EXHIBIT 1015
`
`

`

`(19) United States
`(12) Patent Application Publication (10) Pub. No.: US 2004/0254125 A1
`Saito et al.
`(43) Pub. Date:
`Dec. 16, 2004
`
`US 20040254125A1
`
`(54) ANTHELMINTIC COMPOSITION
`
`(57)
`
`ABSTRACT
`
`(76)
`
`Inventors: Akio Saito, Tokyo (JP); Yoko
`Sugiyama, Shinagawa—ku (JP);
`Toshimitsu Toyama, Shinagawa—ku
`(JP); Toshihjko Nanba, Shinagawa-ku
`(JP)
`
`Correspondence Address:
`FRISHAUF, HOLTZ, GOODMAN & CHICK,
`PC
`767 THIRD AVENUE
`25TH FLOOR
`NEW YORK, NY 10017-2023 (US)
`
`(21) Appl. No.:
`
`10/493,716
`
`(22) PCT Filed:
`
`Oct. 24, 2002
`
`(86) PCT No.2
`
`PCT/JP02/11067
`
`(30)
`
`Foreign Application Priority Data
`
`Oct. 25, 2001
`
`(JP) ...................................... 2001-327198
`
`Publication Classification
`
`Int. 01.7 ........................... A01N 43/04; A01N 43/02
`(51)
`(52) vs. C].
`.............................................. 514/28; 514/450
`
`An anthelmintic composition containing as the active ingre-
`dients the first component consisting of one or more mem-
`bers selected from among compounds represented by the
`general forrnura (I):
`
`
`
`(wherein R1 is methyl or the like; and R2 is acetyl or the like)
`and so on and the second component consisting of one or
`more members selected from among praziquantels and so
`on,
`
`

`

`US 2004/0254125 A1
`
`Dec. 16, 2004
`
`ANTHELMINTIC COMPOSITION
`
`TECHNICAL FIELD
`
`[0001] The present invention relates to an anthelmintic
`composition comprising milbemycin derivatives (the first
`ingredient) and the second ingredient as active ingredient;
`and a method for using it.
`
`BACKGROUND ART
`
`It has been known that Milbemycin or avermectin
`[0002]
`series compounds have excellent
`insecticidal activities
`against wide range of parasites, however, the milbemycin
`derivative represented by the following formula (I) of the
`present invention, a composition comprising thereof and
`bioactivities thereof have been conventionally unknown.
`
`DISCLOSURE OF THE INVENTION
`
`[0003] The inventors of the present invention have con-
`ducted extensive studies in order to obtain a composition
`exhibiting a strong activity against a wide range of endo- and
`eetoparasites including eetoparasites such as fleas, parasites
`in the digestive tract such as ascarides, and endoparasites
`such as filariae in both oral administration and local admin-
`istration, and as a result, they have found that a composition
`comprising a specific milbemycin derivative and a second
`ingredient such as praziquantel, as active ingredients, exhib-
`its an excellent anthelmintic activity, and the present inven-
`tion has been accomplished.
`
`[0004] The present invention relates to an anthelmintic
`composition comprising one or more kinds of first ingredient
`selected from milbemycin derivatives composed of a com-
`pound represented by forrnula a):
`
`
`
`[wherein, R1 represents a methyl group, an ethyl
`[0005]
`group, an isopropyl group or an s-butyl group; and R2
`represents an acetyl group, a methoxyacetyl group, a trif-
`luoroacetyl group, a cyano acetyl group or a methanesulfonyl
`group] or salts thereof, and, one or more kinds of second
`ingredients selected from the group consisting of praziqu-
`antels, fipronils, benzimidazoles and neonicotinoides, as
`active ingredients; and a method for using it.
`
`ides, and endoparasites such as filariae both in a oral
`administration and in a local administration.
`
`In compound (I) which is an active ingredient of
`[0007]
`the present invention, (1) R1 is preferably a methyl group or
`an ethyl group, and (2) R2 is preferably a methoxyacetyl
`group.
`
`In compound (I) which is an active ingredient of
`[0008]
`the present invention, it is preferable that R1 is a methyl
`group or an ethyl group, and R2 is a methoxyacetyl group.
`[0009] As representative compounds of compound (I)
`which are active ingredients of the present invention, for
`example, the compounds described in the following Table
`can be cited, however, compound (I) is not limited to these
`compounds.
`
`Incidentally, in the Table, “Me” represents a methyl
`[0010]
`group, “Et” represents an ethyl group, “iPr” represents an
`isopropyl group and “sBu” represents a s-butyl group,
`respectively.
`
`TABLE 1
`
`2
`R \ N
`|H
`
`O
`
`
`
`Compound
`No.
`R1
`R2
`
`1
`Et
`MeCO
`2
`Et
`CF3CO
`3
`Et
`NCCHZCO
`4
`Et
`MeOCHZCO
`5
`Et
`MeSO2
`6
`Me
`MeCO
`7
`Me
`CFSCO
`8
`Me
`NCCHZCO
`9
`Me
`MeOCHZCO
`
`e
`iPr
`11
`1136?):
`Me
`10
`CF3CO
`iPr
`12
`NCCH2CO
`iPr
`13
`MeOCHZCO
`iPr
`14
`MeSOz
`iPr
`15
`MeCO
`sBu
`16
`CFSCO
`sBu
`17
`NCCHZCO
`sBu
`18
`MeOCHZCO
`sBu
`19
`
`
`sBu20 MeSO2
`
`In compound (I) which is an active ingredient of
`[0011]
`the present invention, preferable compounds are compounds
`of
`
`[0006] The compound which is an active ingredient of the
`present invention and is represented by the above formula
`(I), is a compound exhibiting a strong activity against a wide
`range of endo- and eetoparasites including eetoparasites
`such as fleas, parasites in the digestive tract such as ascar-
`
`exemplified compound number ,1, 2, 3, 4, 5, 6, 7, 9,
`[0012]
`10, 13, 14, 15, 16, 17, 19 and 20, more preferably com-
`pounds of exemplified compound number 1, 3, 4, 5, 6, 7, 9
`and 10, and further more preferably compounds of exem-
`plified compound number 1, 3, 4, 5, 9 and 10.
`
`

