WO2015027040A2

WO2015027040A2 - Methods and compositions for treating schistosome infection

Info

Publication number: WO2015027040A2
Application number: PCT/US2014/052049
Authority: WO
Inventors: Patrick SKELLY; Akram DA'DARAH
Original assignee: Tufts University
Priority date: 2013-08-21
Filing date: 2014-08-21
Publication date: 2015-02-26
Also published as: WO2015027040A3

Abstract

The present invention relates to compositions and methods for the treatment of Schistosome infection. In particular, the present invention provides therapeutic agents for the treatment of Schistosome infection.

Description

METHODS AND COMPOSITIONS FOR TREATING SCHISTOSOME INFECTION

The present application claims priority to United States Provisional Patent

Application Serial Number 61/868,394, filed August 21, 2013, the disclosure of which is herein incorporated by reference in its entirety.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR

DEVELOPMENT

This invention was made with government support under AI056273 awarded by the National Institutes of Health. The government has certain rights in the invention.

FIELD OF THE INVENTION

BACKGROUND OF THE INVENTION

Schistosomes are intravascular parasitic worms commonly called blood flukes that are of enormous public health importance and cause the disease schistosomiasis. Schistosomiasis (also known as bilharzia, bilharziosis or snail fever) is a collective name of parasitic diseases caused by several species of trematodes belonging to the genus Schistosoma. Snails serve as the intermediary agent between mammalian hosts. Individuals within developing countries who cannot afford proper water and sanitation facilities are often exposed to contaminated water containing the infected snails.

Although it has a low mortality rate, schistosomiasis often is a chronic illness that can damage internal organs and, in children, impair growth and cognitive development. The urinary form of schistosomiasis is associated with increased risks for bladder cancer in adults. Schistosomiasis is the second most socioeconomically devastating parasitic disease after malaria.

Schistosomiasis is most commonly found in Asia, Africa, and South America, especially in areas where the water contains numerous freshwater snails, which may carry the parasite.

Schistosomiasis affects many people in developing countries, particularly children who may acquire the disease by swimming or playing in infected water. When children come into contact with a contaminated water source, the parasitic larvae easily enter through their skin and further mature within organ tissues. As of 2009, 74 developing countries statistically identified epidemics of Schistosomiasis within their respective populations.

Schistosomiasis is currently treated using a single oral dose of the drug praziquantel annually. However, there has been emergence of resistance to praziquantel. The WHO and others have called for the urgent development of new drugs to treat infection. Additional treatments suitable for use in the developing world and elsewhere are needed.

SUMMARY

The present invention relates to compositions and methods for the treatment of

Schistosome infection. In particular, the present invention provides therapeutic agents for the treatment of Schistosome infection.

Embodiments of the present invention provide compositions, methods, and uses for treating or preventing infection by a schistosome (e.g., including but not limited to

Schistosoma mansoni, Schistosoma haematobium, or Schistosoma japonicum), comprising administering 2-(2-fluorophenyl)ethyl] {3-methoxy-4-[2-oxo-2-(l- piperidinyl)ethoxy]benzyl} amine hydrochloride to a subject infected with or at risk of infection by a schistosome. In some embodiments, the subject has been diagnosed with schistosome infection or lives in or is traveling to an area where schistosome infection is endemic. In some embodiments, the administration treats or prevents infection by the schistosome. In some embodiments, 2-(2-fluorophenyl)ethyl] {3-methoxy-4-[2-oxo-2-(l- piperidinyl)ethoxy]benzyl} amine hydrochloride inhibits an acetylcholinesterase enzyme of the schistosome.

In further embodiments, the present invention provides compositions, uses, and methods for inhibiting an acetylcholinesterase enzyme in a cell, comprising administering 2- (2-fluorophenyl)ethyl] {3-methoxy-4-[2-oxo-2-(l-piperidinyl)ethoxy]benzyl} amine hydrochloride to the cell. In some embodiments, the cell is in a subject. In some

embodiments, the subject has been diagnosed with a disorder related to acetylcholinesterase activity (e.g., including but not limited to, Alzheimer's disease, glaucoma, or myasthenia gravis) and the administration treats the disorder. In some embodiments, the cell is a schistosome (e.g., Schistosoma mansoni, Schistosoma haematobium, or Schistosoma japonicum).

