Novel chemotypes for treating parasitic diseases and methods of use

Abstract

The present disclosure provides novel chemotypes, pharmaceutical compositions thereof, and method of use thereof for treating parasitic infection, including infection caused by parasitic flatworms.

Description

MCW C2364 Attorney Docket No. 650053.01223NOVEL CHEMOTYPES FOR TREATING PARASITIC DISEASES AND METHODS OF USECROSS REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims priority to U.S. Provisional Patent Application No. 63 / 689.419, filed August 30, 2024, the content of which is hereby incorporated by reference in its entirety.FIELD OF THE INVENTION

[0002] The disclosed technology is generally directed to antiparasitic agents. More particularly the technology7is directed to flatworm TRPMPZQ (transient receptor potential melastatin- praziquantel) channel activators.BACKGROUND

[0003] Infectious diseases caused by parasitic flatworms (e.g. trematodes, cestodes, and monogeneans) cause significant clinical (human) and veterinary (animal) disease worldwide. There are limited drug options for treating these infections which have significant healthcare and economic impact. Given the worldwide burden of neglected tropical diseases, there is ongoing need to develop novel agents to strengthen the pipeline of drugs to combat these burdensome infections. Many diseases caused by parasitic flatworms are treated using the anthelmintic drug praziquantel (PZQ), employed for decades as the key clinical agent to treat schistosomiasis. PZQ activates a flatworm transient receptor potential (TRP) channel within the melastatin family (TRPMPZQ) to mediate sustained Ca2+influx and worm paralysis.

[0004] There remains a need to develop novel agents, such as alternatives to PZQ, to help battle parasitic infections.SUMMARY OF THE INVENTION

[0005] The present disclosure describes novel chemotypes that are broad-spectrum flatworm TRPMPZQ (transient receptor potential melastatin-praziquantel) channel activators, and methods of use thereof for treatment of parasitic infections, particularly parasitic flatworm infections.

[0006] In one aspect, the present disclosure provides new chemotypes that activate a parasite ion channel. Specifically, the present compounds includeMCW C2364 Attorney Docket No. 650053.01223derivative or pharmaceutically acceptable salt thereof. The present compounds may be used as new anti-infective agents for diseases caused by parasitic flatworms. The effectiveness of the new chemotypes can be demonstrated, for example, by results derived from a target-based screen using a Ca2+reporter assay.

[0007] In another aspect, the present disclosure provides a composition comprising the compound as described herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0008] In another aspect, the present disclosure provides a method of treating a parasitic infection in a subject in need thereof, the method comprises administering to the subject an effective amount of the compound as described herein, or derivative or a pharmaceutically acceptable salt thereof. In some embodiments, the parasitic infection is a parasitic flatworm infection. For example, the parasitic flatworm can be trematode, cestode, monogenean, schistosome, tapeworm, or a combination thereof.

[0009] In another aspect, the present disclosure provides a method of treating schistosomiasis in a subject in need thereof, the method comprises administering to the subject an effective amount of the compound as described herein, or derivative or a pharmaceutically acceptable salt thereof.

[0010] In another aspect, the present disclosure provides a method of activating a transient receptor potential channel of a parasitic flatworm, the method comprising introducing to the parasitic flatworm an effective amount of the compound as described herein, or derivative or a salt thereof.

[0011] In another aspect, the present disclosure provides a method of causing paralysis or inhibiting grow th of a parasitic flatworm, the method comprising introducing to the parasitic flatworm an effective amount of the compound as described herein, or derivative or a salt thereof, or the composition as described herein.

[0012] In some embodiments, the parasitic flatworm is in an infected subject. In some embodiments, the subject is a human.MCW C2364 Attorney Docket No. 650053.01223BRIEF DESCRIPTION OF THE DRAWINGS

[0013] FIG. 1 illustrates a dose response curve for compounds praziquantel (PZQ), N-(4- fluorophenyl)-2-{[3-(hydroxymethyl)phenyl]amino}-2-phenylacetamide (22J14) and 2-[({2- [4-(thiolan-3-yl)piperazine-l-carbonyl]phenyl}methyl)sulfanyl]-IH-l,3-benzodi azole (28H14).

[0014] FIG. 2 illustrates the extent of worm damage and worm contraction caused by 22J14 and 28H14 as a percentage of the damage and contraction caused by PZQ.

[0015] The drawings illustrate only example embodiments and are therefore not to be considered limiting of the scope of the embodiments described herein, as other embodiments are within the scope of the disclosure.DETAILED DESCRIPTION OF THE INVENTION

[0016] Before the present materials and methods are described, it is understood that this invention is not limited to the particular methodology, protocols, materials, and reagents described, as these may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention which will be limited only by the appended claims.

[0017] As used in this specification and the claims, the singular forms “a,” "an." and “the” include plural forms unless the context clearly dictates otherwise. For example, the term “a compound” should be interpreted to mean “one or more compounds” unless the context clearly dictates otherwise. As used herein, the term “plurality” means “two or more.”

[0018] As used herein, the terms “about”, “approximately,” “substantially,” and “significantly” will be understood by persons of ordinary7skill in the art and will vary7to some extent on the context in which they are used. If there are uses of the term which are not clear to persons of ordinary skill in the art given the context in which it is used, “about” and “approximately” will mean up to plus or minus 10% of the particular term and “substantially” and “significantly” will mean more than plus or minus 10% of the particular term.

