Title of Invention

A SPOT-ON FORMULATION USEFUL FOR THE TREATMENT OF PARASITE INFESTATION

Abstract This invention provides for; inter alia, spot-on compositions for the treatment or prophylaxis of parasite infestations in mammals or birds which comprise: (1) at least one nodulisporic acid derivative; (2) an acceptable liquid carrier vehicle; and (3) optionally, a crystallization inhibitor. The invention also provides for a method of treating parasitic infestations or for the prophylaxis of parasite infestations in mammals or birds which comprise topically applying to said mammal or bird an effective amount of a composition according to the present invention. Further, this invention relates to an amidation process to prepare amide derivatives of nodulisporic acid and to intermediate formed in this process.
Full Text A SPOT-ON FORMULATION USEFUL FOR THE TREATMENT OF PARASITE
INFESTATION
RELATED APPLICATIONS
This application claims priority to Provisional Application USSN 60/415627 entitled"A Method for the Synthesis of Nodulisporamide"filed October 2,2002 and to Non-Provisional Application USSN 10/618975 entitled:"Nodulisporic Acid Derivative Spot-On Formulations For Combating Parasites"Filed July 14,2003.
This application Reference is also made to copending application USSN 10/279,356, filed October 24,2002, which in turn is a continuation-in-part of application USSN 10/155, 397, filed May 24, 2002, now pending, which in turn is a divisional of application 09/376,736, filed August 17,1999, now U. S. Patent No. 6,426, 333 issued on July 30,2002, which in turn is a continuation in-part of application USSN 09/271,470, filed March 17,1999, now allowed, which in turn is a continuation-in-part of copending International Application PCT/FR97/01548 having an international filing date of 15 September 1997, and designating the U. S. and claiming priority from French Application No. 96/11446, filed 19 September 1996. Reference is also made to: U. S. Applications Serial Nos. 10/052,597, filed January 17,2002, 10/120,691, filed April 11,2002, 08/719, 942, filed September 25,1996, 08/692,430, filed August 5,1996, 08/863,182, filed May 27,1997, 08/692,113, filed August 5,1996, 08/863,392, filed May 27,1997, and 08/891,047, filed July 10,1997 ; French Application No. 97 03709, filed March 26, 1997; and PCT/FR98/00601. All of the above-mentioned applications, as well as all documents cited therein, including parent applications if available, and documents referenced or cited in documents cited herein, are hereby incorporated herein by reference.
FIELD OF THE INVENTION
This invention relates to inter alia solonjormulations for combating parasites in birds and mammals. In p articular, this invention provides for spot-on formulations comprising a composition comprising at least one nodulisporic acid derivative and a pharmaceutically or veterinary acceptable liquid carrier vehicle. This invention also provides for an improved method for eradicating, controlling, and preventing parasite infestation in birds and mammals. Further, this invention relates to an amidation process
to prepare amide derivatives of nodulisporic acid and to jntermediates formed in the process.
BACKGROUND OF THE INVENTION
Animals such as mammals and birds are often susceptible to parasite infestations. These parasites may be ectoparasites, such as insects, and endoparasites such as filariae and worms.
Domesticated animal, such as cats and dogs, are often infested with one or more of the following ectoparasites:
- cat and dog fleas (Ctenocephalides felis, Ctenocephalides sp. and the like),
- ticks (Rhipicephalus sp., Ixodes sp., Dermacentor sp., Amblyomma sp. and the like), and

- mites (Demodex sp., Sarcoptes sp., Otodectes sp. and the like),
- lice (Trichodectes sp., Cheyletiella sp., Lignonathus sp., and the like), and
- flies (Hematobia sp., Musca sp., Stomoxys sp., Dermatobia sp., Coclyomia sp., and the like).
Fleas are a particular problem because not only do they adversely affect the health of the animal or human, but they also cause a great deal of psychological stress. Moreover, fleas are also vectors of pathogenic agents in animals, such as dog tapeworm (Dipylidium caninum), and humans.
Similarly, ticks are also harmful to the physical and psychological health of the animal or human. However, the most serious problem associated with ticks is that they are the vector of pathogenic agents, agents which cause diseases in both humans and animal. Major diseases which are caused by ticks include borreliosis (Lyme disease caused by Borrelia burgdorferi), babesiosis (or piroplasmosis caused by Babesia sp.) and rickettsiosis (also known as Rocky Mountain spotted fever). Ticks also release toxins which cause inflammation or paralysis in the host. Occasionally, these toxins are fatal to the host, such as in the case of the Australian paralysis tick, Ixodes holocyclus.
Moreover, mites and lice are particularly difficult to combat since there are very few active substances which act on these parasites and they require frequent treatment.
Likewise, farm animals are also susceptible to parasite infestations. For example, cattle are affected by a large number of parasites. Likewise, arthropod pests, such as flea, lice and ticks, infest poultry. A parasite, which is very prevalent among farm animals, is
a tick genus Boophilus, especially those of the species microplus (cattle tick), decoloratus and anulatus. Ticks, such as Boophilus microplus, are particularly difficult to control because they live in the pasture where the farm animals graze. Other important parasites of cattle and sheep are listed as follows in order of decreasing importance:
- myiases such as Dermatobia hominis (known as Berne in Brazil) and Cochlyomia hominivorax (greenbottle); sheep myiases such as Lucilia sericata, Lucilia cuprina (known as blowfly strike in Australia, New Zealand and South Africa). These are flies whose larva constitutes the animal parasite;
- flies proper, namely those whose adult constitutes the parasite, such as Haematobia irritans (horn fly);
lice such as Linognathus vitulorum, etc.; and
- mites such as Sarcoptes scabiei and Psoroptes ovis.
The above list is not exhaustive and other ectoparasites are well known in the art to be harmful to animals and humans. These include, for example migrating dipterous larvae.
Animals and humans also suffer from endoparasitical infections including, for example, helminthiasis, which is most frequently caused by a group of parasitic worms described as nematodes or roundworms. These parasites cause severe economic losses in pigs, sheep, horses, and cattle as well as affecting other domestic animals, such as dogs, cats and poultry. Other parasites which occur in the gastrointestinal tract of animals and humans include Ancylostoma, Anecator, Ascaris, Strongyloides, Trichinella, Capillaria, Toxocara, Toxascaris, Trichiris, Enterobius and parasites which are found in the blood or other tissues and organs such as filarial worms and the extra intestinal stages of Strogyloides, Toxocara and Trichinella.
Many insecticides exist in the art for treating parasites. These insecticides vary in their effectiveness to a particular parasite as well as their cost. However, the results of these insecticides is not always satisfactory because of, for example, the development of resistance by the parasite to the therapeutic agent, as is the case, for example, with carbamates, organophosphorous compounds and pyrethroids. Moreover, there is at the present time no truly effective method for controlling the set of parasites indicated above. Thus, there is a need in the art for more effective antiparasitic formulation treatment and protection of animal, e.g. mammals, fish and birds for a wide range of parasites. Moreover, there is a need in the art for an antiparasitic formulation which is easy to use on any type of domestic animal, irrespective of its size and the nature of its coat and
which do not need to be sprinkled over the entire body of the mammal, fish or bird. Further, the formulation should be effective for a long period of time thereby reducing the number of times it has to be applied.
Compounds, which exhibit a degree of activity against a wide range ectoparasites, are known in the art. The arylpyrazoles as a class are known in the .art .arid are described, for example, in copending applications USSN 07/719,942; 08/933,016; 09/174,598; 08/863,182; and 08/863,692, as well as in U.S. Patent No. 5,576,429; U.S. Patent No. 5,122,530, EP-A-295,217, EP-A-352,944 and EP 295 177, the disclosures of which, as well as the references cited herein, are incorporated by reference. Reference is made to, for example, U.S. patent No. 5,567,429, U.S. Patent No. 5,122,530, EP 295,117, and EP 846,686 Al (or Banks GB 9,625,045, filed November 30, 1996 also believed to be equivalent to USSN 309,229, filed November 17, 1997). This class of insecticides is known to possess excellent activity against insects, such as ticks and fleas. Fipronil is a 1-N-aryl pyrazole that is particularly effective against fleas and ticks
Other compounds parasiticides that are known in the art to be effective are those which possess a macrocyclic lactone ring. These compounds are particularly effective against ectoparasites, including lice, blowflies, flies, mosquitoes, mites, migrating dipterous larvae, and ticks, as well as endoparasites, such as nematodes and roundworms. Compounds of this group include avermectins, milbemycins, and derivatives of these compounds, for example, ivermectin or emamectin. Such substances are described, for example, in U.S. Patents 3,950,360; 4,199,569; 4,879,749; and 5,268,710.
These compounds are well known to a person skilled in the art and are easily obtained either commercially or through techniques know in the art. Reference is made to the widely available technical and commercial literature. For avermectins, ivermectin and abamectin, reference may be made, for example, to the work "Ivermectin and Abamectin", 1989, by M.H. Fischer and H. Mrozik, William C. Campbell, published by Springer Verlag., or Albers-SchSnberg et al. (1981), "Avermectins Structure Determination", J. Am. Chem. Soc, 103, 4216-4221. For doramectin, "Veterinary Parasitology", vol. 49, No. 1, July 1993, 5-15 may in particular be consulted. For milbemycins, reference may be made, inter alia, to Davies H.G. et al., 1986, "Avermectins and Milbemycins", Nat. Prod. Rep., 3, 87-121, Mrozik H. et al., 1983, Synthesis of Milbemycins from Avermectins , Tetrahedron Lett., 24, 5333-5336, U.S. Patent 4, 134, 973 and EP 677,054. These compounds are either natural products or are semi-synthetic derivatives thereof. The structure of these compounds is closely related,
e.g., by sharing a complex 16-member macrocyclic lactone ring. The natural product avermectins are disclosed in U.S. Patent 4,310,519 to Albers-Schonberg, et al., and the 22,23-dihydro avermectin compounds are disclosed in Chabala, et al., U.S. Patent No. 4,199,569. Mention is also made of Kitano, U.S. Patent No. 4,468,390, Beuvry et al., U.S. Patent No. 5,824,653, European Patent Application 0 007 812 Al, published June 2, 1980, U.K. Patent Specification 1 390 336, published April 9, 1975, European Patent Application 0 002 916 A2, and Ancare New Zealand Patent No. 237 086, inter alia. Naturally occurring milbemycins are described in Aoki et al., U.S. Patent No. 3,950,360 as well as in the various references cited in "The Merck Index" 12th ed., S. Budavari, Ed., Merck & Co., Inc. Whitehouse Station, New Jersey (1996). Semisynthetic derivatives of these classes of compounds are well known in the art and are described, for example, in U.S. Patent 5,077,308, U.S. Patent 4,859,657, U.S. Patent 4,963,582, U.S. Patent 4,855,317, U.S. Patent 4,871,719, U.S. Patent 4,874,749, U.S. Patent 4,427,663, U.S. Patent 4,310,519, U.S. Patent 4,199,569, U.S. Patent 5,055,596, U.S. Patent 4,973,711, U.S. Patent 4,978,677, U.S. Patent 4,920,148 and EP 667 054,
Other classes of compounds include insect growth regulating (IGR) compounds, which
either mimic juvenile hormones or the inhibit synthesis of chintin. IGR compounds that
mimic juvenile hormones include, for example, azadirachtin, diofenolan, fenoxycarb,
hydroprene, kinoprene, methoprene, pyriproxyfen, tetrahydroazadirachtin, and
4-chloro-2-(2-chloro-2-methylpropyl)-5-(6-iodo-3-pyridylmethoxy)pyridizine-3(2H)-one.
Chintin-synthesis inhibitors include, for example, chlorfluazuron, cyromazine,
diflubenzuron, fluazuron, flucycloxuron, flufenoxuron, hexaflumuron, lufenuron,
tebufenozide, teflubenzuron, triflumuron, l-(2,6-difluorobenzoyl)-3-(2-
fluoro-4-(trifluoromethyl)phenylurea, 1 -(2,6-difluorobenzoyl) -3-(2-fluoro-4-(l, 1,2,2-tetrafluoroethoxy)phenylurea, and 1 -(2,6-difluorobenzoyl)-3-(2-fluoro-4-trifluoromethyl) phenylurea.
Another class of compounds, which are known in the art as potent endo- and
ectoantiparasitic agents, are nodulisporic acid derivatives. These compounds are based
upon three structures, A, B or C, which have the following structures: Nodulisporic acid (compound A)
(Formula Removed)
29,30-dihydro-20,30-oxa-nodulisporic acid (compound B)
(Formula Removed)

and
31-hydroxy-20,30-oxa-29,30,31,32-tetrahydro-nodulisporic acid (compound C)
(Formula Removed)

