JP6388162B2 - Antitrypanosoma protozoan active substance actinoallolides and method for producing the same - Google Patents

Description

Cross reference

  This application claims the priority from Japanese Patent Application No. 2013-007684, which was filed with the Japan Patent Office on January 18, 2013. The contents described in Japanese Patent Application No. 2013-007684 are all incorporated herein by reference. In addition, all documents cited throughout the present application are incorporated herein by reference in their entirety.

  The present invention belongs to the field of growth inhibition of Trypanosoma protozoa. Specifically, the present invention relates to an animal drug, a compound having a growth inhibitory activity of Trypanosoma protozoa that is effective as a pharmaceutical, a production method thereof, and use thereof. In particular, the present invention relates to a therapeutic or prophylactic agent for a disease caused by infection with Trypanosoma protozoa containing the compound of the present invention as an active ingredient (sometimes referred to herein as an “antitrypanosoma drug”). About.

  Trypanosomiasis includes African sleeping sickness (human African trypanosomiasis) prevalent in the African region and Chagas disease prevalent in the South American region. African sleeping sickness is a re-emerging protozoal infection, and the risk of infection is estimated to be 70 million, and the annual death toll is about 50,000. In particular, Trypanosoma protozoa parasitizing humans are classified into two types: Trypanosoma brucei gambiense and Rhodesia protozoa (T. rhodesiens), the former being chronic sleep disease and the latter Causes acute sleep sickness. At the end of infection, protozoa are transferred to the central nervous system, and more than 80% of Trypanosoma protozoa suffer from coma, resulting in death. These trypanosomes are mediated by tsetse flies that only inhabit Africa.

  Furthermore, Gambia trypanosomes and Rhodesia trypanosomes are common animal parasites, and infest livestock such as cattle, horses and sheep, and wild animals such as gazelles and dogs. These animals serve as reservoirs but do not develop. There are also other types of protozoan parasites that do not infest humans and infest livestock animals. T. is a protozoan of the subgenus Trypanosoma brucei brucei (protozoan caused by Nagana disease), a protozoan of the subgenus Duttonella. There are vivax, vivax (Zuma disease-causing protozoa), etc., and when these protozoa are infected, a fatal infection course is followed depending on the animal species. These are also mediated by tsetse flies.

  The tsetse flies live in a wide area ranging from the east coast south of the Sahara Desert on the African continent to 10 million square kilometers in 36 countries on the west coast. Over 150 million livestock animals are threatened with various trypanosomiasis The current situation is being exposed. Furthermore, as a Tsetse fly non-mediated trypanosomatid protozoan that infects animals by mechanical propagation of blood-sucking insects such as abs and flies, T. is a protozoan of the subgenus Trypanosoma. evansi (protozoan causing Sura disease), T. et al. and equiperdum (protozoan caused by pesticidal disease). In particular, Sura disease has a worldwide epidemic such as Africa, Central and South America, Southeast Asia, China, Middle East and India. In recent years, the epidemic has been on the rise, and it is animal trypanosomiasis that should be most wary of entering Japan.

  Existing human antitrypanosoma protozoan drugs against these trypanosomes include suramin (developed in 1923), pentamidine (developed in 1939), melarsoprole (developed in 1953), etc. for the past half century. Chemical drugs such as classic drugs and eflonitin (developed in 1978) have long been used. Suramin is effective early in the infection of Gambia trypanosomes and Rhodesia trypanosomes but is nephrotoxic.

  Pentamidine is effective in the early infection of Gambia trypanosoma protozoa, but is ineffective in rhodesia trypanosoma protozoa, and has side effects of lowering blood pressure and reducing blood sugar. The arsenic agent, meralsoprole, is effective in the end stage of infection (central neurosis) of Gambia trypanosoma protozoa and rhodesia trypanosoma protozoa by crossing the blood-brain barrier, but it has side effects on the central nervous system. Strongly causes encephalopathy. In addition, resistant protozoan strains to meralsoprole have also appeared.

  Ephronitin is effective at the end of infection with resistant Gambia trypanosoma protozoa to which melalsoprol does not work by passing through the blood-brain barrier, but is ineffective against rhodesia trypanosoma protozoa. These drugs are old and their effectiveness is gradually declining. Further, diminazen, suramin, isometadium, mutagenic substance fomidium, and the like have been used for the treatment of trypanosomiasis in animals. In particular, since these drugs have been used in large quantities for a long period of time, drug-resistant protozoa have emerged in various places, and their usefulness as anti-trypanosoma protozoan drugs has significantly decreased, which has become a major problem. Yes.

  Chagas disease or human American trypanosomiasis is a protozoal infection with an estimated estimated 10 million or more annual deaths and approximately 10,000 deaths annually. Trypanosoma cruzi It is caused by being mediated by the turtle, a blood-sucking insect. In the acute phase, fever, diffuse lymphadenopathy, and splenomegaly are seen. Most incentives remit spontaneously, but some transition to a chronic phase after several years to decades of asymptomatic periods. Chronic symptoms are angina pectoral pain, arrhythmia, heart failure such as heart failure or giant organ disease, no effective treatment, and prognosis is very severe.

  As existing Chagas disease therapeutic agents, chemical synthetic drugs such as nifurtimox (developed in 1964) and benznidazole (developed in 1966) have long been used for the past half century. Nifurtimox is ineffective in the chronic phase and has serious side effects such as tremor, insomnia, neuritis, and anemia. Benznidazole is also ineffective in the chronic phase and has side effects such as photosensitivity and neuritis. Both drugs require long-term administration.

  In Africa sleep disease and Chagas disease, due to delays in the development of new drugs, existing drugs with strong classic side effects are currently used for treatment, both of which are effective globally Development is required. Some of the existing therapeutic drugs for African sleeping sickness have different efficacy depending on the type of protozoa and the stage of infection, and some have emerged drug-resistant protozoan strains. For this reason, African sleeping sickness with a new skeleton that is effective regardless of the type of protozoa and the stage of infection, and is effective against rhodesia trypanosoma protozoa and end-stage infection (central neuropathy) and has few side effects The development of therapeutic agents is desired on a global scale. On the other hand, existing Chagas disease drugs are ineffective in the chronic phase and have serious side effects. For this reason, the development of a Chagas disease therapeutic agent having a novel skeleton that is effective for chronic patients and has few side effects is desired on a global scale.

  So far, therapeutic drugs aimed at treating trypanosomiasis have been reported to solve such problems, but no effective compound has been found yet (see Patent Document 1).

