FIELD OF THE INVENTION

The present invention relates to novel dione compounds having pharmacological activity, processes for their preparation, pharmaceutical compositions and their use in the treatment of certain parasitic protozoal infections such as malaria, in particular infection by Plasmodium falciparum.

BACKGROUND OF THE INVENTION

Parasitic protozoal infections are responsible for a wide variety of diseases of medical and veterinary importance, including malaria in man and various coccidioses in birds, fish and mammals. Many of the diseases are life-threatening to the host and cause considerable economic loss in animal husbandry, such as diseases caused by infection by species of Eimeria, Theileria, Babesia, Cryptosporidium, Toxoplasma (such as Toxoplasma brucei, African sleeping sickness and Toxoplasma cruzi, Chagas disease) and Plasmodium (such as Plasmodium falciparum), and the Mastigophora such as species of Leishmania (such as Leishmania donovani). Another parasitic organism of increasing concern is Pneumocytis carinii, which can cause an often fatal pneumonia in immunodeficient or immunocompromised hosts, including those infected with HIV.

Malaria is a mosquito-borne disease that, in humans, can be caused by five species of Plasmodium parasite, of which Plasmodium falciparum is the most virulent. In 2013, there were an estimated 128 million of people infected with malaria worldwide and malarial disease was responsible for an estimated 584,000 deaths (90% of them in sub-saharian Africa), young children and pregnant women being the most affected groups. In 2013, malaria killed an estimated 437,000 children under five years of age (WORLD HEALTH ORGANIZATION. ( 2014). World malaria report. Geneva, Switzerland, World Health Organization).

Resistance to classical treatments and emerging resistance to the current treatment of choice (artemisinins-based combination therapies) reveals the urgent need for new therapeutic agents with novel mechanisms of action (WORLD HEALTH ORGANIZATION. Joint assessment of the response to artemisinin resistance in the greater Mekong sub-region. November 201 1-February 2012. Summary report.). In 2010, GSK released details of more than 13,500 chemical compounds that have already shown to inhibit Plasmodium falciparum parasite growth in the phenotypic screening approach (Gamo, F.J. et al. (2010) Thousands of chemical starting points for antimalarial lead identification. Nature 465, 305-310). Molecular structures and descriptions of these compounds were made publicly available in accessible databases under the name of TCAMS (Tres Cantos Antimalarial set) (http://www.ebi.ac.uk/chemblntd). SUMMARY OF THE INVENTION

The present invention is directed to novel dione compounds for use in the chemotherapy of certain parasitic infections such as malaria, and in particular infection by Plasmodium falciparum, processes for their preparation and pharmaceutical compositions comprising such a compound.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides compounds of Formula (I):

R-i is morpholinyl or pyridyl;

R ₂ is fluoro or hydrogen when R-i is morpholinyl; and

R ₂ is hydrogen when R-i is pyridyl

or a pharmaceutically acceptable salt thereof.

Also included in the present invention are pharmaceutically acceptable salts. In certain embodiments of the invention, pharmaceutically acceptable salts of compounds of Formula (I) may be preferred over the respective free base because such salts impart greater stability or solubility to the molecule thereby facilitating formulation into a dosage form. Therefore, the present invention also covers the pharmaceutically acceptable salts of compounds of Formula (I). As used herein, the term "pharmaceutically acceptable salt" refers to a salt that retains the desired biological activity of the subject compound and exhibits minimal undesired toxicological effects. For a review on suitable salts see Berge et al, J. Pharm. Sci., 1977, 66, 1 -19. The term "pharmaceutically acceptable salt" includes any pharmaceutically acceptable acid or basic addition salts. These pharmaceutically acceptable salts may be prepared in situ during the final isolation and purification of the compound, or by separately reacting the purified compound in its free base form with a suitable acid or base, respectively. The salt may precipitate from solution and be collected by filtration or may be recovered by evaporation of the solvent.

Therefore, according to a further aspect, the invention provides a pharmaceutically acceptable salt of compounds of Formula (I) thereof.

The compounds of Formula (I) contain a basic functional group and are therefore capable of forming pharmaceutically acceptable acid addition salts by treatment with a suitable acid. A pharmaceutically acceptable acid addition salt may be formed by reaction of a compound of Formula (I) with a suitable strong inorganic or organic acid (such as hydrobromic, hydrochloric, sulfuric, nitric, phosphoric, perchloric, p-toluenesulfonic, benzenesulfonic, methanesulfonic, ethanesulfonic, 2-hydroxyethanesulfonic, naphthalenesulfonic (e.g. 2- naphthalenesulfonic), optionally in a suitable solvent such as an organic solvent, to give the salt which is usually isolated for example by crystallisation and filtration. Pharmaceutically acceptable acid addition salts include a hydrobromide, hydrochloride, hydroiodide, sulfate, bisulfate, nitrate, phosphate, perchlorate, p-toluenesulfonate, benzenesulfonate, methanesulfonate, ethanesulfonate, 2-hydroxyethanesulfonate, hemi-edisylate naphthalenesulfonate (e.g. 2-naphthalenesulfonate) salt. In one embodiment, a pharmaceutically acceptable acid addition salt of a compound of Formula (I) is a salt of a strong acid, for example a hydrobromide, hydrochloride, hydroiodide, sulfate, nitrate, perchlorate, phosphate p-toluenesulfonic, benzenesulfonic or methanesulfonic salt.

Suitable pharmaceutically acceptable salts of compounds of Formula (I) include mono- or di- basic salts with the appropriate base. A pharmaceutically acceptable basic addition salt may be formed by reaction of a compound of Formula (I) with a suitable inorganic or organic base. Pharmaceutically acceptable basic addition salt include sodium, potassium, calcium, magnesium, ammonium, N-methylglucamine, ammonium and choline salts The invention includes within its scope all possible stoichiometric and non-stoichiometric forms of the salts of compounds of Formula (I).

Compounds of Formula (I) or a pharmaceutically acceptable salt thereof, may exist as solids or liquids, both of which are included in the invention. In the solid state, compounds of Formula (I) or pharmaceutically acceptable salts thereof, may exist as either amorphous material or in crystalline form, or as a mixture thereof. Compounds of Formula (I) or a pharmaceutically acceptable salt thereof, may exist in solvated or unsolvated forms and may be formed wherein solvent molecules are incorporated into the crystalline lattice during crystallisation. Solvates may involve non-aqueous solvents such as ethanol, isopropanol, dimethylsulfoxide (DMSO), acetic acid, ethanolamine, and ethyl acetate, or they may involve water as the solvent that is incorporated into the crystalline lattice. Solvates wherein water is the solvent that is incorporated into the crystalline lattice are typically referred to as "hydrates." Salts of a compound of Formula (I) may be prepared by contacting appropriate stoichiometric amounts of the free base/acid with the appropriate acid/base in a suitable solvent. The free base/acid of a compound of Formula (I) may for example be in solution with the appropriate acid/base added as a solid or both the free base/acid of a compound of Formula (I) and the appropriate acid/base may independently be in solution.

Suitable solvents for solubilising a compound of Formula (I) free base include for example alcohols such as isopropanol; ketones such as acetone; acetonitrile or toluene. If the base is to be added as a solution in a solvent, the solvent used may include acetone, methanol or water.

The salts of a compound of Formula (I) may be isolated in solid form by conventional means from a solution thereof obtained as above. For example, a non-crystalline salt may be prepared by precipitation from solution, spray drying or freeze drying of solutions, evaporating a solution to a glass, or vacuum drying of oils, or solidification of melts obtained from reaction of the free base and the acid. The salts of a compound of Formula (I) may be prepared by directly crystallising from a solvent in which the salt has limited solubility, or by triturating or otherwise crystallising a noncrystalline salt. For example, organic solvents such as acetone, acetonitrile, butanone, 1 - butanol, ethanol, 1 -propanol or tetrahydrofuran or mixtures of such solvents may be used. An improved yield of the salts may be obtained by the evaporation of some or all of the solvent or by crystallisation at elevated temperature followed by controlled cooling, for example in stages. Careful control of the precipitation temperature and seeding may be used to improve the reproducibility of the production process and the particle size distribution and form of the product. Salts and solvates of a compound of Formula (I) which are suitable for use in medicine are those wherein the counterion or associated solvent is pharmaceutically acceptable. However, salts having non-pharmaceutically acceptable counterions or associated solvents are within the scope of the present invention, for example, for use as intermediates in the preparation of a compound of Formula (I) or their pharmaceutically acceptable salts.

