JP2011037814A - Combination of 5-heterocyclic ring-substituted imino-9-dialkylaminobenzo[a] phenoxathiin compound or salt thereof and artemisinin derivative for prevention or treatment of malaria plasmodium species disease - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a disease prevention or a treatment for a new strain of malaria Plasmodium having a highly therapeutic effect and selective toxicity toward malaria Plasmodium-based infectious disease and capable of suppressing the emergence of a resistant malaria Plasmodium strain as compared with a case of administering an active ingredient in a monotherapy. <P>SOLUTION: As an active ingredient, 5-heterocyclic ring-substituted imino-9-dialkyl aminobenzo[a]phenoxathiin compound or its salt shown by Formula (I) and an artemisinin derivative are combined. In the formula, R<SP>1</SP>and R<SP>2</SP>are each independently a methyl group, ethyl group, propyl group, 2-hydroxyethyl group or 2-methanesulfoneamideethyl group; R<SP>3</SP>represents a 2-pyridyl group or 4-pyridyl group; R<SP>4</SP>represents a fluorine atom, chlorine atom, methoxy group or methyl group; R<SP>5</SP>is a fluorine atom, chlorine atom, methoxy group or methyl group; m represents an integer of 0 or 1; n represents an integer of 0 or 1; and an negative ion represented by X<SP>-</SP>is a chlorine atom, bromine atom, nitric acid ion, sulfonic acid ion, p-toluenesulfonic acid ion or oxalic acid ion. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to novel combinations of 5-heterocyclic substituted imino-9-dialkylaminobenzo [a] phenoxatinates or salts thereof and artemisinin derivatives as active ingredients for preventing or treating malaria parasite diseases, and pharmaceuticals containing them Relates to the composition.

  Malaria caused by malaria parasite infection is the most serious parasitic infection in human history. Malaria-causing pathogens are protozoa belonging to the genus Plasmodium, such as P. falsiparum distributed throughout the tropical regions of Africa, Asia and Latin America, and one of the tropical and temperate zones of the world. Protozoa such as Plasmodium falciparum (P. vivax) distributed in Japan, egg-shaped malaria protozoa (P. ovale) mainly distributed in tropical West Africa, and protozoan malaria parasite (P. malariae) distributed in various parts of the world Infects humans through the mosquitoes. About 300 million new people are infected with malaria every year in the world, and more than 1 million people die each year mainly from infants with weak immunity. Ninety percent of the deaths are children under the age of 5 in Sub-Saharan Africa (south of the Sahara Desert). Malaria has had a major impact on the economy of the Sub-Saharan Africa, and the economic loss is said to be over 1 trillion yen every year. In addition, the habitat of anopheles mosquitoes that mediate this has expanded with global warming, and the development of malaria parasite disease prevention or treatment should be noted from the perspective of future Japanese safety and security It is considered a problem.

  Similar to the emergence of malaria parasites that are resistant to chloroquine, which has been widely used as a specific drug for malaria, malaria parasites that are resistant to drugs such as mefloquine, atobacon, and pyrimesamine have also emerged. Currently, the most effective antimalarial drug is an artemisinin derivative. Artemisinin is an extracted drug from Artemisia annua and has been approved for use as an antimalarial drug since at least the second century BC. Artemisinin and its derivatives have begun to be used as a treatment for malaria mainly in China since the 1970s, and are currently in the spotlight for diseases caused by drug-resistant malaria parasites.

According to the World Health Organization (WHO), the emergence of resistant protozoa to malaria drugs such as chloroquine is considered to be caused by monotherapy, which is a combination therapy of malaria drugs to reduce the risk of drug resistance. Is recommended. In particular, a combination of an artemisinin derivative and another effective antimalarial drug is called a combination therapy based on artemisinin (Artemisinin-based combination therapies) and is currently the most effective antimalarial drug (Non-patent Document 1). Examples of combination therapies that are on the market include artemisinin artesunate and amodiaquine combination, artemeter and lumefantrine combination.

