JPH06345768A - New milbemycin compound - Google Patents

Abstract

PURPOSE:To obtain the subject compound having excellent miticidal, insecticidal or anthelmintic activity against mites, plant vermin and animal parasites by culturing a microorganism belonging to the genus Streptomyces. CONSTITUTION:The objective compound of formula (R is methyl or ethyl; R<1> is methylene when the bond containing the dotted line between 4/26 sites is double bond or R<1> is methyl when the bond is single bond) is produced by culturing a microorganism belonging to the genus Streptomyces (FERM BP-1072) by conventional method for the culture of microorganism of the genus Streptomyces.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a novel milbemycin compound having excellent insecticidal, acaricidal or anthelmintic activity, a novel milbemycin compound useful as a synthetic intermediate, and a method for producing the milbemycin compound.

[0002]

2. Description of the Related Art B-41-of Streptomyces microorganisms
A group of 16-membered macrolide antibiotics isolated from 146 strains is disclosed in Japanese Patent Application Laid-Open No. 50-29742, B-41.
It _{called, A 1, A 2, A} 3, A 4, B 1, B 2, B
Nine kinds of C ₃ , C _{1 and} C ₂ were mentioned. Four more compounds have been isolated and the structures of 13 compounds have been determined. Journal of Antibiotics, Vol. 33, 1
120-1127 (1980) [J. Antibiotics, 33 ,
1120 (1980)]. And further, B-41
D, E and F were isolated, Journal of Antibiotics, 36, 980-990, 1983.
[J. Antibiotics, 36 , 980 (1983)].

These B-41 antibiotics were also named milbemycin. And, milbemycin is reported to have insecticidal activity and acaricidal activity in the above-mentioned documents, and it is also reported in JP-A-54-89041 that it has anthelmintic activity.

Many production examples of milbemycins in which the methyl group at the 26th position is converted, which are related to the present invention, are disclosed, but none of them is the same as the present invention. The bond between the 2/3 positions is 2
There are no reports on milbemycins, which are heavy bonds.

[0005]

The object of the present invention is to synthesize a novel milbemycin compound or a useful milbemycin compound having excellent acaricidal, insecticidal or anthelmintic activity against mites, plant pests or animal parasites. Is to develop an intermediate of.

[0006]

[Constitution of the invention]

[0007]

In order to solve the above-mentioned problems, the present inventors have focused their attention on cultures of Streptomyces microorganisms, particularly cultures of the B-41-146 strain, and have conducted intensive studies. As a result, a new series of milbemycin compounds was found and the present invention was completed.

That is, the present invention has the general formula

[0009]

[Chemical 7]

[In the formula, R represents a methyl or ethyl group,
R ¹ represents a methylene group when the bond containing the dotted line between the 4/26 positions is a double bond, and R ¹ represents a methyl group when the bond containing the dotted line between the 4/26 positions is a single bond. ] Specifically, (1) in the above general formula, R is a methyl group, the bond containing a dotted line between the 4/26 positions is a double bond, and R ¹ is a methylene group. In the general formula of a compound Ia1 (2), R is an ethyl group, 4/2
A compound Ia2 in which the bond including the dotted line between the 6-positions is a double bond and R ¹ is a methylene group, (3) In the above general formula,
A compound Ib ¹ in which R is a methyl group, a bond containing a dotted line between the 4/26 positions is a single bond, and R ¹ is a methyl group, (4) In the above general formula, R is an ethyl group, or between the 4/26 positions. The present invention relates to a compound Ib2 in which a bond including a dotted line is a single bond and R ¹ is a methyl group, and (5) a method for culturing a Streptomyces genus microorganism or a production method by known chemical conversion from milbemycin. The above compounds Ia1 and Ia
The names 2, Ib1 and Ib2 are tentative names.

The details will be described below.

1) Production of Compound (I) by Culture Method Regarding the mycological properties of the Streptomyces sp. Strain B-41-146 producing compound (I) by the culture method of the present invention, JP-A-50-29742. The Streptomyces B-41-146 strain described in the official gazette is deposited in the Institute of Biotechnology, Institute of Industrial Science, and the deposit number is FERM BP-1.
072.

As is well known, actinomycetes are naturally occurring in
Further, mutation is easily caused by an artificial operation (for example, ultraviolet irradiation, irradiation, chemical treatment, etc.), and the B-41-146 strain of the present invention has the same point. The B-41-146 strain referred to in the present invention includes all mutants thereof. That is, in the present invention, (I) is produced and B-
Bacteria that are not clearly distinguished from the 41-146 strain and its mutant strain are all included in the B-41-146 strain.