International Publication WO2009 / 004899

  Trypanosomiasis such as African sleeping sickness and Chagas disease is caused by the growth of protozoan trypanosomes in the host. Substances that inhibit the growth of Trypanosoma protozoa are expected as antitrypanosomal drugs that suppress the onset or alleviate the symptoms of Trypanosomiasis. Therefore, an object of the present invention is to provide a novel antitrypanosome drug in order to solve the above-mentioned problems of the prior art.

  Therefore, as a result of continuing to search for substances that inhibit the growth of Trypanosoma protozoa from the microorganism culture, the present inventors have found that actinoallolides A, B, C, which are novel compounds produced by actinomycetes MK10-036 strain The substance D and / or E was found to have the inhibitory activity, and the present invention was completed based on this finding.

  The present invention has been completed based on such findings, and provides a novel compound represented by the following formulas I to V, actinoallide A, B, C, D and / or E. is there.

(Actinoallolide A substance)
In the present specification, the actinoallide A substance is a compound represented by the following formula I.

  In addition, the actinoallide A substance in the present specification is preferably a compound having the following three-dimensional structure.

(Actino allolide B substance)
In this specification, the actinoallide B substance is a compound represented by the following formula II.

  In addition, the actinoallide B substance in the present specification is preferably a compound having the following three-dimensional structure.

(Actino allolide C substance)
In this specification, the actinoallide C substance is a compound represented by the following formula III.

  In addition, the actinoallide C substance in the present specification is preferably a compound having the following three-dimensional structure.

(Actino allolide D substance)
In the present specification, the actinoallide D substance is a compound represented by the following formula IV.

  In addition, the actinoallide D substance in the present specification is preferably a compound having the following three-dimensional structure.

(Actinoallolide E substance)
In the present specification, the actinoallide E substance is a compound represented by the following formula V.

  In addition, the actinoallide E substance in the present specification is preferably a compound having the following three-dimensional structure.

(Production method)
The present invention also cultivates a microorganism belonging to actinomycetes having the ability to produce the actinoallide A, B, C, D and / or E substance in a medium, and the actinoallide A, B, C in the culture. , D and / or E substances, and actinoallolide A, B, C, D and / or E substances are collected from the culture. The present invention relates to a method for producing substance E. In the production method, as a microorganism belonging to actinomycetes having an ability to produce actinoallolides A, B, C, D and / or E, preferably, Actinoallomulus sp. MK10-036 (Reception number NITE ABP-1208) stock.

(Microorganism)
In another aspect, the present invention further relates to a novel actinomycete, Actinoallomurus sp. MK10-036 strain.

  In this specification, Actinoallomulus sp. (Actinoallomulus sp.) MK10-036 is a microorganism newly isolated from a plant in the Kingdom of Thailand by the present inventors, and is known as Actinoallomulus esp. sp.) The strain of MK10-036 strain deposited on January 24, 2012 at the National Institute of Technology and Evaluation Microbiology Depositary Center (2-5-8 Kazusa Kamashichi, Kisarazu City, Chiba Prefecture) (Reception number NITE ABP-1208). The Actinoallomurus sp. MK10-036 strain of the present invention has the mycological properties described in Example 1 of the present specification.

(Pharmaceutical composition)
Moreover, the actinoallolide A, B, C, D and / or E substance of the present invention can be used as a pharmaceutical composition. Therefore, in still another aspect, the present invention relates to a pharmaceutical composition containing the above-mentioned actinoallide A, B, C, D and / or E substance as an active ingredient. Specifically, since the Actinoallolide A, B, C, D and / or E substance of the present invention has an inhibitory action on the growth of Trypanosoma protozoa, it is a therapeutic drug for diseases caused by Trypanosoma protozoa infection. Alternatively, it can be used as a prophylactic agent (antitrypanosome drug). In another aspect, the present invention also relates to actinoallolides A, B, C, D and / or E for use in the treatment or prevention of diseases resulting from infection with Trypanosoma protozoa. Alternatively, the present invention treats a disease resulting from infection with Trypanosoma protozoa, comprising administering an effective amount of the actinoallolide A, B, C, D and / or E substance to a patient in need thereof. It relates to a method or a preventive method.

  As used herein, the term “trypanosoma protozoa” refers to Trypanosoma cruzi, Trypanosoma cruzi, Trypanosoma cruzi, and subgenus of Trypanosoma protozoa・ Brucei (T. brucei), Trypanosoma congolens (T. convolense), Trypanosoma baivax (T. vivax), Trypanosoma omavansi (T. evani), Trypanosoma tyreli (T. Theilei), including the Trypanosoma Exadam (T. equiperdum).

  Further, in the present specification, a disease derived from infection with Trypanosoma protozoa is a disease or injury that develops or worsens due to infection of the Trypanosoma protozoa. Diseases known as “massis” are included, for example, African trypanosomiasis, Chagas disease, Nagana disease, Surra disease and the like. Since the pharmaceutical composition of the present invention solves the problems of side effects and drug resistance in existing drugs, it can also be used as a therapeutic agent in the chronic phase of these diseases.

  Since the actinoallide A, B, C, D and / or E substance of the present invention can inhibit the growth of Trypanosoma protozoa, it becomes a novel antitrypanosome drug. In addition, since the Actinoallomurus sp. MK10-036 strain of the present invention produces Actinoallorides A, B, C, D and / or E substances, Actinoallorides A, B, It can be used for the production of C, D and / or E substances.

  The actinoallolide A, B, C, D and / or E substance of the present invention is a culture of microorganisms belonging to actinomycetes having the ability to produce actinoallide A, B, C, D and / or E substance in a medium. Then, actinoallide A, B, C, D and / or E substance is accumulated in the culture, and actinoallide A, B, C, D and / or E is collected from the culture (separation / extraction / purification). ).

  In the method for producing an actinoallide A, B, C, D and / or E substance of the present invention, “microorganisms belonging to actinomycetes having the ability to produce an actinoallide A, B, C, D and / or E substance “” Is not particularly limited as long as it is a bacterium belonging to actinomycetes and has the ability to produce actinoallolides A, B, C, D and / or EE substances. In addition to the strains described above, the strains that can be used in the method for producing the actinoallide A, B, C, D and / or E substance of the present invention include not only the above strains but also mutants thereof, actinoallide A, All bacteria having the ability to produce B, C, D and / or E substances are included. Whether or not a microorganism is a “microorganism belonging to actinomycetes having the ability to produce actinoallide A, B, C, D and / or E” can be determined, for example, by the following method. Yeast extract (produced by Oriental Yeast Co., Ltd.) 1%, glucose culture medium (pH 6.0) consisting of 1% glucose, 7000 mL jar fermenter containing 5000 mL test microorganism 100 mL cultured in liquid medium After inoculating and shaking culture at 27 ° C. for 10 days, the obtained seed culture solution was added to a liquid medium (pH 6.0) consisting of 1% yeast extract (produced by Oriental Yeast Co., Ltd.) and 1% glucose. In a culture obtained by inoculating 100 mL each of two 7000 mL jar fermenters containing 5000 mL) and stirring and culturing at 27 ° C., aeration rate 0.2 MMV, and stirring 200 rpm for 10 days. In addition, if an actinoallide A, B, C, D and / or E substance is present, the microorganism has a release capacity having the ability to produce the actinoallide A, B, C, D and / or E substance. It can be determined that it is a microorganism belonging to the filamentous fungus. Preferably, the microorganism belonging to actinomycetes having the ability to produce actinoallide A, B, C, D and / or E substance is the above-mentioned Actinoallomurus sp. MK10-036 strain .

