasites. Moreover, the chemotherapeutic index, which is an index of selective toxicity to malaria parasites, was equivalent to or higher than that of the above-mentioned existing antimalarial agents. Therefore, the bispyridinium salt compound of the present invention has low toxicity and is safe, and is useful as an antimalarial agent for the prevention and / or treatment of malaria infection. Also in in vivo studies, the bispyridinium salt compound of the present invention showed a malaria parasite growth inhibitory action and an increased survival rate.

  When the bispyridinium salt compound of the present invention is actually used as an antimalarial agent, the bispyridinium salt compound of the present invention is orally or parenterally (for example, intravenously, intramuscularly) to humans or other mammals. , Subcutaneous, rectal) and formulated into a dosage form according to the route of administration, such as tablets, capsules, ampoules, powders, granules, injections, drops, suppositories, etc. be able to.

  For formulation, the bispyridinium salt compound of the present invention is prepared into a desired dosage form according to a method known per se, together with additives (for example, excipients, binders, disintegrating agents, flavoring agents, emulsifying agents, etc.). Can be done. Examples of additives that can be used in this case include starch, lactose, sucrose, glucose, mannitol, silica, carboxymethylcellulose, gelatin, polyvinylpyrrolidone, calcium carbonate, kaolin, bentonite, calcium stearate, magnesium stearate, polyethylene glycol. , Calcium silicate, water, ethyl alcohol, isopropyl alcohol, propylene glycol, benzyl alcohol, surfactants and the like.

  The preparation containing the bispyridinium salt compound of the present invention can contain other antimalarial agents and other pharmacologically active substances.

  The daily dose of the bispyridinium salt compound of the present invention can be varied within a wide range according to the patient's symptoms, age, weight, doctor's judgment, etc., but generally 0.1 to 0.1 per day for an adult. It is preferable to administer in the range of 5000 mg, preferably 1-1000 mg.

  The dose can be administered once a day or divided into several times.

  In addition, the malaria parasites to which the antimalarial agent of the present invention is applied are not limited to specific ones, such as Plasmodium falciparum, Plasmodium falciparum, egg-shaped malaria parasite, Plasmodium falciparum, etc. It can be widely applied against infection by

  EXAMPLES Hereinafter, the present invention will be described more specifically with reference to examples. However, these examples are merely illustrative and do not limit the scope of the present invention.

  The structure of the bispyridinium salt (4-6) according to general formula (3), which was evaluated for antimalarial activity in the following examples, is shown in FIG. And the synthetic scheme of those pyridinium salts is shown in FIG. Further, FIG. 3 shows the structure of the bispyridinium salt (11-14) according to the general formula (1), which was evaluated for antimalarial activity in the following examples. And the synthetic scheme of those pyridinium salts is shown in FIG. More specifically, these compounds were synthesized as follows.

Synthesis of N-butyl-4-pyridinecarboxamide (1)
n-Butylamine (0.73 g, 10 mmol), dry DMF (20 mL), isonicotinic acid (1.23 g, 10 mmol), HOBt (1.49 g, 11 mmol), EDC (2.30 g, 12 mmol) were added in order. The mixture was heated and stirred at 60 ° C. for 1 hour. The reaction _{TLC (Merck, Kieselgel F 254,} Solvent system; CH 2 Cl 2 -MeOH, 10: 1, v / v) was followed by. After completion of the reaction, the reaction mixture was poured into sat. NaHCO ₃ (50 mL) and extracted with AcOEt (50 mL × 3). The organic layer was washed with water (50 mL) and brine (50 mL), dried over anhydrous MgSO ₄ , and the solvent was evaporated under reduced pressure. The obtained crystalline residue was recrystallized from THF-cyclohexane to obtain the title compound (1) as colorless plates.
Yield; 48%. Mp 47-47.5 ℃. ¹ H NMR (300 MHz, CDCl ₃ ) δ: 8.74 (2H, dd, J = 4.5, 1.6 Hz), 7.62 (2H, dd, J = 4.5, 1.6 Hz) , 6.22 (1H, br s), 3.48 (2H, q, J = 7.3 Hz), 1.62 (2H, m), 1.42 (2H, m), 0.97 (3H, t, J = 7.3 Hz). FAB-MS m / z; 179 (MH ⁺ ).

Synthesis of N-hexyl-4-pyridinecarboxamide (2)
n-Hexylamine (1.01 g, 10 mmol), dry DMF (20 mL), isonicotinic acid (1.23 g, 10 mmol), HOBt (1.49 g, 11 mmol), EDC (2.30 g, 12 mmol) in this order The mixture was further stirred at room temperature for 2.5 hours. The progress of the reaction was checked by TLC (Merck, Kieselgel F ₂₅₄ , Solvent system; CH ₂ Cl ₂ -MeOH, 10: 1, v / v). Stir for hours. _{TLC (Merck, Kieselgel F 254,} Solvent system; CH 2 Cl 2 -MeOH, 10: 1, v / v) completion of the reaction was confirmed by. After completion of the reaction, the reaction mixture was poured into sat. NaHCO ₃ (50 mL) and extracted with AcOEt (30 mL × 3). The organic layer was washed with water (30 mL) and brine (30 mL), dried over anhydrous MgSO ₄ , the solvent was evaporated under reduced pressure, and 2.40 g of the pale yellowish white crystalline residue obtained was subjected to flash column chromatography. Purification by chromatography (elution solvent; CH ₂ Cl ₂ -MeOH, 10: 1, v / v) gave the title compound (2) as colorless plates.
Yield; 62%. Mp 56-58 ℃. ¹ H NMR (300 MHz, CDCl ₃ ) δ: 8.75 (2H, d, J = 5.9 Hz), 7.65 (2H, d, J = 5.9 Hz), 6.26 (1H , br s), 3.47 (2H, q, J = 6.9 Hz), 2.03 (2H, m), 1.63 (6H, m), 0.90 (3H, t, J = 6.9 Hz). FAB-MS m / z; 207 (MH ⁺ ).

Synthesis of N-octyl-4-pyridinecarboxamide (3)
n-Octylamine (1.29 g, 10 mmol), dry DMF (20 mL), isonicotinic acid (1.23 g, 10 mmol), HOBt (1.49 g, 11 mmol), EDC (2.30 g, 12 mmol) in this order In addition, the mixture was heated to 60 ° C. and stirred for 1 hour. _{TLC (Merck, Kieselgel F 254,} Solvent system; CH 2 Cl 2 -MeOH, 10: 1, v / v) was checked the progress of the reaction by. After completion of the reaction, the reaction solution was poured into sat. NaHCO ₃ (50 mL) and extracted with AcOEt (50 mL × 4). The organic layer is washed with water (50 mL), brine (50 mL), dried over anhydrous MgSO ₄ , the solvent is evaporated under reduced pressure, and the resulting crystalline residue is recrystallized from Et ₂ O. Gave the title compound (3) as colorless plates.
Yield, 82%. Mp 61-62.5 ℃. ¹ H NMR (300 MHz, CDCl ₃ ) δ: 8.75 (2H, d, J = 6.0 Hz), 7.63 (2H, d, J = 6.0 Hz), 6.22 (1H , br s), 3.47 (2H, q, J = 7.1 Hz), 1.86 (2H, br s), 1.63 (2H, quin, J = 7.1 Hz), 1.30 (8H, m), 0.88 (3H, t, J = 7.1 Hz) .FAB-MS m / z; 235 (MH ⁺ ).

Synthesis of 1,1 '-(1,6-hexamethylene) bis [4-{(n-butylamino) carbonyl} pyridinium bromide] (4a)
1,6-Dibromohexane (97.6 mg, 0.4 mmol) and 1a (178 mg, 1.0 mmol) were put in a test tube with rubbing, replaced with Ar, sealed with a septum, and heated and stirred at 100 ° C. for 10 hours. Since the reaction solution was depleted, it was considered that the reaction was completed. The crystalline solid in the reaction solution was recrystallized from MeOH-AcOEt to obtain 168 mg of the title compound (4a) as colorless plates.
Yield; 70%. Mp 194-195 ℃. ¹ H NMR (300 MHz, DMSO-d ₆ ) δ: 9.27 (4H, d, J = 6.4 Hz), 8.45 (4H, d, J = 6.4 Hz), 4.65 (4H, t, J = 7.3 Hz), 3.30 (4H, t, J = 8.4 Hz), 1.93 (4H, br s), 1.55 (4H, quin, J = 7.3 Hz), 1.33 (8H, quin, J = 7.2 Hz), 0.91 (6H, t, J = 7.2 Hz). FAB-MS m / z; 519 ([MH ⁷⁹ Br] H ⁺ ), 521 ([(M + 2) -H ⁷⁹ Br] H ⁺ ).