The compound (I) of the present invention is B-41-146.
It is obtained by culturing the strain in a suitable medium and then harvesting it. As a nutrient source, a known source conventionally used for culturing a bacterium of the genus Streptomyces can be used.
For example, glucose, sucrose, starch, glycerin, starch syrup, molasses, soybean oil and the like can be used as the carbon source. As the nitrogen source, soybean flour, wheat germ,
Meat extract, peptone, yeast, corn steep liquor, ammonium sulfate, sodium nitrate and the like can be used. In addition, if necessary, inorganic salts such as calcium carbonate, sodium chloride, potassium chloride, and phosphate are added, and organic and inorganic substances that help the growth of the bacterium and promote the production of the compound (I) of the present invention are suitable. Can be added to.

As the culturing method, the liquid culturing method, particularly the submerged culturing method, is most suitable as in the method for producing general antibiotics. Culturing is carried out under aerobic conditions, and a suitable temperature for culturing is 22-30 ° C, but in most cases, culturing is performed at around 28 ° C. The compound (I) of the present invention is produced by shaking culture,
The maximum value is reached in 5-15 days for both tank culture.

The following method is used for assaying the compound (I) of the present invention. That is, 1 ml of the culture is placed in a small test tube, 9 ml of 80% methanol water is added, and the mixture is shaken for extraction and centrifuged. High performance liquid chromatography was performed using C ₁₈ reverse phase column (Tosoh), 0.46 × 20 cm),
Using a pump (Hitachi, model 635A) and a sample injector (water, model U6K), 5 μl of the above sample was injected, and a solvent system of acetonitrile-water (75:25) was run at a flow rate of 1.6 ml / min.

The compound (I) of the present invention is an ultraviolet detector (24
0 nm), and quantified using a data processor (Shimadzu Corporation Chromatopack C-R3A).

In collecting the compound (I) from the culture, an adsorbent such as activated carbon, alumina, silica gel, a synthetic adsorbent such as Diaion HP-20 (Mitsubishi Kasei), a fixative such as Avicel (Asahi Kasei), filter paper, Ion exchange resin,
Although an ion exchange gel filter or the like can be used, the following collection method is most effective.

The target product is dissolved in methanol water by filtering the culture using a filter aid such as diatomaceous earth and extracting the cake obtained here with methanol.
Water is added to this, extraction is performed with n-hexane, and this is concentrated under reduced pressure to obtain an oily substance containing the target substance. This is adsorbed on a silica gel column, and impurities are washed and the target substance is eluted with a mixture of organic solvents such as n-hexane, acetone, ethyl acetate, methanol and ethanol. The target compound is further subjected to column chromatography, thin layer chromatography, high performance liquid chromatography, etc. by combining various carriers and solvents to isolate the present compound. The structure of the isolated compound is determined by detailed analysis of various spectral properties of the compound, especially its nuclear magnetic resonance, mass, ultraviolet and infrared spectra.

2) Production of Compound (I) by Chemical Transformation Method The compound (I) of the present invention can also be produced by the chemical transformation of known milbemycins shown by the following Method A and Method B.

[0021]

[Chemical 8]

[0022]

[Chemical 9]

In the above formula, R, R ¹ , TBDMS, Ms, Ia and
Ib has the same meaning as above.

Milbemycin represented by II is described in JP-A-64
-31776 is a known compound, and milbemycin represented by III is a known compound described as milbemycin A ₃ or milbemycin A _{4 in} JP-B-56-45890.

In Method A, the step A1 is the compound (II)
Is a step of methanesulfonylating the hydroxyl group at the 26-position.

The reaction is carried out in the presence of methanesulfonyl chloride or methanesulfonic anhydride. The amount used is usually 1 to 5 equivalents, preferably 1 to 2 equivalents. The reaction is preferably carried out in the presence of a base, and suitable bases include, for example, triethylamine, diisopropylethylamine, N, N-dimethylaniline, pyridine,
4-dimethylaminopyridine, 1,5-diazabicyclo [4,3,0] nonene-5 (DBN) or 1,8-diazabicyclo [5,4,0] undecane-7 (DBU)
Organic bases such as sodium carbonate, potassium carbonate,
Examples thereof include inorganic bases such as sodium hydrogen carbonate, and more preferable are organic bases such as triethylamine, diisopropylethylamine and pyridine.