[0019] As used herein, the terms “include” and “including” have the same meaning as the terms “comprise” and “comprising.” The terms “comprise” and “comprising” should be interpreted as being “open” transitional terms that permit the inclusion of additional components further to those components recited in the claims. The terms “consist” and “consisting of’ should be interpreted as being “closed” transitional terms that do not permit the inclusion of additional components other than the components recited in the claims. The term “consisting essentially of’ should be interpreted to be partially closed and allowing the inclusion only of additional components that do not fundamentally alter the nature of the claimed subject matter.MCW C2364 Attorney Docket No. 650053.01223

[0020] Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this disclosure pertains. All publications and patents specifically mentioned herein are incorporated by reference for all purposes, including those describing and disclosing the chemicals, cell lines, vectors, animals, instruments, statistical analysis and methodologies which might be useful in connection with the present disclosure. In the case of conflict, the present specification, including definitions, will control.

[0021] Definitions

[0022] Chemical entities and the use thereof may be disclosed herein and may be described using terms known in the art and defined herein. Derivatives of the compounds as described herein include structural variants that can be obtained by modifications of the synthesis of such compounds, as generally understood in the art of organic synthesis. As an example, esters and amides of a compound having a reactive group (e.g., COOH, OH, or NH2) can be viewed as derivatives of the original compounds.

[0023] As used herein, "salt" refers to acid addition salts and basic addition salts. It may also refer to those salts that may be prepared in situ during the final isolation and purification of the present compounds.

[0024] The compounds of the disclosure may contain one or more chiral centers and / or double bonds and, therefore, exist as stereoisomers, such as geometric isomers, enantiomers or diastereomers. The term “stereoisomers"’ when used herein consist of all geometric isomers, enantiomers or diastereomers. These compounds may be designated by the symbols “R” or “S,” depending on the configuration of substituents around the stereogenic carbon atom. The present disclosure encompasses various stereo isomers of these compounds and mixtures thereof. Stereoisomers include enantiomers and diastereomers. Mixtures of enantiomers or diastereomers may be designated "(±)" in nomenclature, but the skilled artisan will recognize that a structure may denote a chiral center implicitly. It is understood that graphical depictions of chemical structures, e.g., generic chemical structures, encompass all stereoisomeric forms of the specified compounds, unless indicated otherwise.

[0025] Pharmaceutically acceptable salts of the present compounds are contemplated and may be utilized in the methods disclosed herein. The term “pharmaceutically acceptable salt” as used herein, refers to salts of the compounds, which are substantially non-toxic to living organisms. Typical pharmaceutically acceptable salts include those salts prepared by reaction of the compounds as disclosed herein with a pharmaceutically acceptable mineral or organic acid or an organic or inorganic base. Such salts are known as acid addition and base additionMCW C2364 Attorney Docket No. 650053.01223 salts. It will be appreciated by the skilled reader that most or all of the compounds as disclosed herein are capable of forming salts and that the salt forms of pharmaceuticals are commonly used, often because they are more readily crystallized and purified than are the free acids or bases.

[0026] Acids commonly employed to form acid addition salts may include inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, and the like, and organic acids such as p-toluenesulfonic, methanesulfonic acid, oxalic acid, p- bromophenylsulfonic acid, carbonic acid, succinic acid, citric acid, benzoic acid, acetic acid, and the like. Examples of suitable pharmaceutically acceptable salts may include the sulfate, pyrosulfate, bisulfate, sulfite, bisulfate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, bromide, iodide, acetate, propionate, decanoate, caprylate, acrylate, formate, hydrochloride, dihydrochloride, isobutyrate, caproate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleat-, butyne-. 1,4-dioate, hexyne-l,6-dioate, benzoate, chlorobenzoate, methylbenzoate, hydroxybenzoate, methoxy benzoate, phthalate, xylenesulfonate, phenyl acetate, phenylpropionate, phenylbutyrate, citrate, lactate, a-hydroxybutyrate, glycolate, tartrate, methanesulfonate, propanesulfonate, naphthalene- 1 -sulfonate, naphthal ene-2-sulfonate, mandelate, and the like.

[0027] Base addition salts include those derived from inorganic bases, such as ammonium or alkali or alkaline earth metal hydroxides, carbonates, bicarbonates, and the like. Bases useful in preparing such salts include sodium hydroxide, potassium hydroxide, ammonium hydroxide, potassium carbonate, sodium carbonate, sodium bicarbonate, potassium bicarbonate, calcium hydroxide, calcium carbonate, and the like.

[0028] The particular counter-ion forming a part of any salt of a compound disclosed herein may not be critical to the activity of the compound, so long as the salt as a whole is pharmacologically acceptable and as long as the counter-ion does not contribute undesired qualities to the salt as a whole. Undesired qualities may include undesirably solubility or toxicity.

[0029] Pharmaceutically acceptable esters and amides of the compounds can also be employed in the compositions and methods disclosed herein. Examples of suitable esters include alkyl, aryl, and aralkyl esters, such as methyl esters, ethyl esters, propyl esters, dodecyl esters, benzy l esters, and the like. Examples of suitable amides include unsubstituted amides, monosubstituted amides, and disubstituted amides, such as methyl amide, dimethyl amide, methyl ethyl amide, and the like.MCW C2364 Attorney Docket No. 650053.01223

[0030] Compounds

[0031] In one aspect, the present disclosure provides a compound, which is(22H14); or a derivative or a pharmaceutically acceptable salt thereof.