Synthesis of 1,1 '-(1,8-octamethylene) bis [4-{(n-butylamino) carbonyl} pyridinium bromide] (4b)
1,8-Dibromooctane (109 mg, 0.4 mmol) and 1a (178 mg, 1.0 mmol) were put in a test tube with rubbing, substituted with Ar, sealed with a septum, and heated and stirred at 60 ° C. for 4 hours. Since the reaction solution was depleted, it was considered that the reaction was completed. The crystalline solid in the reaction solution was recrystallized from MeOH-AcOEt to obtain 98.5 mg of the title compound (4b) as a colorless crystalline solid.
Yield; 39%. Mp 224-225.5 ℃. ¹ H NMR (300 MHz, DMSO-d ₆ ) δ: 9.25 (4H, d, J = 6.9 Hz), 8.44 (4H, d, J = 6.9 Hz), 4.63 (4H, t, J = 7.3 Hz), 3.33 (4H, m), 1.91 (4H, br s), 1.55 (4H, quin, J = 7.3 Hz), 1.35 (4H, m), 1.28 (12H, br s), 0.91 (6H, t, J = 7.4 Hz). FAB-MS m / z; 547 ([MH ⁷⁹ Br] H ⁺ ), 549 ([(M + 2) -H ⁷⁹ Br] H ⁺ ).

Synthesis of 1,1 '-(1,10-decamethylene) bis [4-{(n-butylamino) carbonyl} pyridinium bromide] (4c)
1,10-Dibromodecane (120 mg, 0.4 mmol) and 1a (178 mg, 1.0 mmol) were placed in a test tube with rubbing, and after Ar substitution, sealed with a septum, and heated and stirred at 60 ° C. for 19 hours. Thereafter, 0.5 mL of dry DMF was added, and the mixture was further stirred with heating at 60 ° C for 28 hours. After completion of the reaction, AcOEt was added to the reaction solution, and the precipitated white crystalline solid was collected by filtration. This solid was recrystallized from MeOH-AcOEt to obtain 200 mg of the title compound (4c) as a colorless crystalline solid.
Yield; 76%. Mp 154-155.5 ℃. ¹ H NMR (300 MHz, DMSO-d ₆ ) δ: 9.24 (4H, d, J = 6.8 Hz), 8.42 (4H, d, J = 6.8 Hz), 4.62 (4H, t, J = 7.3 Hz), 3.33 (4H, m), 1.89 (4H, br s), 1.53 (4H, quin, J = 7.3 Hz), 1.32 (4H, m), 1.24 (12H, br s), 0.89 (6H, t, J = 7.3 Hz). FAB-MS m / z; 575 ([MH ⁷⁹ Br] H ⁺ ), 577 ([(M + 2) -H ⁷⁹ Br] H ⁺ ).

Synthesis of 1,1 '-(1,12-dodecamethylene) bis [4-{(n-butylamino) carbonyl} pyridinium bromide] (4d)
1,12-dibromododecane (131 mg, 0.4 mmol) and 1a (178 mg, 1.0 mmol) were put in a test tube with rubbing, purged with Ar, sealed with a septum, and heated and stirred at 80 ° C. for 10 hours. Since the reaction solution was depleted, it was considered that the reaction was completed. The crystalline solid in the reaction solution was recrystallized from MeOH-AcOEt to obtain 265 mg of the title compound (4d) as a colorless crystalline solid.
Yield; 97%. Mp 184-185 ℃. ¹ H NMR (300 MHz, DMSO-d ₆ ) δ: 9.25 (4H, d, J = 6.8 Hz), 8.44 (4H, d, J = 6.8 Hz), 4.63 (4H, t, J = 7.4 Hz), 3.30 (4H, t, J = 8.9 Hz), 1.91 (4H, m), 1.55 (4H, quin, J = 7.4 Hz), 1.28 (20H, m), 0.91 (6H, t, J = 7.3 Hz). FAB-MS m / z; 603 ([MH ⁷⁹ Br] H ⁺ ), 605 ([(M + 2) -H ⁷⁹ Br] H ⁺ ).

Synthesis of 1,1 '-(1,6-hexamethylene) bis [4-{(n-hexylamino) carbonyl} pyridinium bromide] (5a)
1,6-Dibromohexane (122 mg, 0.5 mmol) and 1b (248 mg, 1.2 mmol) were put in a test tube with rubbing, substituted with Ar, sealed with a septum, and heated and stirred at 80 ° C. for 11 hours. Since the reaction solution was depleted, it was considered that the reaction was completed. The crystalline solid in the reaction solution was recrystallized from MeOH-AcOEt to obtain 301 mg of the title compound (5a) as colorless needles.
Yield; 92%. Mp 203.5-205 ℃. ¹ H NMR (300 MHz, DMSO-d ₆ ) δ: 9.25 (4H, d, J = 6.7 Hz), 8.44 (4H, d, J = 6.7 Hz), 4.64 (4H, t, J = 7.4 Hz), 3.31 (8H, m), 1.92 (4H, m), 1.55 (4H, m), 1.29 (12H, m), 0.87 (6H, t, J = 6.8 Hz) FAB-MS m / z; 575 ([MH ⁷⁹ Br] H ⁺ ), 577 ([(M + 2) -H ⁷⁹ Br] H ⁺ ).

Synthesis of 1,1 '-(1,8-octamethylene) bis [4-{(n-hexylamino) carbonyl} pyridinium bromide] (5b)
1,8-Dibromooctane (109 mg, 0.4 mmol) and 1b (206.3 mg, 1.0 mmol) were placed in a test tube with rubbing, sealed with a septum after Ar substitution, and heated and stirred at 80 ° C. for 15 hours. Since the reaction solution was depleted, it was considered that the reaction was completed. The crystalline solid in the reaction solution was recrystallized from MeOH-AcOEt to obtain 226 mg of the title compound (5b) as colorless prisms.
Yield; 83%. Mp 217-218.5 ℃. ¹ H NMR (300 MHz, DMSO-d ₆ ) δ: 9.26 (4H, d, J = 6.4 Hz), 8.45 (4H, d, J = 6.4 Hz), 4.64 (4H, t, J = 7.3 Hz), 3.32 (4H, m), 1.92 (4H, br s), 1.55 (4H, m), 1.29 (16H, m), 0.87 (6H, t, J = 6.6 Hz FAB-MS m / z; 603 ([MH ⁷⁹ Br] H ⁺ ), 605 ([(M + 2) -H ⁷⁹ Br] H ⁺ ).

Synthesis of 1,1 '-(1,10-decamethylene) bis [4-{(n-hexylamino) carbonyl} pyridinium bromide] (5c)
1,10-Dibromodecane (120 mg, 0.4 mmol) and 1b (206 mg, 1.0 mmol) were put in a test tube with rubbing, and after Ar substitution, sealed with a septum, and heated and stirred at 80 ° C. for 9 hours. Since the reaction solution was depleted, it was considered that the reaction was completed. The crystalline solid in the reaction solution was recrystallized from MeOH-AcOEt to obtain 210 mg of the title compound (5c) as colorless needles.
Yield; 74%. Mp 176-177.5 ℃. ¹ H NMR (300 MHz, DMSO-d ₆ ) δ: 9.25 (4H, d, J = 6.8 Hz), 8.43 (4H, d, J = 6.8 Hz), 4.63 (4H, t, J = 7.3 Hz), 1.91 (4H, br s), 1.55 (4H, m), 1.27 (26H, m), 0.87 (6H, t, J = 6.8 Hz). FAB-MS m / z; 631 ([MH ⁷⁹ Br] H ⁺ ), 633 ([(M + 2) -H ⁷⁹ Br] H ⁺ ).