It will be appreciated by those skilled in the art that certain protected derivatives of a compound of Formula (I), which may be made prior to a final deprotection stage, may not possess pharmacological activity as such, but may, in certain instances, be administered orally or parenterally and thereafter metabolised in the body to form compounds defined in the first aspect which are pharmacologically active. Such derivatives may therefore be described as "prodrugs". Examples of suitable prodrugs for the compounds of the present invention are described in Drugs of Today, Volume 19, Number 9, 1983, pp 499 - 538 and in Topics in Chemistry, Chapter 31 , pp 306 - 316 and in "Design of Prodrugs" by H. Bundgaard, Elsevier, 1985, Chapter 1 (the disclosures in which documents are incorporated herein by reference). It will further be appreciated by those skilled in the art, that certain moieties, known to those skilled in the art as "pro-moieties", for example as described by H. Bundgaard in "Design of Prodrugs" (the disclosure in which document is incorporated herein by reference) may be placed on appropriate functionalities when such functionalities are present within a compound of Formula (I). Suitable prodrugs for compounds of Formula (I) or salts or solvates thereof include : amides, carbamates, azo-compounds, phosphamides, glycosides. As described above, compounds of Formula (I) may be in the form of their free base or a pharmaceutically acceptable salt, solvate, or prodrug of compounds of Formula (I), which upon administration to the recipient is capable of providing (directly or indirectly) a compound of Formula (I), or an active metabolite or residue thereof. Such pharmaceutically acceptable salts, solvates, and prodrugs are recognizable to those skilled in the art, without undue experimentation. Nevertheless, reference is made to the teaching of Burger's Medicinal Chemistry and Drug Discovery, 5th Edition, Vol 1 : Principles and Practice, which is incorporated herein by reference to the extent of teaching such derivatives. Furthermore, some of the crystalline forms of compounds of Formula (I) or salts thereof may exist in one or more polymorphic form, which are included in the present invention.

It will further be appreciated that compounds of Formula (I) may exist in different tautomeric forms. All possible tautomers are contemplated to be within the scope of the present invention.

Compounds of Formula (I) or a pharmaceutically acceptable salt thereof can be useful in the treatment of certain parasitic infections such as parasitic protozoal infections by the malarial parasite Plasmodium falciparum, species of Eimeria, Pneumocytis carinii, Trypanosoma cruzi, Trypanosoma brucei or Leishmania donovani. In particular, compounds of Formula (I) or pharmaceutically acceptable salts thereof can be useful for treatment of infection by Plasmodium falciparum. Accordingly, the invention is directed to methods of treating such infections. Therefore, there is provided compounds of Formula (I) or pharmaceutically acceptable salts thereof, for use in therapy.

In one aspect of the invention there is provided compounds of Formula (I) or pharmaceutically acceptable salts thereof for use in the treatment of a parasitic protozoal infection. In another aspect of the invention there is provided compounds of Formula (I) or pharmaceutically acceptable salts thereof for use in the treatment of malaria.

In another aspect of the invention, there is provided compounds of Formula (I) or pharmaceutically acceptable salts thereof for use in the treatment of an infection by Plasmodium falciparum.

In another aspect of the invention, there is provided the use of compounds of Formula (I) or pharmaceutically acceptable salts thereof in the manufacture of a medicament for the treatment of a parasitic protozoal infection. In another aspect of the invention, there is provided the use of compounds of Formula (I) or pharmaceutically acceptable salts thereof in the manufacture of a medicament for the treatment of malaria. In another aspect of the invention, there is provided the use of compounds of Formula (I) or pharmaceutically acceptable salts thereof in the manufacture of a medicament for the treatment of an infection by Plasmodium falciparum.

In another aspect of the invention, there is provided a method for the treatment of a human or animal subject suffering from a parasitic protozoal infection, which method comprises administering to said human or animal subject an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.

In another aspect of the invention, there is provided a method for the treatment of a human or animal subject suffering from malaria, which method comprises administering to said human or animal subject an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.

In another aspect of the invention, there is provided a method for the treatment of a human or animal subject suffering from a parasitic protozoal infection by Plasmodium falciparum, which method comprises administering to said human or animal subject an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.

The methods of treatment of the invention comprise administering a safe and effective amount of a compound according to Formula (I), and a pharmaceutically acceptable salt thereof to a patient in need thereof.

As used herein, "treatment" means: (1 ) the amelioration or prevention of the condition being treated or one or more of the biological manifestations of the condition being treated, (2) the interference with (a) one or more points in the biological cascade that leads to or is responsible for the condition being treated or (b) one or more of the biological manifestations of the condition being treated, or (3) the alleviation of one or more of the symptoms or effects associated with the condition being treated. The skilled artisan will appreciate that "prevention" is not an absolute term. In medicine, "prevention" is understood to refer to the prophylactic administration of a drug to substantially diminish the likelihood or severity of a condition or biological manifestation thereof, or to delay the onset of such condition or biological manifestation thereof.

As used herein, an "effective amount" or a "safe and effective amount" means an amount of the compound sufficient to significantly induce a positive modification in the condition to be treated but low enough to avoid serious side effects (at a reasonable benefit/risk ratio) within the scope of sound medical judgment. A safe and effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof will vary with the particular compound chosen (e.g. depending on the potency, efficacy, and half-life of the compound); the route of administration chosen; the nature of the infection and/or condition being treated; the severity of the infection and/or condition being treated; the age, size, weight, and physical condition of the patient being treated; the medical history of the patient to be treated; the duration of the treatment; the nature of concurrent therapy; the desired therapeutic effect; and like factors, but can nevertheless be routinely determined by the skilled artisan.

As used herein, "patient" refers to a human or other animal subject.

A compound of Formula (I) or a pharmaceutically acceptable salt thereof may be administered by any suitable route of administration, including systemic administration. Systemic administration includes oral administration, parenteral administration, transdermal administration, rectal administration, and administration by inhalation. Parenteral administration refers to routes of administration other than enteral, transdermal, or by inhalation, and is typically by injection or infusion. Parenteral administration includes intravenous, intramuscular, and subcutaneous injection or infusion. Inhalation refers to administration into the patient's lungs whether inhaled through the mouth or through the nasal passages.

A compound of Formula (I) or a pharmaceutically acceptable salt thereof may be administered once only, or according to a dosing regimen wherein a number of doses are administered at varying intervals of time for a given period of time. For example, doses may be administered one, two, three, or four times per day. Doses may be administered until the desired therapeutic effect is achieved or indefinitely to maintain the desired therapeutic effect. The dosage will also vary according to the nature of the intended treatment, wherein "treatment" is as herein defined, for example a greater dose of compound may be given for amelioration as compared with prevention of a condition being treated. Suitable dosing regimens for a compound of the invention depend on the pharmacokinetic properties of that compound, such as absorption, distribution, and half-life, which can be determined by the skilled artisan. In addition, suitable dosing regimens including the duration such regimens are administered, depend on the route of administration of the compound, on the condition being treated, the severity of the condition being treated, the age and physical condition of the patient being treated, the medical history of the patient to be treated, the nature of any concurrent therapy, the desired therapeutic effect, and like factors within the knowledge and expertise of the skilled artisan. It will be further understood by such skilled artisans that suitable dosing regimens may require adjustment given an individual patient's response to the dosing regimen or over time as individual patient needs change. It will also be appreciated that if a compound of Formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with one or more additional active therapeutic agents as discussed further hereinbelow, the dosing regimen of the compound of the invention may also vary according to the nature and amount of the one or more additional active therapeutic agents as necessary.

Typical dosages of a compound of Formula (I) may vary depending upon the particular route of administration chosen. Typical dosages for oral administration are predicted to be in a range from about 25 to about 1000mg. Typically a compound of Formula (I) may be administered once a day, once every two days or even up to once weekly.