[0032] In some embodiments, the compound is N-(4-fluorophenyl)-2-{[3-(hydroxymethyl)phenyl]amino)-2-phenylacetamide, which has a formulaor a derivative or a pharmaceutically acceptable salt thereof.

[0033] In some embodiments, the compound is 2-[({2-[4-(thiolan-3-yl)piperazine-l- carbonyl]phenyl}methyl)sulfanyl]-lH-l,3-benzodiazole, which has a formulaor a derivative or a pharmaceutically acceptable salt thereof.

[0034] Pharmaceutical CompositionsMCW C2364 Attorney Docket No. 650053.01223

[0035] The compounds employed in the compositions and methods disclosed herein may be administered as pharmaceutical compositions. Such compositions may take any physical form which is pharmaceutically acceptable; illustratively, they can be orally administered pharmaceutical compositions. Such pharmaceutical compositions contain an effective amount of a disclosed compound, which effective amount is related to the dose of the compound to be administered.

[0036] In one aspect, the present disclosure provides a composition comprising the compounds as described herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0037] The compositions may contain from about 0.5% to about 50% of the compound in total, depending on the desired doses and the type of composition to be used. The amount of the compound may be defined as the “effective amount”, which is the amount of the compound that provides the desired dose to the patient in need of such treatment. The activity of the compounds employed in the compositions and methods disclosed herein are not believed to depend greatly on the nature of the composition, and, therefore, the compositions can be chosen and formulated primarily or solely for convenience and economy. In some embodiments, the pharmaceutical composition includes a compound as described herein in a range from about 0. 1 to about 2000 mg, such as from about 0.5 to 500 mg or from about 1 to about 100 mg. The pharmaceutical composition may be administered to provide the compound at a daily dose of about 0.01 mg / kg to about 1000 mg / kg body weight, such as about 0.01 mg / kg to about 100 mg / kg, about 0. 1 mg / kg to about 1000 mg / kg, about 0.1 to about 500 mg / kg, about 0. 1 to about 100 mg / kg, or about 50 to about 100 mg / kg body weight.

[0038] In some embodiments, after the pharmaceutical composition is administered to a subject (e.g.. after about 1. 2, 3, 4. 5, or 6 hours post-administration), the concentration of the compound at the site of action may be within a concentration range bounded by end-points selected from 0.001 pM, 0.005 pM, 0.01 pM, 0.5 pM, 0.1 pM, 1.0 pM, 10 pM, and 100 pM (e.g., 0.1 pM - 1.0 pM).

[0039] The term "pharmaceutically acceptable" means approved by a regulatory agency of the Federal or a state government or listed in the U.S. Examples of suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by E.W. Martin. Suitable pharmaceutically acceptable carriers include, but are not limited to, for example, suitable diluents, vehicles, excipients, preservatives, solubilizers, emulsifiers, liposomes, or nanoparticles, among others. Additionally, such pharmaceutically acceptable carriers may be aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of nonaqueousMCW C2364 Attorney Docket No. 650053.01223 solutions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include isotonic solutions, alcoholic / aqueous solutions, emulsions or suspensions, including saline and buffered media. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid carriers, particularly for injectable solutions. Suitable excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel. sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like.

[0040] The formulation should be selected according to the mode of administration. The compositions may include a pharmaceutical carrier, excipient, or diluent, which are nontoxic to the subject being exposed thereto at the dosages and concentrations employed. Examples of pharmaceutical carriers include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid; low molecular weight (less than about 10 residues) polypeptide; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and / or nonionic surfactants such as polysorbate (e.g., a TWEEN™ brand surfactant), polyethylene glycol (PEG), and poloxamers (e.g. PLURONICS™ surfactants, which include a central hydrophobic block of polypropylene oxide flanked by two hydrophilic blocks of polyethylene oxide).

[0041] Oral administration is an illustrative route of administering the compounds employed in the compositions and methods disclosed herein. Other illustrative routes of administration include transdermal, percutaneous, intravenous, intramuscular, intranasal, buccal, intrathecal, intracerebral, or intrarectal routes. The route of administration may be varied in any way, limited by the physical properties of the compounds being employed and the convenience of the subj ect and the caregiver.

[0042] Suitable formulations include those that are suitable for more than one route of administration. For example, the formulation can be one that is suitable for both oral and intravenous administration. Alternatively, suitable formulations include those that are suitable for only one route of administration as well as those that are suitable for one or more routes of administration, but not suitable for one or more other routes of administration. For example, the formulation can be one that is suitable for oral, topical, transdermal. percutaneous.MCW C2364 Attorney Docket No. 650053.01223 intravenous, intramuscular, intranasal, buccal, and / or intrathecal administration but not suitable for intracerebral administration.

[0043] The inert ingredients and manner of formulation of the pharmaceutical compositions may be selected from conventional technologies. The usual methods of formulation used in pharmaceutical science may be used here. Suitable types of compositions include, but are not limited to, tablets, chewable tablets, capsules, solutions, parenteral solutions, intranasal sprays or powders, troches, suppositories, transdermal patches, and suspensions.