Synthesis of 1,1 '-(1,12-dodecamethylene) bis [4-{(n-hexylamino) carbonyl} pyridinium bromide] (5d)
1,12-dibromododecane (131 mg, 0.4 mmol) and 1b (206.2 mg, 1.0 mmol) were put in a test tube with rubbing, purged with Ar, sealed with a septum, and heated and stirred at 80 ° C. for 14 hours. Since the reaction solution was depleted, it was considered that the reaction was completed. The crystalline solid in the reaction solution was recrystallized from MeOH-AcOEt to obtain 260 mg of the title compound (5d) as a colorless crystalline solid.
Yield; 88%. Mp 151-152.5 ℃. ¹ H NMR (300 MHz, DMSO-d ₆ ) δ: 9.25 (4H, d, J = 6.8 Hz), 8.43 (4H, d, J = 6.8 Hz), 4.63 (4H, t, J = 7.3 Hz), 3.30 (4H, m), 1.90 (4H, br s), 1.55 (4H, m), 1.26 (28H, m), 0.87 (6H, t, J = 6.8 Hz FAB-MS m / z; 659 ([MH ⁷⁹ Br] H ⁺ ), 661 ([(M + 2) -H ⁷⁹ Br] H ⁺ ).

Synthesis of 1,1 '-(1,6-hexamethylene) bis [4-{(n-octylamino) carbonyl} pyridinium bromide] (6a)
1,6-Dibromohexane (97.6 mg, 0.4 mmol) and 1c (234 mg, 1.0 mmol) were put in a test tube with rubbing, purged with Ar, sealed with a septum, and heated and stirred at 80 ° C. for 10 hours. Since the reaction solution was depleted, it was considered that the reaction was completed. The crystalline solid in the reaction solution was recrystallized from MeOH-AcOEt to obtain 155 mg of the title compound (6a) as colorless scales.
Yield; 64%. Mp 205-207 ° C. ¹ H NMR (300 MHz, DMSO-d ₆ ) δ: 9.27 (4H, d, J = 6.3 Hz), 8.45 (4H, d, J = 6.3 Hz), 4.65 (4H, t, J = 7.3 Hz), 3.29 (8H, t, J = 8.1 Hz), 1.92 (4H, br s), 1.55 (4H, m), 1.26 (20H, m), 0.86 (6H, t , J = 7.0 Hz). FAB-MS m / z; 631 ([MH ⁷⁹ Br] H ⁺ ), 633 ([(M + 2) -H ⁷⁹ Br] H ⁺ ).

Synthesis of 1,1 '-(1,8-octamethylene) bis [4-{(n-octylamino) carbonyl} pyridinium bromide] (6b)
1,8-Dibromooctane (109 mg, 0.4 mmol) and 1c (234 mg, 1.0 mmol) were put into a test tube with rubbing, purged with Ar, sealed with a septum, and heated and stirred at 80 ° C. for 15 hours. Since the reaction solution was depleted, it was considered that the reaction was completed. The crystalline solid in the reaction solution was recrystallized from MeOH-AcOEt to obtain 234 mg of the title compound (6b) as colorless scales.
Yield; 79%. Mp 234.5-235.5 ℃. ¹ H NMR (300 MHz, DMSO-d ₆ ) δ: 9.25 (4H, d, J = 6.6 Hz), 8.44 (4H, d, J = 6.6 Hz), 4.63 (4H, t, J = 7.6 Hz), 3.29 (8H, m), 1.90 (4H, br s), 1.55 (4H, m), 1.27 (30H, m), 0.86 (6H, t, J = 7.6 Hz FAB-MS m / z; 659 ([MH ⁷⁹ Br] H ⁺ ), 661 ([(M + 2) -H ⁷⁹ Br] H ⁺ ).

Synthesis of 1,1 '-(1,10-decamethylene) bis [4-{(n-octylamino) carbonyl} pyridinium bromide] (6c)
1,10-Dibromodecane (120 mg, 0.4 mmol) and 1c (234 mg, 1.0 mmol) were put in a test tube with rubbing, purged with Ar, sealed with a septum, and heated and stirred at 60 ° C. for 14 hours. Since the reaction solution was depleted, it was considered that the reaction was completed. The crystalline solid in the reaction solution was recrystallized from MeOH-AcOEt to obtain 298 mg of the title compound (6c) as a colorless crystalline solid.
Yield; 97%. Mp 134.5-136 ° C. ¹ H NMR (300 MHz, DMSO-d ₆ ) δ: 9.26 (4H, d, J = 6.3 Hz), 8.44 (4H, d, J = 6.3 Hz), 4.64 (4H, t, J = 7.3 Hz), 1.91 (4H, br s), 1.55 (4H, t, J = 6.4 Hz), 1.26 (16H, br s), 0.86 (6H, t, J = 6.6 Hz) FAB-MS m / z; 687 ([MH ⁷⁹ Br] H ⁺ ), 689 ([(M + 2) -H ⁷⁹ Br] H ⁺ ).

Synthesis of 1,1 '-(1,12-dodecamethylene) bis [4-{(n-octylamino) carbonyl} pyridinium bromide] (6d)
1,12-dibromododecane (131 mg, 0.4 mmol) and 1c (234 mg, 1.0 mmol) were put into a test tube with rubbing, purged with Ar, sealed with a septum, and heated and stirred at 80 ° C. for 10 hours. Since the reaction solution was depleted, it was considered that the reaction was completed. The crystalline solid in the reaction solution was recrystallized from MeOH-AcOEt to obtain 297 mg of the title compound (6d) as a colorless crystalline solid.
Yield; 93%. Mp 162.5-163 ℃. ¹ H NMR (300 MHz, DMSO-d ₆ ) δ: 9.24 (4H, d, J = 6.5 Hz), 8.43 (4H, d, J = 6.5 Hz), 4.65 (4H, t, J = 7.3 Hz), 3.33 (12H, m), 1.91 (4H, br s), 1.55 (4H, m), 1.22 (28H, m), 0.86 (6H, t, J = 6.8 Hz ). FAB-MS m / z; 715 ([MH ⁷⁹ Br] H ⁺ ), 717 ([(M + 2) -H ⁷⁹ Br] H ⁺ ).

Synthesis of N, N'-dimethyl-N, N'-hexamethylenebis (4-carbamoyl-1-n-hexylpyridinium bromide) (11a)
To N, N'-dimethyl-N, N'-hexamethylenebis (4-carbamoylpyridine) (9a) (532 mg, 1.50 mmol), add dry DMF 1.0 mL and 1-bromohexane (0.42 mL, 3.00 mmol). The mixture was heated and stirred at 100 ° C. for 11 hours. _{TLC (Merck, Kieselgel F 254,} Solvent system; AcOEt-MeOH, 3: 2, v / v) disappearance of the starting materials was confirmed by, The reaction was terminated. After completion of the reaction, the reaction mixture was triturated with AcOEt-MeOH and the insoluble part was recrystallized from AcOEt-MeOH to obtain 589 mg (58%) of the title compound (11a) as colorless granular crystals. Mp 222.5-225.5 ° C.
IR (KBr) cm ^-1 ; 1627 (C = O). ¹ H-NMR (300 MHz, DMSO-d ₆ ) δ: 9.19 (br d, 16 H, J = 6.5 Hz, syn- and anti-pyridyl- 2,6-H), 8.21-8.15 (m, 16 H, syn- and anti-pyridyl-3,5-H), 4.61 (br t, 16 H, J = 7.2 Hz, syn- and anti-N ⁺ -CH ₂ ), 3.49, 3.41, 3.12 and 3.10 (each t, 16 H, J = 7.2 Hz, syn- and anti-CH ₂ CH ₂ N), 3.03, 2.99, 2.87 and 2.84 (each s, 24 H, syn- and anti-CH ₃ N), 1.98-1.88 (m, 16 H, alkyl-H), 1.68-0.98 (m, 80 H, alkyl-H), 0.87 (br t, 24 H, J = 7.2 Hz , 8 × CH ₃ ). FAB-MS m / z: 605 ([(M + 2) -H ⁷⁹ Br] H ⁺ ), 603 ([MH ⁷⁹ Br] H ⁺ ). IR (KBr) cm ^-1 ; 1627.
Anal.Calcd.for C ₃₂ H ₅₂ Br ₂ N ₄ O ₂ 1 / 10HBr: C, 55.49; H, 7.58; N, 8.09.Found: C, 55.62; H, 7.44; N, 8.03.