The compounds of Formula (I) or pharmaceutically acceptable salts thereof may also be used in combination with other active therapeutic agents. The invention thus provides, in a further aspect, a combination comprising a compound of Formula (I) or a pharmaceutically acceptable salt thereof together with a further active therapeutic agent. When a compound of Formula (I) or a pharmaceutically acceptable salt thereof is used in combination with a second active therapeutic agent which is active against the same disease state the dose of each compound may differ from that when the compound is used alone. Appropriate doses will be readily appreciated by those skilled in the art. It will be appreciated that the amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof required for use in treatment will vary with the nature of the condition being treated and the age and the condition of the patient and will be ultimately at the discretion of the attendant physician or veterinarian.

A compound of Formula (I) or a pharmaceutically acceptable salt thereof may be used alone or in combination with one or more additional active therapeutic agents, such as other antiparasitic drugs, for example antimalarial drugs.

Such other active therapeutic agents include antimalarial drugs such as (e.g. chloroquine, mefloquine, primaquine, pyrimethamine, quinine, artemisinin, halofantrine, doxycycline, amodiaquine, atovaquone, tafenoquinedapsone, proguanil, sulfadoxine, cycloguanil) and fansidar.

The combinations referred to above may conveniently be presented for use in the form of a pharmaceutical formulation and thus pharmaceutical formulations comprising a combination as defined above together with a pharmaceutically acceptable carrier and/or excipient comprise a further aspect of the invention. The individual components of such combinations may be administered either sequentially or simultaneously in separate or combined pharmaceutical formulations by any convenient route.

When administration is sequential, either a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or the one or more additional active therapeutic agent(s) may be administered first. When administration is simultaneous, the combination may be administered either in the same or different pharmaceutical composition. When combined in the same formulation it will be appreciated that a compound of Formula (I) or a pharmaceutically acceptable salt thereof, and the one or more additional active therapeutic agent(s) must be stable and compatible with each other and the other components of the formulation. When formulated separately a compound of Formula (I) or a pharmaceutically acceptable salt thereof, and the one or more additional active therapeutic agent(s) may be provided in any convenient formulation, conveniently in such manner as are known for such compounds in the art.

Compositions

A compound of Formula (I) or a pharmaceutically acceptable salt thereof will normally, but not necessarily, be formulated into pharmaceutical compositions prior to administration to a patient. In one aspect, the invention is directed to pharmaceutical compositions comprising a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In another aspect the invention is directed to a pharmaceutical composition comprising (a) a compound of Formula (I) or a pharmaceutically acceptable salt thereof and (b) one or more pharmaceutically acceptable carriers and/or excipients. In another aspect, the invention provides a pharmaceutical composition comprising a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In a further aspect, the invention provides a pharmaceutical composition comprising (a) a compound of Formula (I) or a pharmaceutically acceptable salt thereof and (b) one or more pharmaceutically acceptable carriers. In a further aspect, the invention provides a pharmaceutical composition comprising (a) a compound of Formula (I) or a pharmaceutically acceptable salt thereof and (b) a pharmaceutically acceptable carrier.

The carrier and/or excipient must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

The pharmaceutical compositions of the invention may be prepared and packaged in bulk form wherein a safe and effective amount of a compound of the invention can be extracted and then given to the patient such as with powders or syrups. Alternatively, the pharmaceutical compositions of the invention may be prepared and packaged in unit dosage form wherein each physically discrete unit contains a safe and effective amount of a compound of the invention. When prepared in unit dosage form, the pharmaceutical compositions of the invention typically contain from about 0.1 to 1000mg, in another aspect 0.1 mg to about 500 mg of a compound of the invention. The pharmaceutical compositions of the invention typically contain one compound of Formula (I) or a pharmaceutically acceptable salt thereof. However, in certain embodiments, the pharmaceutical compositions of the invention contain more than one compound of Formula (I) or a pharmaceutically acceptable salt thereof. For example, in certain embodiments the pharmaceutical compositions of the invention contain two compounds of Formula (I). In addition, the pharmaceutical compositions of the invention may optionally further comprise one or more additional active therapeutic compounds. The pharmaceutical compositions of the invention typically contain more than one pharmaceutically acceptable excipient. However, in certain embodiments, the pharmaceutical compositions of the invention contain one pharmaceutically acceptable excipient.

A compound of Formula (I) or a pharmaceutically acceptable salt thereof, and the pharmaceutically acceptable excipient or excipients will typically be formulated into a dosage form adapted for administration to the patient by the desired route of administration. For example, dosage forms include those adapted for (1 ) oral administration such as tablets, capsules, caplets, pills, troches, powders, syrups, elixirs, suspensions, solutions, emulsions, sachets, and cachets; (2) parenteral administration such as sterile solutions, suspensions, and powders for reconstitution; (3) transdermal administration such as transdermal patches; (4) rectal administration such as suppositories; and (5) inhalation such as aerosols and solutions.

Suitable pharmaceutically acceptable excipients will vary depending upon the particular dosage form chosen. In addition, suitable pharmaceutically acceptable excipients may be chosen for a particular function that they may serve in the composition. For example, certain pharmaceutically acceptable excipients may be chosen for their ability to facilitate the production of uniform dosage forms. Certain pharmaceutically acceptable excipients may be chosen for their ability to facilitate the production of stable dosage forms. Certain pharmaceutically acceptable excipients may be chosen for their ability to facilitate the carriage or transport of compounds of Formula (I) or pharmaceutically acceptable salts thereof from one organ, or portion of the body, to another organ, or portion of the body, once administered to the patient. Certain pharmaceutically acceptable excipients may be chosen for their ability to enhance patient compliance.

Suitable pharmaceutically acceptable excipients include the following types of excipients: binders, disintegrants, lubricants, glidants, granulating agents, coating agents, wetting agents, solvents, co-solvents, suspending agents, emulsifiers, sweeteners, flavouring agents, flavour masking agents, coloring agents, anticaking agents, humectants, chelating agents, plasticizers, viscosity increasing agents, antioxidants, preservatives, stabilizers, surfactants, and buffering agents. The skilled artisan will appreciate that certain pharmaceutically acceptable excipients may serve more than one function and may serve alternative functions depending on how much of the excipient is present in the formulation and what other ingredients are present in the formulation.

Skilled artisans possess the knowledge and skill in the art to enable them to select suitable pharmaceutically acceptable excipients in appropriate amounts for use in the invention. In addition, there are a number of resources that are available to the skilled artisan which describe pharmaceutically acceptable excipients and may be useful in selecting suitable pharmaceutically acceptable excipients. Examples include Remington's Pharmaceutical Sciences (Mack Publishing Company), The Handbook of Pharmaceutical Additives (Gower Publishing Limited), and The Handbook of Pharmaceutical Excipients (the American Pharmaceutical Association and the Pharmaceutical Press).

The pharmaceutical compositions of the invention are prepared using techniques and methods known to those skilled in the art. Some of the methods commonly used in the art are described in Remington's Pharmaceutical Sciences (Mack Publishing Company).

In one aspect, the invention is directed to a solid or liquid oral dosage form such as a liquid, tablet, lozenge or a capsule, comprising a safe and effective amount of a compound of the invention and a carrier. The carrier may be in the form of a diluent or filler. Suitable diluents and fillers in general include lactose, sucrose, dextrose, mannitol, sorbitol, starch (e.g. corn starch, potato starch, and pre-gelatinized starch), cellulose and its derivatives (e.g. microcrystalline cellulose), calcium sulfate, and dibasic calcium phosphate. A liquid dosage form will generally consist of a suspension or solution of the compound or pharmaceutically acceptable salt in a liquid carrier for example, ethanol, olive oil, glycerine, glucose (syrup) or water (e.g. with an added flavouring, suspending, or colouring agent). Where the composition is in the form of a tablet or lozenge, any pharmaceutical carrier routinely used for preparing solid formulations may be used. Examples of such carriers include magnesium stearate, terra alba, talc, gelatin, acacia, stearic acid, starch, lactose and sucrose. Where the composition is in the form of a capsule, any routine encapsulation is suitable, for example using the aforementioned carriers or a semi solid e.g. mono di-glycerides of capric acid, Gelucire™ and Labrasol™, or a hard capsule shell e.g. gelatin. Where the composition is in the form of a soft shell capsule e.g. gelatin, any pharmaceutical carrier routinely used for preparing dispersions or suspensions may be considered, for example aqueous gums or oils, and may be incorporated in a soft capsule shell.

