WO2013174931A1

WO2013174931A1 - 7-oxo-4,7 -dihydro- pyrazolo [1, 5 -a] pyrimidine derivatives which are useful in the treatment, amelioration or prevention of a viral disease

Info

Publication number: WO2013174931A1
Application number: PCT/EP2013/060634
Authority: WO
Inventors: Andrea Wolkerstorfer; Oliver Szolar; Norbert Handler; Stephen Cusack; Thibault SAUVAÎTRE; Céline SIMON; Christophe Morice; Bruno Giethlen; Thierry Langer; Mark Smith; Sung-Sau So; Dirk Classen-Houben; Helmut Buschmann
Original assignee: Savira Pharmaceuticals Gmbh; F. Hoffmann-La Roche Ag; European Molecular Biology Laboratory
Priority date: 2012-05-23
Filing date: 2013-05-23
Publication date: 2013-11-28
Also published as: BR112014029006A2; JP2015521189A; CN104507481B; HK1204987A1; CA2874253A1; US20160367557A1; MX2014014109A; KR20150014506A; US20130317021A1; EP2861232A1; CN104507481A; RU2014146778A

Abstract

The present invention relates to a compound having the general formula (C), optionally in the form of a pharmaceutically acceptable salt, solvate, polymorph, codrug, cocrystal, prodrug, tautomer, racemate, enantiomer, or diastereomer or mixture thereof, which are useful in treating, ameloriating or preventing a viral disease. Furthermore, specific combination therapies are disclosed.

Description

-OXO-4,7-DIHYDRO-PYRAZOLO [1 ,5-A] PYRIMIDINE DERIVATIVES WHICH ARE USEFUL IN THE TREATMENT, AMELIORATION OR PREVENTION OF A VIRAL DISEASE

Field of the invention

The present invention relates to a compound having the general formula (C), optionally in the form of a pharmaceuticaiiy acceptable salt, solvate, polymorph, codrug, cocrystal, prodrug, tautomer, racemate, enantiomer, or diastereomer or mixture thereof,

(C) which is useful in treating, ameioriating or preventing a viral disease. Furthermore, specific combination therapies are disclosed.

Background of the invention

In recent years the serious threat posed by Influenza virus to worldwide public health has been highlighted by, firstly, the ongoing low level transmission to humans of the highly pathogenic avian H5N1 strain (63% mortality in infected humans, http://www.who.int/ csr/disease/avtan_influenza/en/) and secondly, the unexpected emergence in 2009 of a novel pandemic strain A/H1 N1 that has rapidly spread around the entire world (http://www.who.int/csr/disease/swineflu/en/). Whilst the new strain is highly contagious but currently only generally gives mild illness, the future evolution of this virus is unpredictable. In a much more serious, but highly plausible scenario, H5N1 could have been more easily transmissible between humans or the new A/H1 N1 could have been more virulent and could have carried the single point mutation that confers Tamiflu resistance (Neumann et al., Nature, 2009 (18; 459(7249) 931-939)), as many seasonal H1 N1 strains have recently done (Dharan et al., The Journal of the American Medical Association, 2009 Mar 11 ; 301 (10), 1034-1041; Moscona et al., The New England Journal of Medicine, 2009 (Mar 5;360( 0) pp 953-956)). In this case, the delay in generating and deploying a vaccine (-6 months in the relatively favourable case of A/H1 N1 and still not a solved problem for H5N1 ) could have been catastrophically costly in human lives and societal disruption.

It is widely acknowledged that to bridge the period before a new vaccine becomes available and to treat severe cases, as well as to counter the problem of viral resistance, a wider choice of anti-influenza drugs is required. Development of new anti-influenza drugs has therefore again become a high priority, having been largely abandoned by the major pharmaceutical companies once the anti-neuramtnidase drugs became available.

An excellent starting point for the development of antiviral medication is structural data of essential viral proteins. Thus, the crystal structure determination of e.g. the influenza virus surface antigen neuraminidase (Von Itzstein, M. et al., (1993), Nature, 363, pp. 418-423) led directly to the development of neuraminidase inhibitors with anti-viral activity preventing the release of virus from the cells, however, not the virus production. These and their derivatives have subsequently developed into the anti-influenza drugs, zanamivir (Glaxo) and oseltamivir (Roche), which are currently being stockpiled by many countries as a first line of defence against an eventual pandemic. However, these medicaments provide only a reduction in the duration of the clinical disease. Alternatively, other anti-influenza compounds such as amantadine and rimantadine target an ion channel protein, i.e., the M2 protein, in the viral membrane interfering with the uncoating of the virus inside the cell. However, they have not been extensively used due to their side effects and the rapid development of resistant virus mutants (Magden, J. et al., (2005), App!. Microbiol. BiotechnoL, 66, pp. 612-621). In addition, more unspecific viral drugs, such as ribavirin, have been shown to work for treatment of influenza and other virus infections (Eriksson, B. et al., (1977), Antimicrob. Agents Chemother., 11 , pp. 946-951). However, ribavirin is only approved in a few countries, probably due to severe side effects (Furuta et al., ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 2005, p. 981-986). Clearly, new antiviral compounds are needed, preferably directed against different targets. Influenza virus as well as Thogotovirus belong to the family of Orthomyxoviridae which, as well as the family of the Bunyaviridae, including the Hantavirus, Nairovirus, Orthobunyavirus, and Phlebovirus, are negative stranded RNA viruses. Their genome is segmented and comes in ribonucleoprotein particles that include the RNA dependent RNA polymerase which carries out (i) the initial copying of the single-stranded virion RNA (vRNA) into viral mRNAs and (ii) the vRNA replication. This enzyme, a trimeric complex composed of subunits PA, PB1 and PB2, is central to the life cycle of the virus since it is responsible for the replication and transcription of viral RNA. In previous work the atomic structure of two key domains of the polymerase, the mRNA cap-binding domain in the PB2 subunit (Guilligay et al., Nature Structural & Molecular Biology 2008; May; 15(5): 500-506) and the endonuclease-active site in the PA subunit (Dias et al., Nature 2009, 458, 914-918) have been identified and determined. These two sites are criticai for the unique cap-snatching mode of transcription that is used by influenza virus to generate viral mRNAs. For the generation of viral mRNA the polymerase makes use of the so called "cap-snatching" mechanism (Piotch, S. J. et al., (1981 ), Cell, 23, pp. 847-858; Kukkonen, S. K. et al (2005), Arch. Virol., 150, pp. 533-556; Leahy, M. B. et al, (2005), J. Virol., 71 , pp. 8347-8351 ; Noah, D. L et al„ (2005), Adv. Virus Res., 65, pp. 121-145). A 5' cap (also termed an RNA cap, RNA 7-methylguanosine cap or an RNA m7G cap) is a modified guanine nucleotide that has been added to the 5' end of a messenger RNA. The 5' cap consists of a terminal 7-methylguanosine residue which is linked through a 5'-5'- triphosphate bond to the first transcribed nucleotide. The viral polymerase binds to the 5' RNA cap of cellular mRNA molecules and cleaves the RNA cap together with a stretch of 10 to 15 nucleotides. The capped RNA fragments then serve as primers for the synthesis of viral mRNA.

The polymerase complex seems to be an appropriate antiviral drug target since it is essential for synthesis of viral mRNA and viral replication and contains several functional active sites likely to be significantly different from those found in host cell proteins (Magden, J. et al., (2005), Appl. Microbiol. BiotechnoL, 66, pp. 612-621 ), Thus, for example, there have been attempts to interfere with the assembly of polymerase subunits by a 25-amino-acid peptide resembling the PA-binding domain within PB1 (Ghanem, A. et al., (2007), J. Virol., 81 , pp. 7801 -7804). Furthermore, the endonuc!ease activity of the polymerase has been targeted and a series of 4-substituted 2,4-dioxobutanoic acid compounds has been identified as selective inhibitors of this activity in influenza viruses (Tomassini, J. et al., (1994), Antimicrob. Agents Chemother., 38, pp. 2827-2837). In addition, flutimide, a substituted 2,6-diketopiperazine, identified in extracts of Delitschia confertaspora, a fungal species, has been shown to inhibit the endonuciease of influenza virus (Tomassini, J. et al., (1996), Antimicrob. Agents Chemother., 40, pp. 1189-1193). Moreover, there have been attempts to interfere with virai transcription by nucleoside analogs, such as 2'-deoxy-2^!-fiuoroguanosine (Tisdaie, M. et al., (1995), Antimicrob. Agents Chemother., 39, pp. 2454-2458).

V. L. Rusinov et al. described the synthesis and antiviral activity of nucleoside analogs based on 1 ,2,4-triazoio[3,2-c][1 ,2,4]triazsn-7-ones in the Russian Chemical Bulletin, international Edition, 59(1), 2010, 136-143.

H. A. Ai-Khamees et al. discussed the synthesis of 2-substttuted-1 ,2,4-triazolo[1 ,5-a}- pyrimidine and 1 ,2,4-triazoio[4,3-a3pyrimidine derivatives as potential antimicrobial agents (Indian Journal of Heterocyclic Chemistry, 2, 1993, 237-244). It is an object of the present invention to identify further compounds which are effective against viral diseases and which have improved pharmacological properties.

Summary of the invention

The present invention relates a compound having the general formula (C) wherein the compound is for use in the treatment, amelioration or prevention of a virai disease.

It is understood that throughout the present specification the term "a compound having the genera! formula (C)" encompasses pharmaceutically acceptable salts, solvates, polymorphs, prodrugs, tautomers, racemates, enantiomers, or diastereomers or mixtures thereof unless mentioned otherwise.

Detailed description of the invention

Before the present invention is described in detail below, it is to be understood that this invention is not limited to the particular methodology, protocols and reagents described herein as these may vary. It is a!so 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 wi!l be limited only by the appended claims. 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. Preferably, the terms used herein are defined as described in "A multilingual glossary of biotechnological terms: (lUPAC Recommendations}", Leuenberger, H.G.W, Nagel, B. and Kolbl, H. eds. (1995), Helvetica Chimica Acta, CH-4010 Basel, Switzerland.

Throughout this specification and the claims which follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps. In the following passages different aspects of the invention are defined in more detail. Each aspect so defined may be combined with any other aspect or aspects unless clearly indicated to the contrary. In particular, any feature indicated as being preferred or advantageous may be combined with any other feature or features indicated as being preferred or advantageous.

Several documents are cited throughout the text of this specification. Each of the documents cited herein (including all patents, patent applications, scientific publications, manufacturer's specifications, instructions, etc.), whether supra or infra, are hereby incorporated by reference in their entirety. Nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.

Definitions

The term "alky!" refers to a saturated straight or branched carbon chain.

The term "cycioalkyl" represents a cyclic version of "alkyi". The term "cycloalkyl" is aiso meant to include bicyc!ic, tricyclic and poiycyclic versions thereof. Uniess specified otherwise, the cycioalkyl group can have 3 to 12 carbon atoms.

"Hal" or "halogen" represents F, CI, Br and I. The term "aryl" prefer^teJy refers to an aromatic monocyclic ring containing 6 carbon atoms, an aromatic bicyclic ring system containing 10 carbon atoms or an aromatic tricyclic ring system containing 14 carbon atoms. Examples are phenyl, naphthyl or anthraceny!, preferably phenyl. The term "heteroaryi" preferably refers to a five or six-membered aromatic ring wherein one or more of the carbon atoms in the ring have been replaced by 1 , 2, 3, or 4 (for the five membered ring) or 1 , 2, 3, 4, or 5 (for the six membered ring) of the same or different heteroatoms, whereby the heteroatoms are selected from O, N and S. Examples of the heteroaryi group include pyrrole, pyrrolidine, oxolane, furan, imidazolidine, imidazole, pyrazole, oxazotidine, oxazole, thiazole, piperidine, pyridine, morphoiine, piperazine, and dioxolane.

The term "hydrocarbon group which contains from 5 to 20 carbon atoms and optionally 1 to 4 heteroatoms selected from O, N and S and which contains at least one ring" refers to any group having 5 to 20 carbon atoms and optionally 1 to 4 heteroatoms selected from O, N and 2 as long as the group contains at least one ring. The term is also meant to include bicyclic, tricyclic and poiycyciic versions thereof. If more than one ring is present, they can be separate from each other or be annelated. The ring(s) can be either carbocycfic or heterocyclic and can be saturated, unsaturated or aromatic. The carbon atoms and heteroatoms can either all be present in the one or more rings or some of the carbon atoms and/or heteroatoms can be present outside of the ring, e.g., in a linker group (such as -(CH₂)_P- with p = 1 to 6). Examples of these groups include -(optionally substituted C₃_₇ cycloalkyl), -(optionally substituted aryl) wherein the aryl group can be, for example, phenyl, -(optionally substituted biphenyl), adamantyl, -{C_3-7 cycloafkyl)-aryl as well as the corresponding compounds with a linker.

The term "(optionally substituted mono- or poiycyciic group containing 3 to 20 carbon atoms and optionally 1 to 4 heteroatoms selected from O, N and S)" refers to any mono- or poiycyciic group containing 3 to 20 carbon atoms and optionally 1 to 4 heteroatoms selected from O, N and S. This term includes monocyclic, bicyclic, tricyclic and poiycyciic versions thereof. If more than one ring is present, they can be separate from each other or be annelated. The ring(s) can be either carbocyclic or heterocyclic and can be saturated, unsaturated or aromatic. The carbon atoms and heteroatoms can either all be present in the one or more rings or some of the carbon atoms and/or heteroatoms can be present outside of the ring, e.g., in a linker group (such as ~(CH₂)_P- with p = 1 to 6). Examples of these groups include -(optionally substituted cycloalkyl), and -(optionally substituted aryi) wherein the aryl group can be, for example, phenyl or anthracenyi as well as the corresponding compounds with a linker.

If a compound or moiety is referred to as being "optionally substituted", it can in each instance include 1 or more of the indicated substituents, whereby the substituents can be the same or different.