  As used herein, the term “mutant strain” refers to a strain having a mycological property or gene different from that of Actinoallomulus sp. MK10-036 by artificial or natural mutagenesis stimulation. In addition to the strains derived from the Actinoallomurus sp. MK10-036 strain, such mutants are derived from the Actinoallomurus spi (Actinoallomurus sp.) MK10-036 strain. The original strain is also included. In the present specification, the mutant strain does not ask for the presence or absence of actual derivation. For example, the mutant strain is highly homologous to the Actinoallomulus sp. MK10-036 strain gene (for example, 16S rRNA gene). A strain having a gene having sex (for example, 80% or more, 85% or more, 90% or more, 95% or more, etc.) is also included in the mutant strain. In addition, such mutant strains are artificially produced or collected from nature as long as the ability to produce actinoallolides A, B, C, D and / or E is maintained. It doesn't matter if there is.

  A medium for cultivating microorganisms belonging to actinomycetes having the ability to produce actinoallolides A, B, C, D and / or E should be used as a nutrient source and as a nutrient source for actinomycetes Can be contained. For example, commercially available peptone, meat extract, cone steep liquor, cottonseed flour, peanut flour, soy flour, yeast extract, NZ-amine, casein hydrate, soda nitrate, ammonium nitrate, Nitrogen sources such as ammonium sulfate, carbohydrates such as glycerin, starch, glucose, galactose, mannose, or carbon sources such as fat, and inorganic salts such as sodium chloride, phosphate, calcium carbonate, magnesium sulfate Can be used alone or in combination. In addition, if necessary, a trace amount of a metal salt, a moving / planting / mineral oil or the like as an antifoaming agent can be added to the medium. Any of these may be used for the production of actinoallolides A, B, C, D and / or E using the producing bacteria, and all known actinomycete culture materials can be used.

  In addition, the cultivation of microorganisms belonging to actinomycetes having the ability to produce actinoallolides A, B, C, D and / or E substances can be obtained by growing the producing bacteria and actinoallolides A, B, C, D and / or It can be carried out by shaking culture for several days to 2 weeks in a temperature range in which substance E can be produced (for example, 10-40 ° C., preferably 25-30 ° C.). The culture conditions can be appropriately selected and performed according to the properties of the bacterium producing actinoallide A, B, C, D and / or E used with reference to the description of the present specification.

  The actinoallide A, B, C, D and / or E substance can be collected by extraction from the culture solution using a water-immiscible organic solvent such as ethyl acetate. In addition to this extraction method, known methods used for collecting fat-soluble substances such as adsorption chromatography, distribution chromatography, gel filtration chromatography, thin layer chromatography, centrifugal countercurrent distribution chromatography, high performance liquid It can purify | clean until it becomes pure by combining chromatography etc. suitably or repeating.

  The therapeutic agent or prophylactic agent for diseases caused by infection with Trypanosoma protozoa of the present invention can be used in oral administration forms or parenteral administration forms such as injections and infusions. When this compound is administered to mammals or the like, it may be administered orally as tablets, powders, granules, syrups, etc., or may be administered parenterally as injections or drops. Thus, the present invention, in another aspect, involves infection with Trypanosoma protozoa, comprising administering to a patient in need thereof an effective amount of the actinoallolide A, B, C, D and / or E substance. The present invention relates to a method for treating or preventing a disease derived from the disease. The dose of Actinoallolide A, B, C, D and / or E substance varies depending on the degree of symptoms, age, type of disease, etc., but usually 50-500 mg per adult is divided into 1 to several times a day Administer.

  The pharmaceutical composition of the present invention can be formulated by a conventional method using a normal pharmaceutically acceptable carrier. When preparing a solid preparation for oral administration, add excipients to the active ingredient and, if necessary, binders, disintegrants, lubricants, etc., and then add solvents, granules, powders, capsules, etc. by conventional methods. And When preparing an injection, a pH adjuster, a buffer, a stabilizer, a solubilizing agent, etc. may be added to the main drug as necessary to obtain a subcutaneous or intravenous injection by a conventional method.

  Hereinafter, the present invention will be described in detail with reference to examples, but the present invention is not limited thereto. It should be noted that all documents cited throughout this application are incorporated herein by reference in their entirety.

(Example 1) Mycological properties of Actinoallomurus sp. MK10-036 strain A new strain of Actinoallomurus sp. MK10 was newly obtained by the present inventors from plants in the Kingdom of Thailand. -036 strain was isolated. The mycological characteristics of Actinoallomulus sp. MK10-036 strain were as follows.
(I) Morphological properties This strain grows well on starch / inorganic salt agar, yeast extract / malt extract agar, yeast extract / starch agar, etc. Life was seen. The spores were about 1 μm × 0.5 μm in size, and 6-8 spores were connected from the aerial hyphae to form a short helical spore chain.
(II) Properties on various media: EB Shirring and D. Gottlieb's method (International Journal of Table 1 shows the culture properties of the production bacteria examined by Cistimatic Bacteriology, 16, 313, 1966). The color tone is determined by using the Color Harmony Manual 4th edition (Container-Corporation of America Chicago, 1958) as a standard color, and is enclosed in parentheses with the color chart name. The code is also described. The following are the results of observation in each medium at 27 ° C. for 3 weeks unless otherwise specified.