Additional embodiments are described herein. DEFINITIONS

To facilitate an understanding of the present invention, a number of terms and phrases are defined below.

As used herein, the term "inhibits at least one biological activity of

acetylcholinesterase" refers to any agent that decreases any activity of a acetylcholinesterase polypeptide (e.g., Schistosome acetylcholinesterase) (e.g., including, but not limited to, the activities described herein), via directly contacting a acetylcholinesterase protein, contacting a acetylcholinesterase m NA or genomic DNA, causing conformational changes of acetylcholinesterase polypeptides, decreasing acetylcholinesterase protein levels, or interfering with acetylcholinesterase interactions with signaling partners, and affecting the expression of acetylcholinesterase target genes. Inhibitors also include molecules that indirectly regulate acetylcholinesterase biological activity by intercepting or otherwise influencing upstream or downstream signaling molecules.

As used herein, the term "subject" refers to any animal (e.g., a mammal), including, but not limited to, humans, non-human primates, rodents, and the like, which is to be the recipient of a particular treatment. Typically, the terms "subject" and "patient" are used interchangeably herein in reference to a human subject.

As used herein, the term "non-human animals" refers to all non-human animals including, but are not limited to, vertebrates such as rodents, non-human primates, ovines, bovines, ruminants, lagomorphs, porcines, caprines, equines, canines, felines, aves, etc.

As used, the term "eukaryote" refers to organisms distinguishable from "prokaryotes." It is intended that the term encompass all organisms with cells that exhibit the usual characteristics of eukaryotes, such as the presence of a true nucleus bounded by a nuclear membrane, within which lie the chromosomes, the presence of membrane-bound organelles, and other characteristics commonly observed in eukaryotic organisms. Thus, the term includes, but is not limited to such organisms as fungi, protozoa, and animals (e.g., humans).

As used herein, the term "in vitro" refers to an artificial environment and to processes or reactions that occur within an artificial environment. In vitro environments can consist of, but are not limited to, test tubes and cell culture. The term "in vivo" refers to the natural environment (e.g., an animal or a cell) and to processes or reaction that occur within a natural environment.

The terms "test compound" and "candidate compound" refer to any chemical entity, pharmaceutical, drug, and the like that is a candidate for use to treat or prevent a disease, illness, sickness, or disorder of bodily function (e.g., Schistosome infection). Test compounds comprise both known and potential therapeutic compounds. A test compound can be determined to be therapeutic by screening using the screening methods of the present invention. In some embodiments of the present invention, test compounds include antisense compounds.

As used herein, the term "sample" is used in its broadest sense. In one sense, it is meant to include a specimen or culture obtained from any source, as well as biological and environmental samples. Biological samples may be obtained from animals (including humans) and encompass fluids, solids, tissues, and gases. Biological samples include blood products, such as plasma, serum and the like. Such examples are not however to be construed as limiting the sample types applicable to the present invention.

The term "chemical moiety" refers to any chemical compound containing at least one atom (e.g., carbon atom). Examples of chemical moieties include, but are not limited to, aromatic chemical moieties, chemical moieties comprising sulfur, chemical moieties comprising nitrogen, hydrophilic chemical moieties, and hydrophobic chemical moieties. As used herein, the term "aliphatic" represents the groups including, but not limited to, alkyl, alkenyl, alkynyl, alicyclic.

As used herein, the term "alkyl" refers to an unsaturated carbon chain substituent group. In general, alkyls have the general formula CnH₂n+l . Exemplary alkyls include, but are not limited to, methyl (CH₃), ethyl (C₂H₅), propyl (C₃H₇), butyl (C₄H₉), pentyl (C₅Hn), etc.

As used herein, the term "aryl" represents a single aromatic ring such as a phenyl ring, or two or more aromatic rings (e.g., bisphenyl, naphthalene, anthracene), or an aromatic ring and one or more non-aromatic rings. The aryl group can be optionally substituted with a lower aliphatic group (e.g., alkyl, alkenyl, alkynyl, or alicyclic). Additionally, the aliphatic and aryl groups can be further substituted by one or more functional groups including, but not limited to, chemical moieties comprising N, S, O, -NH₂, -NHCOCH₃, -OH, lower alkoxy (C1-C4), and halo (-F, -CI, -Br, or -I).