  On the other hand, the combination of chloroquine and artesunate does not show a good therapeutic effect and seems to induce the emergence of resistant strains, particularly chloroquine resistant strains. When considering the current situation where malaria parasites that are resistant to drugs such as chloroquine, mefloquine, atobacon, and pyrimesamine are emerging and becoming a major threat, the development of new drugs is urgently needed (Non-patent Document 2). .

  Masataka Ihara, one of the inventors of the present invention, has so far used a pharmaceutical composition for preventing or treating protozoan diseases containing a phenoxanidium compound as an active ingredient (Patent Document 1) and an azarodacyanine compound as an active ingredient. A pharmaceutical composition for preventing or treating protozoan diseases (Patent Document 2) has been researched and developed.

So far, a large number of benzo [a] phenoxazonium compounds, which are benzo [a] phenoxytin and salts thereof, have been synthesized, and antitumor properties have been reported for these compounds (Non-Patent Documents 3 to 7). In addition, examples in which R ³ which does not belong to the 5-heterocyclic-substituted imino-9-dialkylaminobenzo [a] phenoxytin compound represented by the general formula (I) used in the combination of the present invention is a hydrogen atom or an ethyl group are described. ing. Although these compounds have been reported to have growth inhibitory activity against protozoa causing tropical diseases such as malaria, their in vitro and in vivo activities are low (Non-patent Documents 8 and 9).

International Publication WO2006 / 087935 Pamphlet International Publication WO2006 / 137258 Pamphlet

WHO briefing on Maria Treatment Guidelines and artemisinin monotherapies, Geneva, 19 April 2006 Journal of Japanese Pharmacology 132, 288-291 (2008) M.M. L. Crossley, P.M. F. Dreisbach, C.I. M.M. L. C. Hofman, R.A. P. Parker, J .; Am. Chem. Soc. 74, 573-578 (1952) M.M. L. Crossley, R.M. J. et al. Turner, C.I. M.M. Hofmann, P.M. F. Dreisbach, R.M. P Parker, J .; Am. Chem. Soc. , 74, 578-584 (1952) M.M. L. Crossley, C.I. M.M. Hofmann, P.M. F. Dreisbach, J. et al. Am. Chem. Soc. 74,584-586 (1952) N. Motohashi, Yakugaku Zashi, 102, 646-650 (1982) N. Motohashi, Medicinal Research Reviews, 11, 239-294 (1991) K. Takasu, T .; Shimogama, C.I. Satoh, M .; Ihara, J .; Med. Chem. , 50, 2281-2284 (2007) J. et al. L. Vennerstrom, M.M. T.A. Makler, C.I. K. Angelhofer, J. et al. A. Williams, Antimicrob. Agents J. Chemother. , 39, 2671-2777 (1995)

  The object of the present invention is to provide a new treatment that has a high therapeutic effect and selective toxicity against parasitic malaria parasite infections, and that can suppress the appearance of resistant malaria parasites compared to when the active ingredient is administered as a single agent therapy. It is an object to provide a combination of various therapeutic and / or prophylactic drugs and a pharmaceutical composition containing them.

  In order to solve the above problems, the present inventors have combined a 5-heterocyclic-substituted imino-9-dialkylaminobenzo [a] phenoxatin compound represented by the following general formula (I) or a salt thereof and an artemisinin derivative: As a result of using malaria-infected mice as a host model and administering them in any amount or form and evaluating their therapeutic effects, they were useful for the prevention or treatment of protozoan diseases, and these active ingredients were administered as monotherapy It has been found that the emergence of resistant protozoa can be prevented or delayed as compared with cases, and the present invention has been completed based on these findings.