`

`US 2004/0254125 A1
`
`Dec. 16, 2004
`
`vaniliprole (see, Japanese Provisional Patent Pub-
`[0018]
`lication No. 10—338678) represented by the following for-
`mula; and
`
`NC
`
`S—CF3
`
`Cl
`
`C]
`
`OH
`
`OMe
`
`CF3
`
`[0019]
`
`a compound represented by the following formula.
`
`[wherein when R is a methyl group and X is a
`[0020]
`chlorine atom, the compound is acetprole, and when R is an
`ethyl group and X is a trifluoromethyl group, the compound
`is ethiprole] are known. Other than these, the compounds
`described in WO98/40359, WO98/02042, WO98/45274,
`WO98/28277, WO98/28278, WO98/28279, WO99/31070,
`WOOD/62616, W001/07413, or W001/00614 can be raised.
`Among these, preferred are fipronil and vaniliprole.
`
`[0021] Benzimidazoles which are active ingredients of the
`present invention, are used as anthelrnintics against parasites
`in the digestive tract and,
`for example, albendazole,
`flubendazole and mebendazole are known (The benzimida—
`zole anthelmintics-chemistry and biological activity, S.
`Sharma and S. Auzar, Progress in Drug Research Vol 27,
`85—161).
`
`[0022] As for neonicotinoides which are active ingredients
`of the present invention, for example, imidacloprid, thia—
`methoxam, nitenpyram and acetamiprid are known. Among
`these, imidacloprid is used as a flea exterminator for the
`local administration similarly to fipronils.
`
`[0023] The second ingredient of the present invention is
`preferably praziquantels or fipronils, more preferably prazi-
`quantel, epsiprantel or fipronil.
`
`[0024] Milbemycin derivatives which are active ingredi-
`ents of the present invention can be prepared by the method
`of the steps A to D shown below.
`
`[0013] When compound (I) which is an active ingredient
`of the present invention or the salt thereof has an asymmetric
`carbon atom within the molecule, a stereoisomer which is
`the R configuration or the S configuration may be present,
`and both the individual isomer and compound thereof in any
`proportion is included in the present
`invention. Such a
`stereoisomer can be, for example, synthesized by using an
`optically-separated starting compound or optically separat-
`ing synthesized compound (I) using a usual optical separa-
`tion method or isolation method, if desired.
`
`[0014] The compound (I) which is an active ingredient of
`the present invention or the salt thereof can be formed into
`a solvate, and such a solvate salt is also included in the active
`ingredient of the present invention. For example, compound
`(I) may absorb water when left in the atmosphere or when
`recrystallized to get absorbed water or to form a hydrate, and
`salt of such compound containing water is also included in
`the milbemycin derivatives which are active ingredients of
`the present invention.
`
`[0015] The praziquantels which are active ingredients of
`the present invention are compound effective against Ces-
`toidea on which milbemycins or avermectins have less effect
`(Parasitorogy Today, V01 1, No 1, 10-17, 1985), and are
`compounds represented by the following formula:
`
`(CH2):
`
`| N
`
`N
`
`/Vro -
`
`[wherein when n is 1, the compound is a praziqu-
`[0016]
`antel, and when n is 2, the compound is an epsiprantel].
`
`[0017] The fipronils which are active ingredients of the
`present invention are compounds having an instantaneous
`etfect against arthropod such as insects and mites, for
`example, flpronil (see, Japanese Provisional Patent Publica-
`tion No. 63316771) represented by the following formula:
`
`0
`
`\
`\s——-c1=3
`
`NC
`
`N
`
`\
`
`\N
`
`C1
`
`NHz
`C1
`
`CF3
`
`