Synthesis of N, N'-di-n-butyl-N, N'-hexamethylenebis (4-carbamoyl-1-n-hexylpyridinium bromide) (11b)
N, N'-di-n-butyl-N, N'-hexamethylenebis (4-carbamoylpyridine) (9b) (431 mg, 0.98 mmol) in dry DMF 0.2 mL, 1-bromohexane (0.28 mL, 1.96) mmol) was added, and the mixture was heated and stirred at 100 ° C. for 7 hours. _{TLC (Merck, Kieselgel F 254,} Solvent system; AcOEt-MeOH, 5: 1, v / v) disappearance of the starting materials was confirmed by, The reaction was terminated. After completion of the reaction, the reaction mixture was repeatedly triturated with hot AcOEt to obtain 662.3 mg (88%) of the title compound (11b) as an insoluble part as colorless granular crystals. This gave a single spot in TLC using several different solvent systems. Mp 103-105 ° C.
IR (KBr) cm ^-1 ; 1639 (C = O). ¹ H-NMR (300 MHz, DMSO-d ₆ ) δ: 9.18 (br d, 16 H, J = 6.5 Hz, syn- and anti-pyridyl- 2,6-H), 8.18 and 8.15 (each br d, 16 H, J = 6.5 Hz, syn- and anti-pyridyl-3,5-H), 4.61 (br t, 16 H, J = 7.5 Hz, syn- and anti-N ⁺ -CH ₂ ), 3.45 (br t, 16 H, J = 7.5 Hz, syn- and anti-NCH ₂ ), 3.12-2.99 (m, 16 H, alkyl-H), 1.98- 1.88 (m, 16 H, alkyl-H), 1.69-1.04 (m, 112 H, alkyl-H), 0.94 (br t, 12 H, J = 7.5 Hz, 4 × CH ₃ ), 0.86 (br t, 24 H, J = 7.5 Hz, 8 × CH ₃ ), 0.75 (br t, 12 H, J = 7.5 Hz, 4 × CH ₃ ).
Anal.Calcd.for C ₃₈ H ₆₄ Br ₂ N ₄ O ₂・ 2 / 5HBr: C, 56.97; H, 8.10; N, 6.99. Found: C, 56.95; H, 8.11; N, 6.92.

Synthesis of N, N'-dibenzyl-N, N'-hexamethylenebis (4-carbamoyl-1-n-hexylpyridinium bromide) (11c)
Dry DMF 0.6 mL and 1-bromohexane (0.30 mL, 2.1 mmol) were added to N, N'-dibenzyl-N, N'-hexamethylenebis (4-carbamoylpyridine) (9c) (478 mg, 0.94 mmol) The mixture was heated and stirred at 100 ° C. for 6 hours. _{TLC (Merck, Kieselgel F 254,} Solvent system; AcOEt-MeOH, 5: 2, v / v) disappearance of the starting materials was confirmed by, The reaction was terminated. After completion of the reaction, the reaction mixture was repeatedly triturated with hot AcOEt to obtain 515.9 mg (66%) of the title compound (11c) as an insoluble part as colorless granular crystals. This gave a single spot in TLC using several different solvent systems. Mp 166.5-167.5 ° C.
IR (KBr) cm ^-1 ; 1640 (C = O). ¹ H-NMR (300 MHz, DMSO-d ₆ ) δ: 9.20, 9.14 (each br d, 16 H, J = 6.0 Hz, syn- and anti -pyridyl-2,6-H), 8.30-8.10 (m, 16 H, syn- and anti-pyridyl-3,5-H), 7.41-7.18 (m, 40 H, syn- and anti-phenyl-H ), 4.73 and 4.69 (each br s, 8 H, syn- or anti-benzyl-H), 4.61 and 4.56 (each br t, 16 H, J = 7.5 Hz, syn- and anti-N ⁺ -CH ₂ ) , 4.41 and 4.38 (each br s, 8 H, syn- or anti-benzyl-H), 3.30-2.92 (m, 16 H, alkyl-H), 2.03-1.82 (m, 16 H, alkyl-H), 1.64-1.03 (m, 16 H, alkyl-H), 0.86 (m, 24 H, 8 × CH ₃ ). FAB-MS m / z: 757 ([(M + 2) -H ⁷⁹ Br] H ⁺ ) , 755 ([MH ⁷⁹ Br] H ⁺ ).
Anal.Calcd.for C ₄₄ H ₆₀ Br ₂ N ₄ O ₂ 1 / 8HBr: C, 62.40; H, 7.16; N, 6.62. Found: C, 62.59; H, 7.00; N, 6.70.

Synthesis of N, N'-dimethyl-N, N'-hexamethylenebis (4-carbamoyl-1-n-octylpyridinium bromide) (12a)
Add dry DMF 0.8 mL, 1-bromooctane (0.70 mL, 4.00 mmol) to N, N'-dimethyl-N, N'-hexamethylenebis (4-carbamoylpyridine) (9a) (709 mg, 2.00 mmol) The mixture was heated and stirred at 100 ° C. for 14 hours. _{TLC (Merck, Kieselgel F 254,} Solvent system; AcOEt-MeOH, 3: 2, v / v) disappearance of the starting materials was confirmed by, The reaction was terminated. After completion of the reaction, the reaction mixture was triturated with AcOEt-MeOH, and the insoluble part was recrystallized from AcOEt-MeOH to obtain 866 mg (59%) of the title compound (12a) as colorless granular crystals. Mp 136-137 ° C.
IR (KBr) cm ^-1 ; 1631 (C = O). ¹ H-NMR (300 MHz, DMSO-d ₆ ) δ: 9.20 (br d, 16H, J = 6.0 Hz, syn- and anti-pyridyl-2 , 6-H), 8.20 and 8.18 (each br d, 16H, J = 6.0 Hz, syn- and anti-pyridyl-3,5-H), 4.61 (br t, 16H, J = 7.2 Hz, syn- and anti-N ⁺ -CH ₂ ), 3.49, 3.42, 3.12 and 3.10 (each t, 16H, J = 7.2 Hz, syn- and anti-CH ₂ CH ₂ N), 3.03, 2.99, 2.87 and 2.84 (each s, 24H, syn- and anti-CH ₃ N), 1.98-1.88 (m, 16H, alkyl-H), 1.69-0.98 (m, 112H, alkyl-H), 0.86 (br t, 24H, J = 7.2 Hz, 8 × CH ₃ ). FAB-MS m / z: 661 ([(M + 2) -H ⁷⁹ Br] H ⁺ ), 659 ([MH ⁷⁹ Br] H ⁺ ).
Anal.Calcd.for C ₃₆ H ₆₀ Br ₂ N ₄ O ₂ : C, 58.38; H, 8.16; N, 7.56. Found: C, 58.16; H, 7.91; N, 7.55.

Synthesis of N, N'-di-n-butyl-N, N'-hexamethylenebis (4-carbamoyl-1-n-octylpyridinium bromide) (12b)
N, N'-di-n-butyl-N, N'-hexamethylenebis (4-carbamoylpyridine) (9b) (147 mg, 0.34 mmol) in dry DMF 0.3 mL, 1-bromooctane (0.12 mL, 0.68 mmol) was added, and the mixture was stirred at 100 ° C. for 15 hours. _{TLC (Merck, Kieselgel F 254,} Solvent system; AcOEt-MeOH, 10: 1, v / v) disappearance of the starting materials was confirmed by, The reaction was terminated. After completion of the reaction, the reaction mixture was repeatedly triturated with hot AcOEt and hot MeOH to obtain 204 mg (73%) of the title compound (12b) as an insoluble part as colorless granular crystals. This gave a single spot in TLC using several different solvent systems. Mp 153-154.5 ° C.
IR (KBr) cm ^-1 ; 1640 (C = O). ¹ H-NMR (300 MHz, DMSO-d ₆ ) δ: 9.17 (br d, 16H, J = 6.8 Hz, syn- and anti-pyridyl-2 , 6-H), 8.17, 8.18 and 8.15 (each br d, 16H, J = 6.8 Hz, syn- and anti-pyridyl-3,5-H), 4.60 (br t, 16H, J = 7.2 Hz, syn -and anti-N ⁺ -CH ₂ ), 3.48-3.40 (m, 16H, alkyl-H), 3.12-3.00 (m, 16H, alkyl-H), 1.98-1.88 (m, 16H, alkyl-H), 1.68-1.01 (m, 144H, alkyl-H), 0.94 and 0.93 (each t, 12H, J = 7.2 Hz, 4 × CH ₃ ), 0.85 (br t, 24H, J = 7.2 Hz, 8 × CH ₃ ) , 0.74 (br t, 12H, J = 7.2 Hz, 4 × CH ₃ ). FAB-MS m / z: 745 ([(M + 2) -H ⁷⁹ Br] H ⁺ ), 743 ([MH ⁷⁹ Br] H ⁺ ).
_{.. Anal Calcd for C 42 H} 72 Br 2 N 4 O 2 · 1 / 3HBr:. C, 59.22; H, 8.56; N, 6.58 Found: C, 59.29; H, 8.57; N, 6.22.