The reaction is preferably carried out in the presence of a solvent, and the solvent used is not particularly limited as long as it does not inhibit the reaction and dissolves the starting material to some extent.
Preferred are hydrocarbons such as hexane, petroleum ether, benzene and toluene, halogenated hydrocarbons such as chloroform and methylene chloride, ethers such as ethyl ether and tetrahydrofuran, N, N-dimethylformamide and the like. Amides, sulfoxides such as dimethylsulfoxide, nitriles such as acetonitrile, and mixtures of these solvents, and more preferably halogenated hydrocarbons.

The reaction temperature is usually -70 to 150 ° C, preferably room temperature to 120 ° C. The reaction time varies depending mainly on the reaction temperature, the reaction reagent or the type of solvent used, but is usually 15 minutes to 1 day and night, preferably 30 minutes to 4 hours.

In Method A, in the step A2, the methanesulfonyl group at the 26th position is eliminated by isomerizing the double bond at the 3rd and 4th positions into the 2nd and 3rd positions in the presence of a base, Four
In this step, a double bond is further introduced at positions 26 and 26.

As the base to be used, the same organic bases or inorganic bases as those used in the step A1 can be used, but inorganic bases such as potassium carbonate and lithium carbonate are preferred. The amount used is usually 1 to 5 equivalents, preferably 2 to 4 equivalents, relative to (II).

The reaction is preferably carried out in the presence of a solvent, and the solvent used is not particularly limited as long as it does not inhibit the reaction and dissolves the starting material to some extent, and preferably, The same hydrocarbons, ethers, amides, sulfoxides or nitriles as the solvent used in step A1 can be used as a solvent, and more preferably,
Amides and sulfoxides.

The reaction temperature is usually room temperature to 150 ° C, preferably 90 to 130 ° C. The reaction time varies depending mainly on the reaction temperature, the reaction reagent or the type of solvent used, but is usually 15 minutes to 1 day and night, preferably 30 minutes to 4 hours.

In the method A, the step A3 is a step of desilylating (deprotecting group) the 5-O-silyl ether compound in the presence of a deprotecting agent to convert the 5-position hydroxyl group.

Examples of the deprotecting agent used include diluted acids such as dilute hydrochloric acid and dilute sulfuric acid, organic acids such as formic acid, acetic acid and trifluoroacetic acid, and sulfonic acids such as p-toluenesulfonic acid and trifluoromethanesulfonic acid. Fluoroions such as tetrabutylammonium fluoride and hydrogen fluoride / pyridine can be used, but sulfonic acids such as p-toluenesulfonic acid are preferable.

The reaction is preferably carried out in the presence of a solvent, but there is no particular limitation as long as it does not inhibit the reaction. In addition to the same hydrocarbons, ethers and nitrites as those used in the step A1, alcohols such as methanol and ethanol, and a mixture of these solvents can be mentioned.

The reaction is carried out over a wide range of temperatures, usually -20 ° C to 70 ° C, more preferably -10 ° C to 30 ° C. The time required for the reaction depends on many factors, but is 30 minutes to 24 hours, preferably 30 minutes to 6 hours.

In the B method, the step B1 is a step of isomerizing the double bond at the 3-position and 4-position to the 2-position and 3-position in the presence of a base.

The base used is the same organic base or inorganic base as used in step A1, preferably an inorganic base such as potassium carbonate or lithium carbonate. The amount used is usually 1 to 5 equivalents, preferably 2 to 4 equivalents, relative to (III).

The reaction is preferably carried out in the presence of a solvent, and the solvent used is not particularly limited as long as it does not inhibit the reaction and dissolves the starting material to some extent.
Preferably, the same hydrocarbons, halogenated hydrocarbons, ethers, amides as those used in Step A1
Sulfoxides and nitriles and mixtures of these solvents, more preferably amides such as N, N-dimethylformamide and sulfoxides such as dimethylsulfoxide.

The reaction temperature is usually 0 ° C to 100 ° C, preferably around room temperature. The reaction time varies depending mainly on the reaction temperature, the reaction reagent or the type of solvent used, but is usually 6 hours to 1 week, and preferably 1 day to 5 days.
It is a day.

After completion of the reaction in each step, the product is recovered from the reaction mixture by a known method and, if necessary, further purified by a known means such as column chromatography.