These compounds were obtained from the fermentation culture of Nodulisporium sp. MF-5954 (ATCC 74245) and the isolation and purification of the three nodulisporic acids are disclosed in US Patent 5,399,582. Derivatives of these compounds are described in WO 96/29073 and US Patent Nos. 5,945,317; 5,962,499; 5,834,260; 6,399,796; 6,221,894; 6,136,838; 5,595,991; and 5,614,546.
Nodulisporic acid derivatives possess potent activity against parasites, particularly helminths, ectoparasites, insects, and acarides, infecting man, animals and plants. These compounds have utility in human and animal health, agriculture and pest control in household and commercial areas.
The disease or group of diseases described generally as helminthiasis is due to infection of an animal host with parasitic worms known as helminths. Helminthiasis is a prevalent and serious economic problem in domesticated animals such as swine, sheep, horses, cattle, goats, dogs, cats, fish, buffalo, camels, llamas, reindeer, laboratory animals, furbearing animals, zoo animals and exotic species and poultry. Among the
helminths, the group of worms described as nematodes causes widespread and often times serious infection in various species of animals. The most common genera of nematodes infecting the animals referred to above are Haemonchus, Trichostrongylus, Ostertagia, Nematodirus, Cooperia, Ascaris, Bunostomum, Oesophagostomum, Chabertia, Trichuris, Strongylus, Trichonema, Dictyocaulus, Capillaria, Habronema, Druschia, Heterakis, Toxocara, Ascaridia, Oxyuris, Ancylostoma, Uncinaria, Toxascaris and Parascaris. Certain of these, such as Nematodirus, Cooperia, and Oesophagostomum attack primarily the intestinal tract while others, such as Haemonchus and Ostertagia, are more prevalent in the stomach while still others such as Dictyocaulus are found in the lungs. Still other parasites may be located in other tissues and organs of the body such as the heart and blood vessels, subcutaneous and lymphatic tissue and the like. The parasitic infections The parasitic infections known as helminthiases lead to anemia, malnutrition, weakness, weight loss, severe damage to the walls of the intestinal tract and other tissues and organs and, if left untreated, may result in death of the infected host. The compounds of this invention have activity against these parasites, and in addition are also active against Dirofilaria in dogs and cats, Nematospiroides, Syphacia, Aspiculuris in rodents, arthropod ectoparasites of animals and birds such as ticks, mites such as scabies lice, fleas, blowflies, and other biting insects in domesticated animals and poultry, such as Tenophalides, Ixodes, Psoroptes, and Hemotobia, in sheep Lucilia sp., biting insects and such migrating dipterous larvae as Hypoderma sp. in cattle, Gasterophilus in horses, and Cuterebra sp. in rodents and nuisance flies including blood feeding flies and filth flies.
Nodulisporic acid derivatives are also useful against parasites which infect mammals, such as cats, dogs and humans. The most common genera of parasites of the gastro-intestinal tract of man are Ancylostoma, Necator, Ascaris, Strongyloides, Trichinella, Capillaria, Trichuris, and Enterobius. Other medically important genera of parasites which are found in the blood or other tissues and organs outside the gastrointestinal tract are the filiarial worms such as Wuchereria, Brugia, Onchocerca and Loa, Dracunuculus and extra intestinal stages of the intestinal worms Strongyloides and Trichinella. The compounds are also of value against arthropods parasitizing man, biting insects and other dipterous pests causing annoyance to man.
Nodulisporic acid derivatives are also active against household pests such as the cockroach, Blatella sp., clothes moth, Tineola sp., carpet beetle, Attagenus sp., the housefly Musca domestica as well as fleas, house dust mites, termites and ants.
Nodulisporic acid derivatives are also useful against insect pests of stored grains such as Tribolium sp., Tenebrio sp. and of agricultural plants such as aphids, (Acyrthiosiphon sp.); against migratory orthopterans such as locusts and immature stages of insects living on plant tissue. The compounds are useful as a nematocide for the control of soil nematodes and plant parasites such as Meloidogyne sp., which may be of importance in agriculture. The compounds are also highly useful in treating acreage infested with fire and nests. The compounds are scattered above the infested area in low levels in bait formulations which are brought back to the nest. In addition to a direct-but-slow onset toxic effect on the fire ants, the compound has a long-term effect on the nest by sterilizing the queen which effectively destroys the nest.
Nodulisporic acid and its derivatives are also effective against arthropod pests, for example fleas, ticks, ice and other biting insects in domesticated animals and poultry, such as Ctenophalides, Ixodes, Psoroptes, Lucilia and Hematobia.
It is known to combine the above-mentioned classes of compounds in order to achieve a broader spectrum of activity or, in some instances synergy. For example, U.S. Patent 5,945,317 discloses co-administering nodulisporic acid derivatives with avermectin or milbemycins, or other antihelmintic agents, such as morantel, pyrantel, or febantel, or benzimidizoles, such as thiabendazole or cambendazole. Other agents described therein include IGR compounds, such as lufenuron, or 1-N-arylpyrazoles, such a fipronil. See also, U.S. Patent 5,962,499 and 6,221,894. While it is known in the art that it is sometimes possible to combine various parasiticides in order to broaden the antiparasitical spectrum, it is not possible to predict, a priori, which combinations will work for a particular animal or disease state. For this reason, the results of various combinations is not always successful and there is a need in the art for more effective formulations which may be easily administered to the animal.
Various methods of formulating antiparasitical formulations are known in the art. These include oral formulations, baits, dietary supplements, powders, shampoos, etc. Formulations for localized topical applications of antiparasitical formulations are also known in the art. For example, pour-on solutions comprising 1-N-phenylpyrazoles, such as fipronil, are known in the art and are described in copending application 08/933,016, herein incorporated by reference. Other methods for formulating antiparasitic agents include spot-on formulations.
Spot-on formulations are well known techniques for topically delivering an antiparasitic agent to a limited area of the host. For example, U.S. Patent 5,045,536
describes such formulations for ectoparasites. Moreover, it is generally known in the art to formulate avermectin and milbemycin derivatives as spot-on formulations. See, e.g. U.S. Patent 5,045,536; EP 677,054; U.S. Patent 5,733,877; U.S. Patent 5,677,332; U.S. Patent 5,556,868; and U.S. Patent 5,723,488. U.S Patent Nos. 5,962,499 and 5,595,998 generally discusses formulating nodulisporic acid derivatives as pour-on or spot-on formulations, with or without additional antiparasitic agents. However, as discussed in U.S. Patent 5,045,536, a large number of solvent systems described in the art provide formulations for localized topical application which cause irritancy or toxicity to the host as well as being effective for a long period of time. Hence, there is a need in the art both for more effective for a longer period of time and less irritant or toxic formulations. Thus, there is a need in the art for a spot-on formulation, which is effective over a long period of time against a wide range of endoparasites and ectoparasites in birds and mammals.
SUMMARY OF THE INVENTION
The invention provides inter alia for spot-on formulations for the treatment or prophylaxis of endoparasites of mammals, fish and birds, and in particular, cats, dogs, horses, chickens, sheep and cattle with the aim of ridding these hosts of all the parasites commonly encountered by birds and mammals. The invention also provides for effective and lasting destruction of ectoparasites, such as fleas, ticks, mites, e.g. itch mites, mosquitoes, flies and lice. Further, the spot-on formulations retain their efficacy over a long period of time, thereby reducing the number of applications of the formulation to the animal.
In particular this invention provides for spot-on formulations for the treatment or prophylaxis of parasite infestations in mammals or birds, which comprise:
(1) a composition comprising an effective amount of at least one nodulisporic acid derivative;
(2) a pharmaceutically or veterinary liquid carrier vehicle; and
(3) optionally, a crystallization inhibitor.
The invention also provides for an easy method of treating parasitic infestations or for the prophylaxis of parasite infestations in mammals or birds which comprises topically applying to said mammal or bird an effective amount of a formulation according to the present invention.
This invention further provides for formulations which, when applied locally, will diffuse over the entire body of the host and then dry, without crystallizing, and which do not affect the appearance of the coat after drying by, for example, leaving crystals or making the coat sticky. This has the further advantage in animals which groom themselves of not being orally ingested, where the therapeutic agent might not be well tolerated orally or might interact with other therapeutic agents.
The very high effectiveness of the method and of the formulations according to the invention provides not only for a high instantaneous effectiveness but also for an effectiveness of very long duration after the treatment of the animal.
This invention further provides for an amidation process to prepare amide derivatives of nodulisporic acid in higher yield with better purity. Novel intermediates in this process also form a part of this invention.
In this disclosure and in appended claims, terms such as"comprising"and "comprises"and the like, have the meanings ascribed to them in U. S. Patent Case Law. The terms"comprises"and"comprising"are open-ended and allow for the inclusion of additional ingredients or steps.
Clearly, a spot-on formulation spot-on comprising at least one nodulisporic acid derivative, advantageously t-butyl nodulisporamide in a veterinary acceptable liquid carrier vehicle is a basic or novel feature of the herein invention, as well as methods for preventing or treating parasites on an animal, e. g. , dog, cat, by applying the formulation, e. g. , monthly, and methods for preparing the formulations, e. g. , by administering the ingredients, are also novel and basic features of the invention. That the invention performs as herein described is surprising, unexpected and nonobvious.
These and other embodiments are disclosed or are obvious from and encompassed by, the following Detailed Description.
DETAILED DESCRIPTION
Accordingly, the present invention provides a spot-on formulation useful for the treatment or prophylaxis of parasite infestation in mammals or birds which comprises:
(a) at least one nodulisporic acid derivative of the kind as herein described;
(b) a pharmaceutically or veterinary acceptable liquid carrier vehicle;
(c) optionally an antioxidant; and
(d) optionally a crystallization inhibitor of the kind as herein described ;
wherein the (w/v) percent ratio of individual component used in the range of 1 -40% : 1 -99% : 0.005-1% : 1-20% respectively.
This invention provides for a spot-on formulation for the treatment and prophylaxis of parasite infestation in mammals or birds which comprises
(1) an effective amount of at least one nodulisporic acid derivative
(2) a pharmaceutically or veterinary acceptable liquid carrier vehicle; and
(3) optionally, a crystallization inhibitor
More especially preferred are spot-on formulations comprising:
(1) an effective amount of at least one nodulisporic acid derivative;
(2) a liquid carrier vehicle comprises a solvent and optionally a cosolvent wherein
the solvent is selected from the group consisting of acetone, acetonitrile, benzyl
alcohol, butyl diglycol, dimethylacetamide, dimethylformamide, dipropylene glycol
n-butyl ether, ethanol, isopropanol, methanol, diethylene glycol monoethyl ether,
ethylene glycol monomethyl ether, monomethylacetamide, dipropylene glycol
monomethyl ether, liquid polyoxyethylene glycols, propylene glycol, 2-
pyrrolidone, in particular N-methylpyrrolidone, diethylene glycol monoethyl ether,
ethylene glycol, diethyl phthalate fatty acid esters, such as the diethyl ester or
diisobutyl adipate, and a mixture of at least two of these solvents and the
cosolvent is selected from the group consisting of absolute ethanol, isopropanol or methanol; and
(3) optionally, a crystallization inhibitor selected from the group consisting of an anionic surfactant, a cationic surfactant, a non-ionic surfactant, an amine salt, an amphoteric surfactant or polyvinylpylTolidone, polyvinyl alcohols, copolymers of vinyl acetate and vinylpyrrolidone, polyethylene glycols, benzyl alcohol, mannitol, glycerol, sorbitol, polyoxyethylenated sorbitan esters; lecithin, sodium carboxymethylcellulose, and acrylic derivatives, or a mixture of these crystallization inhibitors.
This invention includes all nodulisporic acid derivatives know in the art, including all steroisomers, such as those described in the prior publication described above, which are expressly incorporated by reference. Especially preferred are spot-on formulations comprising nordulisporic acid derivatives of the formula:
(Formula Removed)
wherein Rl is (1) hydrogen, (2) optionally substituted alkyl, (3) optionally substituted alkenyl, (4) optionally substituted alkynyl, (5) optionally substituted cycloalkyl, (6) optionally substituted cycloalkenyl, where the substituents on the alkyl, alkenyl, alkynyl, cycloalkyl and cycloalkenyl are 1 to 3 groups independently selected from (i) alkyl, (ii) alkyl, where X is O or S (O) m (iii) cycloalkyl, (iv) hydroxy, (v) halogen, (vi) cyano, (vii) carboxy, (viii) NYIY2, where Y'and Y2 are independently H or alkyl, (ix) alkanoylamino, and (x) aroylamino wherein said aroyl is optionally substituted with 1 to 3 groups independently selected from Rf
(7) aryl or arylalkyl, wherein said aryl is optionally substituted with 1 to 3 groups independently selected from Rf, (8) perfluoroalkyl (9) a 5-or 6-member heterocycle containing from 1 to 4 heteroatoms independently selected from oxygen, sulfur and nitrogen atoms optionally substituted by 1 to 3 groups independently selected from hydroxy, oxo, alkyl and halogen, and which may be saturated or partly unsaturated, R2, R3, and R4 are independently ORa, OC02Rb, OC (O) NRCRd ; or R1 and R2 represent =O, =NORa or =N-NRCRd ; R5 and R6 are H; or R5 and R6 together represent-O-; R7 is (1) CHO, or (2) the fragment
(Formula Removed)
R8 is (1) H, (2) OR', or (3) NRcRd Rg is (1) H, or (2) ORa ; RIO is (1) CN, (2) C (O) ORb, (3) C (O) N (ORb) R, (4) C (O) NR R, (5) NHC (O) ORb, (6) NHC (O) NRCRd, (7) CH2ORa, (8) CH2OCO2Rb, (9) CH20C (O) NRcRd, (10) C (O) NRcNRcRd, or (11) C (O) NRCSO2Rb ; represents a single or a double bond; Ra is (1) hydrogen, (2) optionally substituted alkyl, (3) optionally substituted alkenyl, (4) optionally substituted alkynyl, (5) optionally substituted alkanoyl, (6) optionally substituted alkenoyl, (7) optionally substituted alkynoyl, (8) optionally substituted aroyl, (9) optionally substituted aryl, (10) optionally substituted cycloalkanoyl, (11) optionally substituted cycloalkenoyl, (12) optionally substituted alkylsulfonyl (13) optionally substituted cycloalkyl (14) optionally substituted cycloalkenyl where the substituents on the alkyl, alkenyl, alkynyl, alkanoyl, alkenoyl, alkynoyl, aroyl, aryl, cycloalkanoyl, cycloalkenoyl, alkylsulfonyl, cycloalkyl and cycloalkenyl are from 1 to 10 groups independently selected from hydroxy, alkoxy, cycloalkyl, arylalkoxy, NRgRh, C02Rb, CONRCRd and halogen, (15) perfluoroalkyl, (16) arylsulfonyl optionally substituted with 1 to 3 groups independently selected from alkyl, perfluoroalkyl, nitro, halogen and cyano, (17) a 5-or 6-member heterocycle containing 1 to 4 heteroatoms selected from oxygen, sulfur and nitrogen optionally substituted by 1 to 4 groups independently selected from alkyl, alkenyl,
perfluoroalkyl, amino, C (O) NRCRd, cyano, C02Rb and halogen, and which may be saturated or partly unsaturated ; is (1) H, (2) optionally substituted aryl, (3) optionally substituted alkyl, (4) optionally substituted alkenyl, (5) optionally substituted alkynyl, (6) optionally substituted cycloalkyl, (7) optionally substituted cycloalkenyl, or (8) optionally substituted heterocycle containing from 1 to 4 heteroatoms independently selected from oxygen, sulfur and nitrogen; where the substituents on the aryl, alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocycle, or alkynyl are from 1 to 10 groups independently selected from (i) hydroxy, (ii) alkyl, (iii) oxo, (iv) S02NRgRh, (v) arylalkoxy, (vi) hydroxyalkyl, (vii) alkoxy, (viii) hydroxyalkoxy, (ix) aminoalkoxy, (x) cyano, (xi) mercapto, (xii) alkyl-S (O) m, (xiii) cycloalkyl optionally substituted with 1 to 4 groups independently selected from Re, (xiv) cycloalkenyl, (xv) halogen, (xvi) alkanoyloxy, (xvii) C (O) NRgRh, (xviii) C02R', (xix) formyl, (xx)-NRgRh, (xxi) 5 to 9-member heterocycle, which may be saturated or partially unsaturated, containing from 1 to 4 heteroatoms independently selected from oxygen, sulfur and nitrogen, and optionally substituted with 1 to 5 groups independently selected from Re, (xxii) optionally substituted aryl, wherein the aryl substituents are 1,2-methylenedioxy or 1 to 5 groups independently selected from Re, (xxiii) optionally substituted arylalkoxy, wherein the aryl substituents are 1,2-methylenedioxy or 1 to 5 groups independently selected from Re, and (xxiv) perfluoroalkyl ; Re and Rd are independently selected from Rb ; or Re and together with the N to which they are attached form a 3-to 10-member ring containing 0 to 2 additional heteroatoms selected from O, S (O)"" and N, optionally substituted with 1 to 3 groups independently selected from Rg, hydroxy, thioxo and oxo; Re is (1) halogen, (2) alkyl, (3) perfluoroalkyl, (4) _S (O) mR\ (5) cyano, (6) nitro, (7) Rio (CH2) v-, (8) R'C02 (CH2) v-, (9) R'OCO (CH2) v-, (10) optionally substituted aryl where the substituents are from 1 to 3 of halogen, alkyl, alkoxy, or hydroxy, (11) S02NRgRh, or (12) amino ; Rf is (1) alkyl, (2) X-CI-C4 alkyl, where X is O or S (O) m, (3) alkenyl, (4) alkynyl, (5) perfluoroalkyl, (6) NYIY2, where yl and y2 are independently H or alkyl, (7) hydroxy, (8) halogen, and (9) alkanoylamino, Rg and Rh are independently (1) hydrogen, (2) alkyl optionally substituted with hydroxy, amino, or C02R' (3) aryl optionally substituted with halogen, 1,2-methylenedioxy, alkoxy, alkyl or perfluoroalkyl, (4) arylalkyl, wherein the aryl is optionally substituted with perfluorolkyl or 1, 2-methylenedioxy ; (5) alkoxycarbonyl, (6)
alkanoyl, (7) alkanoylalkyl, (9) aryl alkoxycarbonyl, (10) aminocarbonyl, (11) monoalkylaminocarbonyl (12) dialkylaminocarbonyl; or Rl and together with the N to which they are attached form a 3-to 7-member ring containing 0 to 2 additional heteroatoms selected from O, S (O)m, and N, optionally substituted with 1 to 3 groups independently selected from Re and oxo; R'is (1) hydrogen, (2) perfluoroalkyl, (3) alkyl, (4) optionally substituted aryl or arylalkyl, where the aryl substituents are from 1 to 3 groups independently selected from halogen, alkyl, alkoxy, and hydroxy; m is 0 to 2; and v is 0 to 3 ; or a pharmaceutical^/ acceptable salt thereof
or a fragment
(Formula Removed)
wherein Rx is selected from the group consisting of: H, CH3, CH2CH3, C(CH3)3, CH2CH2CH3, CH2CH20H, CH(C02CH3) CH20H, CH2C02CH3, CH2CH(OCH2CH3)2, CH2CH20CH2CH20H, CH(CH3)(CH2)3C(CH3)20H, (CH2)30H, (CH2)40H, (CH2)SOH, CH (CH20H)CH2CH3, NHC(CH3)3, CH2CN, (CH2)60H, CH2CH(OH)CH3, CH(CH20H) CH2CH2CH3, CH2CH2SCH3, CH2CH2SCH2CH3, CH2CONH2, CH(CH3)(CH20H), CH2CH2NHCH2CH20H, CH (CH20H) (CH2) 3CH3, CH (CH20CH3) CH3, (CH2) 2SH, (CH2) 4NH2, CH2CH2S02CH3, CH2CH2S(0)CH3, CH(CH(CH3)2) CH20H, (CH2)3NH2, (CH2) 3N (CH2CH3) 2, (CH2)3N(CH3) 2, OCH2CH3, CH2CH(OH)CH20H, OCH3, CH2CH20CH3, CH2CH2NHC (O) CH3, C (CH3) 2CH20H, c-C3Hs, CC6H11, (CH2) 30CH2CH3, CH2CH=CH2, C(CH2CH3)(CH20H)2, CH2C=CH, CH2C02CH2CH3, CH2CH2F, (CH2)30(CH2)ICH3, CH2CH2N(CH3)2, CH2CH2IOCH2CH2NH2, CH2CF3, NHCH2C02CH2CH3, CH(CH3) C02CH3, C(CH3)2CH2C(0)CH3, CH(C02CH2CH3)2, CH2CH3, CH(CH2CH2CH3) C02CH3, CH2CH2CH20CH3, C(CH3) 2CH2CSCH, (CH2)4CH3, CH(CH2CH2CH3)2, (CH2) SCH3, CH2CH2C02H, CH(CH(CH3) 2)C02CH3, OCH2C02H, CH(CH(CH3)2) CH20H, CH (CH (CH3)2) CH20H,
CH(CH3)CH20H, CH(CH3)CH20H, CH(CH3)2, (CH2)CH(CH3)2, CH (CH3)CH2CH3, CH2CH(CH3)OH, (CH2)3CH3, (CH2)20CH2CH3, 1-adamantyl, (CH2)8CH3, CH(CH3)CH(CH3)2, (CH2)3NHCH3, (CH2) 2N(CH2CH3)2,
In a preferred embodiment, the present invention provides compounds of Formula I wherein R1 is (1) hydrogen, (2) optionally substituted alkyl, (3) optionally substituted alkenyl, (4) optionally substituted alkynyl, (5) optionally substituted cycloalkyl, (6) optionally substituted cycloalkenyl where the substituents on the alkyl, alkenyl, alkynyl, cycloalkyl and cycloalkenyl are 1 to 3 groups independently selected from (i) alkyl, (ii) X-CI-C6 alkyl, where X is O or S (O) m) (iii) cycloalkyl, (iv) hydroxy, (v) halogen, (vi) cyano, (vii) carboxy, and (viii) NYIY2, where Y'and Y2 are independently H or alkyl, (7) aryl or arylalkyl wherein said aryl is optionally substituted with 1 to 3 groups independently selected from R, (8) perfluoroalkyl, (9) a 5-or 6-member heterocycle containing from 1 to 4 heteroatoms independently selected from oxygen, sulfur and nitrogen atoms optionally substituted by 1 to 3 groups independently selected from hydroxy, oxo, alkyl and halogen, and which may be saturated or partly unsaturated, R8 is (1) H, (2) OH, or (3) NH2 ; R9 is (1) Hor (2) OH; Rio is (1) C (O) ORb, (2) C (O) N (ORb)Rc, (3) C (O) NR R, (4) NHC (O) ORb, (5) NHC (O) NWW, (6) CH20Ra, (7) CH20C02Rb, (8) CH20C (O) NRCRd, (9) C (O) NRCNRcRd or (10) C (O) NRCS02Rb ; Ra is (1) hydrogen, (2) optionally alkyl, (3) optionally substituted alkenyl, (4) optionally substituted alkynyl, (5) optionally substituted alkanoyl, (6) optionally substituted alkenoyl, (7) optionally substituted alkynoyl, (8) optionally substituted aroyl, (9) optionally substituted aryl, (10) optionally substituted cycloalkanoyl, (11) optionally substituted cycloalkenoyl, (12) optionally substituted alkylsulfonyl (13) optionally substituted cycloalkyl (14) optionally substituted cycloalkenyl where the substituents on the alkyl, alkenyl, alkynyl, alkanoyl, alkenoyl, alkynoyl, aroyl, aryl, cycloalkanoyl, cycloalkenoyl, alkylsulfonyl, cyclalkyl and cycloalkenyl are from 1 to 10 groups independently selected from hydroxy, alkoxy, cycloalkyl, aryl alkoxy, NRgRh, C02Rb, CONRcRd and halogen, (15) perfluoroalkyl, (16) arylsulfonyl optionally substituted with 1 to 3 groups independently selected from alkyl, perfluoroalkyl, halogen and cyano, (17) a 5-or 6-member heterocycle containing 1 to 4 heteroatoms selected from oxygen, sulfur and nitrogen optionally substituted by 1 to 4 groups independently selected