As used herein, the term "substituted aliphatic" refers to an alkane, alkene, alkyne, or alicyclic moiety where at least one of the aliphatic hydrogen atoms has been replaced by, for example, a halogen, an amino, a hydroxy, a nitro, a thio, a ketone, an aldehyde, an ester, an amide, a lower aliphatic, a substituted lower aliphatic, or a ring (aryl, substituted aryl, cycloaliphatic, or substituted cycloaliphatic, etc.). Examples of such include, but are not limited to, 1 -chloroethyl and the like. As used herein, the term "substituted aryl" refers to an aromatic ring or fused aromatic ring system consisting of at least one aromatic ring, and where at least one of the hydrogen atoms on a ring carbon has been replaced by, for example, a halogen, an amino, a hydroxy, a nitro, a thio, a ketone, an aldehyde, an ester, an amide, a lower aliphatic, a substituted lower aliphatic, or a ring (aryl, substituted aryl, cycloaliphatic, or substituted cycloaliphatic). Examples of such include, but are not limited to, hydroxyphenyl and the like.

As used herein, the term "cycloaliphatic" refers to an aliphatic structure containing a fused ring system. Examples of such include, but are not limited to, decalin and the like. As used herein, the term "substituted cycloaliphatic" refers to a cycloaliphatic structure where at least one of the aliphatic hydrogen atoms has been replaced by a halogen, a nitro, a thio, an amino, a hydroxy, a ketone, an aldehyde, an ester, an amide, a lower aliphatic, a substituted lower aliphatic, or a ring (aryl, substituted aryl, cycloaliphatic, or substituted cycloaliphatic). Examples of such include, but are not limited to, 1-chlorodecalyl, bicyclo- heptanes, octanes, and nonanes (e.g., nonrbornyl) and the like.

As used herein, the term "heterocyclic" represents, for example, an aromatic or nonaromatic ring containing one or more heteroatoms. The heteroatoms can be the same or different from each other. Examples of heteroatoms include, but are not limited to nitrogen, oxygen and sulfur. Aromatic and nonaromatic heterocyclic rings are well-known in the art. Some nonlimiting examples of aromatic heterocyclic rings include pyridine, pyrimidine, indole, purine, quinoline and isoquinoline. Nonlimiting examples of nonaromatic

heterocyclic compounds include piperidine, piperazine, morpholine, pyrrolidine and pyrazolidine. Examples of oxygen containing heterocyclic rings include, but not limited to furan, oxirane, 2H-pyran, 4H-pyran, 2H-chromene, and benzofuran. Examples of sulfur- containing heterocyclic rings include, but are not limited to, thiophene, benzothiophene, and parathiazine. Examples of nitrogen containing rings include, but not limited to, pyrrole, pyrrolidine, pyrazole, pyrazolidine, imidazole, imidazoline, imidazolidine, pyridine, piperidine, pyrazine, piperazine, pyrimidine, indole, purine, benzimidazole, quinoline, isoquinoline, triazole, and triazine. Examples of heterocyclic rings containing two different heteroatoms include, but are not limited to, phenothiazine, morpholine, parathiazine, oxazine, oxazole, thiazine, and thiazole. The heterocyclic ring is optionally further substituted with one or more groups selected from aliphatic, nitro, acetyl (i.e., -C(=0)-CH₃), or aryl groups.

As used herein, the term "substituted heterocyclic" refers to a heterocylic structure where at least one of the ring carbon atoms is replaced by oxygen, nitrogen or sulfur, and where at least one of the aliphatic hydrogen atoms has been replaced by a halogen, hydroxy, a thio, nitro, an amino, a ketone, an aldehyde, an ester, an amide, a lower aliphatic, a substituted lower aliphatic, or a ring (aryl, substituted aryl, cycloaliphatic, or substituted cycloaliphatic). Examples of such include, but are not limited to 2-chloropyranyl.

The term "derivative" of a compound, as used herein, refers to a chemically modified compound wherein the chemical modification takes place either at a functional group of the compound or backbone.

The term "diagnosed," as used herein, refers to the recognition of a disease by its signs and symptoms (e.g., resistance to conventional therapies), or genetic analysis, pathological analysis, histological analysis, and the like.

As used herein, the term "host cell" refers to any eukaryotic or prokaryotic cell (e.g., mammalian cells, avian cells, amphibian cells, plant cells, fish cells, and insect cells), whether located in vitro or in vivo.