[0042]

Industrial Applicability The compound represented by the general formula (I) according to the present invention has excellent activity against mites, plant pests or animal parasites, and has acaricide, insecticide or It is useful as an active ingredient of an anthelmintic agent.

That is, the compound (I) of the present invention is an adult and an egg such as spider mites (Tetranychus), apple spider mites, citrus spider mites (Panonychus) and rust mites, which parasitize fruit trees, vegetables and flowers, and ticks (Ixodidae), which parasitize animals. Family Arachnidae (Dermanysside) and genus Asteridae (Sarcoptidae)
It has excellent acaricidal activity against the like.

Furthermore, sheep flies (Oestrus) and fly flies (Lu
cilia), Bullflies (Hypoderma), and bots (Gautrophil)
us) etc. and chisel, ectoparasites of animals and birds such as bryophytes;
Sanitary pests such as cockroaches and house flies; other aphids,
It is active against various agricultural and horticultural insects such as Lepidoptera larvae. In addition, root-knot nematodes (Meloidogyne) and mite (Ph
(izoglyphus) and so on.

To use the compound (I) of the present invention as an acaricidal agent, the active compound is diluted with a carrier and, if necessary, other auxiliary agents are added to prepare a powder, coarse powder, granules or fine granules. , A wettable powder, an emulsion, an oil, and the like can be prepared and used.

Further, the compound (I) of the present invention has excellent parasiticidal activity as an anthelmintic agent for animals and humans. Diseases commonly described as parasitic diseases are helminths (Helmint
h) due to infection of the animal host by a protozoa known as. Parasitic diseases are endemic to cattle, poultry and pets such as pigs, sheep, goats, cattle, horses, dogs, cats and chickens and cause serious economic damage. The parasite group described as nematodes in helminths is infested by various animals and often causes serious infections. The most common genus of nematodes that infect the aforementioned animals are Haemonchus, Trichostrongylus, and Ostertergia.
tertagia), Nematodirus, Cooperia, Ascaris, Bunostomum, Eesophagostomum, Chabertia, Trichuris, Trichuris. Genus (Strong
ylus), Triconema (Trichonema), Dictyocaulus (Dictyocaulus), Capillari (Capillari)
a), Heterakis, Toxoc
ara), Ascaridia, Oxyuris, Ancylostoma, Uncinaria, Toxacari
aris) and the genus Parascaris.

Some of the genera Nematodales, Cooperia and Esophagogosthum attack the intestinal tract, while those of the genus Hemonx and Ostertergia parasitize the stomach and parasites of the genus Dichthiocaurus Found in the lungs.

Parasites of the family Filariidae and Setariidae are also found in other tissues and organs of the body such as heart and blood vessels, subcutaneous and lymphatic tissues.

Furthermore, it has a broad spectrum of activity against many endoparasites in a variety of animals, such as the dog genus Dirofilaria, the genus Nematospiroides of the genus Kess, the sifasia Genus (Syphacia) and genus Aspicula (Aspicula
ris) also shows activity.

The compound (I) of the present invention is also useful against parasites that infect humans, and the most common parasites of the human digestive tract are Ancylostoma and Necator. , Ascaris, Strongyloides, Tricinera
hinella), genus Capillaria, genus Trichuris and genus Enterobius
Is.

Other medically important parasites found in blood or other tissues and organs outside the gastrointestinal tract are the genus Wuchereria, genus Bruchia (Bruchia) of the family Filaria.
ugia), Onchocerca and Loa filamentous worms (Loa), and Dracunculidae dracunculus (Dracunculus), in addition to parasites of the intestinal parasite They are the genus Strongiroides and the genus Trichinella.

When the compound (I) of the present invention is used as an anthelmintic in animals and humans, it can be orally administered as a liquid beverage. Beverages are usually solutions, suspensions or dispersions in suitable non-toxic solvents or water with suspending agents such as bentonite and wetting agents or other excipients. Beverages also generally contain an antifoaming agent. Beverage formulations generally contain from about 0.01 to 0.5% by weight of active compound, preferably 0.01 to 0.1.
Contains wt%. When the compound (I) of the present invention is administered by animal feed, it is dispersed homogeneously in the feed,
Used as top dressing or in pellet form. Normally, to achieve the desired antiparasitic effect, 0.0001-0.
Contains 02%.