from alky], alkenyl, perfluoroalkyl, amino, C (O) NRCRd, cyano, C02Rb and halogen, and which may be saturated or partly unsaturated; Rb is (1) H, (2) optionally substituted aryl, (3) optionally substituted alkyl, (4) optionally substituted alkenyl, (5) optionally substituted alkynyl, (6) optionally substituted cycloalkyl, (7) optionally substituted cycloalkenyl, or (8) optionally substituted 5-to 10-member heterocycle containing from 1 to 4 heteroatoms independently selected from oxygen, sulfur and nitrogen; where the substituents on the aryl, alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocycle, or alkynyl are from 1 to 10 groups independently selected from (i) hydroxy, (ii) CI-C3 alkyl, (iii) oxo, (iv) S02NRgRh, (v) aryl alkoxy, (vi) hydroxy alkyl, (vii) alkoxy, (viii) hydroxyalkoxy, (ix) aminoalkoxy, (x) cyano, (xi) perfluoroalkyl, (xii) alkyl-S (O) m, (xiii) cycloalkyl optionally substituted with 1 to 4 groups independently selected from Re, (xiv) cycloalkenyl, (xv) halogen, (xvi) alkanoyloxy, (xvii) C (O) NRgRh, (xviii) C02R', (xix) optionally substituted arylalkoxy, wherein the aryl substituents are 1,2-methylenedioxy or 1 to 5 groups independently selected from Re, (xx) -NRgRh, (xxi) 5 to 6-member heterocycle, which may be saturated or partially unsaturated, containing from 1 to 4 heteroatoms independently selected from oxygen, sulfur and nitrogen, and optionally substituted with 1 to 5 groups independently selected from Re, and (xxii) optionally substituted aryl, wherein the aryl substituents are 1, 2-methylenedioxy or 1 to 5 groups independently selected from Re ; Re is (1) halogen, (2) alkyl, (3) perfluoroalkyl, (4) -S(0)mRi, (5) cyano, (6) amino, (7) R'O (CH2) v-, (8) (CH2)v-, (9) R'OCO (CH2) v-, (10) optionally substituted aryl where the substituents are from 1 to 3 of halogen, alkyl, alkoxy, or hydroxy, or (11) S02NRgRh ; Rf is (1) methyl, (2) X-C1-C2 alkyl, where X is O or S (O) m, (3) halogen, (4) acetylamino, (5) trifluoromethyl, (6) NYIY2, where Yl and Y2 are independently H or methyl, and (7) hydroxy; Rg and Rh are independently (1) hydrogen, (2) alkyl optionally substituted with hydroxy, amino, or C02R' (3) aryl optionally substituted with halogen, 1,2- methylenedioxy, alkoxy, alkyl or perfluoroalkyl, (4) arylalkyl, wherein the aryl is optionally substituted with perfluorolkyl or 1,2-methylenedioxy ; (5) alkoxycarbonyl, (6) alkanoyl, (7) alkanoyl alkyl, (9) arylalkoxycarbonyl, (10) aminocarbonyl, (11) monoalkylaminocarbonyl (12) dialkylaminocarbonyl; or Rg and Rh together with the N to which they are attached form a 5-to 6-member ring containing 0 to 2 additional heteroatoms selected from O, S (O) mn and N, optionally substituted with 1 to 3 groups
independently selected from Re and oxo; R'is (1) hydrogen, (2) perfluoroalkyl, (3) alkyl, (4) optionally substituted arylalkyl, where the aryl substituents are from 1 to 3 groups independently selected from halogen, alkyl, alkoxy, and hydroxy; all other variables are as defined under Formula I.
In another preferred embodiment, the present invention provides compounds of Formula I wherein R'is (1) hydrogen, (2) optionally substituted alkyl, (3) optionally substituted alkenyl, (4) optionally substituted alkynyl, where the substituents on the alkyl, alkenyl, and alkynyl are 1 to 3 groups independently selected from (i) methyl, (ii) X-methyl, where X is O or S (O) m and (iii) halogen, (5) aryl or arylalkyl wherein said aryl is optionally substituted with 1 to 3 groups independently selected from Rf.
(6) trifluoromethyl R8 is (1) H, (2) OH, or (3) NH2 R9 is (1) H, or (2) OH; Rio is (1) C (O) ORb, (2) C (O) N (ORb) RC, (3) C (O) NORD, (4) NHC (O) ORb, (5) NHC (0) NRCRd, (6) CH20Ra, (7) CH20C02Rb, (8) CH20C (O) NRCRd, (9) C (O) NRcNRcRd, or (10) C (0) NRCS02Rb ; Ra is'(1) hydrogen, (2) optionally substituted alkyl, (3) optionally substituted alkenyl, (4) optionally substituted alkynyl, (5) optionally substituted alkanoyl, (6) optionally substituted aroyl, (7) optionally substituted cycloalkanoyl, (8) optionally substituted cycloalkenoyl, (9) optionally substituted alkylsulfonyl where the substituents on the alkyl, alkenyl, alkynyl, alkanoyl, aroyl, cycloalkanoyl, cycloalkenoyl, and alkylsulfonyl, are from 1 to 5 groups independently selected from hydroxy, alkoxy, aryl alkoxy, NRgRh, C02Rb, CONRCRd and halogen, (10) trifluoromethyl, (11) arylsulfonyl optionally substituted with 1 to 3 groups independently selected from methyl, trifluoromethyl and halogen, (12) a 5-or 6-member heterocycle containing 1 to 4 heteroatoms selected from oxygen, sulfur and nitrogen optionally substituted by 1 to 4 groups independently selected from methyl, trifluoromethyl, C (O) NRCRd, Curb and halogen, and which may be saturated or partly unsaturated; Rb is (1) H, (2) optionally substituted aryl, (3) optionally substituted alkyl, (4) optionally substituted alkenyl, (5) optionally substituted alkynyl, (6) optionally substituted cycloalkyl, (7) optionally substituted cycloalkenyl, or (8) optionally substituted 5-to 6-member heterocycle containing from 1 to 4 heteroatoms independently selected from oxygen, sulfur and nitrogen; where the substituents on the aryl,
alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocycle, or alkynyl are from 1 to 10 groups independently selected from (i) hydroxy, (ii) alkyl, (Hi) oxo, (iv) S02NRgRh, (v) arylalkoxy, (vi) hydroxyalkyl, (vii) alkoxy, (viii) hydroxy alkoxy, (ix) amino alkoxy, (x) cyano, (xi) alkyl-S (O) m,.(xii) cycloalkyl optionally substituted with 1 to 4 groups independently selected from Re, (xiii) cycloalkenyl, (xiv) halogen, (xv) alkanoyloxy, (xvi) C (0) NRgRh, (xvii) C02R', (xvii)-NRgRh, (xix) 5 to 6-member heterocycle, which may be saturated or partially unsaturated, containing from 1 to 4 heteroatoms independently selected from oxygen, sulfur and nitrogen, and optionally substituted with 1 to 5 groups independently selected from Re, (xx) optionally substituted aryl, wherein the aryl substituents are 1, 2-methylenedioxy or 1 to 5 groups independently selected from Re (xxi) optionally substituted aryl alkoxy, wherein the aryl substituents are 1,2-methylenedioxy or 1 to 5 groups independently selected from Re, and (xxii) perfluoroalkyl ; Re is (1) halogen, (2) alkyl, (3) perfluoroalkyl, (4) -S (O)mRi, (5) cyano, (6) Riz (CH2)v- (7) RiC02(CH2)v- (8) RioCO (CH2)v-, (9) optionally substituted aryl where the substituents are from 1 to 3 of halogen, alkyl, alkoxy, or hydroxy, (10) S02NRgRh, or (11) amino; Rf is (1) methyl, (2) X-CI-C2 alkyl, where X is O or S (0)m, (3) trifluoromethyl, (4) NYIY2, where Y1 and Y2 are independently H or methyl, (5) hydroxy, (6) halogen, and (7) acetylamino, Rl and Rh are independently (1) hydrogen, (2) alkyl optionally substituted with hydroxy, amino, or COzR' (3) aryl optionally substituted with halogen, 1,2-methylenedioxy, alkoxy, alkyl or perfluoroalkyl, (4) arylalkyl, wherein the aryl is optionally substituted with perfluorolkyl or 1, 2-methylenedioxy ; (5) alkoxycarbonyl, (6) alkanoyl, (7) alkanoylalkyl, (9) arylalkoxycarbonyl, (10) aminocarbonyl, (11) monoalkylaminocarbonyl (12) dialkylaminocarbonyl; or Rl and Rh together with the N to which they are attached form a 5-to 6-membered ring containing 0 to 2 additional heteroatoms selected from O, S (O) m, and N, optionally substituted with 1 to 3 groups independently selected from Re and oxo; is (1) hydrogen, (2) perfluoroalkyl, (3) alkyl, (4) optionally substituted aryl or arylalkyl, where the aryl substituents are from 1 to 3 groups independently selected from halogen, alkyl, alkoxy, and hydroxy ; and all other variables are as defined under Formula I. In another aspect of the present invention there are provided compounds having the formula
(Formula Removed)
where RI-R6, R8 and Rg are as defined under Formula I ; and Rugis (1) COC1, (2) CON3, or (3) NCO.
Most especially preferred are spot-on compositions, wherein the composition comprises nodulisporic acid derivatives which are nodulisporamides, which are compounds of the formula
(Formula Removed)
R1 is (1) hydrogen, (2) optionally substituted CI-CIo alkyl, (3) optionally substituted C2-Clo alkenyl, (4) optionally substituted C2-Clo alkynyl, (5) optionally substituted C3-C8 cycloalkyl, (6) optionally substituted Cs-C8 cycloalkenyl where the substituents on the alkyl, alkenyl, alkynyl, cycloalkyl and cycloalkenyl are 1 to 3 groups independently selected from CI Cs alkyl, CI-CIo alkoxy, CI CIO alkylthio, C1- C10 alkylsulfonyl, C3-C8 cycloalkyl, hydroxy, halogen, cyano, carboxy, amino, C1-C10 maonalkylamino, C1-C10 dialkylamino, C1-C10 alkanoyl amino and benzoyl amino wherein said benzoyl is optionally substituted with 1 to 3 groups independently selected from C1-C4 alkyl, C1-C4 alkoxy, C1-C4 alkylthio, C2-C4 alkenyl, 2-04 alkynyl, C1-C3-perfluoroalkyl, amino, hydroxy, halogen, CI Cs monoalkylamino, Ci. C5 dialkylamino and C1-C5 alkanoyl amino, (7) phenyl Co-CS alkyl wherein said phenyl is optionally substituted with 1 to 3 groups
independently selected from CI-C4 alkyl, CI-C4 alkoxy, C1-C4 alkylthio, C2- C4 alkenyl, 2-04 alkynyl, CI C3 perfluoroalkyl, amino, hydroxy, carboxy, halogen, C1-C5 monoalkylamino, Cl-Cs dialkylamino and Cl-Cs alkanoyl amino, (8) Cl-Cs perfluoroalkyl, (9) a 5-or 6-member ring selected from morpholino, pyridyl and piperazino, optionally substituted by 1 to 3 groups independently selected from hydroxy, oxo, C1-C10alkyl, and halogen, R2, R3, and R4 are independently ORa, OCO2Rb, OC (O) NleR ; or Rl and R2 together represents, =NORa or =N-NRcRd; Rs is NRCRd, Ra is (1) hydrogen, (2) optionally substituted C1-C10 alkyl, (3) optionally substituted C3-C10 alkenyl, (4) optionally substituted C3 CI0 alkynyl, (5) optionally substituted CI CIO alkanoyl, (6) optionally substituted CI-CIo alkenoyl, (7) optionally substituted CI CIO alkynoyl, (8) optionally substituted benzoyl, (9) optionally substituted phenyl, (10) optionally substituted C1-C7 cycloalkanoyl, (11) optionally substituted C4-C7 cycloalkenoyl, (12) optionally substituted CI-CIo alkylsulfonyl (13) optionally substituted Cs-Cg cycloalkyi (14) optionally substituted Cs-Cg cycloalkenyl where the substituents on the alkyl, alkenyl, alkynyl, alkanoyl, alkenoyl, alkynoyl, benzoyl, phenyl, cycloalkanoyl, cycloalkenoyl, alkylsulfonyl, cycloalkyi and cycloalkenyl are from 1 to 5 groups independently selected from hydroxy, C1-C6 alkoxy, C3-C7 cycloalkyi, aryl C1-C3 alkoxy, NRg Rh, C02Rb, CONRcRd and halogen, (15) Ci-Cs perfluoroalkyl, (16) phenylsulfonyl optionally substituted with 1 to 3 groups independently selected from Cl-Cs alkyl, Cl-Cs perfluoroalkyl, nitro, halogen or cyano, (17) a 5-or 6-member ring selected from piperidino, morpholino, pyridyl and piperazino optionally substituted by 1 to 4 groups independently selected from CI Cs alkyl, CI Cs alkenyl, C1-C5 perfluoroalkyl, amino, C (O) Roc Rd, cyano, C02Rb or halogen; R is (1) H, (2) optionally substituted phenyl, (3) optionally substituted C1-C10 alkyl, (4) optionally substituted Cs-Cio alkenyl, or (5) optionally substituted C1-C10 alkynyl, where the substituents on the phenyl, alkyl, alkenyl or alkynyl are from 1 to 5 groups independently selected from hydroxy, C1-C6 alkoxy, C3-c7 cycloalkyi, halogen, Cl-Cs alkanoyloxy, C (O) NRcRd, C02Rb, formyl,-NRg Rh, optionally substituted phenyl, and optionally substituted phenyl C1-C3 alkoxy, wherein the phenyl substituents are 1 to 3 groups independently selected from Re ; R and Rd are independently Rb ; or Re and Rd together with the N to which they are attached form a piperidino, morpholino or piperazino optionally substituted with 1 to 3 groups independently selected from Rg and oxo;
Reis (1) halogen, (2) C1-C7 alkyl, (3) CI-C3 perfluoroalkyl, (4)-S(0)mRi, (5) cyano, (6) nitro, (7) Rio (CH2) v-, (8) RJC02 (CH2)v- (9) RjOCO(CH2)v-, (10) optionally substituted phenyl where the substituents are from 1 to 3 halogen, Cl-C6 alkyl, CI-C6 alkoxy, or hydroxy; v is 0 to 3 ; Rg and Rh are independently (1) hydrogen, (2) C1-C6alkyl, (3) aryl, (4) aryl C1-C6alkyl, (5) Cl-Cs alkoxycarbonyl, (6) Cl-Cs alkylcarbonyl, or (7) Cl-Cs alkanoyl Cl-Cs alkyl; or Rg and Rh together with the N to which they are attached form a piperidino, morpholino or piperazino optionally substituted with 1 to 3 groups independently selected from Rg and oxo; Ri and Rj are independently (1) hydrogen, (2) C1-C3perfluoralkyl, (3) optionally substituted CI-C6 alkyl, where the substituents are aryl or substituted phenyl; (4) phenyl or substituted phenyl where the substituents are from 1 to 3 groups independently selected from halogen, CI-C6 alkyl, CI-C6 alkoxy, or hydroxy ; m is 0 to 2; or a pharmaceutically acceptable salt thereof.
Most especially preferred are compounds of the formula
(Formula Removed)
wherein Rx is selected from the group consisting of: H, CH3, CH2CH3, C (CH3) 3, CH2CH2CH3, CH2CH20H, CH (C02CH3) CH20H, CH2C02CH3, CH2CH (OCH2CH3) 2, CH2CH20CH2CH20H, CH (CH3) (CH2) 3C (CH3) 20H, (CH2) 30H, (CH2) 40H, (CH2) SOH, CH (CH20H) CH2CH3, NHC (CH3) 3, CH2CN, (CH2) 60H, CH2CH (OH) CH3, CH (CH20H) CH2CH2CH3, CH2CH2SCH3, CH2CH2SCH2CH3, CH2CONH, CH (CH3) (CH20H) 2, CH2CH2NHCH2CH20H, CH (CH20H) (CH2) 3CH3, CH (CH20CH3) CH3, (CH2) 2SH, (CH2) 4NH2, CH2CH2S02CH3, CH2CH2S (O) CH3, CH (CH (CH3) 2) CH20H, (CH2) 3NH2, (CH2) 3N (CH2CH3) 2, (CH2) 3N (CH3) 2, OCH2CH3, CH2CH (OH) CH20H, OCH3, CH2CH20CH3, CH2CH2NHC (O) CH3, C (CH3) 2CH20H, C-C3H5, c-C6H11, (CH2) 30CH2CH3, CH2CH=CH2, C (CH2CH3) (CH20H) 2, CH2C#CH, CH2C02CH2CH3, CH2CH2F, (CH2) 30CH2) n CH3, CH2CH2N (CH3) 2, CH2CH20CH2CH2NH2, CH2CF3, NHCH2C02CH2CH3, CH (CH3) C02CH3, C
(CH3) 2CH2C (O) CH3, CH (C02CH2CH3) 2, CH2CH3, CH (CH2CH2CH3) C02CH3, CH2CH2CH20CH3, C (CH3) 2C=CH, (CH2) 4CH3, CH (CH2CH2CH3) 2, (CH2) 5CH3, CH2CH2C02H, CH (CH (CH3) 2) C02CH3, OCH2C02H, CH (CH (CH3) 2) CH20H, CH (CH (CH3) 2) CH20H, CH (CH3) CH20H, CH (CH3) CH20H, CH (CH3) 2, C (CH3) 3, (CH2) CH (CH3) 2, CH (CH3) CH2CH3, CH2CH (CH3) OH, (CH2) 3CH3, (CH2) 20CH2CH3, 1-adamantyl, (CH2) 8CH3, CH (CH3) CH (CH3) 2, (CH2) 3NHCH3, (CH2) 2N (CH2CH3) 2,
(Formula Removed)
A n especially preferred nodulisporamide derivative is one wherein Rx is with t-butyl being most especially preferred.
"Alkyl"as well as other groups having the prefix"alk", such as alkoxy, alkanoyl, alkenyl, alkynyl and the like, means carbon chains which may be linear or branched or combinations thereof. Examples of alkyl groups include methyl,
ethyl, propyl, isopropyl, butyl, sec-and tert-butyl, pentyl, hexyl, heptyl and the like."Alkenyl","alkynyl"and other like terms include carbon chains containing at least one unsaturated C-C bond.
The term"cycloalkyl"means carbocycles containing no heteroatoms, and includes mono-, bi-and tricyclic saturated carbocycles, as well as benzofused carbocycles. Examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, decahydronaphthalene, adamantane, indanyl, indenyl, fluorenyl, 1,2, 3, 4-tetrahydronaphalene and the like. Similarly,"cycloalkenyl"means carbocycles containing no heteroatoms and at least one non-aromatic C-C double bond, and include mono-, bi-and tricyclic partially saturated carbocycles, as well as benzofused cycloalkenes. Examples of cycloalkenyl include cyclohexenyl, indenyl, and the like.
The term"halogen"is intended to include the halogen atoms fluorine, chlorine, bromine and iodine.
The term"heterocycle", unless otherwise specfied, means mono-or bicyclic
compounds that are saturated or partly unsaturated, as well as benzo-or
heteroaromatic ring fused saturated heterocycles or partly unsaturated
heterocycles, and containing from 1 to 4 heteroatoms independently selected
from oxygen, sulfur and nitrogen. Examples of saturated heterocycles include
morpholine, thiomorpholine, piperidine, piperazine, tetrahydropyran,
tetrahydrofuran, dioxane, tetrahydrothiophene, oxazolidine, pyrrolidine; examples
of partly unsaturated heterocycles include dihydropyran, dihydropyridazine,
dihydrofuran, dihydrooxazole, dihydropyrazole, dihydropyridine,
dihydropyridazine and the like. Examples of benzo-or heteroaromatic ring fused heterocycle include 2, 3-dihydrobenzofuranyl, benzopyranyl, tetrahydroquinoline, tetrahydroisoquinoline, benzomorpholinyl, 1, 4-benzodioxanyl, 2, 3-dihydrofuro (2,3-b) pyridyl and the like.
The term"aryl"is intended to include mono-and bicyclic aromatic and heteroaromatic rings containing from 0 to 5 heteroatoms independently selected from nitrogen, oxygen and sulfur. The term"aryl"is also meant to include benzofused cycloalkyl, benzofused cycloalkenyl, and benzofused heterocyclic
groups. Examples of "aryl"groups include phenyl, pyrrolyl, isoxazolyl, pyrazinyl, pyridinyl, oxazolyl, thiazolyl, imidazolyl, triazolyl, tetrazolyl, furanyl, triazinyl, thienyl, pyrimidinyl, pyridazinyl, pyrazinyl, naphthyl, benzoxazolyl, benzothiazolyl, benzimidazolyl, benzofuranyl, furo (2,3-B) pyridyl, 2, 3dihydrofuro (2,3-b) pyridyl, benzoxazinyl, benzothiophenyl, quinolinyl, indolyl, 2, 3-dihydrobenzofuranyl, benzopyranyl, 1,4-benzodioxanyl, indanyl, indenyl, fluorenyl, 1,2, 3,4-tetrahydronaphthalene and the like.
Aroyl means arylcarbonyl in which aryl is as defined above.
Examples of NRCRd or NRgRh forming a 3-to 10-membered ring containing 0 to 2 additional heteroatoms selected from O, S (O) m and N are aziridine, azetidine, pyrrolidine, piperidine, thiomorpholine, morpholine, piperazine, octahydroindole, tetrahydroisoquinoline and the like.
The term"optionally substituted"is intended to include both substituted and unsubstituted ; thus, for example, optionally substituted aryl could represent a pentafluorophenyl or a phenyl ring.
Certain of the above defined terms may occur more than once in the above formula and upon such occurrence each term shall be defined independently of the other; thus, for example, ORa at C4 may represent OH Compounds of formula (I) are available commercially or can be prepared according to one or other of the processes or any other process coming within the competence of a person skilled in the art who is an expert in chemical synthesis. For the chemical preparation of the products of the invention, a person skilled in the art is regarded as having at his disposal, inter alia, the entire contents of'Chemical Abstracts" and of the documents which are cited therein. Semi-synthetic processes are described, for example, in U. S. Patent 6,399, 786 or WO 96/29073, both of which are incorporated by reference.
A particularly effective synthetic method in order to prepare nodulisporamide compounds of the formula:
(Formula Removed)
wherein Ri, R2, R3 and R4 are defined above and R51 is NRCRd where RCRd are independently (1) H, (2) optionally substituted aryl, (3) optionally substituted alkyl, (4) optionally substituted alkenyl, (5) optionally substituted alkynyl, (6) optionally substituted cycloalkyl, (7) optionally substituted cycloalkenyl, or (8) optionally substituted heterocycle containing from 1 to 4 heteroatoms independently selected from oxygen, sulfur and nitrogen; where the substituents on the aryl, alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocycle, or alkynyl are from 1 to 10 groups independently selected from (i) hydroxy, (ii) alkyl, (iii) oxo, (iv) S02NRgRh, (v) arylalkoxy, (vi) hydroxyalkyl, (vii) alkoxy, (viii) hydroxyalkoxy, (ix) aminoalkoxy, (x) cyano, (xi) mercapto, (xii) alkyl-S (O) M, (xiii) cycloalkyl optionally substituted with 1 to 4 groups independently selected from Re, (xiv) cycloalkenyl, (xv) halogen, (xvi) alkanoyloxy, (xvii) C (O) NRgRh, (xviii) Cozy', (xix) formyl, (xx) -NRgRh, (xxi) 5 to 9-member heterocycle, which may be saturated or partially unsaturated, containing from 1 to 4 heteroatoms independently selected from oxygen, sulfur and nitrogen, and optionally substituted with 1 to 5 groups independently selected from Re, (xxii) optionally substituted aryl, wherein the aryl substituents are 1,2-methylenedioxy or 1 to 5 groups independently selected from Re, (xxiii) optionally substituted arylalkoxy, wherein the aryl substituents are 1, 2-methylenedioxy or 1 to 5 groups independently selected from Re, (xxiv) perfluoroalkyl; or Re and Rd together with the N to which they are attached form a 3-to 10-member ring containing 0 to 2 additional heteroatoms selected from O, S (O) m, and N, optionally substituted with 1 to 3 groups independently selected from Rg, hydroxy, thioxo and oxo; Re is (1) halogen, (2) alkyl, (3) perfluoroalkyl, (4) -S(0)mRi, (5) cyano, (6) nitro, (7) Wu (CH2) v-, (8) RiC02(CH2)v-, (9) RiOCO (CH2) v-, (10) optionally substituted aryl where the substituents are from 1 to 3 of halogen, alkyl, alkoxy, or hydroxy, (11) S02NRgRh, or (12) amino; Rf is (1) alkyl, (2) X-alkyl, where X is O or S (O)
m, (3) alkenyl, (4) alkynyl, (5) perfluoroalkyl, (6) NY'Y2, where Y'and Y2 are independently H or alkyl, (7) hydroxy, (8) halogen, and (9) alkanoyl amino, Rg and Rh are independently (1) hydrogen, (2) alkyl optionally substituted with hydroxy, amino, or COUR' (3) aryl optionally substituted with halogen, 1, 2-methylenedioxy, alkoxy, alkyl or perfluoroalkyl, (4) arylalkyl, wherein the aryl is optionally substituted with perfluorolkyl or 1,2-methylenedioxy ; (5) alkoxycarbonyl, (6) alkanoyl, (7) alkanoylalkyl, (9) arylalkoxycarbonyl, (10) aminocarbonyl, (11) monoalkylaminocarbonyl (12) dialkylaminocarbonyl; or Rg and together with the N to which they are attached form a 3-to 7-member ring containing 0 to 2 additional heteroatoms selected from O, S (0) m, and N, optionally substituted with 1 to 3 groups independently selected from Re and oxo ; Ri is (1) hydrogen, (2) perfluoroalkyl, (3) alkyl, (4) optionally substituted aryl, or arylalkyl, where the aryl substituents are from 1 to 3 groups independently selected from halogen, alkyl, alkoxy, and hydroxy; m is 0 to 2 ; and v is 0 to 3 ; or said process comprising : (1) coupling a compound of formula II
(Formula Removed)
wherein RI, R2, R3, and R4 are defined above, with a compound of formula III:
(Formula Removed)
wherein R6 and R7 can be independently selected from alkyl, aminoalkyl or cycloalkyl, in the presence of an organic solvent to produce a first intermediate compound of the formula: v
(Formula Removed)
reacting the first intermediate compound with an activating compound ACT, such as a compound of formula IV:
(Formula Removed)
to produce a second intermediate compound of the formula VI':
(Formula Removed)