(III) Physiological properties (1) Formation of melanin pigment (a) Tyrosine agar: Negative (b) Glucose-peptone gelatin medium: Negative (2) Reduction of nitrate: Positive (3) Liquefaction of gelatin (20 ° C): Negative (glucose, peptone, gelatin medium)
(4) Hydrolysis of starch: Negative (5) Coagulation of skim milk (37 ° C): Negative (6) Peptonization of skim milk (37 ° C): Negative (7) Growth temperature range: 19-40 ° C
(8) Availability of carbon source (Prideham Gotribe agar)
Use: D-glucose, D-xylos, D-mannitol, D-fructose, raffinose
L-rhamnose, myo-inositol, melibiose, D-galactose Not used: L-arabinose, dulcitol, maltose, D-sorbitol, sucrose (9) Cellulose degradation: Negative

(IV) Cell chemical composition The cell wall contains meso-type diaminopimelic acid, alanine, glutamic acid, and lysine, and majurose as the total microbial sugar. The main menaquinones are MK-9 (H ₆ ) and MK-9 (H ₈ ), and the main fatty acids are iso-C16: 0,10-methylC17: 0, C16: 1 and anteiso-17: 0. Cell wall muramic acid is acetyl type. Mycolic acid is not included.
(V) Analysis of 16S rRNA gene The sequence of about 1400 bases of 16S rRNA gene was determined, and data of strains belonging to the genus Actinoalumulus and other actinomycetes registered and published in the DNA database As a result of phylogenetic analysis using the neighbor binding method, it is appropriate to classify this strain into the genus Actinoalomulus, which is closely related to Actinoallomurus fulvus and Actinoallomurus amamiensis. It is a strain.

(VI) Conclusion As described above, the bacteriological properties of this bacterium are summarized as follows. It has meso-type diaminopimelic acid and lysine on the cell wall. Whole cell sugar contains majurose. Cell wall muramic acid is an acetyl type, and main menaquinones are MK-9 (H ₆ ) and MK-9 (H ₈ ). Contains no mycolic acid. Forms a helical spore chain shorter than aerial hyphae. The colony has a white color and does not produce a melanin pigment.
From these results and the analysis result of the 16S rRNA gene, it was determined that it was one bacterial species belonging to the genus Actinoallomulus. This strain was deposited as Actinoallomurus sp. MK10-036 on January 24, 2012 at the National Institute of Technology and Evaluation, Japan (Receipt No. NITE ABP-). 1208).

(Example 2) Acquisition of actinoallolides A, B, C, D and E substances Yeast extract (manufactured by Oriental Yeast Co., Ltd.) 1%, liquid medium (pH 6.0) consisting of glucose 1% 100 mL Inoculate 1 ml each of two Actinoallomurus sp. MK10-036 (reception number NITE ABP-1208) cultured in a liquid medium in a 500 mL Erlenmeyer flask containing 27 mg at 27 ° C. for 7 days. Cultured with shaking. The obtained seed culture solution was 7,000 mL of Jafarmenta containing 5,000 mL of a liquid medium (pH 6.0) consisting of 1% yeast extract (produced by Oriental Yeast Co., Ltd.) and 1% glucose. -100 mL of each was inoculated into 2 groups, and stirred and cultured for 10 days at 27 ° C, aeration rate of 0.2 MMV, and stirring at 200 rpm.

  10 L of the culture solution was centrifuged at 3000 rpm for 15 minutes, and separated into a supernatant and cells. The supernatant was extracted twice with 5 L of ethyl acetate, and the ethyl acetate layer was dried under reduced pressure to obtain 576 mg of crude substance 1. On the other hand, 2 L of acetone was added to the cells and stirred for 1 hour to extract the components of the cells, followed by centrifugation at 3000 rpm for 15 minutes. Acetone was distilled off from the obtained supernatant, followed by extraction twice with 1 L of ethyl acetate, and the ethyl acetate layer was dried under reduced pressure to obtain 170 mg of crude substance 2. The crude substances 1 and 2 were mixed and loaded onto a silica gel column (φ40 × 45 mm) packed with chloroform, eluted with chloroform-methanol (100: 1) and (100: 2), respectively, and the crude substance 3 ( 205 mg) and crude material 4 (161 mg).

  Crude substance 3 was dissolved in methanol and injected into a reverse phase column (Pegasil ODS SP-100, φ20 × 250 mm, manufactured by Senshu Scientific Co., Ltd., Japan) by high performance liquid chromatography, and acetonitrile-water ( 60:40), flow rate of 8 mL / min, and UV 210 nm detection. Peaks having retention times of 17.7 minutes, 26.8 minutes, 30.7 minutes, and 33.6 minutes were collected and dried under reduced pressure to obtain actinoallide E substance (1.8 mg), actinoallo. Ride A substance (81.7 mg), actino allolide D substance (2.7 mg) and actino allolide C substance (4.4 mg) were obtained.

Crude substance 4 was dissolved in methanol, and ODS (φ15 × 20 mm, manufactured by Fuji Silysia Chemical Co., Ltd., Japan) open column chromatography was used for step elution (60:40 in an acetonitrile-water solvent system). , 70:30, 80:20, 90:10, 100: 0), and acetonitrile-water (60:40) was dried under reduced pressure to obtain a crude substance 5. The crude material 5 was injected into a reverse phase column (Pegasil ODS SP-100, φ20 × 250 mm, manufactured by Senshu-Science) by high performance liquid chromatography, acetonitrile-water (60:40), flow rate 8 mL / min, Elution was performed under the condition of UV 210 nm detection. A peak having a retention time of 24.5 minutes was collected and dried under reduced pressure to obtain Actinoallolide B substance (5.9 mg).
The physicochemical properties of the obtained actinoallide A, B, C, D and E substances were measured.

The physicochemical properties of the actinoalloride A substance were as follows.
(1) Property: syrup shape (2) Molecular weight: 564
(3) Molecular formula: C ₃₂ H ₅₂ O ₈
(4) [M + Na] ⁺ theoretical value (m / z) 587.3560, measured value (m / z) 587.3570 by fast atom collision ionization
(5) Specific rotation: [α] _D ²⁵ = 105.2 ° (c = 0.1, methanol)
(6) Ultraviolet absorption maximum λ (in methanol): terminal absorption (7) Infrared absorption maximum ν _max (KBr tablet): 3435, 2968, 2927, 2864, 1749, 1709, 1630, 1458, 1371, 1317 , 1147, 1115, 1057, 1022, 978 cm ⁻¹ .
(8) ¹ H NMR (in deuterated chloroform) δ ppm: 5.40 (1H, dt), 5.13 (1H, t), 4.88 (1H, d), 3.64 (1H, dd), 3.26 (1H, dd), 2.91 (1H, d), 2.83 (1H, dd), 2.77 (1H, d), 2.67 (1H, dq), 2.49 (1H M), 2.47 (1H, m), 2.45 (1H, m), 2.35 (1H, d), 2.34 (1H, m), 2.30 (1H, d), 2 .13 (1H, d), 1.84 (1H, m), 1.81 (1H, m), 1.79 (1H, d), 1.78 (1H, m), 1.63 (3H, brd), 1.61 (1H, m), 1.47 (3H, s), 1.34 (3H, s), 1.14 (3H, t), 1.07 (3H, d),. 05 (3H, d), 1.04 ( H, t), 1.01 (3H, d), 0.88 (3H, d)
(9) ¹³ C NMR (in deuterated chloroform) δ ppm: 217.6, 217.3, 170.2, 133.7, 131.7, 128.8, 126.5, 102.3, 82.2, 76.5, 74.9, 73.6, 53.1, 48.9, 47.5, 46.7, 40.5, 38.8, 36.0, 34.8, 32.2, 29. 9, 26.8, 18.4, 17.9, 17.2, 16.9, 16.1, 12.1, 10.3, 9.4, 7.7
(10) Solubility in solvents: Easily soluble in chloroform, acetone, methanol and dimethyl sulfoxide. Normal hexane, hardly soluble in water.