[0044] Capsules are prepared by mixing the compound with a suitable diluent and fdling the proper amount of the mixture in capsules. The usual diluents include inert powdered substances (such as starches), powdered cellulose (especially crystalline and microcrystalline cellulose), sugars (such as fructose, mannitol and sucrose), grain flours, and similar edible powders, but any suitable capsule formulation can be used.

[0045] Tablets are prepared by direct compression, by wet granulation, or by dry' granulation. Their formulations usually incorporate diluents, binders, lubricants, and disintegrators (in addition to the compounds). Typical diluents include, for example, various types of starch, lactose, mannitol, kaolin, calcium phosphate or sulfate, inorganic salts (such as sodium chloride), and powdered sugar. Powdered cellulose derivatives can also be used. Typical tablet binders include substances such as starch, gelatin, and sugars (e.g., lactose, fructose, glucose, and the like). Natural and synthetic gums can also be used, including acacia, alginates, methylcellulose, polyvinylpyrrolidine, and the like. Polyethylene glycol, ethylcellulose, and waxes can also serve as binders.

[0046] Tablets can be coated with sugar, e.g., as a flavor enhancer and sealant. The compounds also may be formulated as chewable tablets, by using large amounts of pleasant-tasting substances, such as mannitol, in the formulation. Instantly dissolving tablet-like formulations can also be employed, for example, to assure that the patient consumes the dosage form and to avoid the difficulty' that some patients experience in swallowing solid objects.

[0047] A lubricant can be used in the tablet formulation to prevent the tablet and punches from sticking in the die. The lubricant can be chosen from such slippery solids as talc, magnesium and calcium stearate, stearic acid, and hydrogenated vegetable oils.

[0048] Tablets can also contain disintegrators. Disintegrators are substances that swell when wetted to break up the tablet and release the compound. They include starches, clays, celluloses, algins, and gums. As further illustration, com and potato starches, methylcellulose, agar, bentonite, wood cellulose, powdered natural sponge, cation-exchange resins, alginic acid, guar gum, citrus pulp, sodium lauryl sulfate, and carboxymethylcellulose can be used.MCW C2364 Attorney Docket No. 650053.01223

[0049] The composition can be formulated as enteric formulations, for example, to protect the active ingredient from the strongly acid contents of the stomach. Such formulations can be created by coating a solid dosage form with a fdm of a polymer which is insoluble in acid environments and soluble in basic environments. Illustrative films include cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropyl methylcellulose phthalate, and hydroxypropyl methylcellulose acetate succinate.

[0050] Transdermal patches can also be used to deliver the compounds. Transdermal patches can include a resinous composition in which the compound will dissolve or partially dissolve; and a fdm which protects the composition, and which holds the resinous composition in contact with the skin. Other, more complicated patch compositions can also be used, such as those having a membrane pierced with a plurality of pores through w hich the compound is pumped by osmotic action.

[0051] Pharmaceutical compositions adapted for transdermal administration may be presented as discrete patches intended to remain in intimate contact with the epidermis of the recipient for a prolonged period of time. For example, the active ingredient may be delivered from the patch by iontophoresis.

[0052] Pharmaceutical compositions adapted for topical administration may be formulated as ointments, creams, suspensions, lotions, powders, solutions, pastes, gels, impregnated dressings, sprays, aerosols or oils and may contain appropriate conventional additives such as preservatives, solvents to assist drug penetration and emollients in ointments and creams.

[0053] For applications to the eye or other external tissues, for example the mouth and skin, the pharmaceutical compositions are in some embodiments applied as a topical ointment or cream. When formulated in an ointment, the compound may be employed with either a paraffinic or a water-miscible ointment base. Alternatively, the compound may be formulated in a cream with an oil-in-water cream base or a water-in-oil base. Pharmaceutical compositions adapted for topical administration to the eye include eye drops where the active ingredient is dissolved or suspended in a suitable carrier, especially an aqueous solvent.

[0054] The composition can be prepared with materials (e.g, actives excipients, carriers (such as cyclodextrins), diluents, etc.) having properties (e.g., purity) that render the formulation suitable for administration to humans or non-human subjects. In some embodiments, the composition is suitable for use in humans. In some embodiments, the composition is prepared with materials having purity and / or other properties that render it suitable for administration to non-human subjects, but not suitable for administration to humans.MCW C2364 Attorney Docket No. 650053.01223

[0055] Each dosage unit may contain the dose of a given compound, for example, a daily dose, or each dosage unit may contain a fraction of the daily dose, such as one-half or one-third of the dose. The amount of each compound to be contained in each dosage unit can depend, in part, on the identity of the particular compound chosen for the therapy and other factors, such as the indication for which it is given. The pharmaceutical compositions disclosed herein may be formulated so as to provide quick, sustained, or delayed release of the active ingredient after administration to the patient by employing well known procedures.

[0056] The composition may include a single compound or a combination of compounds as described herein for administration. For example, a two or more of the compounds described herein may be administered for an anti-parasitic effect. In addition, the composition may include solvate forms of the compounds or salts, esters, and / or amides, thereof. Solvate forms may include ethanol solvates, hydrates, and the like.