(3) adding an amine of the formula HNRCRd to the second intermediate compound to obtain a compound of formula I'.
This process is described in Provisional Application 60/415627, entitled"Method for the Synthesis of Nodulisporamide,"filed on October 2,2002, herein incorporated by reference.
The advantage to the invention process is that the amidation reaction occurs under mild conditions, thereby reducing the potential of side reactions at the C23-C24 position or epimerization of C7 of the starting material. This in turn increases the overall yield and purity of the final product. It is known that amidation under acidic conditions leads to dehydration of the C23-C24 position and amidation
under basic conditions leads to epimerization of C7 ; the inventive process is mild enough to reduce greatly the occurrence of these side reactions.
The inventive process achieves these results by performing the amidation reaction via an active intermediate by reacting the nodulisporic acid compound with a compound of formula III. Preferred compounds of formula III are N-N'-Diisopropylcarbodiimide (DIPCDI) N-N'-dicyclohexylcarbodimide (DCC), and 1- [ (3-dimethylamino) propyl] -3- ethylcarbodimide HC1 salt (EDC) with DCC being especially preferred. This intermediate may be isolated or it may be reacted in one step with an activating compound such as a 1-hydroxybenzotriazole (HOBT) [Formula IV]. Other compounds which can be used as activating compounds include 2-hydroxypyridine-N-oxide (HOPO), 2-hydroxy pyridine and 1-hydroxysuccinimide.
The amines of the formula HNRCRd are well known to a practioner of this art and are obtainable either commercially or by modification of known synthetic techniques, such as those found in"Organic Synthesis", a source that is well-known and used by a practitioner in the field. Preferred aminos include, for example, amines wherein Re is H and Rd is selected from the group consisting of : H, CH3, CH2CH3, C (CH3) 3, CH2CH2CH3, CH2CH20H, CH (C02CH3) CH20H, CH2C02CH3, CH2CH (OCH2CH3) 2, CH2CH20CH2CH20H, CH (CH3) (CH2) 3C (CH3) 20H, (CH2) 30H, (CH2) 40H, (CH2) SOH, CH (CH20H) CH2CH3, NHC (CH3) 3, CH2CN, (CH2) 60H, CH2CH (OH) CH3, CH (CH20H) CH2CH2CH3, CH2CH2SCH3, CH2CH2SCH2CH3, CH2CONH2, CH (CH3) (CH20H) 2, CH2CH2NHCH2CH20H, CH (CH20H) (CH2) 3CH3, CH (CH20CH3) CH3, (CH2) 2SH, (CH2) 4NH2, CH2CH2S02CH3, CH2CH2S (O) CH3, CH (CH (CH3) 2) CH20H, (CH2) 3NH2, (CH2) 3N (CH2CH3) 2, (CH2) 3N (CH3) 2, OCH2CH3, CH2CH (OH) CH20H, 0CH3, CH2CH20CH3, CH2CH2NHC (O) CH3, C (CH3) 2CH20H, C-C3H5, CC6H11, (CH2) 30CH2CH3, CH2CH=CH2, C (CH2CH3) (CH20H) 2, CH2C#CH, CH2C02CH2CH3, CH2CH2F, (CH2) 30 (CH2) 11CH3, CH2CH2N (CH3) 2, CH2CH20CH2CH2NH2, CH2CF3, NHCH2C02CH2CH3, CH (CH3) C02CH3, C (CH3) 2CH2C (O) CH3, CH (C02CH2CH3) 2, CH2CH3, CH (CH2CH2CH3) C02CH3, CH2CH2CH20CH3, C (CH3) 2CH2C#CH, (CH2)4CH3, CH (CH2CH2CH3) 2, (CH2) SCH3,
CH2CH2C02H, CH (CH (CH3) 2) C02CH3 OCH2C02H, CH (CH (CH3) 2) CH20H, CH (CH (CH3) 2) CH20H, CH (CH3) CH20H, CH (CH3) CH20H, CH (CH3) 2, C (CH3) 3, (CH2) CH (CH3) 2, CH (CH3) CH2CH3, CH2CH (CH3) OH, (CH2) 3CH3, (CH2) 20CH2CH3, 1-adamantyl, (CH2) 8CH3, CH (CH3) CH (CH3) 2, (CH2) 3NHCH3, (CH2) 2N (CH2CH3) 2,
(Formula Removed)
Preferred solvents for this reaction include halogenated hydrocarbons, such as dichloromethane and ethylene chloride, ethers, such as methyl t-butyl ether (MTBE), diethyl ether or tetraydrofuran (THF), or mixtures of the foregoing. Other solvents include polar aprotic solvents including but not limited to dimethoxymethane, 2- methyltetrahydrofuran, methyl iso-butylketone, benzotrifuoride and methylacetate.
Perferably, the inventive process uses a homogeneous solvent system that dissolves both the nodulisporic acid derivatives and the activation agents and avoids the prior process, which perform the activating/coupling reaction two phase water-organic solvent system, thereby permitting the reaction to run faster and obtain conversion more easily. A mixture of MTBE and THF is especially preferred. Preferred ranges of MTBE to THF are about 5: 1 to about 1: 2, with about 4: 1 to about 2: 3 being especially preferred.
The amount of DCC and HOBT should be in molar excess to achieve full conversion. A preferred range would be from about 1.1 to about 2.5, with about 1.2 to about 1.8 equivalents being especially preferred.
Reaction temperatures range from about 10°C to about 50°C with about 20°C to about 30°C being especially preferred.
Optionally, the nodulisporamide derivative may be recrystallized to obtain a product with better purity. Suitable recrystallization solvents include a mixture of a polar solvent such a s water, acetonitrile, acetone and ana polar solvent such as alkanes and cycloalkanes including but not limited to pentane, hexane, cyclohepatane, cyclohexane, cyclohepatane.
Preferred mixture of recrystallization solvent include acetone/heptane with few drops of water. An especially preferred mixture of solvents include first optionally recrystallizing from acetonitrile/water followed by acetone/heptane.
Compounds of formula Vare also novel and are part of this invention.
Administration of the inventive formulation may be intermittent in time and may be administered daily, weekly, biweekly, monthly, bimonthly, quarterly, or even for longer durations of time. The time period between treatments depends upon factors such as the parasite (s) being treated, the degree of infestation, the type of mammal or bird and the environment where it resides. It is well within the skill level of the practitioner to determine a specific administration period for a particular situation. This invention contemplates a method for permanently combating a parasite in an environment in which the animal is subjected to strong parasitic pressure where the administration is at a frequency far below a daily administration in this case. For example, it is preferable for the treatment according to the invention to be carried out monthly on dogs and on cats.
Spot-on formulations may be prepared by dissolving the active ingredients into the pharmaceutically or veterinary acceptable vehicle. Alternatively, the spot-on formulation can be prepared by encapsulation of the active ingredient to leave a residue of the therapeutic agent on the surface of the animal. These formulations
will vary with regard to the weight of the therapeutic agent in the combination depending on the species of host animal to be treated, the severity and type of infection and the body weight of the host. The compounds may be administered continuously, particularly for prophylaxis, by known methods.
Generally, a dose of from about 0.001 to about 100 mg per kg of body weight, with a range of 0.25 to 50 mg/kg being especially preferred, given as a single dose or in divided doses for a period of from about 1 to about 60 days, preferably from about 1 to 30 days, will be satisfactory but, of course, there can be instance where higher or lower dosage ranges are indicated and such are within the scope of this invention. It is well within the routine skill of the practitioner to determine a particular dosing regimen for a specific host and parasite.
It also may be preferable to use controlled-release formulations.
The invention also relates to such a method with a therapeutic aim intended for the treatment and prevention of parasitoses having pathogenic consequences.
This invention also provides for formulations wherein the nodulisporic acid or derivative thereof is combined with a second active agent, such a parasiticide. Examples of classes of these compounds include avermectins, milbemycins, 1-N-arylpyrazoles, IGR compounds, etc. , some of which are discussed above.
The formulations of the present invention provide for the topical administration of a concentrated solution, suspension, microemulsion or emulsion for intermittent application to a spot on the animal, generally between the two shoulders. It has been discovered that the inventive formulations are especially active against parasites when the formulations are applied to mammals and birds, especially poultry, dogs, cats, sheep, pigs, cattle and horses. These formulations comprise a composition of an effective amount of at least one nodulisporic acid derivative dissolved in a pharmaceutical or veterinary acceptable carrier vehicle where a crystallization inhibitor is optionally present. The nodulisporic acid derivative are advantageously resent in the inventive formulation in a proportion of about 1 to about 40%, preferably of about 1 to about 30% and most preferably about 5 to about 15% (percentages as weight by volume = WW).
The liquid carrier vehicle comprises a pharmaceutical^ or veterinary acceptable organic solvent and optionally an organic cosolvent.
Also contemplated are the pharmaceutical^ or veterinary acceptable acid or base salts, where applicable, of the active compounds provided for herein. The term"acid" contemplates all pharmaceutically or veterinary acceptable inorganic or organic acids.
Inorganic acids include mineral acids such as hydrohalic acids, such as hydrobromic and hydrochloric acids, sulfuric acids, phosphoric acids and nitric acids. Organic acids include all pharmaceutically or veterinary acceptable aliphatic, alicyclic and aromatic carboxylic acids, dicarboxylic acids tricarboxylic acids and fatty acids. Preferred acids are straight chain or branched, saturated or unsaturated CI-C20 aliphatic carboxylic acids, which are optionally substituted by halogen or by hydroxyl groups, or C6-CI2 aromatic carboxylic acids. Examples of such acids are carbonic acid, formic acid, fumaric acid, acetic acid, propionic acid, isopropionic acid, valeric acid, a-hydroxy acids, such as glycolic acid and lactic acid, chloroacetic acid, benzoic acid, methane sulfonic acid, and salicylic acid. Examples of dicarboxylic acids include oxalic acid, malic acid, succinic acid, tartaric acid and maleic acid. An example of a tricarboxylic acid is citric acid.
Fatty acids include all pharmaceutically or veterinary acceptable saturated or unsaturated aliphatic or aromatic carboxylic acids having 4 to 24 carbon atoms. Examples include butyric acid, isobutyric acid, sec-butyric acid, lauric acid, palmitic acid, stearic acid, oleic acid, linoleic acid, linolenic acid, and phenylsteric acid. 0 ther acids include gluconic acid, glycoheptonic acid and lactobionic acid.
The term"base"contemplates all pharmaceutically or veterinary acceptable inorganic or organic bases. Such bases include, for example, the alkali metal and alkaline earth metal salts, such as the lithium, sodium, potassium, magnesium or calcium salts.
Organic bases include the common hydrocarbyl and heterocyclic amine salts, which include, for example, the morpholine and piperidine salts.
The organic solvent for the liquid carrier vehicle will preferably have a dielectric constant of between about 10 and about 35, preferably between about 20 and about 30, the content of this solvent in the overall composition preferably representing the remainder to 100% of the composition. It is well within the skill level of the practitioner to select a suitable solvent on the basis of these parameters.
The organic cosolvent for the liquid carrier vehicle will preferably have a boiling point of less than about 100°C, preferably of less than about 80°C, and will have a dielectric constant of between about 10 and about 40, preferably between about 20 and about 30; this cosolvent can advantageously be present in the composition according to a weight/weight (W/W) ratio with respect to the solvent of between about 1/15 and about 1/2; the cosolvent is volatile in order to act in particular as drying promoter and is miscible with water and/or with the solvent. Again, it is well within the skill level of the practitioner to select a suitable solvent on the basis of these parameters.
The organic solvent for the liquid carrier includes the commonly acceptable organic solvents known in the formulation art. These solvents may be found, for example, in Remington Pharmaceutical Science, 16th Edition (1986). These solvents include, for example, acetone, ethyl acetate, methanol, ethanol, isopropanol, dimethylformamide, dichloromethane or diethylene glycol monoethyl ether (Transcutol).
These solvents can be supplemented by various excipients according to the nature of the desired phases, such as Cl-calo caprylic/capric triglyceride (Estasan or Miglyol 812), oleic acid or propylene glycol.
The liquid carrier may also comprise a microemulsion. Microemulsions are also well suited as the liquid carrier vehicle. Microemulsions are quaternary systems comprising an aqueous phase, an oily phase, a surfactant and a cosurfactant. They are translucent and isotropic liquids.
Microemulsions are composed of stable dispersions of microdroplets of the aqueous phase in the oily phase or conversely of microdroplets of the oily phase
in the aqueous phase. The size of these microdroplets is less than 200 nm (1000 to 100,000 nm for emulsions). The interfacial film is composed of an alternation of surface-active (SA) and co-surface-active (Co-SA) molecules which, by lowering the interfacial tension, allows the microemulsion to be formed spontaneously.
The oily phase can in particular be formed from mineral or vegetable oils, from unsaturated polyglycosylated glycerides or from triglycerides, or alternatively from mixtures of such compounds. The oily phase preferably comprises triglycerides and more preferably medium-chain triglycerides, for example Cl-calo caprylic/capric triglyceride. The oily phase will represent, in particular, from about 2 to about 1 5%, more particularly from about 7 to about 10%, preferably from about 8 to about 9%, V/V of the microemulsion.
The aqueous phase includes, for example water or glycol derivatives, such as propylene glycol, glycol ethers, polyethylene glycols or glycerol. Propylene glycol, diethylene glycol monoethyl ether and dipropylene glycol monoethyl ether are especially preferred. Generally, the aqueous phase will represent a proportion from about 1 to about 4% V/V in the microemulsion.
Surfactants for the microemulsion include diethylene glycol monoethyl ether, dipropyelene glycol monomethyl ether, polyglycolysed C8-CI0 glycerides or polyglyceryl-6 dioleate. In addition to these surfactants, the cosurfactants include short- chain alcohols, such as ethanol and propanol.
Some compounds are common to the three components discussed above, i. e., aqueous phase, surfactant and cosurfactant. However, it is well within the skill level of the practitioner to use different compounds for each component of the same formulation.
The cosurfactant to surfactant ratio will preferably be from about 1/7 to about 1/2.
There will preferably be from about 25 to about 75% V/V of surfactant and from about 10 to about 55% V/V of cosurfactant in the microemulsion.
Likewise, the co-solvents are also well known to a practitioner in the formulation art. Preferred co-solvents are those which is a promoter of drying and include, for example, absolute ethanol, isopropanol (2-propanol) or methanol.
If present, it is preferred that the crystallization inhibitor is present in a proportion of about 1 to about 20% (W/V), preferably of about 5 to about 15%. The inhibitor preferably corresponds to the test in which 0.3 ml of a solution comprising 10% (W/V) of the compound of formula (I) in the liquid carrier and 10% of the inhibitor are deposited on a glass slide at 20°C and allowed to stand for 24 hours. The slide is then observed with the naked eye. Acceptable inhibitors are those whose addition provides for few or no crystals, and in particular less than 10 crystals, preferably 0 crystals.
Although this is not preferred, the formulation can optionally comprise water, in particular in a proportion of 0 to about 30% (volume by volume V/V), in particular of 0 to about 5%.
The formulation can also comprise an antioxidizing agent intended to inhibit oxidation in air, this agent being in particular present in a proportion of about 0.005 to about 1% (WA/), preferably of about 0.01 to about 0.05%.
Crystallization inhibitors which can be used in the invention include: - p olyvinylpyrrolidone, polyvinyl alcohols, copolymers of vinyl acetate and of vinylpyrrolidone, polyethylene glycols, benzyl alcohol, mannitol, glycerol, sorbitol or polyoxyethylenated esters of sorbitan; lecithin or sodium carboxymethylcellulose; or acrylic derivatives, such as methacrylates and others, - anionic surfactants, such as alkaline stearates, in particular sodium, potassium orammoniumstearate ; calcium s tearate or t riethanolamine stearate; sodium abietate; alkyl sulphates, in particular sodium lauryl sulphate and sodium cetyl sulphate; sodium dodecylbenzenesulphonate or sodium dioctyl sulphosuccinate; or fatty acids, in particular those derived from coconut oil, - cationic surfactants, such as water-soluble quaternary ammonium salts of formula NFR'R"R'"R""S, in which the R radicals are identical or different optionally hydroxylated hydrocarbon radicals and Y-is an anion of a strong acid, such as halide, sulphate and sulphonate anions; cetyltrimethylammonium bromide is one of the cationic
surfactants which can be used, - amine salts of formula N+R'R"R"', in which the R radicals are identical or different optionally hydroxylated hydrocarbon radicals; octadecylamine hydrochloride is one of the cationic surfactants which can be used, - non-ionic surfactants, such as optionally polyoxyethylenated esters of sorbitan, in particular Polysorbate 80, or polyoxyethylenated alkyl ethers; polyethylene glycol stearate, polyoxyethylenated derivatives of castor oil, polyglycerol esters, polyoxyethylenated fatty alcohols, polyoxyethylenated fatty acids or copolymers of ethylene oxide and of propylene oxide, - amphoteric surfactants, such as substituted lauryl compounds of betaine, - or preferably a mixture of at least two of the compounds listed above.
In a particularly preferred embodiment, a crystallization inhibitor pair will be used. Such pairs include, for example, the combination of a film-forming agent of polymeric type and of a surface-active agent. These agents will be selected in particular from the compounds mentioned above as crystallization inhibitor.
Particularly preferred film-forming agents of polymeric type include: - the various grades of polyvinylpyrrolidone, - polyvinyl alcohols, and - copolymers of vinyl acetate and of vinylpyrrolidone.
Especially preferred surface-active agents, include those made of non- ionic surfactants, preferably polyoxyethylenated esters of sorbitan and in particular the various grades of polysorbate, for example Polysorbate 80.
The film-forming agent and the surface-active agent can in particular be incorporated in similar or identical amounts within the limit of the total amounts of crystallization inhibitor mentioned elsewhere.
The pair thus constituted secures, in a noteworthy way, the objectives of absence of crystallization on the coat and of maintenance of the cosmetic appearance of the fur, that is to say without a tendency towards sticking or towards a sticky appearance, despite the high concentration of active material.
Particularly preferred antioxidizing agents are those conventional in the art and include, for example, butylated hydroxyanisole, butylated hydroxytoluene,
ascorbic acid, sodium metabisulphite, propyl gallate, sodium thiosulphate or a mixture of not more than two of them.
The formulation adjuvants discussed above are well known to the practitioner in this art and may be obtained commercially or through known techniques. These concentrated compositions are generally prepared by simple mixing of the constituents as defined above; advantageously, the starting point is to mix the active material in the main solvent and then the other ingredients or adjuvants are added.
The volume applied can be of the order of about 0.3 to about 1 ml, preferably of the order of about 0.5 ml, for cats and of the order of about 0.3 to about 5 ml for dogs, depending on the weight of the animal.
The formulations according to the invention are extremely effective for long durations of time in the treatment of parasites such as fleas of mammals and, in particular, of small mammals such as dogs and cats. The inventive formulations exhibit a degree of effectiveness against o ther parasitic insects and in particular fleas and ticks.
The inventive formulations further exhibit synergy when treating infestations cause by ectoparasites and endoparasites.
Other advantages and characteristics of the invention will become apparent on reading the following description, given by way of non-limiting examples.
EXAMPLES
EXAMPLE 1: Topical Spot-on Formulation (20% w/v t-butyl nodulisporamide)
A spot-on formulation comprising the following ingredients was prepared by mixing the following ingredients
(Table Removed)