  As a result of studying various physicochemical properties and spectral data of the actinoalloride A substance, it was determined that the actinoalloride A substance has a structure represented by the following formula I.

The physicochemical properties of the actinoalloride B substance were as follows.
(1) Property: syrup shape (2) Molecular weight: 566
(3) Molecular formula: C ₃₂ H ₅₄ O ₈
(4) [M + Na] ⁺ theoretical value (m / z) 589.3716, measured value (m / z) 589.3713 by erocropre-pre-ionization
(5) Specific rotation: [α] _D ²⁵ = 102.7 ° (c = 0.1, methanol)
(6) Ultraviolet absorption maximum λ (in methanol): terminal absorption (7) Infrared absorption maximum ν _max (KBr tablet): 3437, 2972, 1751, 1712, 1628, 1454, 1379, 1319, 1248, 1149 , 1109, 1059, 974 cm ⁻¹ .
(8) ¹ H NMR (in deuterated chloroform) δ ppm: 5.38 (1H, dt), 5.13 (1H, t), 4.88 (1H, d), 3.70 (1H, ddd), 3.51 (1H, dd), 3.26 (1H, dd), 2.91 (1H, d), 2.82 (1H, dd), 2.75 (1H, d), 2.52 (1H , M), 2.52 (1H, m), 2.32 (2H, m), 2.32 (1H, m), 2.12 (1H, d), 1.82 (1H, m), 1 .80 (1H, m), 1.78 (1H, m), 1.77 (1H, d), 1.65 (1H, m), 1.62 (3H, brd), 1.59 (1H, m), 1.52 (1H, m), 1.47 (3H, brd), 1.42 (1H, m), 1.34 (3H, s), 1.13 (3H, t), 1. 03 (3H, d), 1. 0 (3H, d), 0.90 (3H, t), 0.88 (3H, d), 0.84 (3H, d)
(9) ¹³ C NMR (in deuterated chloroform) δ ppm: 217.8, 170.1, 133.2, 131.7, 129.1, 126.5, 102.3, 82.1, 82.0, 79.3, 76.6, 73.6, 53.3, 48.9, 46.6, 40.6, 38.7, 37.8, 36.8, 32.4, 29.8, 28. 1, 26.8, 18.4, 17.9, 17.2, 17.0, 16.2, 12.1, 10.4, 10.3, 4.2
(10) Solubility in solvents: Easily soluble in chloroform, acetone, methanol and dimethyl sulfoxide. Normal hexane, hardly soluble in water.

  As a result of studying various physicochemical properties and spectral data of the actinoallride B substance, it was determined that the actinoallide B substance has a structure represented by the following formula II.

The physicochemical properties of the actinoalloride C material were as follows.
(1) Property: syrup shape (2) Molecular weight: 546
(3) Molecular formula: C ₃₂ H ₅₀ O ₇
(4) [M + Na] ⁺ theoretical value (m / z) 569.3454, measured value (m / z) 569.3444 by electrospray ionization
(5) Specific rotation: [α] _D ²⁵ = 190.0 ° (c = 0.1, methanol)
(6) Ultraviolet absorption maximum λ (in methanol): 278.5 nm (ε value: 9,773)
(7) Infrared absorption maximum ν _max (KBr tablet): Maximum absorption at 3437, 2972, 2927, 1726, 1697, 1620, 1454, 1392, 1277, 1213, 1111, 1057, 984 cm ⁻¹ .
(8) ¹ H NMR (in deuterated chloroform) δ ppm: 5.27 (1H, dt), 5.16 (1H, t), 4.88 (1H, d), 3.64 (1H, dd), 3.63 (1H, d), 3.31 (1H, d), 3.27 (1H, dd), 2.67 (1H, dq), 2.48 (1H, m), 2.46 (2H) M), 2.45 (2H, d), 2.44 (1H, m), 2.30 (1H, m), 2.19 (2H, q), 2.13 (1H, d), 1 .82 (1H, m), 1.80 (1H, m), 1.79 (1H, m), 1.63 (3H, brd), 1.43 (3H, s), 1.41 (3H, s), 1.07 (3H, d), 1.05 (3H, d), 1.04 (3H, t), 1.04 (3H, t), 1.00 (3H, d), 0. 89 (3H, d)
(9) ¹³ C NMR (in deuterated chloroform) δ ppm: 217.2, 206.4, 177.4, 168.3, 133.7, 131.0, 128.8, 126.2, 119.0, 87.6, 76.6, 76.0, 74.9, 47.9, 47.5, 41.1, 38.9, 37.5, 35.9, 34.7, 32.2, 29. 9, 22.2, 17.9, 17.0, 17.0, 16.1, 14.9, 12.3, 10.3, 9.4, 7.7
(10) Solubility in solvents: Easily soluble in chloroform, acetone, methanol and dimethyl sulfoxide. Normal hexane, hardly soluble in water.

  As a result of studying various physicochemical properties and spectral data of the actinoallolide C substance, it was determined that the actinoallolide C substance has a structure represented by the following formula III.