The term "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention. Suitable pharmaceutically acceptable salts include acid addition salts which may, for example, be formed by mixing a solution of compounds of the present invention with a solution of a pharmaceutically acceptable acid such as hydrochloric acid, sulfuric acid, fumaric acid, maleic acid, succinic acid, acetic acid, benzoic acid, citric acid, tartaric acid, carbonic acid or phosphoric acid. Furthermore, where the compound carries an acidic moiety, suitable pharmaceutically acceptable salts thereof may include alkali metal salts (e.g., sodium or potassium salts); alkaline earth metal salts (e.g., calcium or magnesium salts); and salts formed with suitable organic ligands (e.g., ammonium, quaternary ammonium and amine cations formed using counteranions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, alky! sulfonate and aryl sulfonate), illustrative examples of pharmaceutically acceptable salts include, but are not limited to, acetate, adipate, alginate, ascorbaie, aspartate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate, calcium edetate, camphorate, camphorsulfonate, camsylate, carbonate, chloride, citrate, clavulanate, cyclopentanepropionate, digluconate, dihydrochloride, dodecylsuifate, edetate, edisylate, estoiate, esylate, ethanesulfonate, formate, fumarate, gluceptate, glucoheptonate, gluconate, giutamate, glycerophosphate, glycolylarsanilate, hemisulfate, hepianoate, hexanoate, hexyiresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, hydroxynaphthoate, iodide, isothionate, lactate, lactobionate, iaurate, iauryl sulfate, malate, maleate, malonate, mande!ate, mesylate, methanesulfonate, methyisulfate, mucate, 2-naphthalenesulfonate, napsylate, nicotinate, nitrate, N-methylglucamine ammonium salt, oleate, oxalate, pamoate (embonate), patmitate, pantothenate, pectinate, persulfate, 3-phenylpropionate, phosphate/diphosphate, picrate, pivaiate, polygalacfuronate, propionate, salicylate, stearate, sulfate, subacetate, succinate, tannate, tartrate, teoclate, tosylate, trieth^'todide, undecanoate, valerate, and the like {see, for example, S. M. Berge et al., "Pharmaceutical Salts", J. Pharm. Sci., 66, pp. 1-19 (1977)). When the compounds of the present invention are provided in crystalline form, the structure can contain solvent moiecuies. The solvents are typically pharmaceutically acceptable solvents and include, among others, water (hydrates) or organic solvents. Examples of possible solvates include ethanolates and iso-propanolates.

The term "codrug" refers to two or more therapeutic compounds bonded via a covaient chemical bond, A detailed definition can be found, e.g., in N. Das et a!., European Journal of Pharmaceutical Sciences, 41 , 2010, 571-588. The term "cocrystal" refers to a multiple component crystal in which ail components are soiid under ambient conditions when in their pure form. These components co-exist as a stoichiometric or non-stoichiometric ratio of a target molecule or ion (i.e., compound of the present invention) and one or more neutral molecular cocrystal formers. A detailed discussion can be found, for example, in Ning Shan et al., Drug Discovery Today, 13(9/10), 2008, 440-446 and in D. J. Good et al., Cryst. Growth Des., 9(5), 2009, 2252-2264.

The compounds of the present invention can also be provided in the form of a prodrug, namely a compound which is metabolized in vivo to the active metabolite. Suitable prodrugs are, for instance, esters. Specific examples of suitable groups are given, among others, in US 2007/0072831 In paragraphs [0082] to [01 18] under the headings prodrugs and protecting groups. If X¹ is O or S, preferred examples of the prodrug include compounds in which R² is replaced by one of the following groups;

In these formuiae, R⁶ can be the same or different. R⁹ is a cyclic group such as an aryl group or a C_3-_7 cycloalkyl group, p is 2 to 8.

If X¹ is NR⁸, preferred examples of the prodrug include compounds in which R² and R⁸ are not both H.

Compounds having the general formula (C)

The compounds having the general formula (C) are identified in the following.

ίΐ is understood that throughout the present specification the term "a compound having the general formula "(C)" encompasses pharmaceutically acceptable salts, solvates, polymorphs, prodrugs, tautomers, racemates, enantiomers, or diastereomers or mixtures thereof uniess mentioned otherwise.

In the present invention the following definitions apply with respect to the compounds having the genera! formula (C).

V is N, or CR⁶

10 X¹ is O, S, or NR⁸, preferably X¹ is O.

X² is NR⁵, N(R⁵)C(0), C(0)NR⁵, O, C(O), C(0)0, OC(O); S, SO, S0₂, S0₂N(R⁵) or

N(R⁵)S0₂. Preferably X² is NR⁵ or N(R⁵)S0₂.

R^* is -H, -Hal, -(optionally substituted d-e alkyl), -(optionally substituted mono- or polycyclic group containing 3 to 20 carbon atoms and optionally 1 to 4 heteroatoms 5 selected from O, N and S), -d-4 aikyl-(optionaliy substituted mono- or po!ycyclic group containing 3 to 20 carbon atoms and optionally 1 to 4 heteroatoms selected from O, N and S), or -X²-R¹. Preferably R^* is H, -(optionally substituted d-e alkyl),

-(optionally substituted C₃„₇ cycioalkyl) or -X²-R¹.

is -H, -(optionally substituted Ci_e alkyl), -(optionai!y substituted mono- or polycyclic

Of) nroi in nntaininri t i 9Π rorhnn atnmc anrl nnt^'tnnalh/ 1 tn i hpttarnatrslTxi c-iiaptpH

from O, N and S), — Ci_₄ aikyl-(opt natiy substituted mono- or polycyclic group containing 3 to 20 carbon atoms and optionally 1 to 4 heteroatoms selected from O, N and S). Preferably R¹ is -C_1-4 alkyl-(optionaliy substituted mono- or polycyclic group containing 3 to 20 carbon atoms and optionally 1 to 4 heteroatoms selected

25 from O, N and S),

R² is -H, -(optionally substituted Ci_e alkyl), -(optionally substituted C3..7 cycioalkyl), -(optionally substituted ary!), -d-4 alkyl— (optionally substituted C₃_₇ cycioalkyl), or -C_1-J} alkyi-(optionaily substituted aryl) or if X¹ is NR' then R² can also be -OH. Preferably, R² is -H or -d-e alkyl.

30 R³ is -H, -(optionally substituted d-e alkyl), -R⁷, or -X²-R⁷. Preferably R³ is -H, -C^ alkyl— (optionally substituted aryl) or -S0₂-R⁵. Preferably, R³ is -H.

R⁴ is -H, -(optionally substituted d-e alkyl), -(optionally substituted C^₇ cycioalkyl), - (optionally substituted aryl), -d-* alkyl~-(optionally substituted C₃_₇ cycioalkyl), or -d-4 alkyl— (optionally substituted aryl). Preferably, R⁴ is -H, or -(optionally

35 substituted d-e alkyl). R⁵ is -H, -(optionaiiy substituted Ci_6 alkyl), -{optionally substituted C₃_₇ cycloaikyl), -(optionally substituted aryl), -Ci_4 alkyl-(optionatiy substituted cycloaikyl), or -Ci_4 alkyl— (optionally substituted aryl). Preferably R⁵ is -C _-4 alkyl— (optionally substituted aryl) or -(optionally substituted cycloaikyl).

R⁶ H, -Ci_6 alkyl, -aryl, halogen or CN. Preferably R⁶ is H or -aryl.

R⁷ is -(optionally substituted hydrocarbon group which contains from 5 to 20 carbon atoms and optionally 1 to 4 heteroatoms selected from O, N and S and which contains at least one ring). Preferably R⁷ is -C_1-4 alkyl— (optionally substituted aryl).

R⁸ is -H, -Ci_₆ alkyl or -C^ alkyl-(optionally substituted aryl). Preferably R⁸ is -Ci_e alkyl or -C-i_4 alkyi-(optionaiiy substituted aryl).

n is 0 to 4, preferably 0 or 1.

The optional substituent of the alkyl group can be selected from the group consisting of halogen, -CN, -NR⁵R⁵, -OH, and -0-C^₆ alkyl.

The optional substituent of the cycloaikyl group, the aryl group, the mono- or poiycyciic group or the hydrocarbon group can be selected from the group consisting of -Ci_e alkyl, halogen, -CF₃, -CN, -X²-C_6 alkyl and -d_₆ alkyl— aryl. The present inventors have surprisingly found that the compounds of the present invention which have a carbon atom im position 5 have improved pharmacological properties compared to corresponding compounds which have a nitrogen atom in this position. Without wishing to be bound by theory it is assumed that the viral polymerase protein has a pocket for binding and that carbon atom of the compounds of the present invention has improved binding compared to a nitrogen atom. This could not have been predicted or expected based on the art.

The compounds of the present invention can be administered to a patient in the form of a pharmaceutical composition which can optionally comprise one or more pharmaceutically acceptable excipient(s) and/or carrier(s).

The compounds of the present invention can be administered by various well known routes, including oral, rectal, intragastrical, intracranial and parenteral administration, e.g. intravenous, intramuscular, intranasal, intradermal, subcutaneous, and similar administration routes. Oral, intranasal and parenteral administration are particularly preferred. Depending on the route of administration different pharmaceutical formulations are required and some of those may require that protective coatings are applied to the drug formulation to prevent degradation of a compound of the invention in, for example, the digestive tract. Thus, preferably, a compound of the invention is formulated as a syrup, an infusion or injection solution, a spray, a tablet, a capsule, a capslet, lozenge, a liposome, a suppository, a plaster, a band-aid, a retard capsule, a powder, or a stow release formulation. Preferably, the diluent is water, a buffer, a buffered salt solution or a salt solution and the carrier preferably is selected from the group consisting of cocoa butter and vitebesole.

Particular preferred pharmaceutical forms for the administration of a compound of the invention are forms suitable for injectionable use and include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases the final solution or dispersion form must be sterile and fluid. Typically, such a solution or dispersion will include a solvent or dispersion medium, containing, for example, water-buffered aqueous solutions, e.g. biocompatible buffers, ethanoi, polyol, such as glycerol, propylene glycol, polyethylene glycol, suitable mixtures thereof, surfactants or vegetable oils. A compound of the invention can also be formulated into liposomes, in particular for parenteral administration. Liposomes provide the advantage of increased half life in the circulation, if compared to the free drug and a prolonged more even release of the enclosed drug.

Sterilization of infusion or injection solutions can be accomplished by any number of art recognized techniques including but not limited to addition of preservatives like anti-bacterial or anti-fungal agents, e.g. parabene, chlorobutanol, phenol, sorbic acid or thimersaf. Further, isotonic agents, such as sugars or salts, in particular sodium chloride, may be incorporated in infusion or injection solutions.

Production of steriie injectable solutions containing one or several of the compounds of the invention is accomplished by incorporating the respective compound in the required amount in the appropriate solvent with various ingredients enumerated above as required followed by sterilization. To obtain a sterile powder the above solutions are vacuum-dried or freeze-dried as necessary. Preferred diluents of the present invention are water, physiological acceptable buffers, physiological acceptable buffer salt solutions or salt solutions. Preferred carriers are cocoa butter and vitebesole. Excipients which can be used with the various pharmaceutical forms of a compound of the invention can be chosen from the following non-limiting list: a) binders such as lactose, mannitol, crystalline sorbitol, dibasic phosphates, calcium phosphates, sugars, microcrystaSline cellulose, carboxymethyl cellulose, hydroxyethyl celiulose, polyvinyl pyrrolidone and the like;

b) lubricants such as magnesium stearate, talc, calcium stearate, zinc stearate, stearic acid, hydrogenated vegetable oil, leucine, glycerids and sodium siearyl fumarates, c) disintegrants such as starches, croscarrnellose, sodium methyl cellulose, agar, bentonite, aiginic acid, carboxymethyl cellulose, polyvinyl pyrrolidone and the like.

In one embodiment the formulation is for oral administration and the formulation comprises one or more or all of the following ingredients: pregelatinized starch, talc, povidone K 30, croscarrnellose sodium, sodium stearyl fumarate, gelatin, titanium dioxide, sorbitol, monosodtum citrate, xanthan gum, titanium dioxide, flavoring, sodium benzoate and saccharin sodium.

If a compound of the invention is administered intranasaliy in a preferred embodiment, it may be administered in the form of a dry powder inhaler or an aerosol spray from a pressurized container, pump, spray or nebulizer with the use of a suitable prope!lant, e.g., dichiorodifluoromethane, trichlorofluoromethane, dichloroteirafluoroethane, a hydrofluoro- a!kane such as 1 , 1 ,1 ,2 etrafluoroethane (HFA 134A™) or 1 , 1 ,1 ,2,3,3, 3-heptafiuoropropane (HFA 227EA™), carbon dioxide, or another suitable gas. The pressurized container, pump, spray or nebulizer may contain a solution or suspension of the compound of the invention, e.g., using a mixture of ethanol and the propeliant as the solvent, which may additionally contain a lubricant, e.g., sorbitan trioleate.

Other suitable excipients can be found in the Handbook of Pharmaceutical Excipients, published by the American Pharmaceutical Association, which is herein incorporated by reference.

It is to be understood that depending on the severity of the disorder and the particular type which is treatable with one of the compounds of the invention, as well as on the respective patient to be treated, e.g. the general health status of the patient, etc., different doses of the respective compound are required to elicit a therapeutic or prophylactic effect. The determination of the appropriate dose lies within the discretion of the attending physician, !t is contemplated that the dosage of a compound of the invention in the therapeutic or prophylactic use of the invention should be in the range of about 0.1 mg to about 1 g of the active ingredient (i.e. compound of the invention) per kg body weight. However, in a preferred use of the present invention a compound of the invention is administered to a subject in need thereof in an amount ranging from 1.0 to 500 mg/kg body weight, preferably ranging from 1 to 200 mg/kg body weight. The duration of therapy with a compound of the invention will vary, depending on the severity of the disease being treated and the condition and idiosyncratic response of each individual patient. In one preferred embodiment of a prophylactic or therapeutic use, from 10 mg to 200 mg of the compound are orally administered to an adult per day, depending on the severity of the disease and/or the degree of exposure to disease carriers.

As is known in the art, the pharmaceutically effective amount of a given composition will also depend on the administration route. In general the required amount will be higher, if the administration is through the gastrointestinal tract, e.g., by suppository, rectal, or by an intragastric probe, and lower if the route of administration is parenteral, e.g., intravenous. Typically, a compound of the invention will be administered in ranges of 50 mg to 1 g/kg body weight, preferably 10 mg to 500 mg/kg body weight, if rectal or intragastric administration is used and in ranges of 1 to 100 mg/kg body weight if parenteral administration, is used. For intranasal administration, 1 to 100 mg/kg body weight are envisaged.

If a person is known to be at risk of developing a disease treatable with a compound of the invention, prophylactic administration of the biologically active blood serum or the pharmaceutical composition according to the invention may be possible. In these cases the respective compound of the invention is preferably administered in above outlined preferred and particular preferred doses on a daily basis. Preferably, from 0.1 mg to 1 g/kg body weight once a day, preferably 10 to 200 mg/kg body weight. This administration can be continued until the risk of developing the respective viral disorder has lessened. In most instances, however, a compound of the invention will be administered once a disease/disorder has been diagnosed. In these cases it is preferred that a first dose of a compound of the invention is administered one, two, three or four times daily.

The compounds of the present invention are particularly useful for treating, ameliorating, or preventing viral diseases. The type of viral disease is not particularly limited. Examples of possible virai diseases include, but are noi limited to, viral diseases which are caused by Poxviridae, Herpesviridae, Adenoviridae, Papii!omaviridae, Polyomaviridae, Parvoviridae, Hepadnaviridae, Retroviridae, Reoviridae, Filoviridae, Paramyxoviridae, Rhabdoviridae, Orthomyxoviridae, Bunyavi idae, Arenaviridae, Coronavindae, Picornaviridae, Hepeviridae, Caliciviridae, Astroviridae, Togaviridae, Fiaviviridae, Deltavirus, Bornaviridae, and prions. Preferably viral diseases which are caused by Herpesviridae, Retroviridae, Filoviridae, Paramyxoviridae, Rhabdoviridae, Orthomyxoviridae, Bunyaviridae, Arenaviridae, Coronaviridae, Picornaviridae, Togaviridae, Fiaviviridae, more preferably viral diseases which are caused by orthomyxoviridae.