EXAMPLE 2: Topical Spot-on Formulation (10% w/v t-butyl nodulisporamide)
A spot-on formulation comprising t-butyl nodulisporamide was prepared by moving the following ingredients Ingredient
(Table Removed)
EXAMPLE 3: Long Term Efficacy
The spot-on formulation according to Example 1 was applied to four dogs infested with fleas for a thirty-five day period. The results are summarized below
(Table Removed)

The data demonstrate that the spot-on formulation retained its efficacy for the thirty-five day period.
EXAMPLE 4: Long Term Efficacy
The spot-on formulation according to the first Example 1 (40 mg/kg) was applied to four cats infested with fleas and its efficacy for thirty-five days was measured.
(Table Removed)
The
results are summarized below Day 1 7 14 21 28 35 % effic 100 100 100 100 100 99. 2 The data demonstrate that the spot-on formulation retained its efficacy for the thirty-five day period.
EXAMPLE 5: Synthesis of t-butyl nodulisporamide
A reactor fitted with a stirrer was charged with 4. 8 L of MTBE and 1. 2 L of THF.
Stirring was started and after mixing, 1.11 kg (1.5 mol) of nodulisporic acid A, used as a solvate prepared by crystallizing nodulisporic acid A from methanol and acetonitrite in a molar ratio of 1: 1 : 1, was added to the to the reactor and allowed to dissolve. Next, 0.26 kg HOBT. H20 (1,70 mol) was charged to the reactor followed by the addition of 0.35 kg of a melt of DCC (1,70 mol) over a period of 1.5 h, keeping the temperature of the reaction between 20-30°C.
The mixture was then stirred for 4 h at 20-30°C. Following this, 0.27 kg of TBA (3.69 mol) was added over a period of 1 h, again keeping the temperature of the reaction between 20-30°C. The mixture was then stirred for 1 h at 20-30°C.
Following stirring, the reaction mixture was filtered over filter cloth (Dralon) and the cake was washed with 4: 1 (v/v) MTBE/THF (3x1.0 L) by means of a passive wash.
The cake (DCU) is collected as solid organic waste (0.28 kg dry weight).
The filtrates were combined and washed sequentially with 5.0 L of 0.5 N aqueous hydrochloric acid and 5.0 L of aqueous saturated sodium bicarbonate. The
washed filtrate (MTBE/THF) was concentrated in vacuo (30°C, 50-100 mBar) to ca. 6.0 L.
6.0 L of acetonitrile was added and the solution was concentrated in vacuo to 3.6 L. Water (2.4 L) was then added with stirring to the acetonitrile solution over a period of 2 h at 20-25°C. A yellow precipitate formed and this was filtered over filter cloth (Dralon) and then washed with acetonitrile/water (3: 2 v/v) (2x1. 5L). The solid was then air-dried for 3 h at ambient temperature and then in vacuo at 40-45° C to constant weight.
The crude t-butyl nodulisporamide was subsequently recrystallized from acetone/heptane by first dissolving the crude t-butyl nodulisporamide in 2.5 L of acetone and then adding 5.8 L of heptane over a 2 h period of time with mechanical stirring at a temperature 20- 25° C. The mixture was then aged for 2 h at 20° C.
The slurry was filtered over filter cloth and the cake sequentially washed with 1: 4 (v/v) acetone/heptane (2x1. 5L) and heptane (3. 0L). The cake was air-dried for 3 h at ambient temperature and then under vacuum at 4 0-45'C to constant weight to give a molar yield of 75% of t-butyl nodulisporamide based upon nodulisporic acid A.
The above description of the invention is intended to be illustrative and not limiting. V arious changes or modifications in the embodiments described herein may occur to those skilled in the art. These can be made without departing from the scope and spirit of the invention.