（式中、Ｒ^１及びＲ^２はそれぞれ独立にメチル基、エチル基、プロピル基、２−ヒドロキシエチル基、又は２−メタンスルホンアミドエチル基であり、Ｒ^３は２−ピリジル基又は４−ピリジル基を表し、Ｒ^４はフッ素原子、塩素原子、メトキシ基、又はメチル基を表し、Ｒ^５はフッ素原子、塩素原子、メトキシ基又はメチル基を表し、ｍは０又は１の整数、ｎは０又は１の整数を表し、Ｘ⁻で表わされる陰イオンが、塩素原子、臭素原子、硝酸イオン、硫酸イオン、ｐ−トルエンスルホン酸イオン又はシュウ酸イオンを表す。）
＜２＞ 前記アーテミシニン誘導体がアーテスネート、アーテエター、アーテメター又はジヒドロアーテミシニンから成ることを特徴とする前記＜１＞項に記載の組み合わせである。
＜３＞ 一般式（Ｉ）で表わされる５−ヘテロ環置換イミノ−９−ジアルキルアミノベンゾ［ａ］フェノキサチン化合物又はその塩の投与量が１〜５，０００ｍｇ／日／体重７０ｋｇ、アーテミシニン誘導体の投与量が１〜７００ｍｇ／日／体重７０ｋｇであることを特徴とする前記＜１＞項又は＜２＞項に記載の組み合わせである。
＜４＞ マラリア原虫疾患予防又は治療用の活性成分として、以下の構造式１Ａ〜１Ｑのいずれかで示される化合物又はその塩と、アーテミシニン誘導体との組み合わせである。

＜５＞ 活性成分のそれぞれを同時又は逐次的に投与することを特徴とする前記＜１＞〜＜４＞のいずれか１項に記載の組み合わせである。
＜６＞ 前記＜１＞〜＜４＞のいずれか１項に記載の組み合わせであることを特徴とするマラリア原虫疾患予防又は治療用医薬組成物である。Specific means for solving the above problems are as follows.
<1> As an active ingredient for preventing or treating malaria parasite disease, a 5-heterocyclic-substituted imino-9-dialkylaminobenzo [a] phenoxatin compound represented by the following general formula (I) or a salt thereof, and an artemisinin derivative: It is a combination.

Wherein R ¹ and R ² are each independently a methyl group, an ethyl group, a propyl group, a 2-hydroxyethyl group, or a 2-methanesulfonamidoethyl group, and R ³ is a 2-pyridyl group or 4-pyridyl group R ⁴ represents a fluorine atom, a chlorine atom, a methoxy group, or a methyl group, R ⁵ represents a fluorine atom, a chlorine atom, a methoxy group, or a methyl group, m is an integer of 0 or 1, and n is 0 or 1 represents an integer, X ^- an anion represented by is a chlorine atom, a bromine atom, a nitrate ion, sulfate ion, p- toluenesulfonate ion or an oxalate ion).
<2> The combination according to <1>, wherein the artemisinin derivative is composed of artesunate, arteator, artemeta or dihydroartemisinin.
<3> Administration of 5-heterocyclic substituted imino-9-dialkylaminobenzo [a] phenoxatin compound represented by the general formula (I) or a salt thereof is 1 to 5,000 mg / day / 70 kg body weight, administration of artemisinin derivative The combination according to <1> or <2>, wherein the amount is 1 to 700 mg / day / 70 kg of body weight.
<4> A combination of a compound represented by any of the following structural formulas 1A to 1Q or a salt thereof and an artemisinin derivative as an active ingredient for preventing or treating malaria parasite diseases.

<5> The combination according to any one of <1> to <4>, wherein each of the active ingredients is administered simultaneously or sequentially.
<6> A pharmaceutical composition for preventing or treating malaria parasite disease, which is a combination according to any one of <1> to <4>.