An oral solid dosage form may further comprise an excipient in the form of a binder. Suitable binders include starch (e.g. corn starch, potato starch, and pre-gelatinized starch), gelatin, acacia, sodium alginate, alginic acid, tragacanth, guar gum, povidone, and cellulose and its derivatives (e.g. microcrystalline cellulose). The oral solid dosage form may further comprise an excipient in the form of a disintegrant. Suitable disintegrants include crospovidone, sodium starch glycolate, croscarmelose, alginic acid, and sodium carboxymethyl cellulose. The oral solid dosage form may further comprise an excipient in the form of a lubricant. Suitable lubricants include stearic acid, magnesium stearate, calcium stearate, and talc.

There is further provided by the present invention a process of preparing a pharmaceutical composition, which process comprises mixing compounds of Formula (I) or pharmaceutically acceptable salts thereof, together with a pharmaceutically acceptable carrier and/or excipient.

Preparations for oral administration may be suitably formulated to give controlled/extended release of the active compound. PROCESSES

A general procedure for acid chloride formation and amide formation is described in the literature (J. Chem. Res. 2008 (22), 530-533) using appropriate commercially available acids and anilines as starting materials.

The compounds of formula (I) may be synthesised by asymmetric or non-asymmetric routes. A non-asymmetric synthesis of compounds of Formula (I) is shown in Scheme 1 .

An asymmetric synthesis of compounds of Formula (I) is shown in Scheme 2

commercially

available

Intermediate 1 1

Intermediate 12

Scheme 2

It will be readily apparent to those skilled in the art that compounds of Formula (I) may be prepared using methods analogous to those outlined above, or by reference to the experimental procedures detailed in the Examples provided herein. Further details for the preparation of compounds of Formula (I) are found in the Examples.

With appropriate manipulation and protection of any chemical functionality, the synthesis of compounds of Formula (I) is accomplished by methods analogous to those above. In any particular case, particular protecting groups may be required. Suitable protecting groups can be found, but are not restricted to, those found in T W Greene and P G M Wuts 'Protective Groups in Organic Synthesis', 3 ^rd Ed (1999), J Wiley and Sons. EXPERIMENTAL

Abbreviations

In describing the invention, chemical elements are identified in accordance with the Periodic Table of the Elements. Abbreviations and symbols utilised herein are in accordance with the common usage of such abbreviations and symbols by those skilled in the chemical arts. The following abbreviations are used herein: anhydrous

aqueous

Antimalarial whole cell Screening

Catalytic

1 - Cyano-2-ethoxy-2-oxoethylidenaminooxy)dimethylamino-morpholi no- carbenium hexafluorophosphate

concentrated

Cyclohexane

1 ,1 '-Bis(diphenylphosphino)ferrocene

Dicloromethane

Diisopropylethylamine

Dimethoxyethane

Dimethylformamide

Deuterated dimethylsulfoxide

Ethyl acetate

Electrospray mass spectrometry

grams

Hours

N-[(Dimethylamino)-1 H-1 ,2,3-triazolo-[4,5-b]pyridin-1 -ylmethylene]-N- methylmethanaminium hexafluorophosphate N-oxide

High perfomance liquid chromatography

High throughput screening

2- Propanol

Diisopropyl ether

Molar

Methanol

Minuts

mililitres

milimoles

miligrams

Microwave

Normal

N-bromosuccinimide

proton nuclear magnetic resonance spectroscopy

room temperature sat. saturated

TFA Trifluoroacetic acid

THF Tetrahydrofuran

°C degrees centigrade

Compound Preparation

Examples

The following Examples illustrate the invention. These Examples are not intended to limit the scope of the invention, but rather to provide guidance to the skilled artisan to prepare and use the compounds, compositions, and methods of the invention. While particular embodiments of the invention are described, the skilled artisan will appreciate that various changes and modifications can be made without departing from the spirit and scope of the invention. Where materials were commercially available, this is indicated in parentheses after the compound name, in capitals. For example, in the preparation of Intermediate 1 , L-valine tert- butyl ester, hydrochloride was purchased from ALDRICH, so it is stated "L-valine tert-butyl ester, hydrochloride (ALDRICH).

Intermediate 1 (S)-ie f-Butyl-3-methyl-2-ureidobutanoate

To a solution of L-valine tert-butyl ester, hydrochloride (ALDRICH, 10 g, 47.7 mmol) in water (60 mL), potassium cyanate (ALDRICH, 4.1 g, 50.5 mmol) was added and the reaction was stirred at room temperature for 3 hours. The resulting solid was filtered, washed with water (2x20 mL) and dried, first under air stream and then overnight at 35°C under vaccum in presence of P2O ₅ , to give (S)-ie f-butyl-3-methyl-2-ureidobutanoate (8.89 g) as a white solid. The aqueous filtrates were combined and extracted with EtOAc (2x50 mL). The organic layers were combined and washed with 0.5N HCI (30 mL) and brine (2x30 mL), dried (Na ₂ S0 ₄ ), filtered and evaporated to dryness under vacuum to give 0.756 g of a second batch of (S)-ie/f-butyl-3-methyl-2-ureidobutanoate (white solid). ¹ H NMR (400 MHz, DMSO- d6) 5ppm: 6.14 (br.d, 1 H), 5.58 (br.s, 2H), 3.92 (dd, 1 H), 2.04-1 .88 (m, 1 H), 1 .39 (s, 9H), 0.84 (d, 3H), 0.81 (d, 3H). [ES+ MS] m/z 217 (M+H).

Intermediate 2 (S,Z)-Ethyl 3-(3-(1 -(ie/f-butoxy)-3-methyl-1 -oxobutan-2-yl)ureido)but-2- enoate

A suspension of ethyl acetoacetate (3.84 g, 3.75 ml_, 29.5 mmol), (S)-tert-butyl 3-methyl-2- ureidobutanoate (5.8 g, 26.8 mmol) and pyridinium p-toluenesulfonate (0.58 g) in cyclohexane (200 mL) was stirred at room temperature for 1 h, then at 75°C for 2h and finally refluxed for 24 h azeotropically removing the formed water by means of a Dean-Stark. After cooling to rt, the reaction mixture was evaporated to dryness under vacuum to give a crude (10 g) which was purified by chromatography (Merck Si60 15-40μη"ΐ, 2x130 g cartridges, eluting with cyclohexane/ethyl acetate 0% to 10%) to obtain 4.53 g of pure (S,Z)-ethyl 3-(3- (1 -(ie f-butoxy)-3-methyl-1 -oxobutan-2-yl)ureido)but-2-enoate (colorless syrup which solidified in the fridge) and a second batch of slightly impure compound (1 .86 g, colorless syrup which solidified in the fridge, It contains approx 5% of ethyl acetoacetate). ¹ H NMR (400 MHz, DMSO-d6) 5ppm: 10.18 (s, 1 H), 8.04 (br.d, 1 H), 4.77 (d, 1 H), 4.06 (q, 2H), 3.90 (dd, 1 H), 2.21 (d, 3H), 2.07-1.95 (m, 1 H), 1.39 (s, 9H), 1.18 (t, 3H), 0.87 (d, 6H). [ES+ MS] m/z 329 (M+H).