We Claim:
1. A spot-on formulation useful for the treatment or prophylaxis of parasite infestation in mammals or birds which comprises:
(a) at least one nodulisporic acid derivative of the kind as herein described;
(b) a pharmaceutically or veterinary acceptable liquid carrier vehicle;
(c) optionally an antioxidant; and
(d) optionally a crystallization inhibitor of the kind as herein described ;
wherein the (w/v) percent ratio of individual component used in the range of 1 - 40% : 1 -99% : 0.005-1% : 1-20% respectively.
2. A spot-on formulation as claimed in claim 1, wherein the antioxidant is selected from the group consisting of butylated hydroxyanisole, butylated hydroxytoluene, ascorbic acid, sodium metabisulphite, propyl gallate, and sodium thiosulphate.
3. A spot-on formulation as claimed in claim 2, wherein the antioxidant preferably used is butylated hydroxytoluene.
4. A spot-on formulation as claimed in claim 1, wherein the nodulisporic acid derivative used is of the formula I
(Formula Removed)
wherein R| is hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, where the substituents on the alkyl, alkenyl, alkynyl, cycloalkyl and cycloalkenyl are 1 to 3 groups independently selected from alkyl, X-alkyl, where X is O or S(O)m, cycloalkyl, hydroxy, halogen, cyano, carboxy, NY'Y2, where Y1and Y2 are independently H or alkyl, alkanoylamino, and aroylamino wherein said aroyl is optionally substituted with 1 to 3 groups independently selected from Rf and aryl or arylalkyl wherein said aryl is optionally substituted with 1 to 3 groups independently selected from Rf, perfluoroalkyl, a 5-or 6-member heterocycle containing from 1 to 4 heteroatoms independently selected from oxygen, sulfur and nitrogen atoms optionally substituted by 1 to 3 groups independently selected from hydroxy, oxo, alkyl and halogen, and which may be saturated or partly unsaturated,
R2, R3, and R4 are independently ORa, OC02Rb, OC(O)NRcRd ; or
R1 and R2 together represent =0, =NORa or =N-NRcRd;
R5 and R6 are H; or
R5 and R6 together represent -0-;
R7 is CHO, or the fragment
(Formula Removed)