The compound (I) of the present invention dissolved or dispersed in a liquid carrier excipient may be parenterally administered to animals by injection into the forestomach, intramuscular, intratracheal or subcutaneous. it can. For parenteral administration, the active compounds are preferably mixed with suitable vegetable oils such as peanut oil, cottonseed oil.
Such formulations generally contain from 0.05 to 50% by weight of active compound.
contains.

The compound (I) of the present invention may also be administered topically by admixing it with a suitable carrier such as dimethyl sulfoxide or a hydrocarbon solvent. This formulation is applied directly to the external surface of the animal by spraying or direct pouring.

The optimum amount of active compound used for best results depends on the kind of animal to be treated and on the type and degree of parasite infection, but is generally about 0.01 to 100 mg / kg of animal body weight, preferably Is 0.5 to 50.0 mg
Orally administered. Such doses may be given at once or in divided doses over a relatively short period such as 1-5 days.

Next, the present invention will be described in more detail by way of examples.

[0057]

【Example】

Example 1 Δ ^2,3 , Δ ^4,26 -milbemycin A ₃ (Compound No. 1, Ia
1), Δ ^2,3 , Δ ^4,26 -milbemycin A ₄ (Compound number
2, Ia2), Δ ^2,3 -milbemycin A ₃ (Compound No.
3, Ib1) and Δ ^2,3 -milbemycin A ₄ (compound no.
No. 4, Ib2) Sucrose 1%, Polypeptone 0.35% and K ₂ HPO ₄
500 containing 100 ml of pre-culture medium containing 0.05%
Streptomyces B-41-1 in ml Erlenmeyer flask
Forty-six strains were inoculated with one platinum loop and cultured on a rotary shaker at 28 ° C for 48 hours. 40 ml of this culture solution 2 ml
Production medium (sucrose 8%, soybean flour 1%, skim milk 1%, yeast Eski 0.1%, meat extract 0.1%, Ca
_{CO 3 0.3%, K 2 HPO} 4 0.03%, MgSO 4 · 7H 2 O 0.1%, F
The cells were inoculated into a 500 ml Erlenmeyer flask containing eSO ₄ .7H ₂ O (0.005%). Pre-culture medium and production medium should be p
It was adjusted to H7.2 and sterilized in an autoclave at 120 ° C. for 20 minutes and used. Rotary shaker (210
The culture was performed at 28 ° C. for 12 days at 28 rpm.

After culturing for 12 days, the pH of the culture was 5 liters.
Was made to be 3 with sulfuric acid, 300 g of Celite was added, and filtration was performed to obtain a cake of about 530 g. This was extracted with 4 liters of methanol, separated by filtration, 3.5 liters of water was added to 4 liters of the obtained methanol solution, and the mixture was extracted with 8 liters of n-hexane. The obtained n-hexane layer was dehydrated with anhydrous sodium sulfate and then concentrated under reduced pressure in a water bath at 40-45 ° C. to obtain 9 g of oil. This is 20m
1 g of chloroform and adsorbed on a column filled with n-hexane of 300 g of silica gel in advance,
It was developed with n-hexane: acetone (80:20). As a result, the fraction 8 containing the compound (I) of the present invention
00 ml was obtained. This was concentrated under the same conditions as above, and 5
80 mg of oil was obtained.

100 mg of the obtained oil was dissolved in 1 ml of methanol, and the reverse phase column Tosoh ODS-80TM (21.5 mmID
(30 cm) and eluate acetonitrile-water = 70 /
Elution was carried out under the conditions of 30, flow rate 10 ml / min, UV detection wavelength 240 nm.

The peak eluted at 62 minutes was collected and the eluate was concentrated under reduced pressure to ^obtain Δ ^2,3 , Δ ^4,26 -milbemycin A.
1.1 mg of ₃ (Compound No. 1) (Ia1: R = methyl group, dotted line = double bond) was obtained.

Similarly, from the peak eluted at 68 minutes,
Δ ^2,3 -milbemycin A ₃ (Compound No. 3) (Ib1:
R = methyl group, dotted line = single bond) was obtained in an amount of 4.3 mg.
Further, from the peak eluted at 95 minutes, Δ ^2,3 , Δ ^4,26 −
0.9 mg of milbemycin A ₄ (Compound No. 2) (Ia2: R = ethyl group, dotted line = double bond) was obtained. further,
From the peak eluted at 104 minutes, 2.2 mg of Δ ^2,3 -milbemycin A ₄ (Compound No. 4) (Ib2: R = ethyl group, dotted line = single bond) was obtained.