`

`US 2004/0254125 A1
`
`Dec. 16, 2004
`
`
`
`
`
`
`OzN
`
`Step C
`
`(IV)
`
`(V) O! (I)
`
`-continued
`
`HZN
`
`(VI)
`
`
`
`(I)
`
`[0025] wherein R1 and R2 represent the same mean-
`ing as described above, and X represents a nitro
`group or a group represented by formula —NR2H.
`15-Hydroxy milbemycin derivative having for-
`[0026]
`mula (III) as starting compound is a publicly known com-
`pound described in Japanese Provisional Patent Publication
`No. 60-158191.
`
`[0027] Step A
`[0028] Step A is a step of reacting a compound (III) with
`a carboxylic acid represented by formula:
`
`OH
`
`X
`
`in an inactive solvent in the presence of trifluo-
`[0029]
`romethanesulfonic acid which is a strong organic acid or
`trimethylsilyl trifluorosulfonate to produce a compound rep-
`resented by formula (IV).
`[0030] The amount of trifluoromethanesulfonic acid or
`trimethylsilyl
`trifiuorosulfonate to be used is a catalytic
`amount in principle, and it does not need an equivalent
`weight with respect to compound (III). However, the amount
`may vary drastically depending upon the reactivity of the
`carboxylic acid employed.
`
`

`

`US 2004/0254125 A1
`
`Dec. 16, 2004
`
`[0031] Furthermore, when a powder of an inorganic com-
`pound is added to the reaction system, some reaction would
`be accelerated and a good result would be given. As such
`inorganic compounds, metal salts such as copper trifluo-
`romethanesulfonate, cuprous iodide, zinc iodide, cobalt
`iodide and nickel iodide; Celite; silica gel and alumina can
`be cited, a copper salt such as copper trifluoromethane-
`sulfonate or cuprous iodide is preferable, and cuprous iodide
`is the most preferable.
`
`[0032] When the carboxylic acid to be used is hardly
`soluble in a solvent, a silyl ester of a carboxylic acid can be
`used. In this case, there can be used a method where the
`carboxylic acid is reacted with an equivalent amount of
`allyltrimethylsilane in the presence of trifluoromethane—
`sulfonic acid or trimethylsilyl ester thereof as a catalyst and
`compound (III) is added to the obtained solution of trim-
`ethylsilyl ester.
`
`to be used in the reaction is not
`[0033] The solvent
`particularly limited as long as it does not inhibit the reaction
`and it dissolves the starting materials to some extent. Suit-
`ably, aromatic hydrocarbons such as benzene, toluene and
`xylene; and halogenated hydrocarbons such as methylene
`chloride, 1,2-dichloroethane and chloroform can be raised.
`
`[0034] The reaction temperature is from —10° C. to 100°
`C., preferably from 0° C. to 50° C. The reaction time varies
`mainly, depending on the reaction temperature, the starting
`compounds or the sort of the solvent to be used, and it is
`generally from 5 minutes to 6 hours, preferably fiom 10
`minutes to 2 hours.
`
`the reaction, palladium-on-carbon, palladium-on-barium
`sulfate, platinum oxide and so forth can be raised.
`
`[0043] As a suitable solvent to be used in the reaction, an
`alcohol such as methanol or ethanol, an ether such as
`tetrahydrofuran or dioxane, and an ester such as ethyl
`acetate, can be raised.
`
`[0044] The reaction temperature is normally from 10° C.
`to 80° C., and the reaction time is normally from 10 minutes
`to 5 hours.
`
`[0045] As another preferred reduction method, reduction
`with zinc powder in a solvent of acetic acid can be cited.
`
`[0046] The reaction temperature is normally from 0° C. to
`room temperature, and the reaction time is normally from 30
`minutes to 12 hours.
`
`[0047] As a more preferred reduction method, reduction
`vw'th sodium borohydride in the presence of a nickel catalyst
`can be cited.
`
`[0048] As a nickel catalyst, a nickel salt such as nickel