[0057] The disclosed compounds or pharmaceutical compositions comprising the disclosed compounds may be administered with additional therapeutic agents. The additional therapeutic agent may include, for example, one or more known agent for treating parasitic infection. In some embodiments of the disclosed methods, one or more additional therapeutic agents are administered with the disclosed compounds or with pharmaceutical compositions comprising the disclosed compounds, where the additional therapeutic agent is administered prior to, concurrently with, or after administering the disclosed compounds or the pharmaceutical compositions comprising the disclosed compounds. In some embodiments, the disclosed pharmaceutical compositions are formulated to comprise the disclosed compounds and further to comprise the one or more additional therapeutic agents.

[0058] Methods of Use

[0059] The compounds as described herein may effectively inhibit the growth and propagation of parasites in vivo, in particular the growth and propagation of flatworms within a subject. In one aspect, the present disclosure provides a method of treating a parasitic infection in a subject in need thereof, the method comprises administering to the subject an effective amount of the compounds as described herein, or a derivative or a pharmaceutically acceptable salt thereof, or the composition as described herein. In use, the compounds as described herein may be cytotoxic to the parasite but not to the subject being treated. Also provided is a compound, or a derivative or a pharmaceutically acceptable salt thereof, as described herein for use in therapy. In particular, the present disclosure provides a compound, or a derivative or a pharmaceutically acceptable salt thereof as described herein, for use in the treatment of parasitic infection in a subject in need thereof. Also provided is a compound, or a derivative or a pharmaceuticallyMCW C2364 Attorney Docket No. 650053.01223 acceptable salt thereof as described herein, for use in the manufacture of a medicament for the treatment of parasitic infection in a subject in need thereof.

[0060] Suitable subjects include mammals and non-mammals. Mammals include any member of the class Mammalia. Examples of mammals include, but are not limited to, humans, nonhuman primates such as chimpanzees and other apes and monkey species; farm animals such as cattle, horses, sheep, goats, and swine; domestic animals such as rabbits, dogs, and cats; laboratory animals including rodents, such as rats, mice, guinea pigs, and the like. Examples of non-mammals include, but are not limited to, birds, fish, crustaceans, among others. A subject can be any suitable age or sex. In some embodiments, the subject is a human. In some embodiments, the subject is a non-human subject, such as a fish, a bird, a dog, a cat, or a livestock.

[0061] As used herein, “treating” or “treatment” describes the management and care of a subject for combating the disease, condition, or disorder. Treating includes the administration of the compound or composition described herein to reduce, prevent, ameliorate and / or improve the onset of the symptoms or complications, alleviating the symptoms or complications, or reducing or eliminating the disease, condition, or disorder associated with a parasitic infection, such as a flatworm parasitic infection. The term treating as described herein includes the inhibiting or reducing growth and / or propagation / reproduction of parasites within a subject (i.e., in vivo).

[0062] Symptoms of parasitic infections include, but are not limited to, for example, stomach cramps and pain, nausea or vomiting, dehydration, weight loss, swollen lymph nodes, digestive problems including unexplained constipation, diarrhea or persistent gas, skin issues such as rashes, eczema, hives, and itching, continuous muscle and joint pain, fatigue, depression or feeling of apathetic, constant hunger, iron deficiency / anaemia, grinding teeth during sleep, unexplained feelings of anxiety, recurrent yeast infections, itching of the anus or vagina, itching, redness, irritation, and an unusual discharge from the genital area, trouble falling asleep or waking up multiple times during the night.

[0063] As used herein the term “effective amount” or "therapeutically effective amount" refers to the amount or dose of the present compound or composition, which, upon single or multiple dose administration to the subject, provides the desired effect in the subject under diagnosis or treatment. The disclosed methods may include administering an effective amount of the disclosed compounds (e.g., as present in a pharmaceutical composition) for treating a disease or disorder, e.g., a parasitic infection.MCW C2364 Attorney Docket No. 650053.01223

[0064] An effective amount can be readily determined by the attending diagnostician, as one skilled in the art, by the use of known techniques and by observing results obtained under analogous circumstances. In determining the effective amount or dose of compound administered, a number of factors can be considered by the attending diagnostician, such as: the species of the subject; its size, age, and general health; the degree of involvement or the severity’ of the disease or disorder involved; the response of the individual subject; the particular compound administered; the mode of administration; the bioavailability characteristics of the preparation administered; the dose regimen selected; the use of concomitant medication; and other relevant circumstances.

[0065] The term “administering” or “administration” refers to introducing the present compounds or compositions into the body of the subject, such as by oral delivery. Suitable routes of administration include, but are not limited to, oral, topical, transdermal, buccal, sublingual, pulmonary, transdermal, transmucosal, rectal, as well as subcutaneous, intraperitoneal, intravenous, and intramuscular injection.

[0066] The methods of treating a parasitic infection include treating a parasitic flatworm infection. In some embodiments, the parasitic infection is a parasitic flatworm infection. Suitable flatworm infections contemplated to be treated using the compound and compositions described herein include, but are not limited to, trematode (i.e. flukes, such as schistosoma species (blood fluke), Fasciola species (liver fluke). Paragonimus westermani (lung fluke), clonorchis sinensis (liver fluke), Opisthorchis viverrini (liver fluke)), cestode (i.e. tapeworms, such as pork tapeworm (taenia solium), beef tapeworm (taenia saginata), Echinococcus species, Dipylidium caninum (heartworm), Diphyllobothrium species, Gyrodactcylus species, and dwarf tapeworm (hymenolepis nana)). turbellaria (planarians), monogenean (i.e. monopisthocotyleans and the polyopisthocotyleans). and turbellaria (such as temnocephala lamothei, dugesia aenigma, dugesia bifida, planaria simplex, girardia dorotocephala, girardia tigrina). In particular embodiments, the present method can be used to treat a parasitic flatworm infection, in which the parasitic flatworm is trematode, cestode, monogenean, schistosome, tapeworm, or a combination thereof.