Synthesis of N, N'-dibenzyl-N, N'-hexamethylenebis (4-carbamoyl-1-n-octylpyridinium bromide) (12c)
Add dry DMF 0.2 mL, 1-bromooctane (0.38 mL, 2.2 mmol) to N, N'-dibenzyl-N, N'-hexamethylenebis (4-carbamoylpyridine) (9c) (500 mg, 0.99 mmol) The mixture was heated and stirred at 100 ° C. for 6 hours. _{TLC (Merck, Kieselgel F 254,} Solvent system; AcOEt-MeOH, 5: 1, v / v) disappearance of the starting materials was confirmed by, The reaction was terminated. After completion of the reaction, the reaction mixture was repeatedly triturated with hot AcOEt to obtain 731 mg (83%) of the title compound (12c) as an insoluble part as colorless granular crystals. This gave a single spot in TLC using several different solvent systems. Mp 178-181 ° C.
IR (KBr) cm ^-1 ; 1637 (C = O). ¹ H-NMR (300 MHz, DMSO-d ₆ ) δ: 9.21 and 9.15 (each br d, 16H, J = 6.7 Hz, syn- and anti- pyridyl-2,6-H), 8.28, 8.24, 8.21 and 8.18 (each d, 16H, J = 6.7 Hz, syn- and anti-pyridyl-3,5-H), 7.40-7.20 (m, 40H, syn -and anti-phenyl-H), 4.73 and 4.69 (each br s, 8H, syn- or anti-benzyl-H), 4.61 and 4.56 (each br t, 16H, J = 7.5 Hz, syn- and anti-N ⁺ -CH ₂ ), 4.42 and 4.39 (each br s, 8H, syn- or anti-benzyl-H), 3.30, 3.26, 3.04 and 2.97 (each t, 16H, J = 7.5 Hz, syn- and anti-CH ₂ CH ₂ N), 1.99-1.84 (m, 16H, alkyl-H), 1.63-1.43 (m, 16H, alkyl-H), 1.36-1.18 (m, 80H, alkyl-H), 1.14-0.99 (m , 16H, alkyl-H), 0.85 (br t, 24H, J = 7.5 Hz, 8 × CH ₃ ). FAB-MS m / z: 815 ([(M + 4) -H ⁷⁹ Br] H ⁺ ), 813 ([(M + 2) -H ⁷⁹ Br] H ⁺ ), 811 ([MH ⁷⁹ Br] H ⁺ ).
Anal.Calcd.for C ₄₈ H ₆₈ Br ₂ N ₄ O ₂・ 1 / 4HBr: C, 63.14; H, 7.53; N, 6.14.Found: C, 63.19; H, 7.39; N, 6.04.

Synthesis of N, N'-dimethyl-N, N'-octamethylenebis (4-carbamoyl-1-n-hexylpyridinium bromide) (13a)
To N, N'-dimethyl-N, N'-octamethylenebis (4-carbamoylpyridine) (10a) (408 mg, 1.07 mmol) was added 1-bromohexane (0.30 mL, 2.15 mmol), and 5 at 100 ° C. Stir for hours. _{TLC (Merck, Kieselgel F 254,} Solvent system; AcOEt-MeOH, 5: 1, v / v) disappearance of the starting materials was confirmed by, The reaction was terminated. After completion of the reaction, the reaction mixture was triturated with AcOEt-MeOH and the insoluble part was recrystallized from AcOEt-MeOH to obtain 522 mg (68%) of the title compound (13a) as colorless granular crystals. Mp 30-31 ° C.
IR (KBr) cm ^-1 ; 1632 (C = O). ¹ H-NMR (300 MHz, DMSO-d ₆ ) δ: 9.19 (br d, 16H, J = 6.0 Hz, syn- and anti-pyridyl-2 , 6-H), 8.18 (br d, 16H, J = 6.0 Hz, syn- and anti-pyridyl-3,5-H), 4.61 (br t, 16H, J = 7.2 Hz, syn- and anti-N ⁺ -CH ₂ ), 3.47 and 3.42 (each t, 8H, J = 7.5 Hz, syn- or anti-CH ₂ CH ₂ N), 3.10 and 3.08 (each br t, 8H, J = 7.5 Hz, syn- or anti-CH ₂ CH ₂ N), 3.01, 3.00, 2.86 and 2.85 (each s, 24H, syn- and anti-CH ₃ N), 1.99-1.87 (m, 16H, alkyl-H), 1.65-1.44 (m , 16H, alkyl-H), 1.39-0.99 (m, 80H, alkyl-H), 0.87 (br t, 24H, J = 7.5 Hz, 8 × CH ₃ ). FAB-MS m / z: 633 ([( M + 2) -H ⁷⁹ Br] H ⁺ ), 631 ([MH ⁷⁹ Br] H ⁺ ).
Anal.Calcd.for C ₃₄ H ₅₆ Br ₂ N ₄ O ₂・ 1 / 4H ₂ O: C, 56.94; H, 7.94; N, 7.81. Found: C, 56.91; H, 7.67; N, 7.79.

Synthesis of N, N'-di-n-butyl-N, N'-octamethylenebis (4-carbamoyl-1-n-hexylpyridinium bromide) (13b)
To N, N'-di-n-butyl-N, N'-octamethylenebis (4-carbamoylpyridine) (10b) (420 mg, 0.90 mmol) was added 1-bromohexane (0.29 mL, 2.10 mmol), Stir at 100 ° C. overnight. _{TLC (Merck, Kieselgel F 254,} Solvent system; AcOEt-MeOH, 5: 2, v / v) disappearance of the starting materials was confirmed by, The reaction was terminated. After completion of the reaction, the reaction mixture was repeatedly triturated with hot AcOEt to obtain 670 mg (93%) of the title compound (13b) as an insoluble part as colorless granular crystals. This gave a single spot in TLC using several different solvent systems. Mp 122-123 ° C.
IR (KBr) cm ^-1 ; 1636 (C = O). ¹ H-NMR (300 MHz, DMSO-d ₆ ) δ: 9.18 (br d, 16H, J = 6.8 Hz, syn- and anti-pyridyl-2 , 6-H), 8.19-8.13 (m, 16H, syn- and anti-pyridyl-3,5-H), 4.61 (br t, 16H, J = 7.2 Hz, syn- and anti-N ⁺ -CH ₂ ), 3.47-3.39 (m, 16H, alkyl-H), 3.10-3.01 (m, 16H, alkyl-H), 1.98-1.86 (m, 16H, alkyl-H), 1.68-1.02 (m, 128H, alkyl -H), 0.94 (br t, 12H, J = 7.2 Hz, 4 × CH ₃ ), 0.86 (br t, 24H, J = 7.2 Hz, 8 × CH ₃ ), 0.74 (br t, 12H, J = 7.2 Hz, 4 × CH ₃ ). FAB-MS m / z: 719 ([(M + 4) -H ⁷⁹ Br] H ⁺ ), 717 ([(M + 2) -H ⁷⁹ Br] H ⁺ ), 715 ([MH ⁷⁹ Br] H ⁺ ).
_{.. Anal Calcd for C 40 H} 68 Br 2 N 4 O 2 · 1 / 7HBr:. C, 59.43; H, 8.50; N, 6.93 Found: C, 59.66; H, 8.26; N, 6.80.