Examples of the various viruses are given in the following table.

Family Virus (preferred examples)

Poxviridae Smallpox virus

oliuscum contagiosum virus

Herpesviridae Herpes simplex virus

Varicella zoster virus

Cytomegalovirus

Epstein Barr virus

Kaposi's sarcoma-associated herpesvirus

Adenoviridae Human adenovirus A-F

Papil!omaviridae PapiSiomavirus

Polyomaviridae BK-virus

JC-Virsu

Parvoviridae B19 virus

Adeno associated virus 2/3/5

Hepadnaviridae Hepatitis B virus

Retroviridae Human immunodeficiency virus

types 1/2

Human T-cell leukemia virus

Human foamy virus

Reoviridae Reovirus 1/2/3

Rotavirus A B/C

Colorado tick fever virus

Filoviridae Ebola virus

Marburg virus

Paramyxoviridae Parainfluenza virus 1-4

Mumps virus

Measles virus

Respiratory syncytial virus

Hendra virus Rhabdoviridae Vesicular stomatitis virus

Rabies virus

Mokoia virus

European bat virus

Duvenhage virus

Orthomyxoviridae influenza virus types A-C

Bunyaviridae California encephalitis virus

La Crosse virus

Hantaan virus

Puumala virus

Sin Nombre virus

Seoul virus

Crimean- Congo hemorrhagic fever virus

Sakhalin virus

Rift valley virus

Sandfly fever virus

Uukuniemi virus

Arenavirtdae Lassa virus

Lymphocytic choriomeningitis virus

Guanarito virus

Junin virus,

achupo virus

Sabia virus

CoronavSridae Human coronavirus

Picornaviridae Human enterovirus types A-D (Poliovirus, Echovirus,

Coxsackie virus A/B)

Rhinovirus types A/B/C

Hepatitis A virus

Parechovirus

Food and mouth disease virus

Hepeviridae Hepatitis E virus

Calicivirsdae Norwaik virus

Sapporo virus

Astroviridae Human astrovirus 1

Togaviridae Ross River virus

Chikungunya virus

O'nyong-nyong virus

Rubella virus

Flaviviridae Tick-borne encephalitis virus

Dengue virus

Yellow Fever virus

Japanese encephalitis virus

Murray Valley virus

St. Louts encephalitis virus

West Nile virus

Hepatitis C virus

Hepatitis G virus Hepatitis GB virus

Deitavirus Hepatitis deitavirus

Bornaviridae Bornavirus

Prions

Preferably, the compounds of the present invention are employed to treat influenza. Within the present invention, the term "influenza" includes influenza A, B, C, isavirus and thogotovirus and also covers bird flu and swine flu. The subject to be treated is not particularly restricted and can be any vertebrate, such as birds and mammals (including humans).

Without wishing to be bound by theory it is assumed that the compounds of the present invention are capable of inhibiting endonuclease activity, particularly of the influenza virus. More specifically it is assumed that they directiy interfere with the N-terminal part of the influenza PA protein, which harbours endonuclease activity. However, delivery of a compound into a cell may represent a problem depending on, e.g., the solubility of the compound or its capabilities to cross the cell membrane. The present invention not only shows that the claimed compounds have in vitro polymerase inhibitory activity but also in vivo antiviral activity. A possible measure of the in vitro polymerase inhibitory activity of the compounds having the formula (A) and/or (C) is the FRET endonuclease activity assay disclosed herein. Preferably the compounds exhibit a % reduction of at least about 50 % at 25 μΜ in the FRET assay. In this context, the % reduction is the % reduction of the initial reaction velocity (vO) of substrate cleavage of compound-treated samples compared to untreated samples. Preferably the compounds exhibit an IC_S0 of at least about 40 μ , more preferably at least about 20 μΜ, in the FRET assay. The half maximal inhibitory concentration (IC₅₀) is a measure of the effectiveness of a compound in inhibiting biological or biochemical function and was calculated from the initial reaction velocities (vO) in a given concentration series ranging from maximum 100 μΜ to at least 2 nM.

A possible measure of the in vivo antiviral activity of the compounds having the formula (A) and/or (C) is the CPE assay disclosed herein. Preferably the compounds exhibit a % reduction of at least about 30 % at 50 μΜ. In this connection, the reduction in the virus-mediated cytopathic effect (CPE) upon treatment with the compounds was calculated as follows: The ceil viability of infected-treated and uninfected-treated cells was determined using an ATP- based cell viability assay (Promega). The response in relative luminescent units (RLU) of Infected-untreated samples was subtracted from the response (RLU) of the Infected-treated samples and then normalized to the viability of the corresponding uninfected sample resulting in % CPE reduction. Preferably, the compounds exhibit an IC₅₀ of at least about 45 μ , more preferably at least about 10 μ , in the CPE assay. The half maximal inhibitory concentration (IC5₀) is a measure of the effectiveness of a compound in inhibiting biological or biochemical function and was calculated from the RLU response in a given concentration series ranging from maximum 00 μΜ to at least 100 nM.

The compounds having the general formula (C) can be used in combination with one or more other medicaments. The type of the other medicaments is not particularly iimited and wiii depend on the disorder to be treated. Preferably, the other medicament will be a further medicament which is useful in treating, ameloriating or preventing a viral disease, more preferably a further medicament which is useful in treating, ameloriating or preventing influenza. The following combinations of medicaments are envisaged as being particularly suitable:

(i) The combination of endonuclease and cap binding inhibitors (particularly targeting influenza). The endonuclease inhibitors are not particularly limited and can be any endonuclease inhibitor, particularly any viral endonuclease inhibitor. Preferred endonuclease inhibitors are those having the general formula (I) as defined in the US application with the serial number 61/550,045, filed on October 21, 2011 , the complete disclosure of which is incorporated by reference. In particular, all descriptions with respect to the general formula of the compounds according to US 61/550,045, the preferred embodiments of the various substituents as well as the medical utility and advantages of the compounds are incorporated herein by reference.

The compounds having the general formula (I) of this reference can optionally be in the form of a pharmaceutically acceptable salt, solvate, polymorph, codrug, co crystal, prodrug, tautomer, racemate, enantiomer, or diastereomer or mixture thereof. They are defined as follows (wherein the definitions of the various moieties given in this earlier application apply):

wherein

R¹ is selected from -H, -C,_₆ alkyi, -(C₃„₇ cycloalkyl) and -C ₂-(C_:^₇ cycloalkyl);

R² is selected from -H,

, -d_e alkyl, -Hal, -(C₃_₇ cycloalkyl), -CH₂-(C₃_₇ cycloalkyl), -{CH^-ioptionally substituted aryl), -(optionally substituted 5- or 6- membered heterocyclic ring which contains at least one heteroatom selected from N, O and S, wherein the substituent is selected from -C^ alkyl, -halogen, -CN, -CHal₃, -aryl, ~NR⁶R⁷, and -CONR⁶R⁷;

R³ is selected from -H, -C^e alkyl- ~(CH₂}_n-NR⁶R⁸,

-(optionally substituted 5- or 6-membered carbo- or heterocyclic ring wherein the heterocyclic ring contains at least one heteroatom selected from N, O and S), wherein the substituent is selected from -Hal, -C_1-4 alkyl, -NR⁹R¹⁰, -(CH₂)_n-OH, -C(0)-NR⁹R¹⁰, -S0₂-NR^BR¹°, -NH-C(0)-0-R¹¹ , -C(0}-0-R¹¹ , and a 5- or 6-membered heterocyclic ring which contains at least one heteroatom selected from N, O and S; or wherein R¹ and R² together form a phenyl ring or wherein R² and R³ together form a phenyl ring;

R⁴ is -H;

R⁵ is selected from the group consisting of -H or -(CH₂)_n-(optional!y substituted aryl), wherein the substituent is selected from -Hal and -C^ a!kyl; or wherein R⁴ and R⁵ together form a methylene group -CH₂-, ethylene group -CH₂CH₂- or ethyne group -CHCH-, which can be optionally substituted by -C-i_4 aikyl, -halogen, -CHal₃, -R⁶R⁷, -OR⁶, -CONR⁶R⁷, -S0₂R⁶R⁷, aryl or heteroaryS; R⁶ is selected from -H and -C-,^ alkyl;

R⁷ is selected from -H and -C_1- alkyl;

R⁸ is selected from -H, -C^e alkyl, -(CH₂)_n-(optionaily substituted aryt), -S0₂--(CH₂)_n- (optionaily substituted aryl), -S0₂-{CH₂)n-(optionally substituted 5- to 10-membered mono- or bicyciic heteroring which contains at least one heteroatom selected from N, O and S), -(CH₂)n-(opt!onaliy substituted 5- or 6-membered heterocyclic ring which contains at least one heteroatom selected from N, O and S), wherein the substituent is selected from -Hal, -CF₃, -C-M alkyl, and -(CH₂)_n-aryl;

R⁹ is selected from -H, -C_1-4 alkyl, and -C^ alkylene-NR^{1 11}; R^1G is selected from -H,

alkylene-NR¹¹R¹¹;

R¹¹ is selected from -H, -CF₃, and -C_1- alkyl; each m is 0 or 1 ; and each n is independently 0, 1 , 2, or 3.

Further preferred endonuclease inhibitors are those having the general formula (C) as defined in the copending application with attorney's docket number T3450 US which was filed on even date herewith, the complete disclosure of which is incorporated by reference. In particular, all descriptions with respect to the general formula of the compounds having the general formula (C), the preferred embodiments of the various substituents as well as the medical utility and advantages of the compounds are incorporated herein by reference. The compounds having the general formula (C) can be optionally in the form of a pharmaceutically acceptable salt, solvate, polymorph, codrug, cocrystal, prodrug, tautomer, racemate, enantiomer, or diastereomer or mixture thereof. They are defined below.

The cap binding inhibitors are not are not particularly limited either and can be any cap binding inhibitor, particularly any viral cap binding inhibitor. Preferred cap binding inhibitors are those having the general formuia (ii) as defined in US application 61/550,057 and/or the compounds disclosed in WO201 1 /000566, the complete disciosure of which is incorporated by reference, in particular, all descriptions with respect to the general formula of the compounds according to US 61/550,057 or WO201 1/000566, the preferred embodiments of the various substituents as well as the medical utility and advantages of the compounds are incorporated herein by reference.

The compound having the general formuia (II) can be optionally in the form of a pharmaceutically acceptable salt, solvate, polymorph, codrug, cocrystal, prodrug, tautomer, racemate, enantiomer, or diastereomer or mixture thereof. It is defined as follows:

wherein Y is S;

R²¹ is selected from -H, -C^alkyl, -(CH₂)_q-aryl, -{CH₂)_q-heterocyclyl, -(CH₂)_q-cycloalkyl, -(CH^-OR²⁵, and -(CH₂)_P-NR²⁵R²⁶; R²² is selected from -H, -d_₆ alky!, -(CH₂)_q-cycioalkyl, -Hal, -CF₃ and -CN;

R²³ is selected from -aryl, -heterocyclyl, -cycloalkyl, -C(~R²⁸)(-R²⁹)-aryl, -C(-R²⁸)(-R²⁹)-heterocyclyi, and -C(-R²⁸)(-R ⁹)-cyc!oalkyl; R²⁵ is selected from -H, -Ci_e alkyi, and -(CH₂CH₂0)_rH;

R²⁶ is selected from -H, and -Ci_e alkyl;

R²⁷ is independently selected from -C^ alkyl, -~C{0)~C _Q alkyl, -Hal, -CF₃, -CN, -COOR²⁵, -OR²⁵, -(CH₂)_qNR²⁵R²⁶, -C(0)-NR²⁵R²⁶, and -NR²⁵-C(0)-C^ alkyl; R and R are independently selected from -H, -C_1-6 atkyl, -(CH₂)_q-aryi, -(CH₂)_q- heterocyciy!, -(CH₂)_p-cycloalkyi, -OH, -0-C_M alky!, -0-(CH₂)q-aryl, ~~0~~(C ₂)_q- heterocyc!yi, and -0-(CH₂)_q-cycloalkyi; or R²⁸ and R²⁹ are together =0, -CH₂CH₂-, -CH₂CH₂CHr-, or ~CH₂CH₂CH₂CHr-; p is 1 to 4; q is 0 to 4; and r is 1 to 3; wherein the aryl group, heterocyclyl group and/or cycioalkyi group can be optionally substituted with one or more substituents R²⁷.

The compounds of WO2011/000566 have the general formula (XXI):

(ΧΧΪ) or a pharmaceutically effective salt, a solvate, a prodrug, a tautomer, a racemate, an enantiomer or a diastereomer thereof; wherein one of Y and Z is -XR¹² and the other is R^10';

R¹⁰, R^10' and R ^0" are each individuaiiy seiected from the group consisting of hydrogen, d-Ce-alkyl, C₂-C₆-alkenyi, C₂-C₈-aikynyl, -{CH₂)_nC(0)OH,

-(CH₂)_nC(0)OR¹⁶, -(CH₂)_nOH, -{CH₂)_ftOR¹⁶, -CF₃, -(CH₂)_n-cycioalkyl, -(CH₂)_nC(0)NH₂, ~{CH₂)_nC(0)NHR¹⁶, ~(CH₂)_nC(0)NR ^eR¹⁷, -(CH₂)_nS(0)₂NH₂, -(CH₂)_nS(0)₂NHR¹⁶, -(CH₂)_nS(0)₂NR ⁶R¹⁷, -<CH₂)_nS(0)₂R^ie, halogen, -CN, -(CH₂)_n- aryi, -(CH₂)_n-heteroaryl, -(CH₂)_nNH_2l ~(CH₂)_nNHR^ie, and ~{CH₂)_nNR¹⁶R¹⁷; optionally substituted;

R¹¹ is selected from the group consisting of hydrogen, d-Ce-alkyl, -CF₃, d-Ce-aikenyl, C₂-C₈-alkynyI, -(CH₂)n-cycloalkyi, -(CH₂)n-aryl, -(CH₂)_n-heterocycloalkyl and -(CH^- heteroaryl; optionally substituted;

X is selected from the group consisting of CH₂, C(O), C(S), CH(OH), CH{OR¹⁶), S(0)₂, -S(0)₂-N(H)- -S(0)₂-N(R¹⁶)- -N(H)-S(0)₂- _N{R¹⁶)-S(0)2-, C(=NH), C(=N-R¹⁶), CH(NH₂), CHfNHR¹⁶), CH(NR ⁶R¹⁷), -C(0)-N{H)- -C(0)-N(R^ie)- -N(H)-C(0)-, -N(R ⁶)-C{0)- N(H), N(-R¹⁶) and O;

R¹² is selected from the group consisting of Ci-C₆-alkyl, -CF₃, C₂-C₆-alkenyl, C₂-C_e- alkynyl, -(CH₂)_n-cycIoalkyl, -{CH₂)_n-heterocycloalkyl, -{CH₂)_n-aryI, -NR^ieR¹⁷, and -(CH₂)_n-heteroaryl; optionally substituted;

R¹⁶ and R¹⁷ are independently selected from the group consisting of d-Ce-alkyl, C₂-C₆- alkenyl, C₂-C₆-alkynyi, -(CH₂)_n-cycloalkyl, -(CH₂)_n-aryl, -CF₃, -C(0)R¹⁸ and -S{0)₂R¹⁸; optionally substituted;

R is independently selected from the group consisting of d-Ce-alkyl, C₂-C₆-alkenyl, Ca-Cg-alkynyl, -(CH₂)_n-cycloalkyl and -CF₃; optionally substituted; and n is in each instance selected from 0, 1 and 2.