  A combination of a 5-heterocyclic-substituted imino-9-dialkylaminobenzo [a] phenoxatin compound represented by the general formula (I) of the present invention or a salt thereof and an artemisinin derivative caused a parasitic malaria parasite infection. It is possible to provide a novel therapeutic and / or prophylactic pharmaceutical combination or composition thereof which has a low toxicity to a living body and exhibits a significant healing effect upon administration. Moreover, the appearance of resistant protozoa can be suppressed as compared with the case where these active ingredients are administered by monotherapy.

  Hereinafter, the combination by this invention and the pharmaceutical composition containing these are demonstrated concretely. Specific examples and preferred examples of the 5-heterocyclic substituted imino-9-dialkylaminobenzo [a] phenoxatin compound represented by the general formula (I) used in the combination of the present invention or a salt thereof are shown below, but these ranges are included. It is not limited to.

In the compound represented by the general formula (I) used in the combination of the present invention, R ¹ and R ² are each independently a methyl group, an ethyl group, a propyl group, a 2-hydroxyethyl group, or a 2-methanesulfonamidoethyl group. Yes, R ³ represents a substituted or unsubstituted pyridyl group, R ⁴ represents a fluorine atom, a chlorine atom, a methoxy group, or a methyl group, R ⁵ represents a fluorine atom, a chlorine atom, a methoxy group, or a methyl group, m represents an integer of 0 or 1, n represents an integer of 0 or 1, and the anion represented by X ⁻ represents a chlorine atom, bromine atom, nitrate ion, sulfate ion, p-toluenesulfonate ion or oxalate ion. To express. More preferable examples include compounds represented by any one of structural formulas 1A to 1Q. Particularly preferred examples of the pyridyl group for R ³ include a 2-pyridyl group and a 4-pyridyl group.

Artemisinin used in the combination of the present invention is represented by the following structure 2A. Preferred examples of the artemisinin derivative include dihydroartemisinin (the following structural formula 2B) which is a reduced form of artemisinin, and artesunate (the following structural formula 2C) which is a hemisuccinate ester of dihydroartemisinin. A certain artemeta (the following structural formula 2D) and an ethyl ether-based artieter (the following structural formula 2E) can be mentioned.

  The pharmaceutical composition of the present invention is a composition comprising a combination of a 5-heterocyclic-substituted imino-9-dialkylaminobenzo [a] phenoxatin compound of general formula (I) or a salt thereof and an artemisinin derivative. Suitable examples of the pharmaceutical carrier or diluent that can be used in the pharmaceutical composition of the present invention include sodium chloride, magnesium chloride, zinc chloride, glucose, saccharose, lactose, ethyl alcohol, glycerin; Mannitol; sorbitol; pentaerythritol; diethylene glycol, propylene glycol, dipropylene glycol, polyethylene glycol 400, other polyethylene glycols; glyceryl trilaurate, and glyceryl distearate Fatty acid mono-, di- and triglycerides; pectin; starch; arginic acid; xylos; talc; stone matsuko; , Oils and fats such as oilseed oil, sunflower oil and cod liver oil; gelatin; Silica; cellulose; cellulose derivatives such as methyl hydroxypropyl cellulose, methyl cellulose, hydroxyethyl cellulose; calcium stearate, calcium laurate, magnesium oleate, calcium palmitate, calcium behenate and stearic acid Salts of fatty acids having 12 to 22 carbon atoms such as magnesium; cyclodextrins (e.g. [alpha] -cyclodextrin, [beta] -cyclodextrin, [gamma] -cyclodextrin, hydroxyethyl- [beta] -cyclodextrin, hydroxypropyl- [beta] -cyclo Dextrin, dihydroxypropyl-β-cyclodextrin, carboxymethylethyl-β-cyclodextrin, cycloawadolin, dimethyl-β-cyclodextrin, etc .; emulsifiers (eg, Saturated and unsaturated fatty acids and glycols having ˜22 carbon atoms, especially 10-18 carbon atoms, glycol, glycerin, diethylene glycol, pentaerythritol, ethyl alcohol, butyl alcohol, octadecyl alcohol And monovalent aliphatic alcohols having 1 to 20 carbon atoms or esters with polyhydric alcohols, and silicones such as dimethylpolysiloxane. Any additional carrier known to those skilled in the art can be used in the pharmaceutical compositions of the present invention.