[0067] The compounds as disclosed herein may provide treatment for a disease caused by parasitic worms in multiple species (such as humans), including fasciolosis, schistosomiasis, and other diseases. Schistosomiasis is a disease caused by parasitic flatworms, namely, schistosomes. In another aspect, the present disclosure provides a method of treating schistosomiasis in a subject in need thereof, the method comprises administering to the subject an effective amount of the compounds as described herein, or a derivative or a pharmaceuticallyMCW C2364 Attorney Docket No. 650053.01223 acceptable salt thereof, or the composition as described herein. As nonlimiting examples, the present compounds can be used in medications (e.g.. drugs) for treatment of clinical and veterinary diseases caused by parasitic flatworms. The present disclosure also provides a compound, or a derivative or a pharmaceutically acceptable salt thereof as described herein, for use in the treatment of schistosomiasis in a subject in need thereof. Also provided is a compound, or a derivative or a pharmaceutically acceptable salt thereof as described herein, for use in the manufacture of a medicament for the treatment of schistosomiasis in a subject in need thereof.

[0068] The present compounds may activate a flatworm transient receptor potential (TRP) channel, which may in turn mediate sustained Ca2+influx and worm paralysis. Thus, the disclosed compounds may act as ion channel activators to treat parasitic flatform infections. In another aspect, the present disclosure provides a method of activating a transient receptor potential channel of a parasitic flatworm, the method comprising introducing to the parasitic flatworm an effective amount of the compounds as described herein, or a derivative or a salt thereof, or the composition as described herein. In some embodiments, the transient receptor potential channel is a member of the melastatin family (TRPM), such as TRPM1, TRPM2, TRPM3, TRPM4, TRPM5, TRPM6, TRPM7, and TRPM8.

[0069] The present compound may be used to effectively control infection or growth of a parasitic flatworm. In another aspect, the present disclosure provides a method of causing paralysis or inhibiting growth of a parasitic flatworm, the method comprising introducing to the parasitic flatworm an effective amount of the compounds as described herein, or a derivative or a salt thereof, or the composition as described herein.

[0070] For the present methods, the compounds as described herein, or a derivative or a salt thereof, or the composition as described herein may be introduced to a parasitic flatworm that is in an infected subject (e g., to inhibit the growth of, or treat an infection caused by, the parasitic flatworm in the subject). In some embodiments, the subject is ahuman. Alternatively, the compound or composition may be introduced to a parasitic flatworm that is in an environment (outside the body of a subject), such as a river, a lake, a water reservoir, a grassland, or a forest. In these cases, the effective amount of the compound or composition refers to an amount that provides desired effect of activating a TRP channel and / or causing paralysis or inhibiting grow th of the parasitic flatworm in such environment.

[0071] The compounds utilized in the methods disclosed herein may be administered in conventional dosage forms prepared by combining the active ingredient with standard pharmaceutical carriers or diluents according to known procedures, including for exampleMCW C2364 Attorney Docket No. 650053.01223 mixing, granulating, compressing, and / or dissolving various ingredients as appropriate to the desired preparation.

[0072] In some embodiments, the subject may be administered a dose of the disclosed compound as low as 1.25 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg, 12.5 mg, 15 mg, 17.5 mg, 20 mg,22.5 mg, 25 mg, 27.5 mg, 30 mg, 32.5 mg, 35 mg, 37.5 mg, 40 mg. 42.5 mg, 45 mg, 47.5 mg, 50 mg, 52.5 mg, 55 mg, 57.5 mg, 60 mg, 62.5 mg, 65 mg, 67.5 mg, 70 mg, 72.5 mg, 75 mg,77.5 mg, 80 mg, 82.5 mg, 85 mg, 87.5 mg, 90 mg, 100 mg, 200 mg, 500 mg, 1000 mg, or 2000 mg once daily, twice daily, three times daily, four times daily, once weekly, twice weekly, or three times per week in order to treat the disease or disorder in the subject. In some embodiments, the subject may be administered a dose of the disclosed compound as high as 1.25 mg. 2.5 mg. 5 mg, 7.5 mg, 10 mg, 12.5 mg. 15 mg. 17.5 mg, 20 mg, 22.5 mg, 25 mg, 27.5 mg, 30 mg, 32.5 mg, 35 mg, 37.5 mg, 40 mg, 42.5 mg, 45 mg, 47.5 mg, 50 mg, 52.5 mg, 55 mg, 57.5 mg, 60 mg, 62.5 mg, 65 mg, 67.5 mg, 70 mg, 72.5 mg, 75 mg, 77.5 mg, 80 mg, 82.5 mg, 85 mg. 87.5 mg, 90 mg, 100 mg, 200 mg, 500 mg, 1000 mg, or 2000 mg, once daily, twice daily, three times daily, four times daily, once weekly, twice weekly, or three times per week in order to treat the disease or disorder in the subject. Minimal and / or maximal doses of the compounds may include doses falling within dose ranges having as endpoints any of these disclosed doses (e.g., 2.5 mg - 200 mg).