Δ ^2,3 , Δ ^4,26 -milbemycin A ₃ (compound
No. 1, Ia1) Mass spectrum (m / z): 526 (M ⁺ ), 508, 490, 359, 27
6, 249, 209, 181, 164, 153 Nuclear magnetic resonance spectrum (200MHz) δ (CDCl ₃ ) ppm: 6.66
(s, 1H, C (3) -H), 6.12 (dt, 1H, C (9) -H, J _d = 11.1Hz, J _t =
2.4Hz), 5.63 ~ 5.79 (m, 2H, C (10) -H, C (26) -H), 5.32 ~
5.52 (m, 3H, C (11) -H, C (19) -H, C (26) -H), 4.91 ~ 5.02 (m, 1
H, C (15) -H), 4.41 to 4.63 (m, 3H, C (5) -H, C (27) -H ₂ ), 4.1
2 (d, 1H, C (6) -H, J = 2.9Hz), 3.54 to 3.69 (m, 1H, C (17) -H),
3.31 (dq, 1H, C (25) -H, J _d = 3.3Hz, J _q = 9.2Hz), 1.48 (s, 3
H, C (29) -H ₃ ). Δ ^2,3 , Δ ^4,26 -milbemycin A ₄ (Compound No. 2, Ia
2) Mass spectrum (m / z): 540 (M ⁺ ), 522, 504, 276, 19
5, 167, 149 Nuclear magnetic resonance spectrum (200MHz) δ (CDCl ₃ ) ppm: 6.66
(s, 1H, C (3) -H), 6.16 (dt, 1H, C (9) -H, J _d = 11.1Hz, J _t =
2.5Hz), 5.64 ~ 5.78 (m, 2H, C (10) -H, C (26) -H), 5.34 ~
5.50 (m, 3H, C (11) -H, C (19) -H, C (26) -H), 4.91 ~ 4.98 (m, 1
H, C (15) -H), 4.41 to 4.63 (m, 3H, C (5) -H, C (27) -H ₂ ), 4.1
4 (d, 1H, C (6) -H, J = 2.6Hz), 3.61 to 3.65 (m, 1H, C (17) -H),
3.09 (dt, 1H, C (25) -H, J _d = 2.6Hz, J _t = 9.3Hz), 1.48 (s, 3
H, C (29) -H ₃ ), 1.01 (d, 3H, J = 6.6Hz), 1.00 (t, 3H, C (31)-
H ₃ , J = 7.3Hz), 0.83 (d, 3H, J = 6.5Hz). Δ ^2,3 -milbemai
Syn A ₃ (Compound No. 3, Ib1) Mass spectrum (m / z): 528 (M ⁺ ), 510, 181, 153 Nuclear magnetic resonance spectrum (270MHz) δ (CDCl ₃ ) ppm: 6.06〜
6.18 (m, 2H, C (3) -H, C (9) -H), 5.70 (dd, 1H, C (10) -H, J = 11.
2,14.9Hz), 5.31 ~ 5.48 (m, 2H, C (11) -H, C (19) -H), 4.97
(t, 1H, C (15) -H, 7.2Hz), 4.58 (dd, 1H, C (27) -H, J = 2.4,14.
1Hz), 4.50 (dd, 1H, C (27) -H, J = 2.4,14.1Hz), 4.05 (d, 1H,
C (6) -H, J = 2.4Hz), 3.55 to 3.67 (m, 2H, C (5) -H, C (17) -H),
3.29 (dt, 1H, C (25) -H, J _d = 9.7Hz, J _q = 6.4Hz), 1.48 (s,
3H, C (29) -H ₃ ), 1.22 (d, 3H, J = 7.2Hz), 1.15 (d, 3H, J = 6.0H
z), 0.99 (d, 3H, J = 6.5Hz), 0.84 (d, 3H, J = 6.5Hz). Δ ^2,3 -milbemycin A ₄ (Compound No. 4, Ib2) mass spectrum (m / z): 542 (M ⁺ ), 524, 278, 195, 16
7, 149 Nuclear magnetic resonance spectrum (270MHz) δ (CDCl ₃ ) ppm: 6.11〜
6.17 (m, 2H, C (3) -H, C (9) -H), 5.71 (dd, 1H, C (10) -H, J = 11.
3,14.9Hz), 5.34 ~ 5.47 (m, 2H, C (11) -H, C (19) -H), 4.92
~ 4.97 (m, 1H, C (15) -H), 4.59 (dd, 1H, C (27) -H, J = 2.4,14.
1Hz), 4.50 (dd, 1H, C (27) -H, J = 2.4,14.1Hz), 4.05 (d, 1H,
C (6) -H, J = 2.4Hz), 3.55 to 3.66 (m, 2H, C (5) -H, C (17) -H),
3.08 (dt, 1H, C (25) -H, J _d = 2.8Hz, J _t = 9.3Hz), 1.48 (s,
3H, C (29) -H ₃ ), 1.23 (d, 3H, J = 7.3Hz), 1.00 (d, 3H, J = 6.4H
. z), 1.00 (t, 3H, J = 7.3Hz), 0.83 (d, 3H, J = 6.5Hz) Example 2 5-O-t - butyldimethylsilyl -26- Metansuru
Fonyl oxymilbemycin A ₃ (V: R = methyl group) (Step A 1) To a methylene chloride solution of 3.30 g of 5-Ot-butyldimethylsilyl 26-hydroxymilbemycin A ₃ and 0.97 ml of methanesulfonyl chloride. 1.22 ml of triethylamine was added dropwise at 0 ° C., and the mixture was further stirred at room temperature for 2 hours. After completion of the reaction, the reaction solution was poured into an aqueous citric acid solution and extracted with methylene chloride. The extract was washed with saturated brine, dried (MgSO ₄ ), concentrated, and the residue was purified by silica gel column chromatography to give the desired compound (2.6).
0 g (70%) was obtained.