As used herein, the term "cell culture" refers to any in vitro culture of cells. Included within this term are continuous cell lines (e.g., with an immortal phenotype), primary cell cultures, finite cell lines (e.g., non-transformed cells), and any other cell population maintained in vitro, including oocytes and embryos.

As used herein, the term "effective amount" refers to the amount of a compound (e.g., a compound of the present invention) sufficient to effect beneficial or desired results. An effective amount can be administered in one or more administrations, applications or dosages and is not limited intended to be limited to a particular formulation or administration route.

As used herein, the term "co-administration" refers to the administration of at least two agent(s) (e.g., a compound of the present invention) or therapies to a subject. In some embodiments, the co-administration of two or more agents/therapies is concurrent. In some embodiments, a first agent/therapy is administered prior to a second agent/therapy. Those of skill in the art understand that the formulations and/or routes of administration of the various agents/therapies used may vary. The appropriate dosage for co-administration can be readily determined by one skilled in the art. In some embodiments, when agents/therapies are coadministered, the respective agents/therapies are administered at lower dosages than appropriate for their administration alone. Thus, co-administration is especially desirable in embodiments where the co-administration of the agents/therapies lowers the requisite dosage of a known potentially harmful (e.g., toxic) agent(s).

As used herein, the term "toxic" refers to any detrimental or harmful effects on a cell or tissue as compared to the same cell or tissue prior to the administration of the toxicant. As used herein, the term "pharmaceutical composition" refers to the combination of an active agent with a carrier, inert or active, making the composition especially suitable for diagnostic or therapeutic use in vivo, in vivo or ex vivo.

As used herein, the term "pharmaceutically acceptable carrier" refers to any of the standard pharmaceutical carriers, such as a phosphate buffered saline solution, water, emulsions (e.g., such as an oil/water or water/oil emulsions), and various types of wetting agents. The compositions also can include stabilizers and preservatives. For examples of carriers, stabilizers and adjuvants. (See e.g., Martin, Remington's Pharmaceutical Sciences, 15th Ed., Mack Publ. Co., Easton, PA [1975]).

As used herein, the term "pharmaceutically acceptable salt" refers to any

pharmaceutically acceptable salt (e.g., acid or base) of a compound of the present invention which, upon administration to a subject, is capable of providing a compound of this invention or an active metabolite or residue thereof. As is known to those of skill in the art, "salts" of the compounds of the present invention may be derived from inorganic or organic acids and bases. Examples of acids include, but are not limited to, hydrochloric, hydrobromic, sulfuric, nitric, perchloric, fumaric, maleic, phosphoric, glycolic, lactic, salicylic, succinic, toluene-p- sulfonic, tartaric, acetic, citric, methanesulfonic, ethanesulfonic, formic, benzoic, malonic, naphthalene-2-sulfonic, benzenesulfonic acid, and the like. Other acids, such as oxalic, while not in themselves pharmaceutically acceptable, may be employed in the preparation of salts useful as intermediates in obtaining the compounds of the invention and their

pharmaceutically acceptable acid addition salts.

Examples of bases include, but are not limited to, alkali metals (e.g., sodium) hydroxides, alkaline earth metals (e.g., magnesium), hydroxides, ammonia, and compounds of formula NW4+, wherein W is CI -4 alkyl, and the like.

Examples of salts include, but are not limited to: acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate,

flucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, 2- naphthalenesulfonate, nicotinate, oxalate, palmoate, pectinate, persulfate, phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, tosylate, undecanoate, and the like. Other examples of salts include anions of the compounds of the present invention compounded with a suitable cation such as Na+, NH4+, and NW4+ (wherein W is a CI -4 alkyl group), and the like. For therapeutic use, salts of the compounds of the present invention are contemplated as being pharmaceutically acceptable. However, salts of acids and bases that are non- pharmaceutically acceptable may also find use, for example, in the preparation or purification of a pharmaceutically acceptable compound.

DETAILED DESCRIPTION OF THE INVENTION

I. Inhibitors

Embodiments of the present invention provide compositions and methods for treating and preventing Schistosome infection. Experiments conducted during the course of development of embodiments of the present invention identified a novel tegumental (skin) enzyme on Schistosoma mansoni, S. haematobium, and S. japonicum parasites called acetylcholinesterase (and designated SmT-AChE). When expression of this enzyme was suppressed (e.g., using RNAi) the worms were unable to establish a robust infection in experimental animals.