  The pharmaceutically effective amount and administration method or administration means of the compound used in the combination of the present invention include the type of malaria parasite parasite that causes infection, the severity of the disease state, the treatment policy, the patient's age, weight, sex, general Can be appropriately selected by those skilled in the art according to the general health condition and the (genetic) racial background of the patient. Generally, the dosage of the 5-heterocyclic substituted imino-9-dialkylaminobenzo [a] phenoxatin compound or salt thereof of the present invention is 1 to 5,000 mg / day / 70 kg body weight, more typically 25 to 25 kg. The dose of the other artemisinin derivative to be combined is 1 to 700 mg / day / 70 kg body weight, more generally 70 to 400 mg / day / 70 kg body weight.

  In the combination according to the invention, it is desirable to administer each of the two active ingredients once or more daily, preferably continuously for 3 days per day. By limiting the administration period, it is possible to prevent induction of resistance to malaria parasites as compared to 7 days recommended for monotherapy using an artemisinin derivative.

  The pharmaceutical composition of the present invention can have any shape known to those skilled in the art depending on the administration method, administration route and the like. They can be administered by any suitable method. For example, when the liquid is in the form of a liquid, tablet or colloid, it is in the form of a solution dissolved in a 5% glucose aqueous solution or with the above-mentioned carrier or diluent. A method of injecting internally or subcutaneously can be mentioned. In the case of tablets, oral administration can be mentioned, and in the case of colloids, methods such as application to the skin can be mentioned. The compound represented by the general formula (I) and the artemisinin derivative used in the combination of the present invention can be contained in an appropriate amount depending on the purpose of use, the object, the shape, etc. About 10,000 mg, preferably about 10 mg to 3,000 mg is contained.

Hereinafter, a method for synthesizing the 5-heterocyclic-substituted imino-9-dialkylaminobenzo [a] phenoxatin compound represented by the general formula (I) or a salt thereof used in the combination of the present invention will be described.
A 5-heterocyclic-substituted imino-9-dialkylaminobenzo [a] phenoxatin compound represented by the general formula (I) and a salt thereof include a 4-nitrosoaniline derivative represented by the general formula 3 or p-phenylene represented by 5 After obtaining 2 by reacting a diamine with a 2-naphthol derivative represented by the general formula 4 in the presence of an oxidizing agent such as HNO ₃ , ethanol is added in the presence of a primary amine and an oxidizing agent containing oxygen. To give a 5-heterocyclic substituted imino-9-dialkylaminobenzo [a] phenoxatin compound represented by the general formula (I) as a stable substance as a free base. These can also be purified as hydrochlorides.

  In order to elucidate the effectiveness of these compounds and pharmaceutical compositions thereof, the in vivo inhibitory effect using malaria-infected mice was tested and evaluated. The technical scope of the present invention is not limited to these examples.

[Evaluation of inhibition rate during oral administration of malaria-infected mice (in vivo)]
In this experiment, a rodent malaria parasite (Plasmodium berghei NK65 strain) was used. Infected blood is ICR female SPF mice 4-6 weeks old (20% -26%) infected and passed by intraperitoneal or tail vein administration. After collecting blood from the tail vein of a malaria-infected mouse to determine the infection rate and confirming that it was appropriately infected (10-20% infection rate), malaria-infected blood was collected from the mouse heart with a heparin-containing syringe. Thereafter, from the red blood cell count (cells / ml) and the protozoan infection rate, the blood was diluted with physiological saline so as to be 5.0 × 10 ⁻⁶ malaria parasite per 1 ml dose. This was infected from the tail vein by 0.2 ml per 20 g body weight of an uninfected mouse (ICR female 5 weeks old). The compound used for the test was dissolved in a mixed solution of 7 ml of Tween 80, 3 ml of ethyl alcohol and 90 ml of water.