In the context of WO201 1/000566 the term "optionally substituted" in each instance refers to between 1 and 10 substituents, e.g. 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10 substituents which are in each instance preferably independently selected from the group consisting of halogen, in particular F, CI, Br or I; -N0₂, -CN, -OR', -NR'R", -(CO)OR', -{CO)OR"\ -(COJNR'R", -NR'COR"", -NR'COR', -NR"CONR'R",

-NR"S0₂A, -COR"'; -S0₂NR'R", -OOCR'", -CR"'R"OH, -R'OH, =0, and -E;

R' and R" are each independently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, -OE, cycloalkyl, heterocycloaikyl, aryl, heteroaryl, and aralkyl or together form a heteroaryl, or heterocycloaikyl; optionally substituted; FT and R"" are each independently selected from the group consisting of alkyl, aikenyl, aikyny!, cycioa!kyl, heterocyc!oalkyi, alkoxy, aryl, aralky!, heteroaryl, and -NR'R"; and

E is selected from the group consisting of aikyl, aikenyl, cycloalkyi, a!koxy, aikoxyalkyl, heterocycloaikyl, an alicydic system, aryl and heteroaryl; optionally substituted.

Widespread resistance to both classes of licensed influenza antivirals ( 2 ion channel inhibitors (adamantanes) and neuraminidase inhibitors (Oseltamivir)) occurs in both pandemic and seasonal viruses, rendering these drugs to be of marginal utility in the treatment modality. For 2 ion channel inhibitors, the frequency of viral resistance has been increasing since 2003 and for seasonal influenza A H3N2, adamantanes are now regarded as ineffective. Virtually ail 2009 H1 N1 and seasonal H3N2 strains are resistant to the adamantanes (rimantadine and amantadine), and the majority of seasonal H1 1 strains are resistant to oseltamivir, the most widely prescribed neuraminidase inhibitor (NAI). For oseltamivir the WHO reported on significant emergence of influenza A/H1 N1 resistance starting in the influenza season 2007/2008; and for the second and third quarters of 2008 in the southern hemisphere. Even more serious numbers were published for the fourth quarter of 2008 (northern hemisphere) where 95% of all tested isolates revealed no Oseltamivir-susceptibiiity. Considering the fact that now most national governments have been stockpiling Oseltamivir as part of their influenza pandemic preparedness plan, it is obvious that the demand for new, effective drugs is growing significantly. To address the need for more effective therapy, preliminary studies using double or even triple combinations of antiviral drugs with different mechanisms of action have been undertaken. Adamantanes and neuraminidase inhibitors in combination were analysed in vitro and in vivo and found to act highly synergisticaliy. However, it is known that for both types of antivirals resistant viruses emerge rather rapidly and this issue is not tackled by combining these established antiviral drugs.

Influenza virus polymerase inhibitors are novel drugs targeting the transcription activity of the polymerase. Selective inhibitors against the cap-binding and endonuclease active sites of the viral polymerase severely attenuate virus infection by stopping the viral reproductive cycle. These two targets are located within distinct subunits of the polymerase complex and thus represent unique drug targets. Due to the fact thai both functions are required for the so-called "cap-snatching" mechanism mandatory for viral transcription, concurrent inhibition of both functions is expected to act highly synergistically. This highly efficient drug combination would result in lower substance concentrations and hence improved dose-response-reSationships and better side effect profiles.

Both of these active sites are composed of identical residues in all influenza A strains (e.g., avian and human) and hence this high degree of sequence conservation underpins the perception that these targets are not likely to trigger rapid resistant virus generation. Thus, endonudease and cap-binding inhibitors individually and in combination are idea! drug candidates to combat both seasonal and pandemic influenza, irrespectively of the virus strain.

The combination of an endonudease inhibitor and a cap-binding inhibitor or a dual specific polymerase inhibitor targeting both the endonudease active site and the cap- binding domain would be effective against virus strains resistant against adamantanes and neuraminidase inhibitors and moreover combine the advantage of Sow susceptibility to resistance generation with activity against a broad range of virus strains.

The combination of inhibitors of different antiviral targets (particularly targeting influenza) focusing on the combination with (preferably influenza) potymerase inhibitors as dual or multiple combination therapy. Influenza virus polymerase inhibitors are novel drugs targeting the transcription activity of the polymerase. Selective inhibitors against the cap- binding and endonudease active sites of the viral polymerase severely attenuate virus infection by stopping the viral reproductive cycle. The combination of a polymerase inhibitor specifically addressing a viral intracellular target with an inhibitor of a different antiviral target is expected to act highly synergistically. This is based on the fact that these different types of antiviral drugs exhibit completely different mechanisms of action and pharmacokinetics properties which act advantageously and synergistically on the antiviral efficacy of the combination.

This highly efficient drug combination would result in lower substance concentrations and hence improved dose-response-relationships and better side effect profiles. Moreover, advantages described under (i) for polymerase inhibitors would prevail for combinations of inhibitors of different antiviral targets with polymerase inhibitors.

Typical!y, at least one compound selected from the first group of polymerase inhibitors is combined with at least one compound selected from the second group of polymerase inhibitors.

The first group of polymerase inhibitors which can be used in this type of combination therapy includes, but is not limited to, the compounds having the formula (A) or (C).

The second group of polymerase inhibitors which can be used in this type of combination therapy includes, but is not limited to, the compounds having the general formula (I), the compounds having the general formula (II), the compounds disclosed in WO 2011/000566, WO 2010/110231 , WO 2010/1 10409, WO 2006/030807 or US 5,475,109 as well as flutimide and analogues, favipiravir and analogues, epigaliocatechin gallate and analogues, as well as nucleoside analogs such as ribavirine.

The combination of polymerase Inhibitors with neuramidase inhibitors

Influenza virus polymerase inhibitors are novel drugs targeting the transcription activity of the polymerase. Selective inhibitors against the cap-binding and endonuclease active sites of the viral polymerase severely attenuate virus infection by stopping the viral reproductive cycle. The combination of a polymerase inhibitor specifically addressing a viral intracellular target with an inhibitor of a different extracellular antiviral target, especially the (e.g., viral) neuraminidase is expected to act highly synergistica!ly. This is based on the fact that these different types of antiviral drugs exhibit completely different mechanisms of action and pharmacokinetic properties which act advantageously and synergistica!ly on the antiviral efficacy of the combination.

This highly efficient drug combination would result In lower substance concentrations and hence improved dose-response-relationships and better side effect profiles. Moreover, advantages described under (i) for polymerase inhibitors would prevail for combinations of inhibitors of different antiviral targets with polymerase inhibitors. Typically, at least one compound selected from the above-mentioned first group of polymerase inhibitors is combined with at least one neuramidase inhibitor. The neuraminidase inhibitor (particularly influenza neuramidase inhibitor) is not specifically limited. Examples include zanamivir, ose!tamivir, peramivir, KDN, DANA, FANA, and cyclopentane derivatives.

(iv) The combination of polymerase inhibitors with M2 channel inhibitors

Influenza virus polymerase inhibitors are novel drugs targeting the transcription activity of the polymerase. Selective inhibitors against the cap-binding and endonuclease active sites of the viral polymerase severely attenuate virus infection by stopping the viral reproductive cycle. The combination of a polymerase inhibitor specifically addressing a viral intracellular target with an inhibitor of a different extracellular and cytoplasmic antiviral target, especially the viral 2 ion channel, is expected to act highly synergistically. This is based on the fact that these different types of antiviral drugs exhibit completely different mechanisms of action and pharmacokinetic properties which act advantageously and synergistically on the antiviral efficacy of the combination.

This highly efficient drug combination would result in lower substance concentrations and hence improved dose-response-reiationships and better side effect profiles. Moreover, advantages described under (i) for polymerase inhibitors would prevail for combinations of inhibitors of different antiviral targets with polymerase inhibitors.

Typically, at least one compound selected from the above-mentioned first group of polymerase inhibitors is combined with at least one M2 channel inhibitor.

The M2 channel inhibitor (particularly influenza M2 channel inhibitor) is not specifically limited. Examples include amantadine and rimantadine.

(v) The combination of polymerase inhibitors with alpha gtucosidase inhibitors Influenza virus polymerase inhibitors are novel drugs targeting the transcription activity of the polymerase. Selective inhibitors against the cap-binding and endonuclease active sites of the viral polymerase severely attenuate virus infection by stopping the viral reproductive cycle. The combination of a polymerase inhibitor specifically addressing a viral intracellular target, with an inhibitor of a different extracellular target, especially alpha glucosidase, is expected to act highly synergistically. This is based on the fact that these different types of antiviral drugs exhibit completely different mechanisms of action and pharmacokinetic properties which act advantageously and synergistically on the antiviral efficacy of the combination.

Typically at least one compound selected from the above-mentioned first group of polymerase inhibitors is combined with at least one alpha glucosidase inhibitor.

The alpha glucosidase inhibitor (particularly influenza alpha glucosidase inhibitor) is not specifically limited. Examples include the compounds described in Chang et aL, Antiviral Research 2011, 89, 26-34.

The combination of polymerase inhibitors with ligands of other influenza targets influenza virus polymerase inhibitors are novel drugs targeting the transcription activity of the polymerase. Selective inhibitors against the cap-binding and endonuclease active sites of the viral polymerase severely attenuate virus infection by stopping the viral reproductive cycle. The combination of a polymerase inhibitor specifically addressing a viral intraceilular target with an inhibitor of different extracellular, cytoplasmic or nucleic antiviral targets is expected to act highly synergistically. This is based on the fact that these different types of antiviral drugs exhibit completely different mechanisms of action and pharmacokinetic properties which act advantageously and synergistically on the antiviral efficacy of the combination. This highly efficient drug combination would result in lower substance concentrations and hence improved dose-response-relationships and better side effect profiles. Moreover, advantages described under (i) for polymerase inhibitors would prevail for combinations of inhibitors of different antiviral targets with polymerase inhibitors.

Typicaliy at least one compound selected from the above mentioned first group of polymerase inhibitors is combined with at least one ligand of another influenza target.

The ligand of another influenza target is not specifically limited. Examples include compounds acting on the sialidase fusion protein, e.g. Fludase (DAS 181), siRNAs and phosphorothioate oligonucleotides, signal transduction inhibitors (ErbB tyrosine kinase, Abl kinase family, MAP kinases, PKCa-mediated activation of ERK signaling as well as interferon (inducers).

The combination of (preferably influenza) polymerase inhibitors with a compound used as an adjuvance to minimize the symptoms of the disease (antibiotics, anti-inflammatory agents like COX inhibitors (e.g., COX-1/COX-2 inhibitors, selective COX-2 inhibitors), lipoxygenase inhibitors, EP ligands (particularly EP4 !igands), bradykinin ligands, and/or cannabinoid ligands (e.g., CB2 agonists). Influenza virus polymerase inhibitors are novel drugs targeting the transcription activity of the po!ymerase. Selective inhibitors against the cap-binding and endonuclease active sites of the viral polymerase severely attenuate virus infection by stopping the viral reproductive cycle. The combination of a polymerase inhibitor specifically addressing a viral intracellular target with an compound used as an adjuvance to minimize the symptoms of the disease address the causative and symptomatic pathological consequences of viral infection. This combination is expected to act synergistically because these different types of drugs exhibit completely different mechanisms of action and pharmacokinetic properties which act advantageously and synergistically on the antiviral efficacy of the combination.

This highly efficient drug combination would result in lower substance concentrations and hence improved dose-response-relationships and better side effect profiles. Moreover, advantages described under (i) for polymerase inhibitors would prevail for combinations of inhibitors of different antiviral targets with polymerase inhibitors. Compounds having the general formula (A)

The present invention discloses a compound having the general formula (A).

(A)

It is understood that throughout the present specification the term "a compound having the general formula (A)" encompasses pharmaceutically acceptable salts, solvates, polymorphs, prodrugs, iautomers, racemates, enantiomers, or diastereomers or mixtures thereof unless mentioned otherwise.

In the present invention the following definitions apply with respect to the compounds having the general formula (A).

R^* is -H, -Hal, -(optionally substituted d_6 a!kyl), -(optionally substituted C₃_ cycloalkyl), -(optionally substituted aryl), -C_M alkyi— (optionally substituted C₃_₇ cycloalkyl), -C_1-4 aikyl— (optionally substituted aryl) or -X¹-R¹. In a preferred embodiment, R^* is -Hal, -(optionally substituted Ci„₆ alkyl) (wherein the optional substituent of the alkyl group is preferably Hal, more preferably F); -C _-4 alkyl— (optionally substituted aryl) (wherein the optional substituent of the aryl group is preferably halogen) or -X¹-R¹. In a more preferred embodiment R* is X -R¹. X¹ is O, C(O), C(0)0, OC(O); S, SO, S0₂, NR⁴, N(R⁵)C(0), C(0}NR⁵, preferably X¹ is O, or NR⁴, more preferably X¹ is NR⁴, In one preferred embodiment, X¹ is NR⁴ and R and R⁴ are joined together to form a 5- to 7-membered ring, which can optionally contain O, S or further N. In another preferred embodiment, X¹ is NR⁴ and R¹ is -SO2-R⁴. X² is O, S, NR⁴, preferably X² is O. is O or S, preferably X³ is O. is O or S, preferably X⁴ is 0. is -H, -(optionally substituted alkyl), -(optionally substituted C₃^₇ cycloaikyl), - (optionaily substituted aryl), -Ci_4 alkyi-(optionaliy substituted C₃_₇ cycioa!kyl), -C_1→ a!kyl-(optional!y substituted aryl). Preferably, R¹ is -H, -(optionally substituted Ci_6 alkyl), -(optionally substituted benzyl), more preferably, R¹ is -H or -(optionally substituted benzyl). Throughout the present specification, it is understood that the definitions of the substituents of the aryl group apply analogously to the benzyl group. is a hydrocarbon group which contains from 5 to 20 carbon atoms and optionally 1 to 4 heteroatoms selected from O, N and S and which contains at least one ring, wherein the hydrocarbon group can be optionally substituted. Preferably the at least one ring is aromatic such as an aryl or heteroaryl ring. More preferably R^? is a hydrocarbon group which contains from 5 to 20 carbon atoms and optionaily 1 to 4 heteroatoms and which contains at least two rings, wherein the hydrocarbon group can be optionally substituted. Even more preferably, at least one of the at least two rings is aromatic such as an aryl or heteroaryl ring. Preferred examples of R² can be selected from the group consisting of

wherein

X is absent, CH₂, NH, C(0)NH, S or O. Furthermore,

Y is CH₂.