Synthesis of N, N'-dibenzyl-N, N'-octamethylenebis (4-carbamoyl-1-n-hexylpyridinium bromide) (13c)
N, N'-Dibenzyl-N, N'-octamethylenebis (4-carbamoylpyridine) (10c) (540 mg, 1.01 mmol) was added dry AcOH 1.0 mL, 1-bromohexane (0.28 mL, 2.02 mmol) The mixture was heated and stirred at 100 ° C. for 5 hours. _{TLC (Merck, Kieselgel F 254,} Solvent system; AcOEt-MeOH, 5: 1, v / v) disappearance of the starting materials was confirmed by, The reaction was terminated. After completion of the reaction, the reaction mixture was repeatedly triturated with hot AcOEt to obtain 804 mg (92%) of the title compound (13c) as an insoluble part as colorless granular crystals. This gave a single spot in TLC using several different solvent systems. Mp 86.5-87.5 ° C.
IR (KBr) cm ^-1 ; 1637 (C = O). ¹ H-NMR (300 MHz, DMSO-d ₆ ) δ: 9.20 and 9.13 (each br d, 16H, J = 6.0 Hz, syn- and anti- pyridyl-2,6-H), 8.29, 8.26, 8.20 and 8.17 (each d, 16H, J = 6.0 Hz, syn- and anti-pyridyl-3,5-H), 7.40-7.21 (m, 40H, syn -and anti-phenyl-H), 4.73 and 4.72 (each br s, 8H, syn- or anti-benzyl-H), 4.62 and 4.56 (each br t, 16H, J = 7.5 Hz, syn- and anti-N ⁺ -CH ₂ ), 4.41 and 4.40 (each br s, 8H, syn- or anti-benzyl-H), 3.33-3.23 (m, 8H, syn- or anti-CH ₂ CH ₂ N), 3.07-2.97 ( m, 8H, syn- or anti-CH ₂ CH ₂ N), 1.98-1.85 (m, 16H, alkyl-H), 1.58-1.33 (m, 16H, alkyl-H), 1.32-0.92 (m, 80H, alkyl-H), 0.86 (br t, 24H, J = 7.5 Hz, 8 × CH ₃ ). FAB-MS m / z: 785 ([(M + 2) -H ⁷⁹ Br] H ⁺ ), 783 ([ MH ⁷⁹ Br] H ⁺ ).
Anal.Calcd.for C ₄₆ H ₆₄ Br ₂ N ₄ O ₂・ 1 / 2HBr: C, 61.03; H, 7.18; N, 6.19.Found: C, 61.16; H, 7.20; N, 6.15.

Synthesis of N, N'-dimethyl-N, N'-octamethylenebis (4-carbamoyl-1-n-octylpyridinium bromide) (14a)
Add 1-bromooctane (0.29 mL, 1.66 mmol) to N, N'-dimethyl-N, N'-octamethylenebis (4-carbamoylpyridine) (10a) (318 mg, 0.83 mmol) and add 3 at 100 ° C. At the stage of heating and stirring for an hour, since raw materials still remained on TLC (Merck, Kieselgel F ₂₅₄ , Solvent system; AcOEt-MeOH, 5: 1, v / v), dry DMF 0.10 mL, 1-bromooctane ( 0.9 mL) was added, and the mixture was further heated and stirred for 12 hours. _{TLC (Merck, Kieselgel F 254,} Solvent system; AcOEt-MeOH, 5: 1, v / v) disappearance of the starting materials was confirmed by, The reaction was terminated. After completion of the reaction, the reaction mixture was repeatedly triturated with hot AcOEt to obtain 465 mg (73%) of the insoluble part of the title compound (14a) as colorless granular crystals. This gave a single spot in TLC using several different solvent systems. Mp 201-203 ° C.
IR (KBr) cm ^-1 ; 1635 (C = O). ¹ H-NMR (300 MHz, DMSO-d ₆ ) δ: 9.18 (br d, 16H, J = 6.0 Hz, syn- and anti-pyridyl-2 , 6-H), 8.19 and 8.16 (br d, 16H, J = 6.0 Hz, syn- and anti-pyridyl-3,5-H), 4.60 (br t, 16H, J = 7.2 Hz, syn- and anti -N ⁺ -CH ₂ ), 3.47 and 3.44 (each t, 8H, J = 7.5 Hz, syn- or anti-CH ₂ CH ₂ N), 3.10 and 3.08 (each br t, 8H, J = 7.5 Hz, syn -or anti-CH ₂ CH ₂ N), 3.01, 3.00, 2.86 and 2.85 (each s, 24H, syn- and anti-CH ₃ N), 1.97-1.87 (m, 16H, alkyl-H), 1.64-1.41 (m, 16H, alkyl-H), 1.37-0.98 (m, 112H, alkyl-H), 0.86 (br t, 24H, J = 7.5 Hz, 8 × CH ₃ ). FAB-MS m / z: 691 ( [(M + 4) -H ⁷⁹ Br] H ⁺ ), 689 ([(M + 2) -H ⁷⁹ Br] H ⁺ ), 687 ([MH ⁷⁹ Br] H ⁺ ).
Anal.Calcd.for C ₃₈ H ₆₄ Br ₂ N ₄ O ₂・ 1 / 2H ₂ O: C, 58.68; H, 8.42; N, 7.20.Found: C, 58.63; H, 8.14; N, 7.23.

Synthesis of N, N'-di-n-butyl-N, N'-octamethylenebis (4-carbamoyl-1-n-octylpyridinium bromide) (14b)
N, N'-di-n-butyl-N, N'-octamethylenebis (4-carbamoylpyridine) (10b) (474 mg, 1.01 mmol) in dry DMF 0.1 mL, 1-bromooctane (0.35 mL, 2.03 In the stage where the mixture was heated and stirred at 100 ° C. for 6 hours, the raw material still remained on TLC (Merck, Kieselgel F ₂₅₄ , Solvent system; AcOEt-MeOH, 5: 1, v / v). 0.10 mL and 1-bromooctane (0.1 mL) were added, and the mixture was further heated and stirred for 3 hours. _{TLC (Merck, Kieselgel F 254,} Solvent system; AcOEt-MeOH, 5: 1, v / v) disappearance of the starting materials was confirmed by, The reaction was terminated. After completion of the reaction, the reaction mixture was repeatedly triturated with hot AcOEt to obtain 661 mg (77%) of the title compound (14b) as an insoluble part as colorless granular crystals. This gave a single spot in TLC using several different solvent systems. Mp 130-131 ° C.
IR (KBr) cm ^-1 ; 1636 (C = O). ¹ H-NMR (300 MHz, DMSO-d ₆ ) δ: 9.18 (br d, 16H, J = 6.0 Hz, syn- and anti-pyridyl-2 , 6-H), 8.18 (br d, 16H, J = 6.0 Hz, syn- and anti-pyridyl-3,5-H), 4.61 (br t, 16H, J = 7.5 Hz, syn- and anti-N ⁺ -CH ₂ ), 3.51-3.33 (m, 16H, alkyl-H), 3.10-3.00 (m, 16H, alkyl-H), 1.98-1.88 (m, 16H, alkyl-H), 1.65-1.53 (m , 16H, alkyl-H), 1.49-1.39 (m, 16H, alkyl-H), 1.38-1.15 (m, 112H, alkyl-H), 1.08 (br quin, 16H, J = 7.5 Hz, alkyl-H) , 0.94 (br t, 12H, J = 7.5 Hz, 4 × CH ₃ ), 0.86 (br t, 24H, J = 7.5 Hz, 8 × CH ₃ ), 0.74 (br t, 12H, J = 7.5 Hz, 4 × CH ₃ ). FAB-MS m / z: 773 ([(M + 2) -H ⁷⁹ Br] H ⁺ ), 771 ([MH ⁷⁹ Br] H ⁺ ).
Anal.Calcd.for C ₄₄ H ₇₆ Br ₂ N ₄ O ₂ 1 / 13HBr: C, 61.51; H, 8.93; N, 6.52. Found: C, 61.62; H, 8.66; N, 6.50.