`chloride or nickel bromide can be used. Preferred are
`triphenylphosphine complexes of these nickel salts.
`
`[0049] As suitable solvents to be used in the reaction, for
`example, alcohols such as methanol or ethanol, and ethers
`such as tetrahydrofuran or dioxane can be cited.
`
`[0050] The reaction temperature is normally from 0° C. to
`room temperature, and the reaction time is normally about
`from 10 minutes to 120 minutes.
`
`[0035]
`
`(Step B)
`
`[0051]
`
`(Step D)
`
`[0036] Step B is a step of reacting compound (IV) with a
`reducing agent such as sodium borohydride in an inactive
`solvent to convert the carbonyl group at S-position into a
`hydroxy group to produce a compound represented by
`formula (V) or compound (I) which is an active ingredient
`of the present invention.
`
`[0037] The reducing agent to be used is not particularly
`limited as long as it is a reducing agent known as reducing
`the carbonyl group to a hydroxy group, for example, it can
`be a metal borohydride such as sodium borohydride or
`lithium borohydride, preferably sodium borohydride.
`
`[0038] As a reactive solvent, any can be used without
`particular limitation as long as it is not involved in the
`reaction, and particularly the use of a lower alcohol such as
`methanol, ethanol or propanol; or ethers such as tetrahydro-
`furan or dimethoxyethane is preferable.
`
`[0039] The reaction temperature is normally from 0° C. to
`50° C., and the reaction time is normally from 1 hour to 10
`hours.
`
`[0040]
`
`(Step C)
`
`[0041] Step (C) is a step of reducing the nitro group of
`compound (V) in an inactive solvent to produce a compound
`having an amino group, which is represented by formula
`(VI).
`
`[0042] For the reduction of the nitro group, a method
`usually used can be used. As one of such methods,
`the
`catalytic reduction using a precious metal catalyst can be
`raised. As preferred examples for the catalyst employed in
`
`[0052] Step D is a step of reacting an amino group of the
`compound represented by compound (V1) with an acid
`represented by formula Rz—OH (wherein R2 represents the
`same meaning as mentioned above) or reactive derivatives
`thereof in an inactive solvent
`to produce a compound
`represented by compound (I) which is an active ingredient of
`the present invention.
`
`[0053] As reactive derivatives of the acid represented by
`the formula Rz—OH, what is usually used in a condensation
`reaction, such as acid halides (acid chloride, acid bromide,
`etc.), an acid anhydride, a mixed acid anhydride, an active
`ester or an active amide, can be cited.
`
`[0054] When the acid represented by formula Rz—OH is
`used, a dehydrating agent such as dicyclohexylcarbodiimide
`(DCC), 2—chloro-1-rnethylpyridinium iodide, p-toluene-
`sulfonic acid or sulfuric acid is used, and preferably such a
`dehydrating agent is 2-chloro-1-methylpyridinium iodide.
`The amount of dehydrating agent employed is normally
`from 1 to 5 equivalents, preferably from 1 to 2 equivalents,
`to the acid represented by the formula Rz—OH.
`
`[0055] The solvent to be used is not particularly limited as
`long as it does not inhibit the reaction and it dissolves the
`starting material to some extent, for example, it can be a
`hydrocarbon such as hexane, petroleum ether, benzene or
`toluene; a halogenated hydrocarbons such as chloroform,
`methylene chloride or 1,2-dichlor0ethane; an ether such as
`ethyl ether or tetrahydrofuran; an amide, such as N,N—
`dimethylformamide; a sulfoxide such as dimethyl sulfoxide;
`a nitrite such as acetonitrile; and a mixture of these solvents.
`Preferred is dichloromethane or 1,2—dichloroethane.
`
`