The physicochemical properties of the actinoalloride D substance were as follows.
(1) Property: syrup shape (2) Molecular weight: 548
(3) Molecular formula: C ₃₂ H ₅₂ O ₇
(4) [M + H] by fast atom collision ionization ⁺ theoretical value (m / z) 549.3791, actually measured value (m / z) 549.3792
(5) Specific rotation: [α] _D ²⁵ = 167.7 ° (c = 0.1, methanol)
(6) Ultraviolet absorption maximum λ (in methanol): 278.5 nm (ε value: 9,316)
(7) Infrared absorption maximum ν _max (KBr tablet): maximum absorption at 3438, 2972, 2875, 1726, 1689, 1620, 1454, 1392, 1279, 1213, 1057, 968 cm ⁻¹ .
(8) ¹ H NMR (in deuterated chloroform) δ ppm: 5.27 (1H, dt), 5.17 (1H, t), 4.88 (1H, d), 3.70 (1H, dt), 3.63 (1H, d), 3.52 (1H, dd), 3.31 (1H, d), 3.27 (1H, dd), 2.50 (1H, m), 2.48 (1H M), 2.45 (2H, d), 2.28 (1H, q), 2.28 (1H, m), 2.19 (1H, q), 2.11 (2H, d), 1 .82 (1H, m), 1.80 (1H, m), 1.78 (1H, m), 1.66 (1H, m), 1.63 (3H, brd), 1.50 (1H, m) m), 1.43 (3H, s), 1.41 (3H, s), 1.40 (1H, m), 1.05 (3H, t), 1.04 (3H, d), 1. 00 (3H, d), 0.90 (3 , T), 0.89 (3H, d), 0.84 (1H, d)
(9) ¹³ C NMR (in deuterated chloroform) δ ppm: 206.4, 177.4, 168.3, 133.2, 130.9, 129.1, 126.2, 119.0, 87.6, 82.0, 79.3, 76.9, 76.0, 47.9, 41.1, 38.8, 37.8, 37.5, 36.8, 32.3, 29.9, 28. 1,22.2, 17.9, 17.1, 17.0, 16.2, 14.9, 12.3, 10.4, 10.4, 4.2
(10) Solubility in solvents: Easily soluble in chloroform, acetone, methanol and dimethyl sulfoxide. Normal hexane, hardly soluble in water.

  As a result of studying various physicochemical properties and spectral data of the actinoallide D substance, it was determined that the actinoallide D substance has a structure represented by the following formula IV.

アクチノアロライドＤ物質

Actino allolide D substance

The physicochemical properties of the actinoalloride E substance were as follows.
(1) Property: syrup shape (2) Molecular weight: 546
(3) Molecular formula: C ₃₂ H ₅₀ O ₇
(4) [M + H] ⁺ theoretical value (m / z) 547.3635, measured value (m / z) 547.33641 by fast atom collision ionization
(5) Specific rotation: [α] _D ²⁵ = 120.1 ° (c = 0.1, methanol)
(6) Ultraviolet absorption maximum λ (in methanol): 274.5 nm (ε value: 8,627)
(7) Infrared absorption maximum ν _max (KBr tablet): Maximum absorption at 3437, 2974, 2939, 2879, 1730, 1697, 1626, 1454, 1390, 1255, 1099, 1057, 987 cm ⁻¹ .
(8) ¹ H NMR (in deuterated chloroform) δ ppm: 5.22 (1H, t), 4.97 (1H, dd), 4.81 (1H, d), 3.82 (1H, d), 3.64 (1H, dd), 3.26 (1H, dd), 3.26 (1H, d), 2.66 (1H, dq), 2.64 (1H, m), 2.62 (1H D), 2.54 (1H, m), 2.49 (1H, m), 2.47 (1H, d), 2.46 (1H, m), 2.30 (1H, m), 2 .23 (1H, m), 2.22 (1H, m), 2.12 (1H, dd), 1.96 (1H, m), 1.91 (1H, m), 1.68 (1H, dd), 1.62 (3H, brd), 1.53 (1H, m), 1.53 (3H, s), 1.36 (3H, s), 1.11 (3H, t), 1. 09 (3H, d), 1.06 3H, t), 1.04 (3H, d), 0.94 (3H, d), 0.81 (3H, d)
(9) ¹³ C NMR (in deuterated chloroform) δ ppm: 217.3, 207.0, 177.6, 166.7, 132.9, 130.7, 129.4, 126.6, 117.6, 88.2, 81.4, 74.8, 72.7, 47.5, 46.4, 40.2, 39.4, 35.9, 35.8, 35.2, 34.8, 31. 6, 24.9, 17.9, 17.7, 16.5, 15.9, 15.6, 13.1, 9.5, 8.1, 7.7
(10) Solubility in solvents: Easily soluble in chloroform, acetone, methanol and dimethyl sulfoxide. Normal hexane, hardly soluble in water.

  As a result of studying various physicochemical properties and spectral data of the actinoallride E substance, it was determined that the actinoallide E substance has a structure represented by the following formula V.

(Example 3) In vitro Trypanosoma protozoa growth inhibitory activity of substances Actinoallide A, B, C, D and E (Tb brucei GUTat 3.1 strain)
The in vitro T. of the actinoallolides A, B, C, D and E of the present invention. b. Brucei GUTat 3.1 strain The activity of inhibiting the growth of Trypanosoma protozoa was examined as follows.
As the test protozoa, Trypanosoma protozoa caused by Nagana disease b. Brucei GUTat 3.1 strain (available from Lecturer Yoshiyoshi Sada, Nagoya City University School of Medicine) was used. The maintenance passaging of the protozoa is carried out according to the method of Yabu Y, Koide T, Ohota N, Nose M, Ogihara Y. et al. Continuous growth of bloodstream forms of Trypanosoma brucei brucei in axious culture system concentrating of the consortium. Southeast Asian J. et al. Top. Med. Public Health, 29: 591-595 (1998)] was slightly modified. That is, in each well of a 24-well plate, 10% non-immobilized fetal bovine serum (FBS), Iskov modified Dulbecco (IMDM) medium supplemented with antibiotics and various supplements were used at 37 ° C., 5% CO _2. Cultivation was performed under −95% air, and continuous culture was performed by changing the medium every 1 to 3 days.

In vitro measurement of the anti-trypanosoma protozoan activity of this compound was carried out by the method of Otoguro et al. [Otoguro K, Ishiyama A, Namata M, Nishihara A, Furawa O T, Masuma R, Takahashi H S. Selective and Potent in vitro antipanosomal activities of ten micrometabolites. J. et al. Antibiotics, 61: 372-378 (2008)]. That is, 95 mL of a protozoan suspension preliminarily cultured in each well of a 96-well plate (adjusted to a protozoa number of 2.0-2.5 × 10 ⁴ / mL) and 5 mL of a compound solution (50% ethanol aqueous solution) Was added and mixed, followed by culturing at 37 ° C. and 5% CO ₂ -95% air for 72 hours.

After completion of the culture, protozoan growth was measured by adding 10 μL of Alamar Blue reagent (Sigma-Aldorich, USA) to each well of the 96-well plate and mixing at 37 ° C. under 5% CO ₂ -95% air. After culturing for 3-6 hours, the redox potential of the protozoa is measured with a fluorescence microplate reader (Bio-Tek, USA) at an excitation wavelength of 528/20 nm and a fluorescence wavelength of 590/35 nm. Thus, the presence or absence of protozoan proliferation was colorimetrically determined. The 50% protozoan growth inhibitory concentration (IC ₅₀ value) of this compound was determined from the compound concentration action curve of KC-4 (manufactured by Bio-Tek, USA) of the software attached to the fluorescence microplate reader.