[0073] Kits

[0074] In another aspect, the present disclosure provides a kit comprising a pharmaceutical composition comprising the compounds as disclosed herein and instructional material.

[0075] The term "instructional material" refers to a publication, a recording, a diagram, or any other medium of expression which is used to communicate the usefulness of the present pharmaceutical composition for one of the purposes set forth herein in a human. The instructional material can also, for example, describe an appropriate dose of the present pharmaceutical composition. The instructional material of the present kit can, for example, be affixed to a container which contains a pharmaceutical composition as disclosed herein or be shipped together with a container which contains the pharmaceutical composition. Alternatively, the instructional material can be shipped separately from the container with the intention that the instructional material and the pharmaceutical composition be used cooperatively by the recipient.

[0076] It should be apparent to those skilled in the art that many additional modifications beside those already described are possible without departing from the inventive concepts. Other features and advantages of the invention will be apparent from the present descriptionMCW C2364 Attorney Docket No. 650053.01223 and claims. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, suitable methods and materials are described below. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.EXAMPLES

[0077] The following examples are. of course, offered for illustrative purposes only, and are not intended to limit the scope of the present invention in any way. Indeed, various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description and the following examples and fall within the scope of the appended claims.

[0078] Example 1 Novel chemot pes identified

[0079] Two new screening hits were identified in a target-based screen of Schistosoma mansoni TRPMPZQ. This target is the mode of action of the current clinical therapeutic, praziquantel. As new7activators of this channel they could potentially provide an alternative route for eliminating schistsomiasis and other parasitic infections. These chemotypes and logical derivatives are studied in target-based assays and their activities on parasitic flatworms are examined (e.g., potency in vivo, PK / PD evaluations). These chemotypes may be used, for example, in animal health and treatment of parasitic diseases.

[0080] The exemplary compounds include:N-(4-fluorophenyl)-2-{[3-(hydroxymethyl)phenyl] amino} -2-phenylacetami de, having a formula:2-[({2-[4-(thiolan-3-yl)piperazine-l-carbonyl]phenyl}methyl)sulfanyl]-lH-l,3-benzodi azole, having a formula:MCW C2364 Attorney Docket No. 650053.01223

[0081] Example 2 Ca2+reporter assay

[0082] HEK293 cells (ATCC CRL-1573.3) were cultured in DMEM supplemented with 10% fetal bovine serum (FBS), penicillin (100 units / ml), streptomycin (lOOpg / ml), and L-glutamine (290pg / ml). Cells were authenticated by STR profiling (ATCC) and evaluated for mycoplasma contamination through monthly scheduled testing (LookOut® Mycoplasma PCR Detection Kit, Sigma). For functional profiling assays, cDNAs encoding TRPMMCLZ mutants and TRPMMCLZ orthologs were each transiently transfected into cultured cells (3x106 cells per 100mm petri dish) using Lipofectamine 2000. Subsequent Ca2+reporter assays were performed using a Fluorescence Imaging Plate Reader (FLIPRTETRA, Molecular Devices) as has been previously detailed. In brief, HEK293 cells were seeded (20,000 cells / well) in a black-walled clear-bottomed poly-d-lysine coated 384-well plate (Greiner Bio-One. Germany) using DMEM growth media (supplemented with 10% dialyzed FBS). After 24hrs, this growth medium was exchanged and cells loaded with a synthetic Ca2+indicator (Fluo-4 NW, Invitrogen). Plates were then incubated (20pL per well, 50 mins at 37°C, 10 min at room temperature) in Hank’s balanced salt solution (HBSS, 1.26mM CaCh. 0.49mM MgCb, 0.41mM MgSO4, 5.33mM KCL 0.44mM KH2PO4, 4.17mM NaHCOs, 137.9mM NaCl, 0.34mM Na2HPO4, 5.55mM D- glucose) supplemented with probenecid (2.5mM) and HEPES (20mM). Dilutions of ligands (5x concentration, PZQ, 22 JI 4, 28H14) were prepared in assay buffer, without probenecid and dye. After loading with Ca2+indicator, basal fluorescence values were recorded (20s) then 5 pl of compound was added, and the signal monitored for a further 250s. FLIPR assays were performed at room temperature. Peak fluorescence increases for each well were normalized to maximum-fold increase over baseline. Except where indicated, data are presented as mean ± SEM. The results are shown in FIG. 1.

[0083] Example 3 Schistosoma mansoni assaysMCW C2364 Attorney Docket No. 650053.01223

[0084] Adult schistosomes were isolated from the mesenteric vasculature of female Swiss Webster mice following protocols approved by the Medical College of Wisconsin Institutional Animal Care and Use Committee. These mice were previously infected with S. mansoni cercariae (NMRI strain) at the Schistosomiasis Resource Center at the Biomedical Research Institute. Harvested worms were washed in ABC 169 and incubated overnight (37°C, 5% CO2) in vented petri dishes (100x25mm). Motility assays were performed using three adult male worms per well in a twenty-four well dish. Motility was scored for individual worm on a 0-4 scale, using the WHO-TDR scoring matrix guidelines , with images obtained prior to and 20 minutes after treatment with various agonists (e.g., PZQ, 22J14 and 22H14). Images and videos were captured using a Zeiss Discovery v20 stereomicroscope with a QiCAM 12-bit cooled color CCD camera controlled by Image-Pro imaging software (v. 11.0.4). The contraction and damage observed for 22J14 and 28H14 are shown in FIG. 2 as a percentage of the contraction and damage observed for PZQ.