`

`US 2004/0254125 A1
`
`Dec. 16, 2004
`
`[0056] The reaction temperature is usually from —70° C. to
`90° C., preferably from 0° C. to 60° C. The reaction time
`varies mainly according to the reaction temperature, the
`starting compounds, reaction reagents or the sort of the
`solvent employed, and it is normally from 15 minutes to a
`whole day and night, and preferably from 30 minutes to 6
`hours.
`
`[0057] When the above-described reactive derivative is an
`acid halide,
`the reaction is preferably carried out in the
`presence of a base. As preferred bases, for example, organic
`bases such as triethylamine, N,N—dimethylaniline, pyridine,
`4—dimethy1aminopyridine, 1,5-diazabicyclo[4.3.0]nonene—5
`(DBN) and 1,8-diazabicyclo[5.4.0]undecene-7 (DBU) can
`be raised.
`
`[0058] The amount of the acid halide to be used is nor—
`mally from 1 to 10 equivalents, and that of the base
`employed is normally from 2 to 8 equivalents, to compound
`(VI).
`
`the reaction
`[0059] The solvents used in the reaction,
`temperature, the reaction time and the like each is the same
`as that when a carboxylic acid itself is used.
`
`[0060] The reaction temperature is normally from 0° C. to
`50° C., and the reaction time is normally from 5 minutes to
`2 hours.
`
`[0061] After completion of each step described above,
`each desired compound, the compound (IV), (V) and (I) are
`isolated from the reaction mixture by a commonly known
`method, and, if necessary, purified by a known method, such
`as the column chromatography.
`
`thereof,
`and derivatives
`[0062] Natural milbemycins
`which are starting materials of compound (III), are fermen—
`tation products and may be any of a single compound or a
`mixture thereof. Therefore, compound (I) can also be pre-
`pared as a single compound or as a mixture thereof.
`
`[0063] A milbemycin derivative which is an active ingre-
`dient of the present invention can be mixed with a second
`ingredient which is another active ingredient in a certain
`ratio and can be prepared to a formulation suitable for oral
`administration or local administration.
`
`[0064] As combinations of the milbemycin derivatives
`and the second ingredient, compound (I) and praziquantels,
`compound (I) and fipronils, and compound (I), praziquantels
`and fipronils, can be raised. Further, when a plurality of
`compounds are used as the second ingredient, the blending
`ratio of the compounds can be varied depending on the
`combination method and the intended purpose. For example,
`there can be used a combination that fipronil is 0.2 and
`praziquantel is 0.5 in terms of a weight ratio, when com-
`pound (I) is recognized as 1.
`
`In the composition of the present invention, the
`[0065]
`blending ratio of the second ingredient to compound (I)
`varies depending on the sort and the use of the second
`ingredient to be used. For example, when the second ingre-
`dient is fipronil, the ratio is normally from 0.1 to 5, prefer-
`ably from 0.2 to 1.0. When the second ingredient is prazi—
`quantel, the ratio is normally from 0.1 to 2, preferably from
`0.2 to 0.5. When the second ingredient is benzimidazole, the
`ratio is normally from 1.0 to 100, preferably from 2.0 to 20.
`When the second ingredient are neonicotinoide, the ratio is
`normally from 0.2 to 10, preferably from 0.5 to 2.0. In
`
`addition, when a plurality of compounds are used as the
`second ingredient, the blending ratio of each ingredient can
`be prepared to be Within the above-described range, respec-
`tively.
`
`[0066] When the composition of the present invention is
`used as an endo- and ectoanthelrnintics in an animal or a
`human, it can be administered orally as a liquid drink. The
`liquid drink is an appropriate non-toxic solvent or solution
`in water, suspension or dispersion with a suspending agent
`such as bentonite and a wetting agent or other excipients.
`The liquid drink,
`in general, may also contain an anti-
`foaming agent. The content of the active component in a
`drink formulation such as liquid drink is usually from 0.01
`to 0.5% by mass, preferably from 0.01 to 0.1% by mass.
`
`[0067] When it is desired to administer orally the compo-
`sition of the present invention in a unit dosage form as a
`dried solid, capsules, pills or tablets, containing the desired
`amount of the active ingredient are normally employed.
`These dosage forms are prepared by mixing the active
`ingredient uniformly with suitable pulverized diluents, fill—
`ers, disintegrators and/or binding agents, for example starch,
`lactose, talc, magnesium stearate, vegetable gum and the
`like. Such unit dosage formulations may vary widely con-
`cerning the weight and contents of the insecticide depending
`upon the species of the host animal to be treated, the degree
`of the infection, the species of the parasite and the body
`weight of the host.
`
`[0068] When a composition of the present invention is
`administered by animal feedstuffs, the composition is dis-
`persed uniformly in the feedstulfs, used as a top dressing or
`used in the form of pellets. The content of active ingredient
`in the final feedstulf is usually from 0.0001 to 0.02% by
`mass, in order to achieve the desired anthelmintic effect.
`
`[0069] Further, a product dissolved or dispersed in a liquid
`carrier excipient can be administered to an animal parenter-
`ally into the proventriculus, the muscle or the trachea or by
`a hypodermic injection. For parenteral administration, the
`active ingredient is preferably mixed with a suitable veg—
`etable oil such as peanut oil or cottonseed oil. The content
`of the active ingredient in such formulation is generally from
`0.05 to 50% by weight.
`
`[0070] Further, another desirable administration form is a
`method where preparations are dissolved in a solvent and
`administered directly to a local site. As for such a solvent,
`the use of an alcohol such as ethanol, isopropanol, oleyl
`alcohol or benzyl alcohol; a carboxylic acid such as lauric
`acid or oleic acid; an ester such as ethyl lactate, isopropyl
`myristate or propylene carbonate; a sulfoxide such as dim—
`ethylsulfoxide; or an amide such as N—methylpyrrolidone,
`which are known to heighten percutaneous absorptivity,
`individually or as mixed solvent thereof, is preferable.
`
`In the present invention, the amount of the active
`[0071]
`ingredient to be used is varied depending upon the species
`of the animal to be treated, and the type and degree of
`parasitic infection, and it is usually from 0.01 to 100 mg,
`preferably from 0.5 to 50.0 mg, per 1 kg of body weight of
`the animal, and oral administration is desirable. Such a dose
`can be administrated in a single dose or in divided doses
`over a relatively short period such as from 1 to 5 days.
`
`[0072] The composition of the present invention has a
`high anthelmintic activity against wide range of endo- or
`
`