In an alternative embodiment, X and Y can be joined together to form an annulated, carbo- or heterocyiic 3 to 8 membered ring which can be saturated or unsaturated. Specific examples of X-Y include -CH₂~, -CH₂-CH₂-, -0-, and -NH-.

R is independently selected from H, -Ci_e aikyl, halogen, -CN, -OH, and -0-C _-J3 alkyl. R³ is -H, -(optiona!!y substituted Ci-e aikyf), -(optionally substituted C₃_₇ cycloaikyi), - (optionally substituted aryl), or -C-,_4 alkyl-(optionally substituted aryl) if X² is NR⁴ then R³ can also be -OH, preferably R³ is -H, -C^ alky] or Bz. R⁴ is -H, -(optionally substituted d__e alkyl), -(optionally substituted cycloaikyi), - (optionally substituted aryl), -C^ aikyl-(optionally substituted C₃_₇ cycloaikyi), or -C^ alkyl~-(optionally substituted aryl) or if X¹ is NR⁴ then R⁴ and R¹ can be joined together to form a 5- to 7-membered ring, which can optionally contain O, S or further N or if X² is NR⁴ then R⁴ and R³ can be joined together to form a 5- to 7-membered ring, which can optionally contain O, S or further N. Preferably, R⁴ is -H, -(optionally substituted aryl), or

-(optionally substituted Ci_₆ alkyl), more preferably R⁴ is -H or -(optionally substituted benzyl).

R⁵ is -H, -(optionally substituted Ct_s alkyl), -(optionally substituted C3-7 cycloaikyi), - (optionally substituted aryl), -C^ alkyl-(optiona!ly substituted C3_₇ cycloaikyi), or -C_1-Jt aikyl-(optionatiy substituted aryl). Preferably R⁵ is -H.

R⁶ is -H, or -C^ alkyl. The optional substituent of the alkyl group is selected from the group consisting of halogen, - CN, -NR⁶R^S, -OH, and -0-Ci_₆ alkyl. Preferably the substituent is -halogen, more preferably F.

The optional substituent of the cycloaikyi group, the aryl group or the hydrocarbon group is selected from the group consisting of -C,^ alkyl, halogen, -CF>, -CN, -X -R⁵ and -C1-4 alkyl— aryl. Preferably the substituent is -halogen (preferably F), -OCH₃ or -CN.

The present inventors have surprisingly found that the compounds having the formula (A) which have a bulky moiety R² have improved pharmacological properties compared to corresponding compounds which have a smaller moiety R². Without wishing to be bound by theory it is assumed that the viral polymerase protein has a pocket for binding and that the bulky moiety R² of the compounds of the present invention fills this pocket to a larger extent. It is further assumed that the larger moiety R² is able to provide more hydrophobic interaction with the pocket than smaller moieties such as methyl. Various modifications and variations of the invention will be apparent to those skilled in the art without departing from the scope of the invention. Although the invention has been described in connection with specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention which are obvious to those skilled in the relevant fields are intended to be covered by the present invention. The following examples are merely illustrative of the present invention and should not be construed to limit the scope of the invention as indicated by the appended claims in any way.

EXAMPLES

FRET endonudease activity assay

The influenza A virus (IAV) PA-Nter fragment (amino acids 1 - 209) harbouring the influenza endonudease activity was generated a nd purified as described in Dtas et a!., Nature 2009; Apr 16; 458(7240), 914-918. The protein was dissolved in buffer containing 20mM Tris pH 8.0, 100m NaCI and 10mM β-mercaptoethanol and aliquots were stored at -20 °C.

A 20 bases dual-labelled RNA oligo with 5'-FAM fluorophore and 3'-BHQ1 quencher was used as a substrate to be cleaved by the endonudease activity of the PA-Nter. Cleavage of the RNA substrate frees the fluorophore from the quencher resulting in an increase of the fluorescent signal.

All assay components were diluted in assay buffer containing 20mM Tris-HCI pH 8.0, 100m NaCI, 1mM MnCI₂, 10mM gCI₂ and 10mM β-mercaptoethanol. The final concentration of PA- Nter was 0.5μ and 1.6μΜ RNA substrate. The test compounds were dissolved in DMSO and generally tested at two concentrations or a concentration series resulting in a final plate well DMSO concentration of 0.5 %. In those cases where the compounds were not soluble at that concentration, they were tested at the highest soluble concentration. SAV-6004 was used as a reference in the assay at a concentration of 0.1 μΜ. 5μ! of each compound dilution was provided in the welis of white 384-weil microtiter plates (PerkinE!mer) in eight replicates. After addition of PA-Nter dilution, the plates were sealed and incubated for 30min at room temperature prior to the addition of 1 ,6μΜ RNA substrate diluted in assay buffer. Subsequently, the increasing fluorescence signal of cleaved RNA was measured in a microplate reader (Synergy HT, Biotek) at 485nm excitation and 535nm emission wavelength. The kinetic read interval was 35sec at a sensitivity of 35. Fluorescence signal data over a period of 20min were used to calculate the initial velocity (vO) of substrate cleavage. Final readout was the % reduction of vO of compound-treated samples compared to untreated. The half maximal inhibitory concentration (IC₅₀) is a measure of the effectiveness of a compound in inhibiting biological or biochemical function and was calculated from the initial reaction velocities (vO) in a given concentration series ranging from maximum 100 μΜ to at least 2 nM.

Cytopathic effect (CPE) assay

The influenza A virus (IAV) was obtained from American Tissue Culture Collection (A Aichi/2/68 (H3N2); VR-547). Virus stocks were prepared by propagation of virus on Mardin- Darby canine kidney ( DCK; ATCC CCL-34) ceils and infectious titres of virus stocks were determined by the 50 % tissue culture infective dose (TCID₅₀) analysis as described in Reed, L J., and H, Muench. 1938, Am. J. Hyg. 27:493-497.

MDCK cells were seeded in 96-weIi plates at 2><10⁴ cells/well using DMEM/Ham's F- 2 (1 :1 ) medium containing 10 % foetal bovine serum (FBS), 2 mM L-g!utamine and 1 % antibiotics (all from PAA). Until infection the cells were incubated for 5 hrs at 37 °C, 5.0 % C0₂ to form a -80 % confluent monolayer on the bottom of the well. Each test compound was dissolved in DMSO and generally tested at 25 μΜ and 250 μΜ. In those cases where the compounds were not soluble at that concentration they were tested at the highest soluble concentration. The compounds were diluted in infection medium {DMEM/Ham's F-12 (1 :1 ) containing 5 pg/ml trypsin, and 1 % antibiotics) for a final plate well DMSO concentration of 1 %. The virus stock was diluted in infection medium (DMEM/Ham's F-12 (1 :1 ) containing 5 pg/ml Trypsin, 1 % DMSO, and 1 % antibiotics) to a theoretical multiplicity of infection (MOi) of 0.05. After removai of the culture medium and one washing step with PBS, virus and compound were added together to the ceils.- In the wells used for cytotoxicity determination (i.e. in the absence of viral infection), no virus suspension was added. Instead, infection medium was added. Each treatment was conducted in two replicates. After incubation at 37 °C, 5 % C0₂ for 48 hrs, each weli was observed microscopically for apparent cytotoxicity, precipitate formation, or other notable abnormalities. Then, cell viability was determined using CellTiter- Glo luminescent cell viability assay (Promega). The supernatant was removed carefully and 65 μΙ of the reconstituted reagent were added to each well and incubated with gentle shaking for 15 min at room temperature. Then, 60 μΙ of the solution was transferred to an opaque plate and luminescence (RLU) was measured using Synergy HT plate reader (Biotek).

Relative cell viability values of uninfected-treated versus uninfected-untreated cells were used to evaluate cytotoxicity of the compounds. Substances with a relative viability below 80 % at the tested concentration were regarded as cytotoxic and retested at lower concentrations.

Reduction in the virus-mediated cytopathic effect (CPE) upon treatment with the compounds was calculated as follows: The response (RLU) of infected-untreated samples was subtracted from the response (RLU) of the infected-treated samples and then normalized to the viability of the corresponding uninfected sample resulting in % CPE reduction. The half maximal inhibitory concentration (IC₆₀) is a measure of the effectiveness of a compound in inhibiting biological or biochemical function and was calculated from the RLU response in a given concentration series ranging from maximum 100 μΜ to at least 00 nM.

Compounds having the general formula (A)

Scheme I Series:

Genera! Procedure: Synthesis of 2-aminoacetonitrile (1-1) and 5-aminothiazole-2 -thiol (I -2)

A solution of sodium methoxide, prepared from sodium (23 g, 1.0 moi) in dry meihanol (500 mL), was added dropwise under ice-cooiing to a stirred suspension of aminoacetonitriie hydrochloride (100 g, 1.08 moi) in dry methanol (100 mL) his reaction mixture was stirred for 2 hours at room temperature (r.t.), then the mixture was concentrated in vacuo , the residue was dissolved in dry ethyl acetate (500 mL), the mixture was filtered and the filtrate was dropwise added to the solution of carbon disulfide (136 g, 1.79 moi) in dry ethyl acetate (100 mL).The reaction mixture was stirred overnight while the temperature rose from 0 °C to room temperature. The precipitate was filtered to afford the crude product !-2 as yellow solid 107.4 g, yield 75.6%.

Synthesis of 2-arninoacetoniiri!e (i-3)

A solution of sodium methoxide, prepared from sodium (18.7 g, 0.814 moi) in dry methanol (600 ml), was cooled to -78 °C, compound !-2 was added at -78 °C. To this red-brown soiution, methyl iodide ( 15 g, 0.814 mmol) was dropwise added at -78 °C This reaction mixture was stirred for 3h at -78 °C. The methanol was removed in vacuo and the residue was extracted with ethyl acetate (EA) and water, the organic phase was dried and concentrated in vacuo to afford the crude product I-3 as brown oil 117 g, yield 98 %. Synthesis of 2-aminoacetonitrile (1-4)

The compound 1-3 (117 g, 0.801 mmol) was dissolved in ethanol (400 mi) and diethyl ethoxymethylenemalonate was added. This reaction mixture was stirred for 3 h at reflux. Then the mixture was cooled to r.t.. The precipitate was filtered to afford the product 1-4 as brown solid 163 g, yield 64%.

Synthesis of 2-aminoacetonitrile (I-5)

The compound 1-4 (20 g, 63.6 mmoi) was added to diphenyi ether (150 mL). The mixture was heated to 250 °C for 40 min. Then the mixture was cooled to r.t.. and was added to petrolether (PE). The precipitate was filtered to afford the product I -5 as brown solid 16 g, yield 94 %.

Synthesis of 2-aminoacetonitrile (1-6)

The compound I-5 (6.5 g, 24.07 mmoi), 2-(bromomethyl)biphenyi (6.5 g, 26.48 mmol) and potassium carbonate (6.6 g, 48.14 mmol) were added to methylsulfinylmethane (60 mL). This reaction mixture was stirred overnight at r.t.. The mixture was extracted with EA and water, the organic phase was concentrated in vacuo to afford the crude product which was purified by column chromatography on silica gel with EA to afford the product !-6 as brown solid 7.4 g, yield 70.5 %.

Synthesis of 2-aminoacetonitrile (I-7)

The compound i-6 (3.1 g, 0.711 mmol) and m-CPBA (3.0 g, 17.775 mmol) were added to dichioromethane (DCM) (20 mL). This reaction mixture was stirred for 5 h at r.t.. The mixture was extracted with DCM and a saturated NaHC0₃ solution. The organic phase was concentrated in vacuo to afford the crude product 1-7 as yellow solid 3.2 g, yield 97 %.

Representative synthetic method of 2-aminoacetonitrile (l₄-8)

The compound 1-7 (200 mg, 0.427 mmol), phenylmethanamine (183 mg, 1.709 mmol) and potassium carbonate (1 18 mg, 0.854 mmol) were added to dimethy!sulfoxide (DMSO) (3 mL), This reaction mixture was stirred overnight at r.t.. This mixture was extracted with DCM and water, the organic phase was concentrated in vacuo to afford the crude product I-8 as brown oil 180 mg, yield 85%. Representative synthetic method of 2-aminoacetonitrile (I₄) The compound i₄-8 (62 mg, 0.125 mmo!) was dissolved in EtOH (6 mL), then lithium hydroxide hydrate (21 mg, 0.501 mmol) was added. This reaction mixture was stirred for 4 h at r.t.. The mixture was adjusted to pH=5 with HCI, the precipitate was filtered to afford the product I₄ as pale white solid 32mg, yield 55%.

Example 1

4-(Biphenyl-2- lmethyl)-7-oxo-2-(phenylsulfonamido)-4,7-dihydrothiazoio[5,4- b]pyridine-6-carboxylic acid (F4)

I-7 (ί-7') was treated with phenylsulfonamide according to the representative method to obtain compound F4 as a pale white solid.

Yield: 5%

MS (ESI): 518(M+H)⁺, 105

1H NMR (dg-DMSO, 300 Hz):

δ 8.46 (br, s, 1 H), 7.34-7.73 (m, 14H), 5.35 (s, 2H)

Example 2

4-(Biphenyl-2-ylmethyl)-2-{methylamino)-7-oxo-4,7-dihydrothia2olo[5,4-b]pyridine-6- carboxylic acid (11)

I-7 (Ι-7') was treated with methanamine according to the representative method to obtain compound 11 as a pale white solid.

Yield: 5%

MS (ESi): 392 (M+H}⁺, 157

1H NMR (de-DMSO, 300 Hz): δ 8.39 (s, 1 H), 8.06-8.07 (br, s, 1 H), 7.23-7.51 (m, 9H), 5.58 (s, 2H), 2.84 (d, J - 4.8 Hz, 3H)

Example 3

4-(Biphenyl-2-ylmethyl)-2-(cyclopropylamino)-7-oxo

6-carboxylic acid (12)

1-7 (1-7') was treated with amtnocyctopropane according to the representative method to obtain compound 12 as a pale white solid.