SmT-AChE was then expressed as a functionally active recombinant protein and a library of 1 ,894 small molecules was tested for their ability to inhibit SmT-AChE. The following four novel inhibitors (and the percent inhibition) were identified:

1. [2-(2-fluorophenyl)ethyl] {3-methoxy-4-[2-oxo-2-(l -piperidinyl)ethoxy]benzyl} amine hydrochloride (85%)

2. Pentamidine isethionate salt (69%)

3. 3-(2,3-dihydro- 1 ,4-benzodioxin-6-yl)-6-ethyl-7-hydroxy-8-[(2-methyl-l - piperidinyl)methyl] -4H-chromen-4-one (49%)

4. 6,7-Dimethyl-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinoline-4-carboxylic acid (49%)

The above-described compounds are commercially available (e.g., from ChemBridge Corporation, San Diego, CA). In addition, embodiments of the present invention specifically contemplate variants, derivatives, stereoisomers, and mimetics of the above described compounds. II. Pharmaceutical compositions, formulations, and exemplary administration routes and dosing considerations

Exemplary embodiments of various contemplated medicaments and pharmaceutical or food based compositions are provided below.

Embodiments of the present invention provide methods of using the aforementioned compounds in the prevention and treatment of Schistosome infection.

A. Preparing Medicaments

The compounds of embodiments of the present invention are useful in the preparation of medicaments to treat Schistosome infection. The methods and techniques for preparing medicaments of a compound are well-known in the art. Exemplary pharmaceutical formulations and routes of delivery are described below.

One of skill in the art will appreciate that any one or more of the compounds described herein, including the many specific embodiments, are prepared by applying standard pharmaceutical manufacturing procedures. Such medicaments can be delivered to the subject by using delivery methods that are well-known in the pharmaceutical arts.

B. Exemplary pharmaceutical compositions and formulation

In some embodiments of the present invention, the compositions are administered alone, while in some other embodiments, the compositions are preferably present in a pharmaceutical formulation comprising at least one active ingredient/agent (e.g., compounds described herein), as defined above, together with a solid support or alternatively, together with one or more pharmaceutically acceptable carriers and optionally other therapeutic agents. Each carrier should be "acceptable" in the sense that it is compatible with the other ingredients of the formulation and not injurious to the subject.

Contemplated formulations include those suitable oral, rectal, nasal, topical (including transdermal, buccal and sublingual), vaginal, parenteral (including subcutaneous,

intramuscular, intravenous and intradermal) and pulmonary administration. In some embodiments, formulations are conveniently presented in unit dosage form and are prepared by any method known in the art of pharmacy. Such methods include the step of bringing into association the active ingredient with the carrier which constitutes one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association (e.g., mixing) the active ingredient with liquid carriers or finely divided solid carriers or both, and then if necessary shaping the product. Formulations of the present invention suitable for oral administration may be presented as discrete units such as capsules, cachets or tablets, wherein each preferably contains a predetermined amount of the active ingredient; as a powder or granules; as a solution or suspension in an aqueous or non-aqueous liquid; or as an oil-in- water liquid emulsion or a water-in-oil liquid emulsion. In some embodiments, the active ingredient is presented as a bolus, electuary, or paste, etc.

In some embodiments, tablets comprise at least one active ingredient and optionally one or more accessory agents/carriers are made by compressing or molding the respective agents. In some embodiments, compressed tablets are prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with a binder (e.g., povidone, gelatin, hydroxypropylmethyl cellulose), lubricant, inert diluent, preservative, disintegrant (e.g., sodium starch glycolate, cross-linked povidone, cross-linked sodium carboxymethyl cellulose) surface-active or dispersing agent. Molded tablets are made by molding in a suitable machine a mixture of the powdered compound (e.g., active ingredient) moistened with an inert liquid diluent. Tablets may optionally be coated or scored and may be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropylmethyl cellulose in varying proportions to provide the desired release profile. Tablets may optionally be provided with an enteric coating, to provide release in parts of the gut other than the stomach.