`

`US 2004/0254125 A1
`
`Dec. 16, 2004
`
`ectoparasites and exhibits excellent control effects against
`various diseases caused by insects and the parasites being
`parasitic on animals.
`
`[0073] That is, the composition of the present invention
`exerts an excellent insecticidal elfect against fleas being
`parasitic on a pet or a human, and is extremely effective as
`anthelmintics against them.
`
`[0074] As fleas to be targeted, Ctenocephalides felis,
`Ctenocephalides canis and so forth can be raised.
`
`In the field of veterinary medicines, the composi—
`[0075]
`tion of the present invention can be used against various
`harmful parasites of animals (endo— and ectoparasites), for
`example,
`insects and helminthes. As examples of these
`parasites of animals, Gasterophilus spp., Stomoxys spp.,
`Trichodectes spp., Rhodnius spp., Hypoderma spp., Oestrus
`spp., Haematopinus spp and so forth can be raised. Further,
`the composition of the present invention has an excellent
`miticidal activity against Ixodidae, Dermanyssid-ac, Sarcop-
`tidae, Argasidae, Dermanyssid and Psoroptidae, which are
`parasitic on animals.
`
`invention has
`[0076] The composition of the present
`excellent parasiticidal activity as an anthelmintic for an
`animal and a human, and in particular, is effective against
`nematodes which infect livestock, poultry and pet animals
`such as pigs, sheep, goats, cows, horses, dogs, cats and fowl.
`As such nematodes, for example, Haemonchus, Trichos—
`trongylus, Ostertagia, Nematodirus, Cooperia, Ascaris,
`Bunostomum, Oesophagostomum, Chabertia, Trichuris,
`Storongylus, Trichonema, Dictyocaulus, Capillaria, Heter-
`akis, Toxocara, Ascaridia, Oxyuris, Ancylostoma, Unci-
`narz'a, Toxascaris, and Parascan's can be raised. In particu-
`lar, certain species of the genera Nematodirus, Coaperia and
`Oesophagostomum attack the intestines, while those of the
`genera Haemonchus and Ostertagia parasitize the stomach,
`and parasites of genus Dictyocaulus can be found in the
`lungs, and the composition of the present invention also
`exhibits the activity against these. Further, parasites belong-
`ing to the families Filariidae and Setariidae can be found in
`other tissues or organs such as the heart, blood vessels and
`subcutaneous and lymphatic tissues. The composition of the
`invention also exhibits the activity against these parasites.
`
`[0077] The composition of the present invention exhibits
`activity also against cestoid and flukes. As the cestoids, for
`example,Dipylidium caninum, Taenia taeniaeformis, Taenia
`solium, Taenia saginata, Hymenolepis diminuta, Moniezia
`benedem', Diphyllobothrium latum, Diphyllobothrium eri-
`nacei, Echinococcus granulosus, and Echinococcus mul-
`tilocularis can be raised and as the flukes, Fasciola hepatica,
`nggamica, Paragonimus westermanii, Fasciolopsic bruski,
`Eurytrema pancreaticum, E.coelamaticum, Clonorchis sin-
`ensis, Schistosoma japonicum, S.haematobium, and S.man-
`soni can be raised.
`
`[0078] The composition of the present invention is also
`useful against parasites which infect a human, and as para—
`sites of human digestive, for example, Ancylostoma, Neca-
`tor, Asdaris, Strangyloides, Trichinella, Capillaria, Trichu-
`ris, and Enterobius can be raised.
`
`[0079] The compound of the present invention also exhib-