Intermediate 3 (S)-ieri-Butyl 3-methyl-2-(4-methyl-2,6-dioxo-2,3-dihydropyrimidin-1 (6H)- yl)butanoate

A solution of (S,Z)-ethyl 3-(3-(1 -(tert-butoxy)-3-methyl-1 -oxobutan-2-yl)ureido)but-2-enoate (1 .85 g, 5.63 mmol) in 20 mL of anhydrous tetrahydrofuran under argon atmosphere was cooled to 0°C (ice-water bath) and treated portionwise over 25 min with potassium tert- butoxide (1 .39 g, 12.39 mmol). The mixture was warmed to room temperature and stirred for 2h. The reaction was cooled to 0°C (ice-water bath) and quenched by addition of 0.5N HCI (60 mL). The mixture was extracted twice with tert-butyl methyl ether (75 and 50 mL). The organic phases were combined and washed with water (2x50 mL) and brine (50 mL), dried (Na ₂ S0 ₄ ), filtered and evaporated to dryness under vacuum to give a crude which was purified by chromatography (Merck Si60 15-40μη"ΐ, 50 g cartridge, eluting with cyclohexane/ethyl acetate 5% to 60%) to obtain (S)-ieri-butyl 3-methyl-2-(4-methyl-2,6-dioxo- 2,3-dihydropyrimidin-1 (6H)-yl)butanoate (0.92 g, colorless syrup which solidified in the fridge). ¹ H NMR (400 MHz, DMSO-d6) 5ppm: 1 1 .21 (br.s, 1 H), 5.48 (br.d, 1 H), 4.72 (d, 1 H), 2.55-2.40 (m, 1 H + DMSO-d6 residual peak), 2.04 (d, 3H), 1 .31 (s, 9H), 1.08 (d, 3H), 0.61 (d, 3H). [ES+ MS] m/z 283 (M+H). 94.25/5.75 S:R enantiomeric ratio (Chiralpak IC (25 x 0.46 cm), 5 μηη, Heptane / Isopropanol 90/10 v/v, flow 1 mL/min, UV (254 nm): 5.75 % a/a (1 1.3 min) and 94.25 % a/a (13.5min)).

Intermediate 4 (S)-ieri-Butyl 2-(5-bromo-4-methyl-2,6-dioxo-2,3-dihydropyrimidin-1 (6H)-yl)- 3-methylbutanoate

A solution of (S)-tert-butyl 3-methyl-2-(4-methyl-2,6-dioxo-2,3-dihydropyrimidin-1 (6H)- yl)butanoate (745 mg, 2.64 mmol) in 20 mL of anhydrous dichloromethane under nitrogen atmosphere was cooled to 0°C (ice-water bath) and treated portionwise over 15 min with N- bromosuccinimide (517 mg, 2.9 mmol). The mixture was allowed to reach room temperature and stirred for 1 h. Solvents were evaporated to dryness under reduced pressure to give a crude which was purified by chromatography (Merck Si ₆ 0 15-40μη"ΐ, 30 g cartridge, eluting with cyclohexane/ethyl acetate 0% to 40%) to obtain (S)-ferf-butyl 2-(5-bromo-4-methyl-2,6- dioxo-2,3-dihydropyrimidin-1 (6H)-yl)-3-methylbutanoate (0.74 g, white foam). ¹ H NMR (400 MHz, CDCI3) 5ppm: 10.1 1 (br.s, 1 H), 4.98 (d, 1 H), 2.75-2.60 (m, 1 H), 2.34 (s, 3H), 1.43 (s, 9H), 1 .22 (d, 3H), 0.78 (d, 3H). [ES- MS] m/z 359 and 361 with Br pattern (M-H). 95/5 S:R enantiomeric ratio (Chiralpak IC (25 x 0.46 cm), 5 μηη, Heptane / Isopropanol 95/5 v/v, flow 1 mL/min, UV (254 nm): 4.6 % a/a (8.8 min) and 95.4 % a/a (13.8 min)).

Intermediate 5 (S)-tert-butyl 3-methyl-2-(4-methyl-2,6-dioxo-5-(pyridin-3-yl)-2,3- dihydropyrimidin-1 (6H)-yl)butanoate.

A vial was charged with tert-butyl (2S)-3-methyl-2-[4-methyl-2,6-dioxo-5-(piperidin-1 -yl)- 1 ,2,3,6-tetrahydropyrimidin-1 -yl]butanoate (intermediate 4, 300 mg, 0.831 mmol) and (pyridin-3-yl)boronic acid (ALDRICH, 153 mg, 1 .246 mmol) and placed under nitrogen. DME (10 mL) and EtOH (3 mL) were added to give a solution. A solution of monobasic potassium phosphate (ALDRICH, 1 13 mg, 0.831 mmol) and tribasic potassium phosphate (ALDRICH, 176 mg, 0,831 mmol) in water (5 mL) were added. The mix was deoxygenated via several vacuum/N ₂ cycles before the addition of PdCI ₂ (dbpf) (ALDRICH, 39 mg, 0.083 mmol). The resulting reaction mixture was then stirred at 65 °C for 6 hours. Further PdCI2(dbpf) (39 mg, 0.083 mmol) was added and heating continued overnight. Further PdCI2(dbpf) (39 mg, 0.083 mmol) was added and heating continued for 6 hours. The reaction mixture was partially evaporated under reduced pressure to remove volatiles then the residue was partitioned between half-saturated sodium bicarbonate solution (100 mL) and EtOAc (100 mL).[NB it was necessary to filter the aqueous/organic mixture to help avoid emulsion formation]. The aqueous phase was further extracted with EtOAc (3 x 50 mL). The combined organic phases were washed with brine (50 mL), dried over Na ₂ S0 ₄ and evaporated under reduced pressure. The residue was chromatographed on NH-modified silica gel (SNAP 28) eluting with a gradient of 50-100% EtOAc in cyclohexane but many product containing fractions were mixed. Product containing fractions were evaporated to give -200 mg of partially purified product which was further chromatographed on silica gel (Isolute 10g cartridge) eluting with 50% EtOAc in cyclohexane and then EtOAc to give (S)-tert-butyl 3-methyl-2-(4-methyl-2,6- dioxo-5-(pyridin-3-yl)-2,3-dihydropyrimidin-1 (6H)-yl)butanoate (165 mg, 0.46 mmol, 55% yield) as colourless glass.

¹ H NMR (400 MHz, CDCI3) 5ppm: 10.10 (s, 1 H), 8.61 (d, 1 H), 8.50 (s, 1 H), 7.65 (d, 1 H), 7.37 (dd, 1 H), 5.01 (d, 1 H), 2.72 (m, 1 H), 2.16 (s, 3H), 1 .45 (s, 9H), 1.23 (d, 3H), 0.85 (d, 3H). [ES+ MS] m/z 360 (M+H). Intermediate 6 (2S)-3-methyl-2-[4-methyl-2,6-dioxo-5-(pyridin-3-yl)-1 ,2,3,6- tetrahydropyrimidin-1 -yl]butanoic acid hydrochloride.

(S)-tert-butyl 3-methyl-2-(4-methyl-2,6-dioxo-5-(pyridin-3-yl)-2,3-dihydrop yrimidin-1 (6H)- yl)butanoate (for example as prepared for Intermediate 5, 175 mg, 0.49 mmol) was dissolved in 4M hydrochloric acid solution in 1 ,4-dioxane (ALDRICH, 2.43 mL, 9.74 mmol) and the resulting solution was left to stand for 5 hours. The reaction concentrated under reduced pressure without heating. The beige solid residue was triturated with diethyl ether (10 mL) and dried under reduced pressure to give 213 mg of (2S)-3-methyl-2-[4-methyl-2,6-dioxo-5- (pyridin-3-yl)-1 ,2,3,6-tetrahydropyrimidin-1 -yl]butanoic acid hydrochloride as a beige solid impure with residual solvent and 2-(2-chloroethoxy)ethan-1 -ol. This material was used without further purification. [ES+ MS] m/z 304 (M+H). Intermediate 7 (S)-tert-butyl 3-methyl-2-(4-methyl-5-morpholino-2,6-dioxo-2,3- dihydropyrimidin-1 (6H)-yl)butanoate

tBuOK (ALDRICH, 4.81 g, 42.9 mmol) was added in portions (0.25 eq every 25 min) at 0 °C to a solution of (2£)-3-({[(2S)-1 -(tert-butoxy)-3-methyl-1 -oxobutan-2-yl]carbamoyl}amino)-2- (morpholin-4-yl)but-2-enoate (for example as prepared for Intermediate 1 1 , 17.6 g, 42.6 mmol) in THF (350 mL). Stirring went on at 0 °C throughout the reaction time.

30 min after the last addition, 400 mL of 1 N NH ₄ CI were added at 0 °C followed by 400 mL of EtOAc. The layers were separated and the aqueous layer was extracted with EtOAc (400 mL). Combined organic layers were washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure to give 14.4 g of crude reaction mixture.