Yield: 5%

MS (ESI): 418 {M+H)^*

1 HN R (d6-DMSO, 300 MHz):

δ 8.59 (s, 1 H), 8.48 {s, 1 H), 7.49-7.25 (m, 9H), 5.59 (s, 2H), 2.57 (d, J = 1.8 Hz, 1 H), 0.72 (m, 2H), 0.47 (m, 2H). Example 4

4-(Biphenyl-2-ylmethyl)-2-(cyclopentylamino)-7-oxo-4,7-dihydrothia2olo[5,4-b]pyridine- 6-carboxylic acid (I3)

1-7 (1-7') was treated with aminocyclopentane according to the representative method to obtain compound 13 as a yeliow solid.

Yield: 5%

MS (ESI): 446 (M+H)⁺, 407

1HNMR (cfs-DMSO, 300 MHz):

δ 8.42 (s, 1 H), 8.16 (d, J = 6.0 Hz, 1 H), 7.48-7.24 (m, 10H), 5.57 (s, 2H), 4.03 (d, J = 6.0 Hz, 2H), 1.89-1.85 (m, 2H), 1.63-1.41 (m, 7H) Example 5

2-( Benzylami noH-(bi phenyl -2-yl methyI}-7^

carboxylic acid (14)

i-7 (1-7') was treated with benzylamine according to the representative method to obtain compound 14 as a pale white solid.

Yield: 5%

MS (ESI): 468 <M+H)⁺

1HNMR (c/_e-DMSO, 300 MHz):

δ 8.60 (s, 1 H), 8.43 (s, 1 H), 7.48-7.25 (m, 14H), 5.57 (s, 2H), 4.49 (d, J = 4.5 Hz, 2H)

Example 6

4-(Biphenyl-2-ylmethyl)-7 )xo-2-(pyrrolidin-1 -yl)^J-dihydrothiazolo[5,4-b]pyridine-6- carboxylic acid (15)

[-7 (1-7') was treated with pyrrolidine according to the representative method to obtain compound 15 as a pale white solid.

Yield: 5%

MS (ESI): 433 (M+H)⁺

¹HNMR (de-DMSO, 300 MHz):

5 8.43 (s, 1 H), 7.50-7.269 (m, 9H), 5.54 (d, J = 8.4 Hz, 2H), 3.32 (s, 5H), 1.95 (s, 4H)

Example 7 4-(BiphenyS-2-y!methyf)-2-{4-hydroxypsperidin-1 -yl)-7-oxo-4,7-dihydroihiazo!o[5,4- b]pyridine-6-carboxylic acid (I6)

1-7 (!-7') was treated with piperidin-4-ol according to the representative method to obtain compound Ιβ as a yellow solid.

Yield: 3%

MS (ESI): 462 (M+H)⁺

1HNMR (de-DMSO, 300 MHz):

δ 16.20 (br, s, 1 H), 8.45 (s, 1 H,), 7.51-7.23 (m, 10H), 5.55 (d, J = 7.8 Hz, 2H), 3.76-3.21 (m, 7H), .70-1.78 (m, 2H), 1.39-1.48 (m, 2H)

Example 8

4-(BiphenyI-2-ylmethy!)-7-oxo-2-(pipera∑in -yl)-4,7-dlhydrothiazoIo[5,4-b]pyridine-6- carboxylic acid (I7)

[-7 (i-7') was treated with piperazine according to the representative method to obtain compound 17 as a ye!low solid.

Yield: 3%

MS (ESI): 447 (M÷H)⁺

J'HNMR ( DMSO, 300 MHz):

δ 9.07 (s, 2H), 8.52 (s, 1 H), 7.49-7.23 (m, 10H), 5.61 (s, 2H), 3.63 (s, 4H), 3.22 (s, 4H)

Example 9

2-(4-Benzylpiperazin-1-yl)-4-(biphenyl-2-ylmethyl)-7-oxo-4,7-dihydrothiazolo[5,4- b]pyridine-6-carboxylic acid (I8)

i-7 (1-7') was treated with 1-benzySpiperazine according to the representative method to obtain compound IS as a yellow solid.

Yield: 5%

MS (ESi): 537 {M+H)⁺

¹HNMR (d₆-D SO, 300 MHz):

δ 8.55 (s, 1 H), 7.49-7.22 (m, 14H), 5.61 (s, 2H), 4.30 (s, 2H), 3.16-3.39 (m, 8H)

Example 10

4-(Biphenyl-2-ylmethyl)-7-oxo-2-(piperidin-1 -y!)-4,7-dihydrothiazolo[5,4-b]pyridme-6- carboxylic acid (I9)

I-7 (Ι-7') was treated with piperidine according to the representative method to obtain compound I9 as a pale white solid,

Yield: 5%

MS (ESI): 446 (M+Hf

"HNMR (de-DMSO, 300 MHz):

δ 16.22 (s, 1 H), 8.45 (s, 1 H), 7.49-7.24 (m, 9H), 5.58 (s, 2H), 3.41-3.42 (m, 4H), 1.57 (s, 6H) Example 11

4-(BiphenyJ-2-ylmethyl)-2-(4-methylpiperidin-1 -yl)-7-oxo-4,7-dihydrothiazolo[5,4- b]pyridine-6-carboxylic acid (110)

1-7 (ί-7') was treated with 4-methyipiperidine according to the representative method to obtain compound 110 as a pafe white solid.

Yield: 5%

MS (ESI): 460 (M+H)⁺

¹H NMR (cVDMSO, 300 MHz):

δ 8.44 (s, 1 H), 7.25-7.51 <m, 9H), 5.58 (s, 2H), 3.67-3.71 (m, 2H), 3,02-3.10 (t, J = 12 Hz, 2H), 1.57-1.70 (m, 3H) 1.1 1 -1.17 (m, 2H), 0.90 (d, J = 6.9 Hz, 3H) Example 12

4-(Biphenyl-2-ylmethy!)-2-{isopropylamino)-^

carboxylic acid ((11)

1-7 (!-7^!) was treated with 2-arninopropane according to the representative method to obtain compound 111 as a yeiiow solid.

Yield: 5%

MS (ES!): 420 (M+H)⁺, 105

1H NMR (de-D SO, 300 MHz):

5 8.42 (s, 1 H), 8.06 (d, J = 7.2Hz, 1 H), 7.23-7.51 (m, 9H), 5.56 (s, 2H), 3.85-3.91 (m, 1 H), 1.13 (d, J = 6.6 Hz, 6H)

Example 13

4-{Bipheny1-2-ylmethyl)-2-(2-methoxyethylamino)-7-oxo-4,7-dihydrothiazoio[5,4- b]pyridine-6-carboxylic acid (112)

1-7 {1-7') was treated with 2-methoxyethanamine according to the representative method to obtain compound 112 as a pale white solid.

Yieid: 5%

MS (ES!): 436 (M+H)⁺

¹H NMR (cfe-D SO, 300 MHz):

6 8.43 (s, 1 H), 8.25 (s, 1 H), 7.23-7.51 (m, 9H), 5.57 (s, 2H), 3.45-3.50 (m, 4H), 3.25 (s, 3H) Example 14

4-{Biphenyl-2-yImethyi)-2-(4-methylpiperazin-1 -yl)-7-oxo-4,7-dihydrothiazoio[5,4- b]pyridine-6-carboxylic acid (114)

I-7 ('-7') was treated with 1-methylpiperazine according to the representative method to obtain compound 114 as a yellow solid.

Yield: 2%

MS (ESI): 461 (M+H)⁺, 157, 231

1H NMR (de-OMSQ, 300 MHz):

δ 9.89 (s, 1 H), 8.54 (s, 1 H), 7.21 -7.49 (m, 9H), 5.62 (s, 2H), 3.94-3.97 (br,2H), 3.33-3.48 (m, 4H), 3.12-3.17 (m, 2H), 2.84 (s, 3H).

Example 5

4-(Biphenyl-2-ylmethyl)-2-morpholino-7-oxo-4,7-dihydrothiazolo[5,4-b]pyridine-6- carboxylic acid (115)

1-7 (S-7') was treated with morphoiine according to the representative method to obtain compound 115 as a yellow solid.

Yield: 2%

MS (ESI): 448 (M+H)^*, 157

1H NM (<¾-DMSO, 300 MHz):

δ 8.41 (s, 1 H), 7.24-7.48 (m, 9H), 5.59 (s, 2H), 3.66-3.68 (m, 4H), 3.40-3.41 (m, 4H)

Exampie 16

4-(Biphenyl-2-ylmethyl)-N-methyl-2-(methylamino)-7 >xo^,7-dihydrothia2olo[5,4^ b]pyndine-6-carboxamide (116)

!-7 (Ι-7') was treated with methanamine according to the representative method to obtain compound S16 as a yellow solid.

Yield: 5%

MS (ESI): 405 (M+H)⁺

Ή NMR (ck-DMSO, 300 MHz):

δ 10.16 (br, s, 1 H), 8.39 (s, 1 H), 7.84 (br, s, 1 H), 7.29-7.48 (m, 8H), 7.08-7.10 (d, J = 6.9 Hz, 1 H), 5.43 (s, 2H), 2.83 (s, 3H), 2.81 (s, 3H)

Example 17

2-(Benzy!amino}^-(biphenyl-2-ylmethyl)-N-methyl-7-oxo-4,7-dihydrothiazolo[5,4- b]pyrtdine-6-carboxamide (117)

The ethyl ester precursor of 14 was treated with methanamine according to the representative method to obtain compound 117 as a pale white soiid.

Yield: 5%

MS (ESI): 481 (M+H)⁺

¹H NMR ( /₆-DMSO, 300 MHz):

δ 10.15 (s, 1 H), 8.41 (s, 1 H), 8.37(s, 1 H), 7.68-7.72 (m, 1 H), 7.28-7.48 (m, 13H), 7.09 (d, J = 7.5 Hz, 1 H), 5.42 (s, 2H), 4.45 (d, J = 5.1 Hz, 2H), 2.82 (d, J = 4.2 Hz, 3H)

Example 18

4-(Biphenyl-2-ylmethyl)-N-methyl-7-oxo-2-(pyrrolidin-1-yl)^,7-dihydrothiazolo[5,4- b]pyridine-6-carboxamide (118)

The ethyl ester precursor of IS was treated with methanamine according to the representative method to obtain compound 118 as a pale white solid.

Yield: 5%

MS (ESI): 445 (M+H)\ 157

1H NMR (CDCI₃, 300 MHz):

δ 10.28 (s, 1 H), 8.32 (s, 1 H), 7.22-7.46 (m, 8H), 7.08 (d, J = 7.8 Hz, 1 H), 5.15 (s, 2H), 3.67 (s, 4H), 2.97 (d, J = 4.5 Hz, 3H), 2.02 (s, 4H)

Example 19

N-Benzyl-2-(benzylamino)^(biphenyl-2-ylmethyl)-7-oxo-4,7-dihydrothiazolo[5,4- b}pyridine-6-carboxamide (119)

14 was treated with benzyiamine according to the representative method to obtain compound f 19 as a brown solid.

Yieid: 2%

MS (ESi): 557 {M+H)M05.

H NMR (<_yD SO, 300 MHz):

δ 10.75 (s, 1 H ), 8.42 (s, 1 H), 8.39 (s, 1 H), 7.25-7.46 (m, 18H), 7.14 (d, J = 7.2 Hz, 1 H), 5.44 (s, 2H), 4.52 (d, J = 5.4 Hz, 2H), 4.44 (d, J = 5.7 Hz, 2H) Example 20

4-(Biphenyl-2-ylmethyl)-7-oxo-2-(phenylmethylsulfonamido)-4,7-dihydrothiazolo[5,4- b] pyri d i ne-6-carboxylic acid (I20)

I-7 (ί-7') was treated with benzylsuifonamide according to the representative method to obtain compound 120 as a pale white solid.

Yield: 5%

ivio (,cSi)- o (rvt+n)

¹H NMR (c ₆-DMSO, 300 MHz):

δ 8.51 (s, 1 H), 7.20-7.54 (m, 14H), 5.53 (s, 2H), 4.36 (s, 2H)

ExampSe 21

4-(Biphenyl-2-ylmethyl)-2-(3-fluorophenylsulfonamido)-7-oxo-4,7-dihydrothiazoloi5,4- b]pyridine-6-carboxylic acid (121)

1-7 (1-7') was treated with 3-fluorobenzylsuifonamide according to the representative method to obtain compound 121 as a paie white solid.

Yield: 5%

MS (ESI): 286 (M+H)⁺, 157, 105.

1 H NMR (de-DMSO, 300 MHz):

6 8.53 (s, 1 H), 7.24-7.57 (m, 1 3H), 5.63 (s, 2H)

Example 22

4-(Bipheny!-2-ylmethyl)-2-(methylsulfonamido)-7-oxo-4₎7-dihydrothiazolo[5,4-

i-7 (1-7') was treated with methyisulfonamide according to the representative method to obtain compound 122 as a paie white solid.

Yield: 5%

MS (ESI): 456(M+H)⁺

¹ H NMR (de-DMSO, 300 MHz):

δ 8.55 (s, 1 H), 7.26-7.50 (m, 9H), 5.60 (s, 2H), 2.96 (s, 3H)

Example 23

4-(Biphenyl-2-ylmethyl)-2-(2-chlorobenzylamino)-7-oxo-4,7-dihydrothiazolo[5,4 b]pyridine-6-carboxylic acid (l _A)

1-7 (Ι-7') was treated with 2-chiorobenzylamine according to the representative method to obtain compound l_1A as a pale white solid.

Yield: 4 %

MS (ESI): 502 (M+H)⁺

¹H NMR ( _e-DMSO, 300 Hz):

δ 8.62 (br, s, 1 H), 8.44 (s, 1 H), 7.24-7.49 (m, 13H), 5.59 (s, 2H), 4.57 (d, J = 3.9 Hz, 2H)

Example 24

Eihyl-4-(biphenyi-2-y methyi)-2-(2-chloroben2yiamino)-7-oxo-4,7-dthydrothiazolo[5,4- bJpyridine-S-carboxylate (l_1A-h)

!-7 (!-7') was treated with 2-chlorobenzylamine according to the representative method to obtain compound l_1A-h as a pale white solid.

Yield: 4 %

MS (ESI): 531{M+H)⁺, 169

1H NMR (oVDMSO, 400 Hz):

δ 8.31 (br, s, H), 8.07 (s, H), 7.20-7.46 (m, 13H), 5.36 (s, 2H), 4.51 (d, J = 3.9 Hz, 2H), 4.16 (q, J = 6.8 Hz, 2H), 1.26 (t, J = 7.2Hz, 3H)

Example 25

4-(Biphenyl-2-ylmethyl)-2-(3-chlorobenzylamino)-7-oxo^ ,7-dihydrothiazolo[5,4- b] pyri di ne-6-carboxyl i c acid

i-7 (!-7') was treated with 3-chlorobenzy!amine according to the representative method to obtain compound \_2A as a pale white solid.

Yield: 3%

MS (ESI): 502 (M+H)⁺, 405

1HNMR (d₆-DMSO, 400 MHz):

δ 8.64 (s, 1 H), 8.44 (s, 1 H), 7.50-7.23 (m, 13H), 5.59 (s, 2H), 4.51 (d, J = 9.2 Hz, 2H).