Mass spectrum (m / z): 736 (M ⁺ ), 679,
640, 622, 563. Nuclear magnetic resonance spectrum (200MHz) δ (CDCl ₃ ) ppm: 5.70〜
5.87 (m, 3H, C (3) -H, C (9) -H, C (10) -H), 5.25 ~ 5.47 (m, 2H,
C (11) -H, C (19) -H), 4.54 to 5.07 (m, 6H, C (5) -H, C (15) -H, C
(26) -H ₂ , C (27) -H ₂ ), 3.86 (d, 1H, C (6) -H, J = 5.5Hz), 3.02
(s, 3H, OSO ₂ CH ₃ ). Example 3 Δ ^2,3 , Δ ^4,26 -5- ^Ot -butyldimethylsilyl mill
Bemycin A ₃ (VI: R = methyl group) (Step A2) 5-Ot-butyldimethylsilyl-26-methanesulfonyloxymilbemycin A ₃ To a solution of 2.60 g of dimethylformamide, 0.78 g of lithium carbonate was added. ,
The mixture was heated and stirred at 120 ° C. for 4 hours. After completion of the reaction, the reaction solution was poured into water and extracted with ethyl acetate. Dry the extract (MgSO 4
₄ ), concentrated, and the residue was purified by silica gel column chromatography to obtain 1.08 g (48%) of the desired compound.

Mass spectrum (m / z): 640 (M ⁺ ), 622,
583, 565 Nuclear magnetic resonance spectrum (200MHz) δ (CDCl ₃ ) ppm: 6.65
(s, 1H, C (3) -H), 6.11 (dt, 1H, C (9) -H, J = 11.0Hz, J _t = 2.5
Hz), 5.61 ~ 5.79 (m, 2H, C (10) -H, C (27) -H), 5.31 ~ 5.49
(m, 3H, C (11) -H, C (19) -H, C (27) -H), 4.95 (t, 1H, C (15) -H,
J = 6.7Hz), 4.73 (br.s, 1H, C (7) -OH), 4.61 (d, 1H, C (5) -H,
J = 2.3Hz), 4.53 (dd, 1H, C (26) -H, J = 2.4,14.3Hz), 4.43 (d
d, 1H, C (26) -H, J = 2.4,14.3Hz). Example 4 Δ ^2,3 , Δ ^4,26 Milbemycin A ₃ (Ia: R = methyl group) (Step A3) 5-O -T-Butyldimethylsilyl milbemycin A ₃
0.45 g of p-toluenesulfonic acid monohydrate was added to 1.07 g of a methanol solution, and the mixture was stirred at room temperature for 4 hours. After completion of the reaction, the reaction solution was poured into water and extracted with ethyl acetate. The extract was dried (MgSO _4), concentrated and the residue was purified by silica gel column chromatography, the desired compound, 0.84 g (95%) was obtained.