Formulations suitable for topical administration in the mouth include lozenges comprising the active ingredient in a flavored basis, usually sucrose and acacia or tragacanth; pastilles comprising the active ingredient in an inert basis such as gelatin and glycerin, or sucrose and acacia; and mouthwashes comprising the active ingredient in a suitable liquid carrier.

Pharmaceutical compositions for topical administration according to the present invention are optionally formulated as ointments, creams, suspensions, lotions, powders, solutions, pastes, gels, sprays, aerosols or oils. In alternatively embodiments, topical formulations comprise patches or dressings such as a bandage or adhesive plasters impregnated with active ingredient(s), and optionally one or more excipients or diluents. In some embodiments, the topical formulations include a compound(s) that enhances absorption or penetration of the active agent(s) through the skin or other affected areas. Examples of such dermal penetration enhancers include dimethylsulfoxide (DMSO) and related analogues.

If desired, the aqueous phase of a cream base includes, for example, at least about 30% w/w of a polyhydric alcohol, i.e., an alcohol having two or more hydroxyl groups such as propylene glycol, butane-l,3-diol, mannitol, sorbitol, glycerol and polyethylene glycol and mixtures thereof.

In some embodiments, oily phase emulsions are constituted from known ingredients in a known manner. This phase typically comprises a lone emulsifier (otherwise known as an emulgent), it is also desirable in some embodiments for this phase to further comprise a mixture of at least one emulsifier with a fat or an oil or with both a fat and an oil.

Preferably, a hydrophilic emulsifier is included together with a lipophilic emulsifier so as to act as a stabilizer. It some embodiments it is also preferable to include both an oil and a fat. Together, the emulsifier(s) with or without stabilizer(s) make up the so-called emulsifying wax, and the wax together with the oil and/or fat make up the so-called emulsifying ointment base which forms the oily dispersed phase of the cream formulations.

Emulgents and emulsion stabilizers suitable for use in the formulation of the present invention include Tween 60, Span 80, cetostearyl alcohol, myristyl alcohol, glyceryl monostearate and sodium lauryl sulfate.

The choice of suitable oils or fats for the formulation is based on achieving the desired properties (e.g., cosmetic properties), since the solubility of the active

compound/agent in most oils likely to be used in pharmaceutical emulsion formulations is very low. Thus creams should preferably be a non-greasy, non-staining and washable products with suitable consistency to avoid leakage from tubes or other containers. Straight or branched chain, mono- or dibasic alkyl esters such as di-isoadipate, isocetyl stearate, propylene glycol diester of coconut fatty acids, isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate, 2-ethylhexyl palmitate or a blend of branched chain esters known as Crodamol CAP may be used, the last three being preferred esters. These may be used alone or in combination depending on the properties required. Alternatively, high melting point lipids such as white soft paraffin and/or liquid paraffin or other mineral oils can be used.

Formulations suitable for topical administration to the eye also include eye drops wherein the active ingredient is dissolved or suspended in a suitable carrier, especially an aqueous solvent for the agent.

Formulations for rectal administration may be presented as a suppository with suitable base comprising, for example, cocoa butter or a salicylate.

Formulations suitable for vaginal administration may be presented as pessaries, creams, gels, pastes, foams or spray formulations containing in addition to the agent, such carriers as are known in the art to be appropriate. Formulations suitable for nasal administration, wherein the carrier is a solid, include coarse powders having a particle size, for example, in the range of about 20 to about 500 microns which are administered in the manner in which snuff is taken, i.e., by rapid inhalation (e.g., forced) through the nasal passage from a container of the powder held close up to the nose. Other suitable formulations wherein the carrier is a liquid for administration include, but are not limited to, nasal sprays, drops, or aerosols by nebulizer, an include aqueous or oily solutions of the agents.

Formulations suitable for parenteral administration include aqueous and non-aqueous isotonic sterile injection solutions which may contain antioxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents, and liposomes or other microparticulate systems which are designed to target the compound to blood components or one or more organs. In some embodiments, the formulations are presented/formulated in unit-dose or multi-dose sealed containers, for example, ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets of the kind previously described.

Preferred unit dosage formulations are those containing a daily dose or unit, daily subdose, as herein above -recited, or an appropriate fraction thereof, of an agent.