Example 26

Ethyl 4-(biphenyi-2-ylmethyl)-2-(3-chlorobenzylamino)-7-oxo-4,7- dihydrothiazolo[5,4-b]pyridin -6-carboxylate (l₂A-h)

I-7 (ϊ-7') was treated with 3-chlorobenzylamine according to the representative method to obtain compound l₂A-h as a pale white solid.

Yield: 4%

MS (ESI): 530 ( +H)⁺

HNMR (c/e-DMSO, 400 MHz):

δ 8.39 (s, 1 H), 8.10 (s, 1 H), 7.48-7.21 (m, 13H), 5.39 (s, 2H), 4.47 (d, J - 4.4 Hz, 2H), 4.19 (q, J = 7.2 Hz, 2H), 1.27 (t, J = 7.2 Hz, 2H)

Example 27

4-(Biphenyl-2-ylmethyl)-2-(4^hlorobenzylamino)-7-oxo-4,7-dihydrothiazolo[5,4- b] pyridi n e-6-carboxyl ic acid (l_3A)

i-7 (Ι-7') was treated with 4-chiorobenzyiamine according to the representative method to obtain compound !_3A as a pale white solid.

Yield: 3%

MS (ESI): 502 ( +H)⁺

¹HNMR (dg-DMSO, 400 MHz):

δ 8.63 (s, 1 H), 8.44 (s, 1 H), 7.48-7.23 (m, 13H), 5.58 (s, 2H), 4.48 (d, J = 5.2 Hz, 2H)

Example 28

Ethyl 4-(biphenyl-2-ylmethyl)-2-(4-chlorobenzylamino)

dihydrothiazolo[5,4-b]pyridi -6-carboxylate (l_3A-h)

i-7 (i-7^*) was treated with 4-chlorobenzylamine according to the representative method to obtain compound ^-h as a yellow sotid.

Yield: 3%

MS (ESI): 531 (M+H)⁺

^'HNMR (oVDMSO, 400 MHz), 58.31 (s, 1 H), 8.06 (s, 1 H), 7.48-7.16 (m, 13H), 5 35 (s, 2H), 4.43 (d, J = 5.2 Hz, 2H), 4.16 (q, J = 6.8 Hz, 2H), 1.26 (t, J = 6.8 Hz, 3H) Example 29

4-(Biphenyl-2-ylmethyl)-2-(4-methoxybenzylamino)-7-oxo-4,7-djhydrothiazolo[5,4- b]pyridine-6-carboxylic acid (l_4A)

1-7 (ί-7') was treated with 4-methoxybenzylamine according to the representative method to obtain compound I₄A as a pale white solid.

Yield: 1 1 %

MS (ESI): 498 (M+H)\ 405

HNMR (<¼-D SO, 400 MHz):

δ 16.38 (s, 1 H), 8.44 (s, 1 H), 7.42-7.23 (m, 1 1 H), 6.89 (d, J = 8.0 Hz, 2H), 5.57 (s, 2H), 4.40 (d, J = 5.6 Hz, 2H), 3.73 (s, 3H) Example 30

Ethyl 4-(biphenyl-2-ylmethyl)-2-(4-methoxybenzylamino)-7-oxo-4,7- dihydrothiazolo[5,4-b]pyridine-6-carboxylate (l_4A-h)

S-7 (1-7') was treated with 4-methoxybenzylamine according to the representative method to obtain compound l_4A-h as a pale white solid.

Yield: 5%

MS (ESI): 526 (M+H)⁺, 405

1HNMR (cVDMSO, 400 MHz):

δ 8.26 (s, 1 H), 8.09 (s, 1 H), 7.47-7.18 (m, 1 1 H), 6.87 (d, J - 8.4 Hz, 2H), 5.37 (s, 2H), 4.35 (d, J = 4.8 Hz, 2H), 4.17 (q, J = 6.8 Hz, 2H), 1.27 (t, J - 6.8 Hz, 3H)

Example 31

4-Benzhydryl-2-(4-methoxybenzylamino)-7-oxo-4,7-dihydrothiazolo[5,4-b]pyridine-6- carboxylic acid (l_4D)

The analogue of i-7 (i-7') with diphenylrnethy! substitution was treated with 4- methoxybenzyiamine according to the representative method to obtain compound l_4D as a pale white solid.

Yield: 5%

MS (ES!): 498 (M+H)⁺

HNMR (de-DMSO, 400 MHz):

6 8.63 (s, 1 H), 8.03 (s, 1 H), 7.47-7.49 (m, 6H), 7.25-7.29 (m, 6H), 7.11 (s, 1 H), 6.88 (d, J = 8.0 Hz, 2H), 4.45 (d, J = 5.2 Hz, 2H), 3.73 (s, 3H)

Example 32

4-{Biphenyl-2-ylmethyl)-2-(2,6-dichlorobenzylamino)-7-oxo-4,7-dihydrothiazolo[5,4- b]pyridine-6-carboxylic acid (l

I-7 (S-7^!) was treated with 2,5-dichlorobenzylamine according to the representative method to obtain compound l_5A as a pink solid.

Yield: 2%

¹HNMR (de-DMSO, 400 MHz):

δ 8.48 (s, 1 H), 8.34 (s, 1 H), 7.21 -7.53 (m, 12H), 5.58 (s, 2H), 4.70 (d, J = 4.0 Hz, 2H)

Example 33

Ethyl 4-(biphenyl-2-ylmethyl)-2-(2,6-dichlorobenzylamino)-7-oxo-4,7- di hyd roth^"iazolo[5,4-b] py ri di ne-6-carboxylate (l_5A-h)

i-7 {[-7') was treated with 2,5-dichlorobenzylamine according to the representative method to obtain compound l_5A-h as a yeliow soiid.

Yield: 2%

MS (ESI): 564 ( +Hf

¹HN R c/e-DMSO, 400 MHz):

58.15 (s, 1 H), 8.08 (s, 1 H), 7.21-7.53 (m, 12H), 5.40 (s, 2H), 4.66 (s, 2H), 4.20 (q, J = 6.8 Hz, 2H), 1.29 (t, J = 6.8Hz, 3H) Example 34

4= Biphenyl-2-yimeihyS)_"2-{4-carbamoyibenzylaminio}-7-oxo-4,7-dihydrothiaz !o[5,4- b]pyridine-6-carboxamide (l₆A-h')

i~7 (ί-7') was treated with ethyl 4-(aminomethyl)benzoate according to the representative method and then ammonia to obtain compound !_6A-h' as a pale white solid.

Yield: 1 %

MS (ESI): 5 0 (M+Hf

1HNMR (cfe-DMSO, 400 MHz):

δ 12.99 (s, 1 H), 9.59 (s, 1 H), 8.39-8.44 (m, 2H), 7.83-7.88 (m, 2H), 7.48-7.56 (m, 12H), 7.12 (d, J = 6.8 Hz, H), 5.42 (s, 2H), 4.54 (s, 2H)

Example 35

4-(Biphenyl-2-ylmethyl)-7-oxo-2-(1-phenylethylamino)-4,7-dihydrothiazolo[5,4- b]pyridine-6-carboxylic acid (l_7A)

1-7 {i-7') was treated with 1 -phenylethanamine according to the representative method to obtain compound \_7A as a pale white solid.

Yield: 3%

MS (ESI): 482 (M+H)⁺

¹HNMR (£/₆-DMSO, 400 MHz):

δ 8.68 (d, J = 7.2Hz, 1 H), 8.43 (s, 1 H), 7.23-7.50 (m, 14H), 5.56 (s, 2H), 4.89-4.92 (m, 1 H), 1.41 (d, J = 6.8Hz, 3H) Example 36

Ethyl 4-(biphenyl-2-ylmethy')-7-oxo-2-{1-phenylethy!ammo)-4,7- dihydrothiazolo[5,4-b]pyridine-6-carboxylate (l₇A-h)

I-7 (Ι-7') was treated with 1-phenylethanamine according to the representative method to obtain compound i_7A-h as a pink solid.

Yield: 1 %

MS (ESS): 510 (M+H)^*

1HNMR cVDMSO, 400 MHz):

δ 8.38 (d, J = 6.8Hz, 1 H), 8.07 (s, 1 H), 7.16-7.46 (m, 14H), 5.38 (s, 2H), 4.83-4.84 (m, 1 H), 4.12-4.17 (m, 2H), 1.39 (d, J = 6.0Hz, 3H), 1.26 (i, J - 7.2Hz, 3H)

Example 37

4-(Biphenyl-2-ylmethyl)-2-(4-chloro-2-fluorophenylsulfonamido)-7-oxo-4,7- dihydrothiazolo[5,4-b]pyridine-6-carboxylic acid {i_9A)

1-7 (1-7') was treated with 2~fiuoro-4-chlorophenyisulfonamide according to the representative method to obtain compound i_SA as a pale white solid.

Yield: 2%

MS (ESI): 571 (M+H)⁺

HNMR {d₆-DMSO, 400 MHz):

δ 8.57 (s, 1 H), 7.69-7.77 (m, 2H), 7.25-7.55 (m, 10H), 5.65 (s, 2H) Example 38

Ethyl 4-{biphenyI-2-ylmethyl)-2-(4-chloro-2-fluorophenylsulfonamido)-

7-oxo-4,7_"dlhydroi ia∑oIo[5,4-b]pyridine-6-carboxyfaie (!_9Α-Ι )

1-7 ([.J'} _Was treated with 2-fluoro-4-chlorophenylsuifonamide according to the representative method to obtain compound l_9A-h as a pa!e white solid.

Yield: 7%

MS (ESI): 599 (M+H)⁺

1HNMR (dg-DMSO, 400 MHz):

5 8.21 (s, 1 H), 7.68-7.77 (m, 2H), 7.26-7.54 (m, 10H), 5.48 (s, 2H), 4.20 (q, J = 7.2 Hz, 2H), 1 .27 (t, J = 7.2 Hz, 3H)

Example 39

4-Benzhydryl-2-(4-chloro-2-fluorophenylsulfonamido)-7-oxo-4,7-dlhydrothiazolo[5,4- b]pyridine-6-carboxy1ic acid (l_9D)

The anaiogue of 1-7 (1-7') with dtphenylmethyi substitution was treated with 2-fiuoro-4- chlorophenylsuifonamide according to the representative method to obtain compound l_9D as a pale white solid.

Yield: 1 %

MS (ESi): 570 (M+Hf

HNMR {cfe-DMSO, 400 MHz):

δ 8.14 (s, 1 H), 7.67-7.74 (m, 2H), 7.40-7.50 (m, 7H), 7.27-7.30 (m, 5H) Example 40

Ethyl 4-benzhydryl-2-(4-chloro-2-fluorophenylsulfonamido)-7-oxo-4,7- dihydrothiazolo[5,4-b]pyridi -6-carboxylate (l_9D-h)

The anaiogue of i-7 (Ι-7') with diphenylmethyl substitution was treated with 2-fluoro-4- chlorophenylsuifonamide according to the representative method to obtain compound I_9D-h as a yellow solid.

Yield: 5%

MS (ESI): 598 (M+H)⁺

1HNMR (ds-DMSO, 400 MHz):

δ 8.01 (s, 1 H), 7.68-7.73 (m, 2H), 7.40-7.49 (m, 7H), 7.29 (s, 4H), 7.12 (s, 1 H), 4.10 (q, J = 7.2 Hz, 2H), 1.14 (t, J = 6.8 Hz, 3H)

Example 41

4-(Biphenyl-2-ylmethyl)-2-(4-cyanophenylsulfonamido)-7-oxo-4,7-dihydrothia2olo[5,4- b] py r i di n e-6-car boxyli c acid (l_10A)

1-7 (Ι-7') was treated with 4-cyanophenylsuifonamide according to the representative method to obtain compound a pale white solid.

Yield: 9%

MS (ESI): 543 (M+H)⁺

¹H NMR (c/_fi-DMSO, 400 MHz):

δ 8.54 (s, 1 H), 8.02 (d, J = 8.0 Hz, 2H), 7.83 (d, J = 8.0 Hz, 2H), 7.57 (t, J = 7.2 Hz, 1 H), 7.46- 7.49 (t, J = 7.6 Hz, 1 H), 7.32-7.41 (m, 5H), 7.24 (d, J = 7.2 Hz, 2H), 5.64 (s, 2H), Exampie 42

Ethy! 4-(biphenyl-2-y!methyl)-2-(4-cyanophenyIsulfonamido)-7-oxo-4,7- dihydrothiazolo[5,4-b]pyridine-6-carboxylate (hoA-h)

f-7 {1-7') was treated with 4-cyanophenylsulfonamsde according to the representative method to obtain compound l₁₀A-h as a pale white soiid.

Yield: 2%

MS (ESI): 571 (M+H)⁺

1HNMR (d₆-DMSO, 400 MHz):

δ 8.20 (s, 1 H), 8.03 (d, J = 7.6Hz, 2H), 7.82 (d, J = 8.0 Hz, 2H), 7.26-7.55 (m, 9H), 5.49 (s, 2H), 4.18 (q, J = 7.2 Hz, 2H), 1.27 (t, J = 7.2 Hz, 3H)

Example 43

Ethyl 4-(biphenyl-2-ylmethyl)-2-(4-{ethoxycarbonyl)phenylsulfonamido)- 7-oxo-4,7-dihydrothiazolo[5,4-b]pyridine-6-carboxylate (ΙΙΟΑ-Η')

f-7 (i-7') was treated with ethyl 4-sulfamoylbenzoate according to the representative method to obtain compound oA-h' as a pale white solid.

Yield: 2%

MS (ESS): 618 (M+H)⁺

¹HNMR (cf_frDMSO, 400 MHz):

δ 8.19 (s, 1 H), 8.06 (d, J - 8.0 Hz, 2H), 7.82 (d, J = 7.6 Hz, 2H), 7.53-7.27 (m, 9H), 5.47 (s, 2H), 4.34 (q, J = 6.8 Hz, 2H), 4.18 (q, J = 6.8 Hz, 2H), 4.20 (q, J - 6.8 Hz, 2H), 1.33 (t, J = 7.2 Hz, 3H), 1.26 (t, J = 7.2 Hz, 3H)

Example 44

4-(Biphenyl-2-yimethyI)-2-(4-methoxyphenylsulfonamido)-7-oxo-4,7-dihydrothiazolo[5,4- b] pyrid i ne-6-carboxy I ic aci

I-7 (I -7') was treated with 4-methoxyphenylsulfonamide according to the representative method to obtain compound l^ as a pale white solid.