`its the activity against parasites of the genera Wuchereria,
`Brugia, Onchoceca, Dirofilaria, Lou, and Dracunculidae of
`the family Filariidae which are found in blood or other
`
`tissues and organs other than the digestive tract and are
`medically important, parasites of the genus Dracunclidae of
`the family Deacunculus and parasites of the genera Strongy-
`[aides and Trichinella, which in a particular state may
`parasitize outside the intestinal
`tract, although they are
`essentially intestinal parasites.
`
`BEST MODE FOR CARRYING OUT THE
`INVENTION
`
`[0080] Although the composition of the present invention
`is explained below in more detail by referring to Reference
`Examples and Test Examples, the composition of the present
`invention is not limited thereto.
`
`REFERENCE EXAMPLE 1
`
`13—[1—(4-methoxyaoetylaminophenyl) cyclo-
`[0081]
`pentanecarbonyloxy]-5-hydroxy-milbemycin A4 (I:
`R1=Et, Rz-MeOCHzCO, Compound No. 4)
`[0082]
`(1) 13-[1-(4-Nitrophenyl)cyclopentanecarbo-
`nyloxy]-5-oxomilbemycin A4 (IV: RlsEt, X=N02,
`Step A)
`
`[0083] Trifluoromethanesulfonic acid (0.18 ml) were
`added to a suspension of 1—(4-nitrophenyl)cyclopentanecar-
`boxylic acid (10.13 g, 40.0 mmol) in dichloromethane (150
`ml) and stirred for 20 minutes in a nitrogen stream. To the
`obtained transparent reaction solution, 15 -hydroxy—5-oxom-
`ilbemycin A4 (5.57 g, 10.0 mmol) were added and further
`continuously stirred for 30 minutes. After the completion of
`reaction, the reaction solution was poured into 4% aqueous
`sodium hydrogencarbonate solution and then extracted with
`ethyl acetate. The extracted solution was washed with a 4%
`aqueous sodium hydrogencarbonate solution and water,
`dried over magnesium sulfate and then, the solvent was
`removed. The obtained residue containing 13-[1-(4-nitro-
`phenyl)cyclopentanecarbonyloxy]—5-oxomilbemycin
`A4
`was used in the next step without further purification. (2)
`13-[1-(4-Nitrophenyl)cyclopentanecarbonyloxy]-5-hy-
`droxymilbemycin A4 (V: R1=Et, Step B)
`
`[0084] The S-oxo derivative obtained in the Step A was
`dissolved in methanol (200 ml), to which sodium borohy-
`dride (1.52 g, 40 mmol) and boron trifluoride.ether adduct
`(two drops) were added at —40° C. and stirred for 10
`minutes. After the completion of reaction, ethyl acetate (400
`ml) was added. The mixed solution was washed three times
`with water and then dried over anhydrous sodium sulfate.
`The solvent was removed under a reduced pressure. The
`obtained residue was purified by chromatography on a silica
`gel column (eluant: ethyl acetate/hexane-1/1)to give 6.79 g
`of
`13-[1-(4—nitrophenyl)cyclopentanecarbonyloxy]-5-hy-
`droxymilbemycin A4.
`
`[0085] NMR Spectrum (400 MHz) 5 (ppm): 8.16(2H,
`d,7.9 Hz), 7.53(2H, d, J=7.9 Hz), 5.70-5.90(2H, m),
`5.37(1H, s), 5.25-5.40(3H, m), 4.84(1H, d, J-10.6 HZ), 4.67
`and 4.63(2H, AB—q, J=14.4 Hz), 4.28(1H, m), 4.07(1H, s),
`3.94(1H, d, J=6.4 Hz), 3.55(1H, m), 3.25(1H, m), 3.02(1H,
`m), 1.88(3H, s), 1.29(3H, s), 0.99(3H, t, J=7.3 Hz), 0.82(3H,
`d, J=6.5 Hz), 0.77(3H, d, J=6.4 Hz).
`
`13—[1-(4-Aminophenyl)cyclopentanecarbo-
`(3)
`[0086]
`nyloxy]-5—hydroxymilbemycin A4 (VI: Rl-Et, Step C)
`
`[0087] The nitro derivative obtained in (2) was dissolved
`in ethyl acetate (100 m1), and Methanol (100 ml) and a
`
`