It should be understood that in addition to the ingredients particularly mentioned above, the formulations of this invention may include other agents conventional in the art having regard to the type of formulation in question, for example, those suitable for oral administration may include such further agents as sweeteners, thickeners and flavoring agents. It also is intended that the agents, compositions and methods of this invention be combined with other suitable compositions and therapies. Still other formulations optionally include food additives (suitable sweeteners, flavorings, colorings, etc.), phytonutrients (e.g., flax seed oil), minerals (e.g., Ca, Fe, K, etc.), vitamins, and other acceptable compositions (e.g., conjugated linoelic acid), extenders, and stabilizers, etc.

C. Exemplary administration routes and dosing considerations

Various delivery systems are known and can be used to administer a therapeutic agent (e.g., inhibitor of Schistosome infection), e.g., encapsulation in liposomes, microparticles, microcapsules, receptor-mediated endocytosis, and the like. Methods of delivery include, but are not limited to, intra-arterial, intra-muscular, intravenous, intranasal, and oral routes. In specific embodiments, it may be desirable to administer the pharmaceutical compositions of the invention locally to the area in need of treatment; this may be achieved by, for example, and not by way of limitation, local infusion during surgery, injection, or by means of a catheter.

The agents identified herein as effective for their intended purpose can be

administered to subjects or individuals diagnosed with or at increased risk of Schistosome infection. When the agent is administered to a subject such as a mouse, a rat or a human patient, the agent can be added to a pharmaceutically acceptable carrier and systemically or topically administered to the subject.

In some embodiments, in vivo administration is effected in one dose, continuously or intermittently throughout the course of treatment. Methods of determining the most effective means and dosage of administration are well known to those of skill in the art and vary with the composition used for therapy, the purpose of the therapy, the target cell being treated, and the subject being treated. Single or multiple administrations are carried out with the dose level and pattern being selected by the treating physician.

Suitable dosage formulations and methods of administering the agents are readily determined by those of skill in the art. Preferably, the compounds are administered at about 0.01 mg/kg to about 200 mg/kg, more preferably at about 0.1 mg/kg to about 100 mg/kg, even more preferably at about 0.5 mg/kg to about 50 mg/kg. When the compounds described herein are co-administered with another agent (e.g., as sensitizing agents), the effective amount may be less than when the agent is used alone.

The pharmaceutical compositions can be administered orally, intranasally, parenterally or by inhalation therapy, and may take the form of tablets, lozenges, granules, capsules, pills, ampoules, suppositories or aerosol form. They may also take the form of suspensions, solutions and emulsions of the active ingredient in aqueous or nonaqueous diluents, syrups, granulates or powders. In addition to an agent of the present invention, the pharmaceutical compositions can also contain other pharmaceutically active compounds or a plurality of compounds of the invention.

More particularly, an agent of the present invention also referred to herein as the active ingredient, may be administered for therapy by any suitable route including, but not limited to, oral, rectal, nasal, topical (including, but not limited to, transdermal, aerosol, buccal and sublingual), vaginal, parental (including, but not limited to, subcutaneous, intramuscular, intravenous and intradermal) and pulmonary. It is also appreciated that the preferred route varies with the condition and age of the recipient, and the disease being treated.

In some embodiments, agents are administered intravenously. In some embodiments, agents are formulated in Cremophor (BASF, Parsippany, N.J.)

Ideally, the agent should be administered to achieve peak concentrations of the active compound at sites of disease. This may be achieved, for example, by the intravenous injection of the agent, optionally in saline, or orally administered, for example, as a tablet, capsule or syrup containing the active ingredient.

Desirable blood levels of the agent may be maintained by a continuous infusion to provide a therapeutic amount of the active ingredient within disease tissue. The use of operative combinations is contemplated to provide therapeutic combinations requiring a lower total dosage of each component agent than may be required when each individual therapeutic compound or drug is used alone, thereby reducing adverse effects. D. Exemplary co-administration routes and dosing considerations

The present invention also includes methods involving co-administration of the compounds described herein with one or more additional active agents (e.g., agents useful in the treatment of schistosome infection). Indeed, it is a further aspect of this invention to provide methods for enhancing existing therapies (e.g., praziquantel) and/or pharmaceutical compositions by co-administering a compound of this invention. In co-administration procedures, the agents may be administered concurrently or sequentially. In one embodiment, the compounds described herein are administered prior to the other active agent(s). The pharmaceutical formulations and modes of administration may be any of those described above. In addition, the two or more co -administered chemical agents, biological agents or other treatments may each be administered using different modes or different formulations.