Purification by silica gel chromatography (Biotage SP, 340 g KP-Sil Cartridge, Cy/AcOEt 85:15 to 50:50) yielded 1 1 .02 g of (S)-tert-butyl 3-methyl-2-(4-methyl-5-morpholino-2,6-dioxo- 2,3-dihydropyrimidin-1 (6H)-yl)butanoate (1 1.02 g, 29.9 mmol, 70% yield).

1H NMR (400 MHz, DMSO-d6) 5ppm: 1 1 .09 (s, 1 H), 4.71 (d, 1 H), 3.61 (br. s, 4H), 2.91 (br. s, 4H), 2.52 (m, 1 H), 2.18 (s, 3H), 1 .35 (s, 9H), 1 .12 (d, 3H), 0.66 (d, 3H). [ES+ MS] m/z 368 (M+H).

Intermediate rac-7 ie f-butyl 3-methyl-2-(4-methyl-5-morpholino-2,6-dioxo-2,3- dihydropyrimidin-1 (6H)-yl)butanoate

tert-butyl 2-(5-bromo-4-methyl-2,6-dioxo-2,3-dihydropyrimidin-1 (6H)-yl)-3-methylbutanoate (for example as prepared for Intermediate 3, 363 mg, 1 .005 mmol) was dissolved in Morpholine (ALDRICH, 2 ml, 22.96 mmol). The reaction was stirred at 140°C for 3 h in microwave. The mixture was triturated with diethyl ether to give a white solid and the filtrate was purified on silica gel column and was eluted with CH ₂ CI ₂ /MeOH gradient 0-15%. Collected fractions was evaporated under reduced pressure to give tert-butyl 3-methyl-2-(4- methyl-5-morpholino-2,6-dioxo-2,3-dihydropyrimidin-1 (6H)-yl)butanoate (252 mg, 0.686 mmol, 68% yield) with some AcOEt. The compound was used in the next step of reaction without further purification.

1 H NMR (400 MHz, DMSO-d6) δρρπι: 1 1 .18 (br.s., 1 H), 4.76 (d, 1 H), 3.66 (m, 4 H), 2.51 (m, 4 H), 2.23 (s, 3 H), 1 .40 (s., 9 H), 1 .17 (d, 3 H), 0.71 (d, 3 H).

Intermediate 8 Ethyl 2-(morpholin-4-yl)-3-oxobutanoate

To a mixture of ethyl 2-chloro-3-oxobutanoate (ALDRICH, 12.6 mL, 91 .1 mmol) in anhydrous DMF (30 mL) under N2 atmosphere, morpholine (ALDRICH, 17.5 mL, 200.4 mmol) was added and after stirring for 15 minutes, the resulting mixture was stirred at 50 °C overnight. The reaction was treated with 1 N NH ₄ CI and extracted with AcOEt. Combined organic layers were whashed with water and brine, dired over anhydrous MgS0 ₄ , filtered and concentrated under reduced pressure to give 19.6 g of crude as a orange oil that was purified by silica gel chromatography used Biotage SP, 340 g KP-Sil cartridge, eluting with Cy/AcOEt 100:0 to 80:20 to give 14.7 g of pure ethyl 2-(morpholin-4-yl)-3-oxobutanoate as a pale oil of 27:73 mixture of isomers (14.7 g, 60.8 mmol, 75% yield).

[ES+ MS] m/z 216 (M+H).

Intermediate 9 Ethyl 3-amino-2-(morpholin-4-yl)but-2-enoate

A solution of ammonium acetate (ALDRICH, 26.3 g, 341 .47 mmol) and ethyl 2-(morpholin-4- yl)-3-oxobutanoate (for example as prepared for Intermediate 8, 14.7 g, 68.29 mmol) in MeOH (44 mL) was stirred at room temperature for 2 h. Volatiles were removed under reduced pressure. Aqueous sodium bicarbonate was added to the crude and mixture extracted with EtOAc (x3). Organic layers was washed with brine, dried over sodium sulphate, filtered and concentrated under reduced pressure to give 15 g of Ethyl 3-amino-2- (morpholin-4-yl)but-2-enoate, as a yellow solid (15 g, 69.3 mmol, quant, yield) as 28:71 mixture of geometric isomers.

[ES+ MS] m/z 215 (M+H). Intermediate 10 Ethyl 2-(morpholin-4-yl)-3-[(phenoxycarbonyl)amino]but-2-enoate

Phenyl chloroformate (ALDRICH, 10.53 mL, 84 mmol) was added at 0 °C to a solution of ethyl 3-amino-2-(morpholin-4-yl)but-2-enoate (intermediate 9, 15 g, 70 mmol) and pyridine (ALDRICH, 1 1.32 mL, 140 mmol) in chloroform (73 mL). The resulting mixture was allowed to reach room temperature and stirred overnight. The reaction was diluted with EtOAc and it was washed with 0.5 M HCI, saturated aqueous sodium bicarbonate and brine. The resulting organic layer was dried over anhydrous MgS04, filtered and concentrated under reduced pressure to give 24.5 g of ethyl 2-(morpholin-4-yl)-3-[(phenoxycarbonyl)amino]but-2-enoate (24.5g, 58.61 mmol, 84% yield) as a orange oil (39:41 mixture of geometric isomers).

[ES+ MS] m/z 335 (M+H). Intermediate 11 (S)-ethyl 3-(3-(1 -(tert-butoxy)-3-methyl-1 -oxobutan-2-yl)ureido)-2- morpholinobut-2-enoate

Potassium carbonate (ALDRICH, 15.19 g, 109.9 mmol) was added at 0 °C to a solution of ethyl 2-(morpholin-4-yl)-3-[(phenoxycarbonyl)amino]but-2-enoate (for example as prepared for Intermediate 10, 24.5 g, 73.27 mmol) and L-valine tert-butyl ester hydrochloride (16.13 g, 76.94 mmol) in anhydrous DMF (76 mL). The resulting mixture was stirred at room temperature overnight. The reaction was treated with 1 N NH ₄ CI and the mixture was extracted with EtOAc (2 x). Combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give 40 g of crude as a orange oil. First purification was performed through Biotage SP, 340 g KP-Sil cartridge, Cy/AcOEt from 10% to 20%, to give 12.79 of £-(S)-ethyl 3-(3-(1 -(tert-butoxy)-3- methyl-1 -oxobutan-2-yl)ureido)-2-morpholinobut-2-enoate (12.79 g, 28.61 mmol, 39% yield) as a white foam and 13.78 g of Z-(S)-ethyl 3-(3-(1 -(tert-butoxy)-3-methyl-1 -oxobutan-2- yl)ureido)-2-morpholinobut-2-enoate (13.78 g, 29.86 mmol, 41 % yield) as a white foam. E-isomer: ¹ H NMR (400 MHz, DMSO-d6) 5ppm: 10.22 (s, 1 H), 7.72 (d, 1 H), 4.18 (q, 2H), 3.91 (dd, 1 H) , 3.58 (m, 4H), 2.75 (m, 4H), 2.44 (s, 3H), 2.01 (m, 1 H), 1 .41 (s, 9H), 1.28 (t, 3H), 0.69 (d, 6H). [ES+ MS] m/z 414 (M+H).

Z-isomer: ¹ H NMR (400 MHz, DMSO-d6) 5ppm: 8.55 (s, 1 H), 7.85 (d, 1 H), 4.12 (q, 2H), 3.99 (dd, 1 H) , 3.72 (m, 4H), 2.71 (m, 4H), 2.42 (s, 3H), 2.03 (m, 1 H), 1.42 (s, 9H), 1.24 (t, 3H), 0.91 (d, 3H), 0.89 (d, 3H). [ES+ MS] m/z 414 (M+H).

Intermediate 12 (S)-3-methyl-2-(4-methyl-5-morpholino-2,6-dioxo-2,3-dihydrop yrimid 1 (6H)-yl)butanoic acid.