[0085] Reference:Park et al., Mechanism of praziquantel action at a parasitic flatworm ion channel, Sci TranslMed. 2021, 13(625): eabj5832.

[0086] For reasons of completeness, various aspects and embodiments of the present disclosure are set out in the following numbered clauses:

[0087] Clause 1. A compound, which isor a derivative or pharmaceutically acceptable salt thereof.

[0088] Clause 2. A composition comprising the compound of clause 1, or a derivative or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0089] Clause 3. A method of treating a parasitic infection in a subject in need thereof, the method comprises administering to the subject an effective amount of the compound of clause 1, or a derivative or a pharmaceutically acceptable salt thereof.MCW C2364 Attorney Docket No. 650053.01223

[0090] Clause 4. The method of clause 3, wherein the parasitic infection is a parasitic flatworm infection.

[0091] Clause 5. The method of clause 4, wherein the parasitic flatworm is trematode, cestode, monogenean, schistosome, tapeworm, or a combination thereof.

[0092] Clause 6. The method of any one of clauses 3-5, wherein the compound is. or a derivative or pharmaceutically acceptable salt thereof.

[0093] Clause 7. The method of any one of clauses 3-5, wherein the compound is, or a derivative or pharmaceutically acceptable salt thereof.

[0094] Clause 8. A method of treating schistosomiasis in a subject in need thereof, the method comprises administering to the subject an effective amount of the compound of clause 1, or a derivative or a pharmaceutically acceptable salt thereof.

[0095] Clause 9. The method of clause 8, wherein the compound is. or a derivative or pharmaceutically acceptable salt thereof.

[0096] Clause 10. The method of clause 8, wherein the compound isMCW C2364 Attorney Docket No. 650053.01223, or a derivative or pharmaceutically acceptable salt thereof.

[0097] Clause 11. A method of activating a transient receptor potential channel of a parasitic flatworm, the method comprising introducing to the parasitic flatworm an effective amount of the compound of clause 1, or a derivative or a salt thereof.

[0098] Clause 12. The method of clause 11, wherein the compound is, or a derivative or pharmaceutically acceptable salt thereof.

[0099] Clause 13. The method of clause 11 , wherein the compound is, or a derivative or pharmaceutically acceptable salt thereof.

[0100] Clause 14. A method of causing paralysis or inhibiting growth of a parasitic flatworm, the method comprising introducing to the parasitic flatworm an effective amount of the compound of clause 1 , or a derivative or a salt thereof.MCW C2364 Attorney Docket No. 650053.01223

[0101] Clause 15. The method of clause 14, wherein the compound is, p y p thereof.

[0103] Clause 17. The method of any one of clauses 11-16, wherein the parasitic flatworm is in an infected subject.

[0104] Clause 18. The method of any one of clauses 3-10 and 17, wherein the subject is a human.

Claims

MCW C2364 Attorney Docket No. 650053.01223CLAIMSWhat is claimed is:

1. A compound, which isor a derivative or pharmaceutically acceptable salt thereof.

2. A composition comprising the compound of claim 1, or a derivative or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

3. A method of treating a parasitic infection in a subject in need thereof, the method comprises administering to the subject an effective amount of the compound of claim 1, or a derivative or a pharmaceutically acceptable salt thereof.

4. The method of claim 3. wherein the parasitic infection is a parasitic flatworm infection.

5. The method of claim 4, wherein the parasitic flatworm is trematode, cestode, monogenean, schistosome, tapeworm, or a combination thereof.

6. The method of claim 3, wherein the compound is, or a derivative or pharmaceutically acceptable salt thereof.MCW C2364 Attorney Docket No. 650053.012237. The method of claim 3, wherein the compound is, or a derivative or pharmaceutically acceptable salt thereof.

8. A method of treating schistosomiasis in a subj ect in need thereof the method comprises administering to the subject an effective amount of the compound of claim 1, or a derivative or a pharmaceutically acceptable salt thereof.

9. The method of claim 8, wherein the compound is, or a derivative or pharmaceutically acceptable salt thereof.

10. The method of claim 8, wherein the compound isMCW C2364 Attorney Docket No. 650053.01223, or a derivative or pharmaceutically acceptable salt thereof.

11. A method of activating a transient receptor potential channel of a parasitic flatworm, the method comprising introducing to the parasitic flatworm an effective amount of the compound of claim 1 , or a derivative or a salt thereof.

12. The method of claim 11, wherein the compound is, or a derivative or pharmaceutically acceptable salt thereof.

13. The method of claim 11, wherein the compound is, or a derivative or pharmaceutically acceptable salt thereof.MCW C2364 Attorney Docket No. 650053.0122314. A method of causing paralysis or inhibiting growth of a parasitic flatworm, the method comprising introducing to the parasitic flatworm an effective amount of the compound of claim 1, or a derivative or a salt thereof.

15. The method of claim 14, wherein the compound is, or a derivative or pharmaceutically acceptable salt thereof.

16. The method of claim 14, wherein the compound is, or a derivative or pharmaceutically acceptable salt thereof.

17. The method of claim 11, wherein the parasitic flatworm is in an infected subject.

18. The method of claim 3, wherein the subject is a human.

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