III. Therapeutic Methods

In some embodiments, the present invention provides method for treating and preventing Schistosome infection. In some embodiments, one or more of the compounds described herein (e.g., 2-(2-fluorophenyl)ethyl] {3-methoxy-4-[2-oxo-2-(l- piperidinyl)ethoxy]benzyl} amine hydrochloride) is administered to a subject diagnosed with or exhibiting symptoms of Schistosome infection. In some embodiments, one of more of the compounds are administered to a subject at risk of contracting Schistosome infection (e.g., a subject traveling to or living in an area where Schistosome infection is endemic). In some embodiments, the compounds described herein find use in the treatment of other disorders mediated by actylcholinesterase enzymes (e.g., including but not limited to, Alzheimer's disease and myasthenia gravis).

Claims

CLAIMS We claim:

1. A method of treating or preventing infection by a schistosome, comprising administering 2-(2-fluorophenyl)ethyl] {3-methoxy-4-[2-oxo-2-(l- piperidinyl)ethoxy]benzyl} amine hydrochloride to a subject infected with or at risk of infection by a schistosome.

2. The method of claim 1 , wherein said subject has been diagnosed with schistosome infection.

3. The method of claim 1, wherein said subject lives in or is traveling to an area where schistosome infection is endemic.

4. The method of any one of claims 1 to 3, wherein said administration treats or prevents infection by said schistosome.

5. The method of any one of claims 1 to 4, wherein said schistosome is selected from the group consisting of Schistosoma mansoni, Schistosoma haematobium, and

Schistosoma japonicum

6. The method of any one of claims 1 to 5, wherein said 2-(2- fluorophenyl)ethyl] {3-methoxy-4-[2-oxo-2-(l-piperidinyl)ethoxy]benzyl} amine

hydrochloride inhibits an acetylcholinesterase enzyme of said schistosome.

7. The use of 2-(2-fluorophenyl)ethyl] {3-methoxy-4-[2-oxo-2-(l - piperidinyl)ethoxy]benzyl} amine hydrochloride in the treatment or prevention of schistosome infection in a subject.

8. The use of claim 7, wherein said subject has been diagnosed with schistosome infection.

9. The use of claim 1, wherein said subject lives in or is traveling to an area where schistosome infection is endemic.

10. The use of any one of claims 7 to 9, wherein said administration treats or prevents infection by said schistosome.

11. The use of any one of claims 7 to 10, wherein said schistosome is selected from the group consisting of Schistosoma mansoni, Schistosoma haematobium, and

Schistosoma japonicum.

12. The use of any one of claims 7 to 11, wherein said 2-(2-fluorophenyl)ethyl] {3- methoxy-4-[2-oxo-2-(l-piperidinyl)ethoxy]benzyl}amine hydrochloride inhibits an acetylcholinesterase enzyme of said schistosome.

13. A method of inhibiting an acetylcholinesterase enzyme in a cell, comprising administering 2-(2-fluorophenyl)ethyl] {3-methoxy-4-[2-oxo-2-(l- piperidinyl)ethoxy]benzyl} amine hydrochloride to said cell.

14. The method of claim 13, wherein said cell is in a subject.

15. The method of claim 14, wherein said subject has been diagnosed with Alzheimer's disease or myasthenia gravis and said administration treats said Alzheimer's disease or myasthenia gravis.

16. The method of claim 13, wherein said cell is a schistosome.

17. The method of claim 16, wherein said schistosome is selected from the group consisting of Schistosoma mansoni, Schistosoma haematobium, and Schistosoma japonicum.

18. The use of 2-(2-fluorophenyl)ethyl] {3-methoxy-4-[2-oxo-2-(l- piperidinyl)ethoxy]benzyl} amine hydrochloride in inhibiting an acetylcholinesterase enzyme in a cell.

19. The use of claim 18, wherein said cell is in a subject.

20. The use of claim 19, wherein said subject has been diagnosed with

Alzheimer's disease or myasthenia gravis and said administration treats said Alzheimer's disease or myasthenia gravis.

21. The use of claim 18, wherein said cell is a schistosome.

22. The use of claim 21, wherein said schistosome is selected from the group consisting of Schistosoma mansoni, Schistosoma haematobium, and Schistosoma japonicum.