To (S)-tert-butyl 3-methyl-2-(4-methyl-5-morpholino-2,6-dioxo-2,3-dihydropyrim idin-1 (6H)- yl)butanoate (for example as prepared for intermediate 7, 0.98 g, 2.67 mmol) under N2 atmosphere at 0 °C, 4N HCI in dioxane (14.67 mL, 58.7 mmol) was added and the resultant mixture was stirred allowing to reach room temperature. 26 h later, the reaction was concentrated under reduced pressure. The white solid obtained was treated with 35 mL of water and extracted with EtOAc (2x70 mL). The combination of the organic layers was washed with brine, dried over anhydrous MgS0 ₄ , filtered and concentrated under reduced pressure to give 806 mg of N31741 -6-1 , (S)-3-methyl-2-(4-methyl-5-morpholino-2,6-dioxo- 2,3-dihydropyrimidin-1 (6H)-yl)butanoic acid (806 mg, 2.59 mmol, 97 % yield) as a white solid. ¹ H NMR (400 MHz, DMSO-d6) 5ppm: 12.46 (s, 1 H), 1 1.08 (s, 1 H), 4.74 (d, 1 H), 3.57 (br. s, 4H), 2.53 (br. s, 4H), 2.49 (m, 1 H), 2.13 (s, 3H), 1.09 (d, 3H), 0.62 (d, 3H). [ES+ MS] m/z 312 (M+H).

Intermediate rac-M (S)-3-methyl-2-(4-methyl-5-morpholino-2,6-dioxo-2,3-dihydrop yrimid 1 (6H)-yl)butanoic acid.

Tert-butyl (2S)-3-methyl-2-[4-methyl-5-(morpholin-4-yl)-2,6-dioxo-1 ,2,3,6-tetrahydropyrimidin- 1 -yl]butanoate (for example as prepared for Intermediate rac-1, 185 mg, 0.50 mmol) was treated with 4M hydrochloric acid solution in 1 ,4-dioxane (ALDRICH, 2.50 mL, 10.0 mmol) to give a white suspension. Further 4M hydrochloric acid solution in 1 ,4-dioxane (2.50 mL, 10.0 mmol) was added but the reaction mixture remained a suspension which was then stirred overnight. The reaction was concentrated under reduced pressure and the residue was treated twice with diethyl ether (20 mL) and evaporated under reduced pressure to give 196 mg of a (S)-3-methyl-2-(4-methyl-5-morpholino-2,6-dioxo-2,3-dihydrop yrimidin-1 (6H)- yl)butanoic acid (196 mg, 0.50 mmol, quant, yield) white solid, the desired product with residual solvent. This material was used without further purification.

[ES+ MS] m/z 312 (M+H).

Example 1 (2S)-N-(3-fluorophenyl)-3-methyl-2-[4-methyl-2,6-dioxo-5-(py ridin-3-yl)- 1 ,2,3,6-tetrahydropyrimidin-1 -yl]butanamide.

3-Fluoroaniline (ALDRICH, 60 uL, 0.62 mmol), triethylamine (ALDRICH, 100 uL, 0.72 mmol) and then COMU (ALDRICH, 247 mg, 0.58 mmol) were added to a stirred solution of (2S)-3- methyl-2-[4-methyl-2,6-dioxo-5-(pyridin-3-yl)-1 ,2,3,6-tetrahydropyrimidin-1 -yl]butanoic acid hydrochloride (for example as prepared for Intermediate 6, 210 mg, 0.048 mmol) in EtOAc (2 mL) and DMF (2 mL) at 0°C. The reaction mixture was allowed to warm slowly to room temperature and stir for 4 hours. The reaction mixture was diluted with EtOAc (100 mL), washed with 1 M HCI solution (50 mL), sodium bicarbonate solution (50 mL) and brine (50 mL), dried over sodium sulphate and evaporated under reduced pressure. The residue was chromatographed on NH-modified silica gel (SNAP 28) eluting with a gradient of 0.5-4% MeOH in dichloromethane to give 173 mg of (2S)-N-(3-fluorophenyl)-3-methyl-2-[4-methyl- 2,6-dioxo-5-(pyridin-3-yl)-1 ,2,3,6-tetrahydropyrimidin-1 -yl]butanamide (171 mg, 0.431 mmol, 90% yield) as a white foam ¹ H NMR (400 MHz, CDCI3) 5ppm: 10.00 (br. s, 1 H), 9.1 1 (br. s, 1 H), 8.64 (dd, 1 H), 8.50 (d, 1 H), 7.64 (dt, 1 H), 7.51 (dt, 1 H), 7.40 (dd, 1 H), 7.23 (ddd, 1 H), 7.13 (d, 1 H), 6.79 (ddd, 1 H), 5.17 (d, 1 H), 3.06 (m, 1 H), 2.14 (s, 3H), 1.16 (d, 3H), 0.92 (d, 3H). [ES+ MS] m/z 397 (M+H). 87.2% ee (Chiralpak IC (25 x 0.46 cm), 5 μπι, Isocratic gradient of n-Hexane / Ethanol 85/15 v/v, flow 0.8 mL/min; 6.4% a/a by UV (15.45 min) and 93.6% a/a by UV (20.99 min)).

Example 2A (S)-N-(3,5-difluorophenyl)-3-methyl-2-(4-methyl-5-morpholino -2,6-dioxo-2,3- dihydropyrimidin-1 (6H)-yl)butanamide. i) (S)-tert-butyl 3-methyl-2-(4-methyl-5-morpholino-2,6-dioxo-2,3-dihydropyrim idin-1 (6H)- yl)butanoate (for example as prepared for Intermediate 4, 252 mg, 0.686 mmol) was dissolved in Trifluoroacetic acid (TFA) (5 mL) and Water (0.128 mL) and stirred at room temperature overnight. LCMS showed the desired compound. The solvent was removed to dryness under vacuum and co-evaporated several times with toluene to aford 422 mg of (S)- 3-methyl-2-(4-methyl-5-morpholino-2,6-dioxo-2,3-dihydropyrim idin-1 (6H)-yl)butanoic acid. The residue was taken for the next step as such without any purification. Quantitave yield was considered. ii) The crude mixture was dissolved in anhydrous DMF (4 mL) under N ₂ atmosphere, HATU (CARBOSYNTH, 287 mg, 0.755 mmol) was added and the resulting solution was stirred 5 minutes. Them 3,5-difluoroaniline (ALDRICH, 97 mg, 0.755 mmol) and DIPEA (FLUKA, 0.360 mL, 2.059 mmol) were added and the reaction mixture was stirred at room temperature overnigth. The reaction was treated with H ₂ 0 (10 mL) and EtOAc (10 mL). The layers were separated and the aqueous phase was extracted with 3x5 mL of EtOAc. The combination of the organic layers was washed with 1 N aq. NH ₄ CI (10 mL) and brine (10 mL), then it was dried over anhydrous MgS0 ₄ , filtered and concentrated under reduced pressure and the crude was purified on silica gel column and was eluted with cyclohexane/ gradient 0-70% to give 107 mg of 3-methyl-2-(4-methyl-5-morpholino-2,6-dioxo-2,3-dihydropyrim idin-1 (6H)- yl)butanoic acid (107 mg, 0.253 mmol, 37% yield two steps) as a white solid.

Chiral separation was carried out by semi-preparative chiral HPLC (Column Chiralpack IC 20X250mm, isocratic gradient Hexane:Ethanol 90:10; wavelenght 254 nm, flow 18 mL/min) to obtain the compound, (S)-N-(3,5-difluorophenyl)-3-methyl-2-(4-methyl-5-morpholino -2,6- dioxo-2,3-dihydropyrimidin-1 (6H)-yl)butanamide (29 mg, 0.069 mmol), as the second eluted compound. 1 H NMR (400 MHz, DMSO-d6) δρρπι: 1 1 .17 (s, 1 H), 9.85 (s, 1 H), 7.39 (m, 2 H), 6.94 (m, 1 H), 4.88 (d, 1 H), 3.65 (br.s., 4 H), 3.01 (m, 2 H), 2.72 (m, 1 H), 2.57 (m, 2 H), 2.24 (s, 3 H), 1 .16 (d, 3 H), 0.74 (d, 3 H). [ES+ MS] m/z 423 (M+H).

Example 2B (S)-N-(3,5-difluorophenyl)-3-methyl-2-(4-methyl-5-morpholino -2,6-dioxo- 2,3-dihydropyrimidin-1 (6H)-yl)butanamide.