Example 5 Δ ^2,3 -milbemycin A ₃ (Ib: R = methyl group)
(Step B1) To a solution of 2.0 g of milbemycin A ₃ in methylene chloride,
1,8-diazabicyclo [5,4,0] undecene-7
(DBU) (1.06 ml) was added, and the mixture was left at room temperature for 4 days. After completion of the reaction, the reaction solution was poured into water and extracted with ethyl acetate. The extract was dried (MgSO ₄ ) and concentrated, and the residue was purified by silica gel column chromatography to obtain 1.40 g (70%) of the target compound.

Example 6 Activity Test Example 1: A chemical solution containing 10 ppm of each of the compounds of the present invention and 0.01% of a spreading agent was prepared. Cowpea ( Vigna sinenis Savi)
First the leaves of the organophosphorus insecticides sensitive Tetranychus urticae (Te
transychus urticae ), and one day after the inoculation, 7 ml of the above chemical solution was applied in a Mizuho rotary spraying tower.
Was sprayed so that the amount of spray liquid was 3.5 mg / cm ² leaves. After spraying, store cowpea leaves in a high temperature room at 25 ° C for 3 days.
After a day, the life and death of adults were examined by a stereoscopic microscope, and the mortality rate (%) was calculated. The results are shown in Table 1.

[0067]

[Table 1]

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁵ Identification number Internal reference number FI technical display location C12R 1: 465) 7804-4B (72) Inventor Shinji Yokoi 1041 Sansu, Yasu-cho, Yasu-gun, Shiga Prefecture Stock company in-house (72) Inventor Takeshi Kinoshita 1-25-2 Hiromachi, Shinagawa-ku, Tokyo Sankyo stock company (72) Inventor Noriaki Kudo 1041 Yasu, Yasu-cho, Yasu-gun, Shiga Sankyo stock company In-house

Claims (8)

[Claims] 1. The following general formula: [In the formula, R represents a methyl or ethyl group, and R ¹ is 4/2.
When the bond containing the dotted line between the 6-positions is a double bond, it represents a methylene group, and R ¹ represents a methyl group when the bond containing the dotted line between the 4/26 positions is a single bond. ] The novel milbemycin compound represented by these. 2. A compound Ia1 according to claim 1, wherein R is a methyl group, the bond containing the dotted line between the 4/26 positions is a double bond, and R ¹ is a methylene group. 3. A compound Ia2 according to claim 1, wherein R is an ethyl group, the bond containing the dotted line between the 4/26 positions is a double bond, and R ¹ is a methylene group. 4. A compound Ib1 according to claim 1, wherein R is a methyl group, the bond containing the dotted line between the 4/26 positions is a single bond, and R ¹ is a methyl group. 5. A compound Ib2 according to claim 1, wherein R is an ethyl group, the bond containing a dotted line between the 4/26 positions is a single bond, and R ¹ is a methyl group. 6. The method for producing a compound according to claim 2, wherein the compound (I) -producing bacterium belonging to Streptomyces is produced by a general culturing method in which a microorganism of the genus Streptomyces is conventionally cultivated. . 7. A general formula: [In the formula, R represents a methyl group or an ethyl group. ] In the presence of a base, 26-hydroxymilbemycin represented by
By silylation, the general formula: [In the formula, R has the same meaning as described above, and TBDMS represents a t-butyldimethylsilyl group. ] The 5-silylated product represented by the formula] is produced, and the 5-silylated product is subjected to methanesulfonylation in the presence of a base to give a compound represented by the general formula: [In the formula, R and TBDMS have the same meanings as described above, and Ms represents a methanesulfonyl group. ] The 5-silylated-26-sulfonylated compound represented by the formula] is produced, and this 5-silylated-26-sulfonylated compound is desulphonylated in the presence of an alkali to give a compound of the general formula: [Wherein R and TBDMS have the same meanings as described above. ] The 5-silylated-26-methylenylated product represented by the formula] is produced, and the 5-silylated-26-methylenylated product is treated with a desilylating agent. Method for the synthesis of milbemycin compounds. 8. A general formula: [In the formula, R has the same meaning as described above. Synthesis process according to claim 4 or claim 5 milbemycin compound according milbemycin A ₃ or milbemycin A _4, characterized in that the treatment with alkali represented by.