`

`US 2004/0254125 A1
`
`Dec. 16, 2004
`
`triphenylphosphine complex of nickel (II) chloride (1.12 g,
`1.7 mmol) were added to the solution, to which sodium
`borohydride (100 mg, 2.6 mmol) were gradually added over
`10 minutes under ice cooling. After being stirred for 10
`minutes, ethyl acetate (70 ml) was added to the reaction
`solution, and which was then washed three times with water
`and dried over anhydrous sodium sulfate, and the solvent
`was then removed under reduced pressure. The obtained
`residue was crystallized by adding ethyl acetate hexane to
`give 5.87 g of 13-[1-(4-aminophenyl)cyclopentanecarbo-
`nyloxy]-5-hydroxymilbemycin A4 as a crystal.
`
`[0088] Melting Point: 235-239° C.
`
`[0089] NMR Spectrum (400 MHz) 6 (ppm): 7.16(2H, d,
`J-8.6 Hz), 6.65(2H, d, 8.6 Hz), 5.77-5.84(2H, m), 5.44(1H,
`s), 5.32-5.43(3H, m), 4.84(1H, d, J=10.5 Hz), 4.70 and
`4.73(2H, AB-q, J=14.5 Hz), 4.33(1H, m), 4.13(1H, s),
`4.00(1H, d, J=6.2 Hz), 3.54(1H, m), 3.30(1H, m), 3.09(1H,
`m), 1.92(3H, s), 1.36(3H, s), 1.01(3H, t, J=7.3 Hz), O.87(3H,
`d, J=6.5 Hz), 0.82(3H, d, J=6.5 Hz)
`
`13-[1-(4-Methoxyacetylaminophenyl)cyc-
`(4)
`[0090]
`lopentanecarbonyloxy]-5-hydroxymilbemycin A4 (I: Rl-Et,
`R2=MeOCH2CO, Compound No. 4, Step D)
`
`[0091] The amino derivative obtained in (3) was dissolved
`in tetrahydrofuran (60 m1), followed by adding pyridine
`(0.52 ml, 6.4 mmol) and methoxyacetyl chloride (0.58 ml,
`6.2 mmol) at -30° C., and stirring for 10 minutes. After the
`completion of reaction, ethyl acetate (250 ml) was added to
`the reaction solution, which was washed successively with
`0.5 mol/L citric acid solution, water, 4% aqueous sodium
`hydrogencarbonate solution and water, and then dried over
`anhydrous sodium sulfate, and the solvent was then removed
`under a reduced pressure. The obtained residue was purified
`by chromatography on a silica gel column (eluant: ethyl
`acetate/hexane-Z/l) to give 4.38 g of the desired compound.
`
`[0092] NMR Spectrum (400 MHZ) 5 (ppm): 8.20(1H, s),
`7.49(2H, d, J-8.6 HZ), 7.29(2H, d, J-8.6 HZ), 5.77(1H, m),
`5.74(1H, m), 5.39(1H, s), 5.27-5.37(3H, m), 4.80(1H, d,
`J-10.5 Hz), 4.68 and 4.64(2H, AB-q, J=14.5 Hz), 4.28(1H,
`m), 4.08(1H, s), 4.01(2H, s), 3.95(1H, d, J=6.4 Hz),
`3.54(1H, m),3.51(3H,
`s), 3.25(1H, m), 3.01(1H, m),
`2.64(2H, m), 2.32(1H, d, J=8.1 Hz), 1.87(3H, s), 0.96(3H, t,
`J=7.3 Hz), 0.82(3H, d, J=6.5 Hz), 0.76(3H, d, J=6.5 Hz).
`
`REFERENCE EXAMPLE 3
`
`13-[1-(4-Cyanoacetylaminophenyl)cyclo-
`[0096]
`pentanecarbonyloxy]—5-hydroxymilbemycin A4 (I: RlsEt,
`R2=NCCH2CO, Compound No. 3)
`
`In the same manner as in Reference Example 1 (4)
`[0097]
`except for using cyanoacetyl chloride in place of methoxy-
`acetyl chloride in Reference Example 1 (4), the title com-
`pound was produced (yield in the Step D: 89.8%).
`
`[0098] NMR Spectrum (400 MHz) 6 (ppm): 7.68(1H, s),
`7.42(2H, d, J=8.7 Hz), 7.33(2H, d, 8.7 Hz), 5.72-5.77(2H,
`m), 5.39(1H, s), 5.28-5.37(3H, m), 4.81(1H, d, J=10.6 Hz),
`4.64 and 4.68(2H,AB-q, J-14.5 Hz), 4.29(1H, m), 4.07(1H,
`s), 3.95(1H, d, J=6.2 Hz), 3.55(2H, s), 3.52(1H, m),3.25(1I-I,
`m), 3.01(1H, m), 2.62(2H, 2), 2.32(1H, d, J=8.2 Hz),
`1.87(3H, s), 0.96(3H, t, J=7.3 Hz), 0.82(3H, d, J-6.5 Hz),
`0.76(3H, d, J=6.5 Hz).
`
`REFERENCE EXAMPLE 4
`
`13-[1-(4-Methanesulfonylaminophenyl)cy-
`[0099]
`clopentanecarbonyloxy]-5~hydroxymilbemycin A4
`R1=Et, Rz-MeSOZ, Compound No. 5)
`
`(I:
`
`In the same manner as in Reference Example 1 (4)
`[0100]
`except for using methanesulfonyl chloride in place of meth-
`oxyacetyl chloride in Reference Example 1 (4),
`the title
`compound was produced (yield in the Step D: 88.2%).
`
`[0101] NMR Spectrum (400 MHz) 5 (ppm): 7.32(2H, d,
`J=8.7 Hz), 7.14(2H, d, 8.7 Hz), 6.47(1H, s), 5.70-5.80(2H,
`m), 5.39(1H, s), 5.25-5.37(3H, m), 4.80(1H, d, J=10.6 Hz),
`4.64 and 4.68(2H, AB—q, J=14.0 Hz), 4.28(1H, dd, J=6.4 and
`8.2 Hz), 4.07(1H, s), 3.95(1H, d, J=6.4 Hz), 3.54(1H,
`m),3.25(1H, m), 3.01(1H, m), 2.93(3H, s), 2.62(2H, 2),
`2.33(1H, d, J=8.2 Hz), 1.87(3H, s), 0.96(3H, t, J-7.3 Hz),
`0.82