To a solution of (S)-3-methyl-2-(4-methyl-5-morpholino-2,6-dioxo-2,3-dihydrop yrimidin-1 (6H)- yl)butanoic acid (for example as prepared for Intermediate 12, 800 mg, 2.57 mmol) and pyridine (ALDRICH, 0.278 mL, 3.22 mmol in anhydrous 2-Methyltetrahydrofuran (2-MeTHF) (10 mL) under N2 atmosphere was cooled to 0 °C. T3P (ALDRICH, 50% in EtOAc, 4.1 1 mL, 6.90 mmol) was added to the cold solution and the resulting mixture was stirred for 30 min. Then, 3,5-difluoroaniline (ALDRICH, 0.3342.99 mmol) were added and the resulting solution was stirred at 4°C over night. Then, the reaction was treated with NaHC0 ₃ 0.1 M (35 mL) and extracted with EtOAc (2x60 mL). The combination of the organic layers was washed with brine (35 mL), dried over anhydrous MgS0 ₄ , filtered and concentrated under reduced pressure. The resultant crude was purified by chromatographic column (20g, CyHex:EtOAc gradients from 100:0 to 80:20 to 70:30) to give 735 mg of (S)-N-(3,5-difluorophenyl)-3- methyl-2-(4-methyl-5-morpholino-2,6-dioxo-2,3-dihydropyrimid in-1 (6H)-yl)butanamide (735 mg, 1.74 mmol, 68% yield) as a white solid and another fraction of 198 mg of target compound a little impure by TLC.

¹ H NMR (400 MHz, DMSO-d6) 5ppm: 1 1.08 (s, 1 H), 9.76 (s, 1 H), 7.30 (m, 2H), 6.85 (m, 1 H), 4.79 (d, 1 H), 3.55 (br. s, 4H), 2.70 (br. s, 4H), 2.61 (m, 1 H), 2.14 (s, 3H), 1 .08 (d, 3H), 0.65 (d, 3H). [ES+ MS] m/z 423 (M+H). 87.2% ee (Chiralpak IC (25 x 0.46 cm), 5 μπι, isocratic gradient of Heptane / Ethanol 90/10 % v/v, flow 1 mL/min, 1 .8% a/a by UV (10.6 min) and 98.2% a/a by UV (12.8 min)).

Example 3 (S)-N-(3,5-difluorophenyl)-3-methyl-2-(4-methyl-5-morpholino -2,6-dioxo- 2,3-dihydropyrimidin-1 (6H)-yl)butanamide.

Fluoroaniline (61 uL, 0.639 mmol), N-methylmorpholine (81 uL, 0.737 mmol) and then COMU (252 mg, 0.588 mmol) were added to a stirred solution of (2S)-3-methyl-2-[4-methyl-5- (morpholin-4-yl)-2,6-dioxo-1 ,2,3,6-tetrahydropyrimidin-1 -yl]butanoic acid hydrochloride (for example as prepared for Intermediate rac-12, 192 mg, 0.491 mmol) in EtOAc (2 mL) and DMF (2 mL) at 0°C. The reaction mixture was allowed to warm slowly to room temperature and stirred overnight. Further 3-Fluoroaniline (20 uL, 0.210 mmol), N-methylmorpholine (30 uL, 0.273 mmol) and then COMU (50 mg, 0.177 mmol) were added and the reaction stirred for a further 4 hours. The reaction mixture was diluted with EtOAc (75 ml_), washed with 0.5M HCI solution (50 ml_), sodium bicarbonate solution (50 + 25 mL) and brine (50 ml_), dried over sodium sulphate and evaporated under reduced pressure. The residue was chromatographed on silica gel (SNAP 25) eluting with a gradient of 20-100% EtOAc in cyclohexane to give 192 mg of a pale yellow foam (192 mg, 0.475 mmol, yield 97%).

Chiral separation was carried out by semi-preparative chiral HPLC (Column Chiralpack IC 20X250mm, isocratic gradient Hexane:Ethanol 85:15; wavelenght 254 nm, flow 14 mL/min) to obtain, eluting the second compound.

1H NMR (400 MHz, CDCI3) 5ppm: 9.39 (br. s, 2H), 7.54 (dt, 1 H), 7.27 (ddd, 1 H), 7.16 (m, 1 H), 6.81 (m, 1 H), 5.09 (d, 1 H), 3.79 (br. s, 4H), 3.08 (br. s, 4H), 3.05 (m, 1 H), 2.29 (s, 3H), 1 .15 (d, 3H), 0.85 (d, 3H). [ES+ MS] m/z 405 (M+H).

Biological Assays

A compound of this invention may be tested in one of several biological assays to determine the concentration of compound which is required to have a given pharmacological effect. The assays are described below.

In vitro potency

P. falciparum growth inhibition assay.

The sensitivity of P. falciparum infected erythrocytes to the compound is determined in triplicate using the [3H]hypoxanthine incorporation method with an inoculum of 0.5% parasitemia (ring stage) and 2% hematocrit. The parasites were grown in RPMI 1640, 25 mM HEPES and supplemented with 5% Albumax. Plates are incubated at 37°C, 5% C02, 5% 02, 90% N2. After 24h of incubation, [3H]hypoxanthine is added and plates are incubated for another 24 h. After that period, plates are harvested on a glass fiber filter using a TOMTEC Cell harvester 96. Filters are dried and melt on scintillator sheets and the bound radioactivity is quantified by use of a Wallac Microbeta Trilux (Model 1450 LS- Perkin Elmer). IC50s are determined using Grafit 5 program (Grafit program; Erithacus Software, Horley, Surrey, United Kingdom).

In vivo efficacy

P. falciparum In Vivo efficacy assay.

Antimalarial in vivo efficacy was determined using the P.falciparum mouse model following the procedure described in: Jimenez-Diaz, M.B., Mulet, T., Viera, S., Gomez, V., Garuti, H., Ibafiez, J., Alvarez-Doval, A., Shlutz, D.L., Martinez, A., Improved Murine Model Of Malaria Using Plasmodium falciparum (Competent Strains and Non-Myelodepleted NOD-sc/c/ \L2R_nu\\

Mice Engrafted with Human Erythrocytes) Antimicrob. Agents Chemother 2009, 53 (10), 4533-4536

Results

Solubility assessment in biorelevant media

Compound's equilibrium solubility was measured at 4h room temperature in Fasted State- Simulated Intestinal Fluid (FaSSIF).

Solvents and buffers

Organic solvents of HPLC grade were used. Ultra pure water (Milli-Q grade) was used. Buffers were prepared with ultra pure water and filtered using 0.45 μ nylon filters.

I. Procedure.

Determination of equilibrium solubility (assuming chemical stability in the desired solvent is not a problem).

a) 1 mg of solid compound was weighted in one 4 mL glass vial and 1 mL of the freshly prepared medium was added. All these samples were prepared by duplicate,

b) The samples were stirred (roller mixer) for 4 hr at room temperature. If required, additional solid compound (0.1 mg) was added to maintain excess of it (saturated solutions).

c) After 4 hours, the samples were centrifuged (10000 rpm, 10 min.) and the supernatants were transfer to an HPLC vial and analysed by LC-MS (previous dilution with mobile phase when needed).

d) The pH of the final solution in each sample was measured with a pH-meter (WTW pH330i and a pH-electrode Sentix 41 ).

LC-MS assay for analytical quantification

All supernatants were analysed by LC-MS. Quantification of those samples was carried out against calibration curves obtained from 1 mg/mL DMSO (Aldrich cat. ref.: 27685-5) stock solutions, by dilution with the mobile phase used in the chromatography. Depending on the solubility range, U.V. (1 μg/mL to 100 μg mL) or MS (1 μg/mL to 1 ng/mL ) detector were used in the quantification.

Analysis of data The analysis of all LC-MS data was performed with MassLynx 3.4 software and Analyst 1 .4.2. Statistical and graphic analysis of data was performed using Microsoft Excel. The concentration (μΜ) and solubility ^g/ml) for each compound was calculated using the peak areas from the sample and those from the calibration curve.

Results

All publications, including but not limited to patents and patent applications cited in this specification are herein incorporated by reference as if each individual publication were specifically and individually indicated to be incorporated by reference as though fully set forth.

The application of which this description and claims forms part may be used as a basis for priority in respect of any subsequent application. The claims of such subsequent application may be directed to any feature or combination of features described herein. They may take the form of product, composition, process, or use claims and may include, by way of example and without limitation, the following claims.