R8 is H, 0Ra, or NRcRd
R9isH,orORa;
R10is CN, C(O)ORb, C(O)N(ORb)Rc, C(O)NRcRd, NHC(O)ORb, NHC(O)NRcRd, CH2ORa, CH2OCO2Rb, CH2OC(O)NRcRd, C(O)NRCNRcRd, or C(O)NRcSO2Rb ; represents a single or a double bond;
Ra is hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkanoyl, optionally substituted alkenoyl, optionally substituted alkynoyl, optionally substituted aroyl, optionally substituted aryl, optionally substituted cycloalkanoyl, optionally substituted cycloalkenoyl, optionally substituted alkylsulfonyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl where the substituents on the alkyl, alkenyl, alkynyl, alkanoyl, alkenoyl, alkynoyl, aroyl, aryl, cycloalkanoyl, cycloalkenoyl, alkylsulfonyl, cycloalkyl and cycloalkenyl are from 1 to 10 groups independently selected from the group consisting of hydroxy, alkoxy, cycloalkyl, aryl alkoxy, NR9Rh, CO2Rb, CONRcRd and halogen, and perfluoroalkyl, arylsulfonyl optionally substituted with 1 to 3 groups independently selected from alkyl, perfluoroalkyl, nitro, halogen and cyano, and a 5-or 6-member heterocycle containing 1 to 4 heteroatoms selected from oxygen, sulfur and nitrogen optionally substituted by 1 to 4 groups independently selected from alkyl, alkenyl, perfluoroalkyl, amino, C(O)NRcRd, cyano, CO2Rb and halogen, and which may be saturated or partly unsaturated;
Rb is H, optionally substituted aryl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally
substituted cycloalkenyl, or optionally substituted heterocycle containing from 1 to 4 heteroatoms independently selected from oxygen, sulfur and nitrogen; where the substituents on the aryl, alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocycle, or alkynyl are from 1 to 10 groups independently selected from hydroxy, alkyl, oxo, SO2NRgRh, arylalkoxy, hydroxyalkyl, alkoxy, hydroxyalkoxy, aminoalkoxy, cyano, mercapto, alkyl-S(O)m, cycloalkyl optionally substituted with 1 to 4 groups independently selected from Re, cycloalkenyl, halogen, alkanoyloxy, C(O)NR9Rh, COzR', formyl, -NR9Rh, 5 to 9-member heterocycle, which may be saturated or partially unsaturated, containing from 1 to 4 heteroatoms independently selected from oxygen, sulfur and nitrogen, and optionally substituted with 1 to 5 groups independently selected from Re, optionally substituted aryl, wherein the aryl substituents are 1, 2-methylenedioxy or 1 to 5 groups independently selected from Re, optionally substituted arylalkoxy, wherein the aryl substituents are 1,2-methylenedioxy or 1 to 5 groups independently selected from Re, and perfluoroalkyl;
Rc and Rd are independently selected from Rb; or
Rc and Rd together with the N to which they are attached form a 3-to 10-member ring containing 0 to 2 additional heteroatoms selected from O, S(O)m, and N, optionally substituted with 1 to 3 groups independently selected from R9, hydroxy, thioxo and oxo;
Re is halogen, alkyl, perfluoroalkyl, -SCO^R1, cyano, nitro, R'0(CH2),r, R'CO2(CH2)v, R'OCO(CH2)v-, optionally substituted aryl where the substituents are from 1 to 3 of halogen, alkyl, alkoxy, or hydroxy, SO2NR9Rh, or amino;
Rf is alkyl, X-alkyl, where X is O or S(O)m, alkenyl, alkynyl, perfluoroalkyl, NY1Y2, where Y1and Y2 are independently H or alkyl, hydroxy, halogen, and alkanoyl amino,
R9 and Rh are independently hydrogen, alkyl optionally substituted with hydroxy, amino, or C02R', aryl optionally substituted with halogen, 1, 2-methylenedioxy, alkoxy, alkyl or perfluoroalkyl, arylalkyl, wherein the aryl is optionally substituted with perfluorolkyl or 1, 2-methylenedioxy; alkoxycarbonyl, alkanoyl, alkanoylalkyl, arylalkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl, dialkylaminocarbonyl; or R9 and Rh together with the N to which they are attached form a 3-to 7-member ring containing 0 to 2
additional heteroatoms selected from O, S(O)m and N, optionally substituted with 1 to 3 groups independently selected from Re and oxo;
R' is hydrogen, perfluoroalkyl, alkyl, optionally substituted aryl, or arylalkyl, where the aryl substituents are from 1 to 3 groups independently selected from halogen, alkyl, alkoxy, and hydroxy;
m is 0 to 2 ; and
v is 0 to 3; or a pharmaceutically acceptable salt thereof.
or fragment
(Formula Removed)