  For comparison, known antitrypanosoma protozoan drugs whose effects on cultured Trypanosoma protozoa were measured were pentamidine, suramin, and efronitin [provided from (Prof. R. Brun, Swiss Tropical Institute, Basel, Switzerland)] Using.

  Antitrypanosoma protozoan activity against cultured Trypanosoma protozoa (Tb brucei GUtat3.1 strain) of the substances actinoallide A, B, C, D and E of the present invention and known antitrypanosoma protozoa It was as shown in Table 2 below.

The Actinoallolides A, B, C, D and E substances of the present invention are described in T. b. Antitrypanosoma protozoan activity (IC ₅₀ values) against Brucei GUTat 3.1 strain is 0.0049, 1.01, 0.11, 0.77 and 0.13 μg / mL, respectively. It showed the best antitrypanosoma protozoan activity. The actinoalloride A substance had 322 and 463 times stronger antitrypanosoma protozoan activity than suramin and efronitine, comparable to pentamidine, when compared to existing antitrypanosome protozoan drugs. The actinoallide B, C, D and E substances were 2-14 times stronger antitrypanosoma protozoan activity than suramin and efronitine when compared to existing antitrypanosoma protozoan drugs.

Example 4 In Vitro Trypanosoma protozoa Growth Inhibitory Activity of Actinoalloride A Substance (Tb rhodosense STIB900 Strain)
The in vitro T. elegans of the actinoalloride A material of the present invention. b. The activity of inhibiting the growth of rhodenense STIB900 strain Trypanosoma protozoa was examined as follows.
As a test protozoa, protozoa caused by Rhodesia African sleeping sickness b. A tripomastigote body of Rhodensense STIB900 strain (distributed from (Prof. R. Brun, Swiss Tropical Institute, Basel, Switzerland)) was used. Within each well of a 24-well plate, add 15% non-immobilized fetal bovine serum (FBS), 25 mM N-2-hydroxyethylpiperazine-2-ethanesulfonic acid (HEPES), 0.1% (w / W) Glucose, 1% MEM non-essential amino acid, 0.2 mM 2-mercaptoethanol, 2 mM sodium pyruvate, 0.1 mM hypoxanthine, 37 ° C., 5% CO ₂ -95 Culturing was performed under% air, and continuous culture was performed by changing the medium every few days.

In vitro measurement of the anti-trypanosoma protozoan activity of this compound was performed by measuring the protozoa suspension (final protozoa number 8 × 10 ³ / μL) pre-cultured in each well of a 96-well plate at a predetermined concentration. The compound solution (DMSO solution) was added to a final volume of 100 μL, mixed, and cultured at 37 ° C. under 5% CO ₂ -95% air for 72 hours.

After completion of the culture, protozoan growth was measured by adding 10 μL of Alamar Blue reagent (Sigma-Aldorich, USA) to each well of the 96-well plate and mixing at 37 ° C. under 5% CO ₂ -95% air. After culturing for 2 hours, the redox potential of the protozoa is measured with a fluorescence microplate reader (Bio-Tek, USA) at the excitation wavelength of 528/20 nm and the fluorescence wavelength of 590/35 nm. The colorimetric determination of the presence or absence of protozoan proliferation was performed. The 50% protozoan growth inhibitory concentration (IC ₅₀ value) of this compound was determined from the compound concentration action curve of KC-4 (manufactured by Bio-Tek, USA) of the software attached to the fluorescence microplate reader.

  As a comparison, a known antitrypanosoma protozoan drug whose effect on cultured Trypanosoma protozoa was measured was used as melalsoprole (distributed from (Prof. R. Brun, Swiss Tropical Institute, Basel, Switzerland)]. .

  The antitrypanosoma protozoan activity of the actinoalloride A substance of the present invention and a known antitrypanosoma protozoan drug against cultured trypanosoma protozoa (Tb rhodensense STIB900 strain) was as shown in Table 3 below. .

The actinoalloride A substance of the present invention is described in T.W. b. The antitrypanosoma protozoa activity (IC ₅₀ value) against rhosense STIB900 strain was 0.086 μg / mL, indicating excellent antitrypanosoma protozoa activity. The actinoalloride A substance had 1/43 times the antitrypanosoma protozoan activity of meralsoprole compared to the existing antitrypanosoma protozoan drugs. From this result, it is expected that the actinoalloride A substance will lead to drug development for Rhodesia type African sleeping sickness.

(Example 5) In vitro activity of actinoballide A substance to inhibit growth of American trypanoso-ma protozoa (T. cruzi Tulahuen C4C8 strain)
The in vitro T. elegans of the actinoalloride A material of the present invention. The activity of inhibiting the growth of cruzi Tulahuen C4C8 strain Trypanosoma protozoa was examined as follows.
As a test protozoa, the protozoan caused by Chagas disease Amastigotes and tripomastigotes infected with rat L6 cells of cruzi Tulahuen C4C8 strain (provided from (Prof. R. Brun, Swiss Tropical Institute, Basel, Switzerland)) were used. In each well of a 24-well plate, a medium prepared by adding 10% non-immobilized fetal bovine serum (FBS) and 1% L-glutamine to RPMI1640 medium containing L6 cells was used at 37 ° C., 5% CO _2. Cultivation was performed under −95% air, and continuous culture was performed by changing the medium every few days.
In vitro measurement of the anti-trypanosoma protozoan activity of this compound was performed by adding a protozoa suspension (final protozoa number of 5 × 10 ³ cells / well) to each well of a 96-well plate and then culturing for 48 hours. went. After exchanging the medium, a compound solution (DMSO solution) having a predetermined concentration was added, mixed, and cultured for 96 hours at 37 ° C. under 5% CO ₂ -95% air.
After completion of the culture, protozoan growth was measured by adding 50 μL of CPRG / Nonidet to each well of a 96-well plate. Further, after standing for 2-6 hours, the absorbance at 540 nm was measured with an absorption microplate reader (manufactured by Labosystems, Finland), and the 50% protozoan growth inhibitory concentration of this compound was determined from the compound concentration action curve. It was.
As a comparison object, benznidazole was used as a known Chagas disease therapeutic agent whose effect on cultured Trypanosoma protozoa was measured.