  Three mice per group, 24 hours (1 day) after malaria infection, compound 1B per kg body weight of compound 1B, 10mg of compound 1B, 5mg of artemeter, 10mg of compound 1B and 5mg of artemeta in the combination of compound 1B and artemeta The resultant was orally administered, and was orally administered three times in total: 24 hours (1 day) after malaria infection, 48 hours (2 days), and 72 hours (3 days later).

  Blood was collected from the tail of the mouse at 96 hours (4 days) and 144 hours (6 days) after malaria infection, and a thin smear was prepared. Under the microscope, the compound administration group and the control (no compound administration) group The number of malaria parasite infections was counted and the malaria parasite infection rate (Parasisitemia) was calculated.

The suppression rate (suppression) at the time of drug administration was calculated from the malaria parasite infection rate obtained above by the following formula.
Inhibition rate (%) = (B−A) / B × 100
A: Protozoa infection rate in mice administered with this test compound B: Protozoa infection rate in control (no compound administration) mice

  Furthermore, changes in body weight, diarrhea symptoms, color gloss, etc. of the mice were observed, and the presence or absence of acute toxicity due to compound administration was examined.

As can be seen from Table 1, administration by the combination of Compound 1B, which is a combination of the present invention, and Artemeta (dose is 1B 10 mg / kg, Artemeta 5 mg / kg) is to achieve a suppression rate of 97% or more with each single administration. The amount of inhibition after drug administration can be remarkably increased at a dose lower than the dose required for administration (30 mg / kg for 1B and 5 mg / kg for artemeta). In addition, acute toxicity such as diarrhea and sudden weight loss was not observed in the above combination.
In the present invention, the combination of the 5-heterocyclic-substituted imino-9-dialkylaminobenzo [a] phenoxatin compound represented by the general formula (I) and a salt thereof with an artemisin derivative is compared with each single administration. Each dose can be expected to be drastically reduced, and the occurrence of resistant protozoa can be suppressed as compared to the case where these active ingredients are administered by monotherapy.

Claims (5)

A combination of a 5-heterocyclic-substituted imino-9-dialkylaminobenzo [a] phenoxatin compound represented by the following general formula (I) or a salt thereof and an artemisinin derivative as an active ingredient for preventing or treating malaria parasite diseases.

Wherein R ¹ and R ² are each independently a methyl group, an ethyl group, a propyl group, a 2-hydroxyethyl group, or a 2-methanesulfonamidoethyl group, and R ³ is a 2-pyridyl group or 4-pyridyl group R ⁴ represents a fluorine atom, a chlorine atom, a methoxy group, or a methyl group, R ⁵ represents a fluorine atom, a chlorine atom, a methoxy group, or a methyl group, m is an integer of 0 or 1, and n is 0 or 1 represents an integer, X ^- an anion represented by is a chlorine atom, a bromine atom, a nitrate ion, sulfate ion, p- toluenesulfonate ion or an oxalate ion).   2. A combination according to claim 1, wherein the artemisinin derivative consists of artesunate, arteator, artemeter or dihydroartemisinin.   The dosage of the 5-heterocyclic substituted imino-9-dialkylaminobenzo [a] phenoxatin compound or its salt represented by the general formula (I) is 1 to 5,000 mg / day / 70 kg of body weight, and the dosage of artemisinin derivative is 1 The combination according to claim 1 or 2, characterized in that it consists of ~ 700mg / day / 70kg body weight.   4. The combination according to any one of claims 1 to 3, wherein each of the active ingredients is administered simultaneously or sequentially.   The pharmaceutical composition for the prevention or treatment of the malaria parasite disease containing the combination of any one of Claims 1-3.