wherein Rx is selected from the group consisting of : H, CH3, CH2CH3, C(CH3)3,
CH2CH2CH3, CH2CH20H, CH(C02CH3) CH20H, CH2C02CH3,
CH2CH(OCH2CH3)2, CH2CH20CH2CH20H, CH(CH3)(CH2)3C(CH3)20H, (CH2)30H, (CH2)40H, (CH2)SOH, CH (CH20H)CH2CH3, NHC(CH3)3, CH2CN, (CH2)60H, CH2CH(OH)CH3, CH(CH20H) CH2CH2CH3, CH2CH2SCH3, CH2CH2SCH2CH3, CH2CONH2, CH(CH3)(CH20H), CH2CH2NHCH2CH20H, CH (CH20H) (CH2) 3CH3, CH (CH20CH3) CH3, (CH2) 2SH, (CH2) 4NH2, CH2CH2SO2CH3, CH2CH2S(O)CH3, CH(CH(CH3)2) CH20H, (CH2)3NH2, (CH2) 3N (CH2CH3) 2, (CH2)3N(CH3) 2, OCH2CH3, CH2CH(OH)CH20H, OCH3, CH2CH20CH3, CH2CH2NHC (O) CH3, C (CH3) 2CH20H, c-C3Hs, cC6H11, (CH2) 30CH2CH3, CH2CHHCH2, C(CH2CH3)(CH20H)2, CH2C=CH, CH2C02CH2CH3, CH2CH2F, (CH2)30(CH2)ICH3, CH2CH2N(CH3)2, CH2CH2IOCH2CH2NH2, CH2CF3, NHCH2C02CH2CH3, CH(CH3) C02CH3, C(CH3)2CH2C(O)CH3, CH(C02CH2CH3)2, CH2CH3, CH(CH2CH2CH3)
C02CH3, CH2CH2CH20CH3, C(CH3) 2CH2CSCH, (CH2)4CH3, CH(CH2CH2CH3)2, (CH2) SCH3, CH2CH2C02H, CH(CH(CH3) 2)C02CH3, OCH2C02H, CH(CH(CH3)2) CH20H, CH (CH (CH3)2) CH20H, CH(CH3)CH20H, CH(CH3)CH20H, CH(CH3)2, (CH2)CH(CH3)2, CH (CH3)CH2CH3, CH2CH(CH3)OH, (CH2)3CH3, (CH2)20CH2CH3, 1-adamantyl, (CH2)8CH3, CH(CH3)CH(CH3)2, (CH2)3NHCH3, (CH2) 2N(CH2CH3)2,
(Formula Removed)
5. A spot-on formulation as claimed in claim 4, wherein R| is preferably used in a compound of Formula 1 is selected from the group comprising hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl where the substituents on the alkyl, alkenyl, alkynyl, cycloalkyl and cycloalkenyl are 1 to 3 groups independently selected from the group consisting of alkyl, X-alkyl, where X is O or S(O)mS , cycloalkyl, hydroxy, halogen, cyano, carboxy, and NY'Y2, where Y'and Y2,are independently H or alkyl, and aryl alkyl wherein said aryl is optionally substituted with 1 to 3 groups independently selected from Rf, perfluoroalkyl, a 5-or 6-member heterocycle containing from 1 to 4 heteroatoms independently selected from oxygen, sulfur and nitrogen atoms
optionally substituted by 1 to 3 groups independently selected from hydroxy, oxo, alkyl and halogen, and which may be saturated or partly unsaturated,
R8 is H, OH, or NH2;
R9 is H or OH;
R10 is C(O)ORb, C(O)N(ORb)Rc, C(O)NRcRd, NHC(O)ORb, NHC(O)NRcRd, CH2ORa, CH2OCO2Rb, CH2OC(O)NRcRd; C(O)NRcNRcRd or C(O)NRcSO2Rb;
Ra is hydrogen, optionally alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkanoyl, optionally substituted alkenoyl, optionally substituted alkynoyl, optionally substituted aroyl, optionally substituted aryl, optionally substituted cycloalkanoyl, optionally substituted cycloalkenoyl, optionally substituted alkylsulfonyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl where the substiuents on the alkyl, alkenyl, alkynyl, alkanoyl, alkenoyl, alkynoyl, aroyl, aryl, cycloalkanoyl, cycloalkenoyl, a Ikylsulfonyl, cycloalkyl and cycloalkenyl are from 1 to 10 groups independently selected from hydroxy, alkoxy, cycloalkyl, aryl alkoxy, NR9Rh, C02Rb, CONRcRd and halogen, perfluoroalkyl, arylsulfonyl optionally substituted with 1 to 3 groups independently selected from alkyl, perfluoroalkyl, halogen and cyano, a 5-or 6-member heterocycle containing 1 to 4 heteroatoms selected from oxygen, sulfur and nitrogen optionally substituted by 1 to 4 groups independently selected from alkyl, alkenyl, perfluoroalkyl, amino, C(O)NRcRd, cyano, CO2Rb and halogen, and which may be saturated or partly unsaturated;
Rb is H, optionally substituted aryl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, or optionally substituted 5-to 10-member heterocycle containing from 1 to 4 heteroatoms independently selected from oxygen, sulfur and nitrogen ; where the substituents on the aryl, alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocycle, or alkynyl are from 1 to 10 groups independently selected from hydroxy, alkyl, oxo, SO2NR9Rh, arylalkoxy, hydroxyalkyl, alkoxy, hydroxyalkoxy, aminoalkoxy, cyano, perfluoroalkyl, alkyl-S(O)m, cycloalkyl optionally substituted with 1 to 4 groups independently selected from Re, cycloalkenyl, halogen, alkanoyloxy, C(O)NR9Rh, C02R',
optionally substituted aryl alkoxy, wherein the aryl substituents are 1,2-methylenedioxy or 1 to 5 groups independently selected from Re, -NR9Rh, 5 to 6-member heterocycle, which may be saturated or partially unsaturated, containing from 1 to 4 heteroatoms independently selected from oxygen, sulfur and nitrogen, and optionally substituted with 1 to 5 groups independently selected from Re, and optionally substituted aryl, wherein the aryl substituents are 1,2-methylenedioxy or 1 to 5 groups independently selected from Re;
Re is halogen, alkyl, perfluoroalkyl, -S(O)R1, (5) cyano, amino, (CH2)V-, RiCO2(CH2)v-, R'OCO(CH2)v- optionally substituted aryl where the substituents are from 1 to 3 of halogen, alkyl, alkoxy, or hydroxy, or SO2NR9Rh;
Rf is methyl, X-alkyl, where X is O or S(O)m, halogen, acetylamino, trifluoromethyl, NYV2, where Y1and Y2 are independently H or methyl, and hydroxy;
R9 and Rh are independently hydrogen, alkyl optionally substituted with hydroxy, amino, or C02R1 aryl optionally substituted with halogen, 1,2- methylenedioxy, alkoxy, alkyl or perfluoroalkyl, aryl alkyl, wherein the aryl is optionally substituted with perfluorolkyl or 1, 2-methylenedioxy; alkoxycarbonyl, alkanoyl, alkanoylalkyl, arylalkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl, dialkylaminocarbonyl; or
R9 and Rh together with the N to which they are attached form a 5-to 6membered ring containing 0 to 2 additional heteroatoms selected from O, S(O)m, and N, optionally substituted with 1 to 3 groups independently selected from Reand oxo;
R' is hydrogen, perfluoroalkyl, alkyl, optionally substituted aryl alkyl, where the aryl substituents are from 1 to 3 groups independently selected from halogen, alkyl, alkoxy, and hydroxy.
6. A spot-on formulation as claimed in claim 4, wherein R' used in compound of Formula 1 is preferably selected from the group comprising of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, where the substituents on the alkyl, alkenyl, and alkynyl are 1 to 3 groups independently selected from methyl,
X-methyl, where X is O or S(O)m and halogen, arylalkyl wherein said aryl is optionally substituted with 1 to 3 groups independently selected from Rf, trifluoromethyl
R8isH,OH, orNH2
R9isH, or OH;
R10 is C(O)ORb, C(O)N(ORb)Rc, C(O)NRcRd, NHC(O)ORb, NHC(O)NRcRd, CH2ORa, CH2OC02Rb, CH2OC(O)NRcRd, C(O)NRcNRcRd or C(O)NRcSO2Rb;
Ra is hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkanoyl, optionally substituted aroyl, optionally substituted cycloalkanoyl, optionally substituted cycloalkenoyl, optionally substituted alkylsulfonyl where the substituents on the alkyl, alkenyl, alkynyl, alkanoyl, aroyl, cycloalkanoyl, cycloalkenoyl, and alkylsulfonyl, are from 1 to 5 groups independently selected from hydroxy, alkoxy, aryl alkoxy, NR9Rh, C02Rb, CONRcRd and halogen, trifluoromethyl, arylsulfonyl optionally substituted with 1 to 3 groups independently selected from methyl, trifluoromethyl and halogen, and a 5-or 6-member heterocycle containing 1 to 4 heteroatoms selected from oxygen, sulfur and nitrogen optionally substituted by 1 to 4 groups independently selected from methyl, trifluoromethyl, C(O)NRcRd, CO2Rb and halogen, and which may be saturated or partly unsaturated;
Rb is H, optionally substituted aryl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, or optionally substituted 5-to 6-member heterocycle containing from 1 to 4 heteroatoms independently selected from oxygen, sulfur and nitrogen; where the substituents on the aryl, alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocycle, or alkynyl are from 1 to 10. groups independently selected from hydroxy, alkyl, oxo, SO2NR9Rh, arylalkoxy, hydroxyalkyl, alkoxy, hydroxyalkoxy, aminoalkoxy, cyano, alkyl-S(O)m, cycloalkyl optionally substituted with 1 to 4 groups independently selected from Re, cycloalkenyl, halogen, alkanoyloxy, C(O)NR9Rh, COzR1, -NRgRh, 5 to 6-member heterocycle, which may be saturated or partially unsaturated, containing from 1 to 4 heteroatoms independently selected from oxygen, sulfur and nitrogen, and optionally substituted with 1 to 5 groups independently selected from Re, optionally substituted aryl,
wherein the aryl substitutents are 1, 2-methylenedioxy or 1 to 5 groups independently selected from Re (xxi) optionally substituted aryl alkoxy, wherein the aryl substituents are 1,2-methylenedioxy or 1 to 5 groups independently selected from Re, and perfluoroalkyl;
Re is halogen, alkyl, perfluoroalkyl, -S(O)mR', cyano, R'O(CH2)v-, R'CO2(CH2)v_ R1OCO(CH2)v- optionally substituted aryl where the substituents are from 1 to 3 of halogen, alkyl, alkoxy, or hydroxy, S02NR9Rh, or amino;
Rf is methyl, X-alkyl, where X is O or S(O)m, trifluoromethyl, NY'Y2, where Y'and Y2 are independently H or methyl, hydroxy, halogen, and acetylamino,
R9 and Rh are independently hydrogen, alkyl optionally substituted with hydroxy, amino, or C02R', aryl optionally substituted with halogen, 1, 2methylenedioxy, alkoxy, alkyl or perfluoroalkyl, aryl or arylalkyl, wherein the aryl is optionally substituted with perfluorolkyl or 1, 2-methylenedioxy ; alkoxycarbonyl, alkanoyl, alkanoylalkyl, arylalkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl, dialkylaminocarbonyl; or
R9 and Rh together with the N to which they are attached form a 5-to 6-member ring containing 0 to 2 additional heteroatoms selected from O, S(O)m, and N, optionally substituted with 1 to 3 groups independently selected from Re and oxo;
R' is hydrogen, perfluoroalkyl, alkyl, optionally substituted aryl or arylalkyl, where the aryl substituents are from 1 to 3 groups independently selected from halogen, alkyl, alkoxy, and hydroxy.
7. 8 A spot-on formulation as claimed in claim 45, wherein R10 used in a compound of Formula 1 is most preferably selected from the group consisting of C(O)NRcRd ;
Rb is hydrogen, optionally substituted aryl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, or optionally substituted 5 to 6-member heterocycle containing from 1 to 4 heteroatoms independently selected from oxygen, sulfur and nitrogen; where the substituents on the aryl, alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocycle, or alkynyl are from 1 to 10 groups independently selected from the group
consisting of hydroxy, alkyl, oxo, SO2NR9Rh, arylalkyl, hydroxyalkylfoxy, hydroxyalkoxy, aminoalkoxy, cyano, perfluoroalkyl, alkyl-S(O)m), cycloalkyl optionally substituted with 1 to 4 groups selected from Re, (xiv) cycloalkenyl, halogen, C(O)NR9Rh, CO2R1, -NR9Rh, 5 to 9-member heterocycle containing from 1 to 4 heteroatoms independently selected from oxygen, sulfur and nitrogen, and optionally substituted with 1 to 3 groups independently selected from Re, optionally substituted aryl, wherein the aryl substituents are 1,2-methylenedioxy or 1 to 5 groups independently selected from Re and optionally substituted aryl alkoxy, wherein the aryl substituents are 1,2-methylenedioxy or 1 to 5 groups independently selected from Re;
Rc and Rd are independently selected from Rb; or
Re and Rd together with the N to which they are attached form a 3-to 10-member ring containing 0 to 2 additional heteratoms selected from O, S(0)m, and N, optionally substituted with 1 to 3 groups independently selected from R9, hydroxy, thioxo and oxo;
Re is halogen, alkyl, perfluoroalkyl, RiO(CH2)v., R'CO2(CH2)v., R'OCO (CH2)V-, SO2NR9Rh ; amino v is 0 ;
Rg and Rh are independently hydrogen, alkyl optionally substituted with hydroxy, amino, or CO2R', aryl optionally substituted with halogen, 1,2-methylenedioxy, alkoxy, alkyl or perfluoroalkyl, aryl or arylalkyl, wherein the aryl is optionally substituted with perfluoroalkyl or 1, 2methylenedioxy, alkoxycarbonyl, alkanoyl, arylalkoxycarbonyl, aminocarbonyl, or R9 and Rh together with the N to which they are attached form a 5-to 6-member ring containing 0 to 2 additional heteroatoms selected from O, S(O)m, and N, optionally substituted with 1 to 3 groups independently selected from Re and oxo;
R' is hydrogen or optionally substituted alkyl wherein the substituents are aryl or substituted aryl, and the aryl substituents are from 1 to 3 groups independently selected from halogen, alkyl, alkoxy, and hydroxy.
8. A spot-on formulation as claimed in claim 4, wherein the nodulisporic acid derivative is preferably used is t-butyl nodulisporamide of formula 2
(Formula Removed)
Formula 2
wherein Rx is selected from the group consisting of : H, CH3, CH2CH3, C(CH3)3,
CH2CH2CH3, CH2CH20H, CH(C02CH3) CH20H, CH2C02CH3,
CH2CH(OCH2CH3)2, CH2CH20CH2CH20H, CH(CH3)(CH2)3C(CH3)20H, (CH2)30H, (CH2)40H, (CH2)SOH, CH (CH20H)CH2CH3, NHC(CH3)3, CH2CN, (CH2)60H, CH2CH(OH)CH3, CH(CH20H) CH2CH2CH3, CH2CH2SCH3, CH2CH2SCH2CH3, CH2CONH2, CH(CH3)(CH20H), CH2CH2NHCH2CH20H, CH (CH20H) (CH2) 3CH3, CH (CH20CH3) CH3, (CH2) 2SH, (CH2) 4NH2, CH2CH2S02CH3, CH2CH2S(0)CH3, CH(CH(CH3)2) CH20H, (CH2)3NH2, (CH2) 3N (CH2CH3) 2, (CH2)3N(CH3) 2, OCH2CH3, CH2CH(OH)CH20H, OCH3, CH2CH20CH3, CH2CH2NHC (O) CH3, C (CH3) 2CH20H, c-C3Hs, cC6H11, (CH2) 30CH2CH3, CH2CH=CH2, C(CH2CH3)(CH20H)2, CH2C=CH, CH2C02CH2CH3, CH2CH2F, (CH2)30(CH2)ICH3, CH2CH2N(CH3)2, CH2CH2IOCH2CH2NH2, CH2CF3, NHCH2C02CH2CH3, CH(CH3) C02CH3, C(CH3)2CH2C(0)CH3, CH(C02CH2CH3)2, CH2CH3, CH(CH2CH2CH3) C02CH3, CH2CH2CH20CH3, C(CH3) 2CH2CECH, (CH2)4CH3, CH(CH2CH2CH3)2, (CH2) SCH3, CH2CH2C02H, CH(CH(CH3) 2)C02CH3, OCH2C02H, CH(CH(CH3)2) CH20H, CH (CH (CH3)2) CH20H, CH(CH3)CH20H, CH(CH3)CH20H, CH(CH3)2, (CH2)CH(CH3)2, CH (CH3)CH2CH3, CH2CH(CH3)OH, (CH2)3CH3, (CH2)20CH2CH3, 1-adamantyl, (CH2)8CH3, CH(CH3)CH(CH3)2, (CH2)3NHCH3, (CH2) 2N(CH2CH3)2,
(Formula Removed)
9. A spot-on formulation as claimed in claim 8, wherein Rx is preferably used in Formula
2 is C(CH3)3.
10. A spot-on formulation as claimed in claim 1, wherein the said liquid carrier vehicle is
selected from the group comprising a solvent, microemulsion, excipient and optionally a
cosolvent
11. A spot-on formulation as claimed in claim 10, wherein the solvent is selected from the group consisting of acetone, acetonitrile, benzyl alcohol, butyl diglycol, dimethylacetamide, dimethylformamide, dipropylene glycol n-butyl ether, ethanol, isopropanol, methanol, diethylene glycol monoethyl ether, ethylene glycol monomethyl ether, monomethylaceamide, dipropylene glycol monomethyl ether, liquid polyoxyethylene glycols, propylene glycol, 2-pyrrolidone, diethylene glycol monoethyl ether, ethylene glycol, diethyl phthalate, a mixture of at least two of these solvents.
12. A spot-on formulation as claimed in claim 10, wherein the excipient is selected from the group consisting of C8-C10 caprylic/capric triglycerides, oleic acid or propylene glycol.
13. A spot-on formulation as claimed in claim 10, wherein the cosolvent is selected from the group consisting of absolute ethanol, isopropanol or methanol.
14. A spot-on formulation as claimed in claim 1, wherein the liquid vehicle carrier preferably used is diethylene glycol monoethyl ether and C8-C10 caprylic/capric triglycerides.
15. A spot-on formulation as claimed in claim 1, wherein the said crystallization inhibitor is selected from the group consisting of an anionic surfactant, polymeric film- forming agent, a cationic surfactant, a non-ionic surfactant, an amine salt, an amphoteric surfactant, polyvinylpyrrolidone, polyvinyl alcohols, copolymers of vinyl acetate and vinylpyrrolidone, polyethylene glycols, benzyl alcohol, mannitol, glycerol, sorbitol, p olyoxyethylenated sorbitan esters; lecithin, sodium carboxymethylcellulose, and acrylic derivatives, or a mixture of these crystallization inhibitors.
16. A spot-on formulation as claimed in claim 15, wherein the said anionic surfactant is selected from the group comprising of alkaline stearates, sodium abietate; alkyl sulphates; sodium dodecylbenzenesulphonate, sodium dioctylsulphosuccinate; and fatty acids.
17. A spot-on formulation as claimed in claim 15, wherein the said polymeric film-forming agent is selected from the group comprising polyvinylpyrrolidone, polyvinyl alcohols, or a copolymer of vinyl acetate and polyvinylpyrrolidone and the surfactant is a non-ionic surfactant.
18. A spot-on formulation as claimed in claim 15, wherein the said cationic surfactant is selected from the group comprising water-soluble quaternary ammonium salts of formula N+R'R"R"'R""Y-in which the radicals R independently are hydrocarbon radicals, optionally hydroxylated, and Y is an anion of a strong acid.
19. A spot-on formulation as claimed in claim 15, wherein the said non-ionic surfactant is selected from the group comprising polyoxyethylenated sorbitan esters, polyoxyethylenated alkyl ethers ; polyethylene glycol stearate, polyoxyethylenated
derivatives of castor oil, polyglycerol esters, polyoxyethylenated fatty alcohols, polyoxyethylenated fatty acids, copolymers of ethylene oxide and propylene oxide.
20. A spot-on formulation as claimed in claim 15, wherein the said amine salt is selected
from the group comprising N+R'R"R" " in which the radicals R independently are
optionally hydroxylated hydrocarbon radicals;
21. A spot-on formulation as claimed in claim 15, wherein the said amphoteric surfactant is lauryl-substituted betaine compounds.
22. A spot-on formulation as claimed in claim 1, wherein the crystallization inhibitor preferably used is a mixture of polyvinylpyrrolidone and polyoxethylene (20) sorbitan mono-oleate.
23. A medicament prepared by using the spot-on formulation as claimed in claim 1.
24. A spot on formulations substantially as described and shown with reference to the description, examples and drawings.

Documents:

1334-delnp-2005-abstract.pdf

1334-delnp-2005-claims.pdf

1334-delnp-2005-correspondence- po.pdf

1334-delnp-2005-description (complete).pdf

1334-delnp-2005-form-1.pdf

1334-DELNP-2005-Form-13.pdf

1334-delnp-2005-form-18.pdf

1334-delnp-2005-form-2.pdf

1334-delnp-2005-form-3.pdf

1334-delnp-2005-form-5.pdf

1334-delnp-2005-gpa.pdf


Patent Number 248353
Indian Patent Application Number 1334/DELNP/2005
PG Journal Number 27/2011
Publication Date 08-Jul-2011
Grant Date 06-Jul-2011
Date of Filing 04-Apr-2005
Name of Patentee MERIAL LIMITED
Applicant Address 33239 SATELLITE SHORE ALPHARTTA,GA 30005 U.S.A.
Inventors:
# Inventor's Name Inventor's Address
1 SOLL, MARK,D. 605 NEWPORT SHORE ALPHARETTA, GA 30005 U.S.A
2 BOECKH, ALBERT 7415 TADWORTH TRACE, CUMMING, GA 30041 U.S.A.
3 DE BODE, RONUS HASSELDERHEIDELAAN 26 NL-5941 CV VELDEN NETHERLAND
4 VAN EIJK,PETER,JOHANNES, SERVAS,SAVIO KAREKIETHOF 25 NL-6005 JM WEERT (NL) NETHERLANDS
PCT International Classification Number A01N 43/90
PCT International Application Number PCT/US03/030500
PCT International Filing date 2003-09-29
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 10/618,975 2003-07-14 U.S.A.
2 60/415,627 2002-10-02 U.S.A.