  The antitrypanosoma protozoan activity of the Actinoallolide A substance of the present invention and known antitrypanosomes protozoa against cultured Trypanosoma protozoa (T. cruzi Tulahuen C4C8 strain) was as shown in Table 4 below.

The actinoalloride A substance of the present invention is described in T.W. The antitrypanosoma protozoan activity (IC ₅₀ value) against cruzi Tulahuen C4C8 strain was 0.226 μg / mL, indicating excellent antitrypanosoma protozoa activity. The actinoalloride A substance had an antitrypanosoma protozoan activity that was approximately twice as good as benznidazole compared to existing Chagas disease drugs. From this result, it is expected that the actinoalloride A substance will lead to drug development for Chagas disease.

(Example 6) Cytotoxicity test The cytotoxicity test of the actinoallolides A, B, C, D and E of the present invention was carried out by the method of Otoguro et al. [Otoguro K, Kohana A, Manabe C, Ishiyama A, Ui H, Shiomi. K, Yamada H, Omura S. et al. Potential axial activities of polymer antibiotic, X-206. J. et al. Antibiotics, 54: 658-663, (2001)]. That is, human fetal lung-derived normal fibroblast MRC-5 cells [Dr. L. Maes (available from Tibotec NV, Mechelen, Belgium)] was maintained and subcultured in 10% fetal calf serum (FCS) and MEM medium supplemented with antibiotics.

Adjust the suspension of human fetal lung-derived normal fibroblast MRC-5 cells to 1 × 10 ³ cells / mL with 10% FCS-MEM, and add 100 μL of the suspension to a 96-well plate. After mixing, the cells were cultured for 24 hours at 37 ° C. and 5% CO ₂ -95% air. Thereafter, 90 μL of 10% FCS-MEM and 10 μL of the solution of this compound (50% ethanol aqueous solution) were added to each well. After mixing, the cells were cultured at 37 ° C., 5% CO ₂ -95% air for 7 days. It was. The presence or absence of proliferation of MRC-5 cells was measured by colorimetric determination by the MTT method. The 50% cell growth inhibitory concentration (IC ₅₀ value) of this compound was determined from the compound concentration action curve. Furthermore, selective toxicity ratio (Selectivity Index: SI) is, _{(an IC 50} value of cytotoxicity) / - was calculated by (anti Toripanoso _{an IC 50} value of Ma protozoal activity).

The calculated results for IC ₅₀ and selective toxicity ratio are shown in Table 5 below.

The cytotoxicity (IC ₅₀ value) of the actinoallolides A, B, C, D and E of the present invention against human fetal lung-derived normal fibroblast MRC-5 cells was> 100, 51.83 and 32.44, respectively. , 16.45 and 4.71 μg / mL, and the selective toxicity ratio (Selectivity Index: SI) with antitrypanosoma protozoan activity (Tbb GUAT3.1 strain) is> 20,408, 51.3, 295, 21.4 and 36.2. Among these, the SI of the actinoalloride A substance showed a selective toxicity> 5.4 times that of pentamidine when compared with the existing antitrypanosoma protozoan drugs. In addition, the activity of Actinoallolide A against other Trypanosoma protozoa (Tbr STIB900 strain and T. Cruzi Tulahuen C4C8 strain) showed excellent selective toxicity of> 1163 and> 442. .

(Example 7) Antibacterial activity The antibacterial activity of the Actinoallolide A substance of the present invention was measured by the following method. 10 μL each of 1 mg / mL methanol solution of actinoallolide A substance was immersed in a filter paper disk (Advantech Co., Ltd., diameter 6 mm), air-dried for a certain period of time, and the solvent was removed. After sticking on a fungal agar plate and culturing at 37 ° C. or 27 ° C. for 24 hours, the diameter of the growth inhibition circle formed around the filter paper disc was measured.

  Table 6 shows the diameter of the growth inhibition circle measured as an antibacterial activity of the actinoalloride A substance against the test bacteria.

＊ −：生育阻止円が観測されない

*-: Growth inhibition circle is not observed

  The actinoalloride A material of the present invention showed little antimicrobial activity against the microorganisms listed in Table 7. Therefore, it can be considered that the actinoalloride A substance of the present invention has a mechanism of action specific to Trypanosoma protozoa.

  From the above results, the actinoallide A, B, C, D and E substances of the present invention have the effect of inhibiting the growth of Trypanosoma protozoa, but are low in cytotoxicity and proliferate against other microorganisms. Since it does not show an inhibitory action, it has been shown to have a very excellent effect as an antitrypanosoma protozoan drug.

Claims (14)

A compound represented by the following formula I:

A compound represented by the following formula II.

A compound represented by the following formula III.

The compound represented by the following formula IV.

A compound represented by the following formula V:

A microorganism belonging to Actinoalumulus actinomycetes having the ability to produce the compound according to any one of claims 1 to 5 is cultured in a medium, and the culture according to any one of claims 1 to 5 The compound according to any one of claims 1 to 5, wherein the compound according to any one of claims 1 to 5 is collected from the culture by accumulating the compound according to any one of claims 1 to 5. Method.   A microorganism belonging to actinomycetes having the ability to produce the compound according to any one of claims 1 to 5, wherein Actinoalulomurus sp. MK10-036 (reception number NITE ABP-1208) strain The manufacturing method according to claim 6.   Actinoallomulus sp. MK10-036 (reception number NITE ABP-1208) strain.   An agent for inhibiting the growth of Trypanosoma protozoa comprising the compound according to any one of claims 1 to 5 as an active ingredient.   A therapeutic or prophylactic agent for a disease derived from infection with Trypanosoma protozoa, comprising the compound according to any one of claims 1 to 5 as an active ingredient.   The therapeutic or prophylactic agent according to claim 10, wherein the Trypanosoma protozoa is a human infectious Trypanosoma protozoa.   Human infectious trypanosomes are Gambia trypanosomes, Rhodesia trypanosomes and subgenus Trypanosoma cruzi, Trypanosoma brucei, Trypanoso T. concolense, Trypanosoma baivacs (T. vivax), Trypanosoma evansi (T. evansi), Trypanosoma tyerieri (T. heileri) or Trypanosoma Exadam The therapeutic or prophylactic agent according to claim 11, which is (T. equiperdum).   The therapeutic or prophylactic agent according to claim 10, wherein the Trypanosoma protozoa is a livestock infectious Trypanosoma protozoa.   Liver infectious trypanosomatid protozoa are brucei brucei (Nagana disease-causing protozoa), T. vivax vivax (Zuma disease-causing protozoa), T. Evansi (protozoan causing Sura disease), or T. The therapeutic or prophylactic agent according to claim 13, which is an equiperdum (protozoan causing pesticidal disease).