Synthesis of N, N'-dibenzyl-N, N'-octamethylenebis (4-carbamoyl-1-n-octylpyridinium bromide) (14c)
Dry DMF 0.5 mL, 1-bromooctane (0.31 mL, 1.78 mmol) were added to N, N'-dibenzyl-N, N'-octamethylenebis (4-carbamoylpyridine) (10c) (477 mg, 0.89 mmol) The mixture was heated and stirred at 100 ° C. for 14 hours. _{TLC (Merck, Kieselgel F 254,} Solvent system; AcOEt-MeOH, 5: 1, v / v) disappearance of the starting materials was confirmed by, The reaction was terminated. After completion of the reaction, the reaction mixture was repeatedly triturated with hot AcOEt to obtain 630 mg (77%) of the insoluble part of the title compound (14c) as colorless granular crystals. This gave a single spot in TLC using several different solvent systems. Mp 143-145 ° C.
IR (KBr) cm ^-1 ; 1647 (C = O). ¹ H-NMR (300 MHz, DMSO-d ₆ ) δ: 9.24 and 9.17 (each br d, 16H, J = 6.0 Hz, syn- and anti- pyridyl-2,6-H), 8.29, 8.27, 8.21 and 8.19 (each d, 16H, J = 6.0 Hz, syn- and anti-pyridyl-3,5-H), 7.40-7.21 (m, 40H, syn -and anti-phenyl-H), 4.74 and 4.72 (each br s, 8H, syn- or anti-benzyl-H), 4.64 and 4.57 (each br t, 16H, J = 7.2 Hz, syn- and anti-N ⁺ -CH ₂ ), 4.42 and 4.41 (each br s, 8H, syn- or anti-benzyl-H), 3.26-3.35 (m, 8H, alkyl-H), 3.08-2.98 (m, 8H, alkyl-H ), 1.99-1.85 (m, 16H, alkyl-H), 1.60-0.99 (m, 128H, alkyl-H), 0.85 (br t, 24H, J = 7.2 Hz, 8 × CH ₃ ). FAB-MS m / z: 845 ([(M + 4) -H ⁷⁹ Br] H ⁺ ), 841 ([(M + 2) -H ⁷⁹ Br] H ⁺ ), 839 ([MH ⁷⁹ Br] H ⁺ ).
Anal.Calcd.for C ₅₀ H ₇₂ Br ₂ N ₄ O ₂ 1 / 10HBr: C, 64.64; H, 7.82; N, 6.03.Found: C, 64.82; H, 7.58; N, 6.00.

  The synthesized compound was evaluated in vitro for antimalarial activity by the following method.

(1) Cultivation of P. falciparum P. falciparum FCR-3 strain (ATCC-30932) was used as the malaria parasite, and the filter sterilized RPMI 1640 medium (pH 7.4, supplemented with 10% human serum) ) Was used under the conditions of oxygen concentration 5.0%, carbon dioxide concentration 5.0%, nitrogen concentration 90%, and temperature 37 ° C. The hematocrit value (ratio of red blood cell volume in the red blood cell suspension) was 5.0%, and the initial infection rate of P. falciparum at the start of the culture was 0.3%. The medium was changed using a 24-well culture plate (the medium was changed when the infection rate exceeded 1%), and transplantation was performed at an infection rate of 2.0-3.5%. The infection rate was measured under a microscope (oil immersion, 1,000 x) after making a thin-layer smear and stained with Diff-Quik, and the malaria parasite infection rate was calculated by the following formula.
Malaria parasite infection rate (%) = {(number of infected red blood cells) / (total number of red blood cells)} × 100

(2) Malaria parasite growth inhibition screening test Cultivated malaria parasite-infected erythrocytes are collected by centrifugation, washed with serum-containing medium (RPMI (+) 1640 medium), non-infected erythrocytes are added, and the initial infection rate is determined. 0.3%. The hematocrit value at this time was 3.0%. Each sample was dissolved in sterilized water or dimethyl sulfoxide (DMSO) to prepare a test solution having a predetermined concentration.

5 μL of the test solution was added to each 24-well culture plate, and two or three tests were performed for one compound. As a control, 5 μL / well of sterilized water or DMSO was added. Next, the P. falciparum culture solution, which had been adjusted to a predetermined concentration in advance, was gently pipetted and uniformly suspended in the medium, and 995 μL was added to each well. After culturing the culture plate in an incubator (oxygen concentration: 5.0%, carbon dioxide concentration: 5.0%, nitrogen concentration: 90%) for 72 hours, a thin-layer smear was prepared for each well, stained with Diff-Quik, The malaria parasite infection rate of the test solution addition group and the control group was calculated. From the malaria parasite infection rate determined above, the growth rate was calculated according to the following formula to determine the 50% growth inhibitory concentration (IC ₅₀ ).
Growth rate (%) = {(b−a) / (c−a)} × 100
[In the formula, a is the initial infection rate, b is the infection rate when the test solution is added, and c is the control infection rate.]

(3) Mouse FM3A cell growth inhibition test The F28-7 strain, a wild strain of mouse breast cancer-derived FM3A cells, was used. The medium was supplemented with 2% fetal calf serum inactivated in ES medium and cultured at a carbon dioxide concentration of 5.0% at 37 ° C. Under these conditions, the doubling time of FM3A cells was about 12 hours. Preculture and dilute cells that have entered the logarithmic growth phase to 5.0 × 10 ⁴ cells / ml. Prepare the sample solution in a 24-well culture plate using the sample prepared at the time of measuring malaria activity. 5 μL each was added (final concentration with medium etc. added 1 × 10 ⁻⁴ to 1 × 10 ⁻⁹ M). Two or three tests were conducted for one compound, and wells containing 5 μL of sterilized water or DMSO were prepared simultaneously as controls. Next, 995 μL each of the prepared cultured cell suspension was added and gently pipetted to suspend uniformly in the medium. After culturing for 48 hours, the number of cells in each well was counted with an upright microscope using a hemocytometer, and the growth rate was calculated by the following formula to calculate the 50% growth inhibition rate (IC ₅₀ ).
Growth rate (%) = {(C−A) / (B−A)} × 100
[In the formula, A is the initial cell number, B is the control cell number after 2 days, and C is the cell number 2 days after adding the sample.]

The cell proliferation activity was calculated from the number of cells in the well to which the sample was added and the number of control cells, and the cytotoxicity of the sample was evaluated. The anti-malarial activity of the sample is evaluated from the IC ₅₀ value of each sample against P. falciparum and mouse FM3A cells, and the chemotherapeutic coefficient used as an indicator of selective toxicity to P. falciparum is calculated using the following formula to determine the efficacy. I did it.
Chemotherapeutic coefficient = (IC ₅₀ value of sample for mouse FM3A cells) ÷ (IC ₅₀ value of sample for Plasmodium falciparum)

  As is clear from the results in Tables 1 and 2, the compound according to the present invention showed excellent malaria parasite growth inhibitory activity and a high chemotherapy index. In particular, the compounds 4d, 5c, 12a, 12b, and 12c showed high antimalarial activity, and the chemotherapeutic index, which is an indicator of selective toxicity, also showed a high value comparable to chloroquine and quinine. The compounds 4d, 5c, 12a, 12b, and 12c were further evaluated for antimalarial activity in vivo. The in vivo evaluation method is as follows.

(1) Acute toxicity test Compounds of various concentrations dissolved in PBS (-) buffer containing 1% DMSO were administered intraperitoneally to 4-5 week-old male ICR mice weighing 25-30 g (dosage was 10 mL / kg) And was reared. Drug administration every 24 hours was performed for 4 days. Mice were weighed daily from the start of drug administration, and the results were used to determine the possibility of acute toxicity of the compound and to serve as a guideline for determining drug doses in in vivo studies.

(2) 4-Day Suppressive Test
As a test method for evaluating the sensitivity of the test substance, a 4-Day Suppressive Test using mice was performed. Infected red blood cells 1 × 10 ⁶ cells / 200 μL suspended in PBS (-) buffer are inoculated into the tail vein of mice, and dissolved 2 hours later in PBS (-) buffer containing DMSO 1% and adjusted to each concentration Was fixed at a dose of 10 mL / kg and administered intraperitoneally. The drug administration was continued every 24 hours for 4 days, and the malaria infection rate of the mice was determined the day after the drug administration was completed. The malaria infection rate was determined by collecting a few drops of blood from the tail of infected mice, dropping them on a slide, preparing a thin-layered sample, performing Diff-Quik staining, and observing under an inverted microscope. The protozoan infection rate was calculated. As a control, using mice without drug administration (administered with PBS (-) buffer containing 1% DMSO instead of drug), the ratio of malaria infection rate of control to that of drug-administered mice was The malaria growth rate and 50% effective drug amount (ED ₅₀ ) were determined using the following formula.
Malaria parasite infection rate (%) = {(number of infected red blood cells) / (total number of red blood cells)} × 100
Growth rate (%) = {(b−a) / (c−a)} × 100
[Where a is the initial infection rate, b is the infection rate when the test solution is added, and c is the control infection rate]

(3) How to calculate the survival rate and malaria infection rate after drug administration To investigate whether mice infected with malaria are completely cured from malaria or how the survival rate changes after drug administration, after 4-Day Suppressive Test The body weight of each mouse was measured, the survival rate of the mouse was calculated using the following formula, and blood was collected from the tail vein of the mouse to calculate the malaria infection rate.
Life extension rate (%) = {(d × e) / (f × g)} × 100
[Where d is the number of days of survival after administration of the test substance in the test substance administration group, e is the number of surviving animals after administration of the test substance in the test substance administration group, and f is the survival after administration of the vehicle in the vehicle administration group. (Days, g indicates the number of surviving animals after vehicle administration in the vehicle administration group)

  As shown in Table 3, when the compounds 4d, 5c, 12a, 12b, and 12c were administered to mice, a clear decrease in the growth rate of malaria parasites was observed particularly at a dose of 15 mg / kg. Although the life extension rate shown in Table 4 was not as high as that of chloroquine, 15 mg / kg showed a life extension effect.

  From the above results, the bispyridinium salt compound of the present invention is useful as an antimalarial agent for the prevention and / or treatment of infectious diseases caused by malaria parasites, and is expected to have a clinical effect. Among them, the compounds 4d, 5c, 12a, 12b, and 12c synthesized in the examples showed high antimalarial activity and chemotherapeutic index, and thus can be a novel antimalarial agent having excellent antimalarial effect and high safety. . In addition, since the compound according to the present invention has a structure not found in existing antimalarial drugs, it can be expected to have an effect on malaria parasites having resistance to existing drugs.

  The bispyridinium salt compound according to the present invention is useful for the prevention and / or treatment of infectious diseases caused by malaria parasites as an antimalarial agent. Therefore, the pharmaceutical composition containing the bispyridinium salt compound according to the present invention has high utility and safety, and can be used clinically as an excellent antimalarial agent. In addition, it is also possible to further search for a substance having antimalarial activity using the bispyridinium salt compound of the present invention as a lead compound.

FIG. 1 is a diagram showing the structural formula of a bispyridinium salt (4-6). FIG. 2 is a diagram showing a synthesis scheme of a bispyridinium salt (4-6). FIG. 3 is a diagram showing the structural formula of a bispyridinium salt (11-14). FIG. 4 is a diagram showing a synthesis scheme of a bispyridinium salt (11-14).

Claims (29)

An antimalarial agent comprising a bispyridinium salt compound represented by the following general formula (1) as an active ingredient.
General formula (1)

In the formula, two R _{1 s} are the same or different and each represents an alkyl group having 1 to 10 carbon atoms, or an alkyl group having 1 to 10 carbon atoms terminated with an aromatic ring which may have a substituent,
R ₂ represents a linear alkylene group having 1 to 20 carbon atoms, or a phenylene group that has an alkylene group having 0 to 5 carbon atoms at both ends and may have a substituent on the benzene ring;
Two R _{3 s} are the same or different and each represents an alkyl group having 1 to 20 carbon atoms, or an alkyl group having 1 to 10 carbon atoms having an aromatic ring which may have a substituent;
X ⁻ represents an anion. An antimalarial agent comprising a bispyridinium salt compound represented by the following general formula (2) as an active ingredient.
General formula (2)

In the formula, two R _{1 s} are the same or different and each represents an alkyl group having 1 to 10 carbon atoms, or an alkyl group having 1 to 10 carbon atoms terminated with an aromatic ring which may have a substituent,
R ₂ represents a linear alkylene group having 1 to 20 carbon atoms, or a phenylene group that has an alkylene group having 0 to 5 carbon atoms at both ends and may have a substituent on the benzene ring;
Two R _{3 s} are the same or different and each represents an alkyl group having 1 to 20 carbon atoms, or an alkyl group having 1 to 10 carbon atoms having an aromatic ring which may have a substituent;
X ⁻ represents an anion. An antimalarial agent comprising a bispyridinium salt compound represented by the following general formula (3) as an active ingredient.
General formula (3)

In the formula, R ₁ has a linear alkylene group having 1 to 20 carbon atoms, a branched alkylene group having 3 to 20 carbon atoms, or an alkylene group having 0 to 5 carbon atoms at both ends, and a substituent on the benzene ring. Represents a phenylene group which may have
Two R _{2 s} are the same or different and each represents an alkyl group having 0 to 20 carbon atoms, or an alkyl group having 1 to 15 carbon atoms having an aromatic ring which may have a substituent;
X ⁻ represents an anion. An antimalarial agent comprising a bispyridinium salt compound represented by the following general formula (4) as an active ingredient.
General formula (4)

In the formula, R ₁ has a linear alkylene group having 1 to 20 carbon atoms, a branched alkylene group having 3 to 20 carbon atoms, or an alkylene group having 0 to 5 carbon atoms at both ends, and a substituent on the benzene ring. Represents a phenylene group which may have
Two R _{2 s} are the same or different and each represents an alkyl group having 0 to 20 carbon atoms, or an alkyl group having 1 to 15 carbon atoms having an aromatic ring which may have a substituent;
X ⁻ represents an anion. An antimalarial agent comprising a bispyridinium salt compound represented by the following general formula (5) as an active ingredient.
General formula (5)

In the formula, R ₁ has a linear alkylene group having 1 to 20 carbon atoms, a branched alkylene group having 3 to 20 carbon atoms, or an alkylene group having 0 to 5 carbon atoms at both ends, and a substituent on the benzene ring. Represents a phenylene group which may have
Four R _{2 s} are the same or different and each represents an alkyl group having 0 to 20 carbon atoms or an alkyl group having 1 to 15 carbon atoms having an aromatic ring which may have a substituent;
X ⁻ represents an anion.   1,1 '-(1,6-hexamethylene) bis [4-{(n-butylamino) carbonyl} pyridinium bromide].   1,1 '-(1,8-octamethylene) bis [4-{(n-butylamino) carbonyl} pyridinium bromide].   1,1 '-(1,10-Decamethylene) bis [4-{(n-butylamino) carbonyl} pyridinium bromide].    1,1 '-(1,12-dodecamethylene) bis [4-{(n-butylamino) carbonyl} pyridinium bromide].   1,1 '-(1,6-hexamethylene) bis [4-{(n-hexylamino) carbonyl} pyridinium bromide].   1,1 '-(1,8-octamethylene) bis [4-{(n-hexylamino) carbonyl} pyridinium bromide].   1,1 '-(1,10-Decamethylene) bis [4-{(n-hexylamino) carbonyl} pyridinium bromide].   1,1 '-(1,12-dodecamethylene) bis [4-{(n-hexylamino) carbonyl} pyridinium bromide].   1,1 '-(1,6-hexamethylene) bis [4-{(n-octylamino) carbonyl} pyridinium bromide].   1,1 '-(1,8-octamethylene) bis [4-{(n-octylamino) carbonyl} pyridinium bromide].   1,1 '-(1,10-Decamethylene) bis [4-{(n-octylamino) carbonyl} pyridinium bromide].   1,1 '-(1,12-dodecamethylene) bis [4-{(n-octylamino) carbonyl} pyridinium bromide].   N, N'-dimethyl-N, N'-hexamethylenebis (4-carbamoyl-1-n-hexylpyridinium bromide).   N, N'-di-n-butyl-N, N'-hexamethylenebis (4-carbamoyl-1-n-hexylpyridinium bromide).   N, N'-dibenzyl-N, N'-hexamethylenebis (4-carbamoyl-1-n-hexylpyridinium bromide).   N, N'-dimethyl-N, N'-hexamethylenebis (4-carbamoyl-1-n-octylpyridinium bromide).   N, N'-di-n-butyl-N, N'-hexamethylenebis (4-carbamoyl-1-n-octylpyridinium bromide).   N, N'-dibenzyl-N, N'-hexamethylenebis (4-carbamoyl-1-n-octylpyridinium bromide).   N, N'-dimethyl-N, N'-octamethylenebis (4-carbamoyl-1-n-hexylpyridinium bromide).   N, N'-di-n-butyl-N, N'-octamethylenebis (4-carbamoyl-1-n-hexylpyridinium bromide).   N, N'-dibenzyl-N, N'-octamethylenebis (4-carbamoyl-1-n-hexylpyridinium bromide).   N, N'-dimethyl-N, N'-octamethylenebis (4-carbamoyl-1-n-octylpyridinium bromide).   N, N'-di-n-butyl-N, N'-octamethylenebis (4-carbamoyl-1-n-octylpyridinium bromide).   N, N'-dibenzyl-N, N'-octamethylenebis (4-carbamoyl-1-n-octylpyridinium bromide).

Images