Yield: 7%

MS (ESI): 548 ( +H)⁺

1H NMR (de-DMSO, 400 MHz):

δ 8.56 (s, 1 H), 7.59-7.65 (m, 3H), 7.55 (t, J - 7.2 Hz, 1 H), 7.44 (d, J = 7.2 Hz, 1 H), 7.29-7.49 (m, 4H), 7.20 (d, J = 6.8 Hz, 2H), 7.07 (d, J = 7.6 Hz, 2H), 5.63 (s, 2H), 3.83 (s, 3H)

Example 45

Ethyl 4-(biphenyl-2-ylmethyl}-2-(4-methoxyphenylsulfonamido)-7-oxo- 4,7-dihydrothiazolo[5,4-b]pyridine-6-carboxylate (l_11A-h)

1-7 {1-7') was treated with 4-methoxyphenyisuffonamide according to the representative method to obtain compound ! _1A-h as a pale white solid.

Yield: 3%

MS (ESI): 576(M+H)^÷, 169

1H NMR (c/e-DMSO, 400 MHz):

5 8.19 (s, 1 H), 7.60 (d, J = 8.4 Hz, 2H}, 7.55 (t, J = 7.2 Hz, 1 H), 7.48 (t, J = 7.2 Hz, 1 H), 7.33- 7.43 (m, 5H), 7.27 (d, J = 6.8 Hz, 2H), 7.04 (d, J = 8.8 Hz, 2H), 5.48 (s, 2H), 4.18 (q, J = 7.2 Hz, 2H), δ 3.82 (s, 3H), 1.26 (t, J = 7.2 Hz, 3H)

Example 46

4-BenzhydrYl-2-(4-methoxyphenylsulfonamido)-7-oxo-4,7-dihydrothia20lo[5,4- b]pyridine-6-carboxylic acid (li_1D)

The analogue of I-7 {!-?') with diphenylmethyl substitution was treated with 4- methoxyphenylsulfonamide according to the representative method to obtain compound l₁₁₀ as a pale white solid.

Yield: 1 %

MS (ESI): 548(M+H}⁺, 169

1H NMR {de-DMSO, 400 MHz):

δ 8.12 (s, 1 H), 7.60 (d, J = 8.4 Hz, 2H), 7.52 (br, s, 6H), 7.32 (br, s, 4H), 7.26 (s, 1 H), 7.02 (d, J = 8.4 Hz, 2H), 3.83 (s, 3H)

Example 47

Ethyl 4-benzhydryl-2-(4-methoxyphenylsulfonamido)-7-oxo-4,7-dihydro

thiazolo[5,4-b]pyridine-6-carboxylate (l_11D-h)

The analogue of I-7 (1-7') with diphenylmethy! substitution was treated with 4- methoxyphenylsulfonamide according to the representative method to obtain compound ^ -h as a pale white solid,

Yield: 2%

MS (ESI): 576(M+H)\ 169

1H NMR {d₆-DMSO, 400 MHz):

δ 7.98 (s, 1 H), 7.59 (d, J = 8.8 Hz, 2H), 7.51 (br, s, 6H), 7.30 (br, s, 4H), 7.1 1 (s, 1 H), 7.02 (d, J = 8.4 Hz, 2H), 4.10 (q, J ~ 7.2 Hz, 2H), 3.83 (s, 3H), 1.13 (t, J = 7.2 Hz, 3H)

Example 48

4-(Biphenyl-2-ylmethyl)-2-((4-chlorophenyI)methylsulfonamido)-7-oxo-4,7- dihydrothiazolo[5,4-b]pyridine-6-carboxylic acid (I-I2A)

i-7 (1-7') was treated with 4-chlorophenylsulfonamide according to the representative method to obtain compound l^ as a pale white soiid.

Yield: 1 % H NMR (de-DMSO, 400 MHz):

δ 8.51 (s, 1 H), 7.19-7.54 (m, 13H), 5.57 (s, 2H), 4.39 (s, 2H)

Example 49

Ethyl 4-(biphenyl-2-ylmethyl)-2-((4-chlorophenyl)methylsulfonamido)-7

dihydrothiazolo{5,4-b]pyridine-6-carboxylate(l₁₂A-h)

1-7 (1-7') was treated with 4-ch!orophenylsulfonamide according to the representative met to obtain compound !i₂A-h as a pale white solid.

Yield: 2%

MS (ESI): 594 (M+Hf

¹H NMR (de-DMSO, 400 MHz):

6 8.24 (s, 1 H), 8.17 (s, 1 H), 7.25-7.50 (m, 13H), 5.36 (s, 2H), 4.30 (s, 2H), 4.21 (q, J = 7.2 2H), 1.30 (t, J = 7.2 Hz, 3H) Example 50

Ethyl 4-(biphenyl-2-ylmethyl)-2-({2,4-dichlorophenyl)methyisulfonamido)-7-oxo-4,7-di hydrothiazolo[5,4-b]pyridine-6-carboxylate (l_13A-h)

S-7 (Ι-7') was treated with 2,4-dichlorophenylsulfonamide according to the representative method to obtain compound l_13A-h as a pale white solid.

Yield: 1 %

MS (ESI): 628 (M+H)^÷, 169

1H NMR (d DMSO, 400 MHz):

δ 8.16 (s, 1 H), 7.23-7.57 (m, 12H), 5.41 (s, 2H), 4.45 (s, 2H), 4.19 (q, J = 6.8 Hz, 2H), 1 .28 (t, J = 7.2 Hz, 3H)

Example 51

Ethyl 7-(benzhydryloxy)-2-(methylthio)thiazolo[5₎4-b]pyridine-6-carboxylate (l_d-f)

f-5 was treated with (bromomethylene)dibenzene according to the general procedure to obtain compound l_d-f as a pale white solid.

Yield: 1 %

MS (ESI): 437 ( +H)⁺, 105

1H NMR (de-DMSO, 400 MHz):

δ 8.66 (s, 1 H), 7.98 (s, 1 H), 7.57-7.59 (m, 4H), 7.32-7.36 (m, 4H), 7.23-7.26 (m, 2H), 4.42 (q, J = 6.8 Hz, 2H), 2.90 (s, 3H), 1.35-1.38 (t, J = 7.2 Hz, 3H) Example 52

Ethyl 4-(biphenyl-2-ylmethyl)-7-oxo-4,7-dihydrothia2olo[5,4-b]pyridine- 6-carboxylate (l-f-a)

I-6 was treated with zinc in acetic acid to obtain compound l-f-a as a yellow solid.

Yield: 5%

MS (ESI): 391 (M+H)⁺, 130, 105

1H NMR (CDCl₃, 400 MHz):

δ 8.57 (s, 1 H), 8.00 (s, 1 H), 7.32-7.50 (m, 7H), 7.12 (d, J = 6.4 Hz, 2H), 5.28 (s, 2H), 4.35 (q, J = 6.8 Hz, 2H), 1.38 (t, J = 7.2 Hz, 3H)

Example 53

7-Hydroxy-2-(methylthio)thiazolo[5,4-b]pyridine-6-carboxylic acid (l-e-1 )

i-5 was treated with LiOH in ethanoi and water to obtain compound i-e-1 as a paie white solid. Yield: 5%

MS (ESI): 243 ( +H)⁺, 157

1H NMR (c/e-DMSO, 300 MHz):

δ 8.77 (s, 1H), 2,79 (s, 3H)

Example 54

7-(Biphenyl-2-yimethoxy)-2-(methylthio)thia2olo[5,4-b]pyridine-6-carboxylic acid(l-f'-l)

f-5 was treated with 2-(bromomethyl)biphenyl and then LiOH to obtain compound f-f-1 as a pale white solid.

Yield: 5%

MS (ESI): 409 ( +H)⁺, 157

1H NMR (d₆-DMSO, 300 MHz):

δ 13.24 (br.s, 1 H), 8.62 (s, 1 H), 7.87-7.90 (m, 1H), 7.28-7.46 (m, 8H), 5.92 (s, 2H), 2.54 (s, 3H)

Example 55

4-(Biphenyl-2-ylmethyl)-2-(methylthio)-7-oxo-4J-dihydrothiazolo[5,4-b]pyridine^- carboxylic acid (f-f-2)

I-5 was treated with 2-(bromomethyl)bipheny! and then LiOH to obtain compound l-f-2 as a pale white solid.

Yield: 5%

MS (ESI): 409(M÷H)⁺, 157 ¹H NMR (ofe-DMSO, 300 MHz):

δ 15.46 (s, 1 H), 8.57 (s, 1 H), 7.21-7.50 (m, 9H), 5.67 (s, 2H), 2.69 (s, 3H)

Example 56

4-(Bip enyl-2-yimethyi)-2-hydroxy-7-oxo-4,7-dihydrothiazo!o[5,4-fa]pyridine-6-carboxyiic acid(l-h')

f-6 was treated with sodium hydroxide to obtatn compound l-h' as a pale white solid.

Yield: 2%

MS (ESI): 379(M+Hf

¹H NMR (ck-DMSO, 400 MHz):

δ 15.23 (s, 1 H), 12.71 (s, 1 H)_t 8.53 (s, 1 H), 7.26-7.51 (m, 9H), 5.55 (s, 2H)

All of the compounds listed in the following table have been prepared as set out above or by analogous methods.

Activity data for compounds having the general formula (A)

Compounds having the general formula (C)

Key Intermediate !

2-Formyl -succinic acid diethyl ester

To a suspension of sodium (333 mg, 14 mmoi, 1.2 eq) in diethyl ether (7 mL) were added succinic acid diethyl ester (2.1 g, 12 mmoi, 1 eq) and formic acid ethyl ester (1.7 mL, 20 mmoi, 1.7 eq). The mixture was stirred at 40°C for 5 h. Water (10 mL) was added and the aqueous layer was washed with diethyl ether (2 x 10 mL). The aqueous layer was then acidified with a 6N solution of hydrochloric acid and extracted with diethyl ether (3 x 10 mL). The organic layers were dried over magnesium suifate, filtered and evaporated in vacuo to afford the expected compound as orange oil (2.6 g, quant, yield).

Key Intermediate II

-Cyclopropyl-7-oxo-4,7-dihydro-pyrazolo[1,5-a]pyrimidine-6-carboxylic acid ethyl ester

sealed tube, 120°C

To a solution of 5-cyclopropyi-2H-pyrazol-3-ylamine (280 mg, 2.3 mmol, 1 eq) in acetic acid (3 mL) was added 2-ethoxymethyIene-malonic acid diethyl ester (500 pL, 2.5 mmol, 1.1 eq). The mixture was heated at 120°C for 2 h in a sealed tube. After cooling, the precipitate was filtered and washed with ethanol to afford the expected compound as white powder (420 mg, 75% yield).

Key Intermediate III

-lsopropyl-7-oxo- ,7-dihydro-pyrazoloi1,5-a]pyrimidine-6-carboxylic acid ethyl ester

sealed tube, 120°C

To a solution of 5-isopropyl-2H-pyrazol-3-ylamine (2.5 g, 20 mmol, 1 eq) in acetic acid (20 mL) was added 2-ethoxymethylene-maIonic acid diethyl ester (4.4 mL, 22 mmoi, 1.1 eq). The mixture was heated at 120 °C for 3 h in a sealed tube. After cooling, the precipitate was filtered and washed with ethanol to afford the expected compound as beige powder (3.2 g, 65% yield).

Key Intermediate IV

-Cyclopentyl-7-oxo-4,7-dihydro-pyrazolo[1,5-a]pyrimidine-6-carboxylic acid ethyl ester

°C

Step 1 : To a suspension of sodium hydride (350 mg, 8.8 mrnoi, 1.2 eq) in 1 ,4-dioxane (10 mL) was added acetonitrile (450 μΙ_, 8.8 mmol, 1.2 eq). The mixture was stirred at room temperature for 30 min. Then cyclopentanecarboxy!ic acid ethyl ester (660 μΙ_, 7.3 mmol, 1 eq) was added. After stirring for 30 min at room temperature, the mixture was heated at 105°C during 16 h. After cooling, the solvent was evaporated to dryness and water was added (30 mL). The mixture was extracted with dichloromethane (3 x 30 mL) to get rid of the starting material and the aqueous phase was acidified with a 1 solution of hydrochloric acid and extracted with dichloromethane (3 x 30 mL). The combined organic phases were dried over magnesium sulfate, fiitered and dried in vacuo to afford 3-cyclopentyl-3-oxo-propionitrile as very volatile yellow oil (1.0 g, quant, yield)

Step 2:

To a solution of 3-cyclopentyl-3-oxo-propionitrile (1.0 g, 7.3 mmol, 1 eq) in ethanol (10 mL) was added a 64 wt.-% solution of hydrazine hydrate (1.1 mL, 14.6 mmol, 2 eq). The mixture was heated at 80 °C for 16 h and was evaporated to dryness. The residue was purified by flash chromatography using dichloromethane and methanol (100/0 to 90/ 0) to afford 5- cyciopentyi-2H-pyrazol-3-ylamine as yellow oil (510 mg, 46 % yield).

Step 3:

To a solution of 5-cyclopentyl-2H-pyrazol-3-y!amine (510 mg, 3.4 mmol, 1 eq) in acetic acid (4.8 mL) was added 2-ethoxymethylene-malonic acid diethyl ester (750 pL, 3.7 mmol, 1.1 eq). The mixture was heated at 120 °C for 3 h in a sealed tube. After cooling, the precipitate was filtered and washed with ethanol and diethyl ether and recrystaliised from methanol to afford the expected compound as white powder (657 mg, 71 % yield).

MS: 276.1

Mp: decomposes at 300°C

Key Intermediate V

7-Oxo-4,7-dihydro-pyrazolo[1,5-a]pyrimidine-2,6-dicarboxylic acid 6-ethyl ester

HO. Ν-_ΝΛΛ O, Et

To a solution of 5-amino-1 H-pyrazo!e-3-carboxy!ic acid (600 mg, 4.7 mmol, 1 eq) in acetic acid (30 mL) was added 2-ethoxymethylene-maionic acid diethyl ester (1.1 g, 5.2 mmol, 1.1 eq). The mixture was heated at 120 °C for 4 h in a sealed tube. After cooling, the precipitate was filtered and washed with ethanoi to afford the expected compound as grey powder (353 mg, 30% yield).

Genera! Procedure

^MeSY^N*CN R'^{NHs S0}°^C

SMe ste 1

step 3

Step 1 :

To a solution of the appropriate amine (4.3 mmol, 1 eq) in ethanoi (10 mL) was added dimethyl N-cyanodithioiminocarbonate (1.0 g, 6.8 mmol, 1.6 eq). The mixture was stirred at 80 °C for 20 h. After cooling, the precipitate was filtered and rinsed with ethanoi to afford the expected compound (from 25% to 70% yield).

Step 2:

To a solution of the compound from step 1 (1.1 mmol, 1 eq) in ethanoi (10 mL) was added a 1 M soiution of hydrazine in tetrahydrofuran (2.3 mL, 2.3 mmol, 2 eq). The mixture was heated at 80 °C for 20 h and was evaporated to dryness. The product was then triturated with diethyl ether, filtered and washed with diethyl ether to afford the expected compound (from 75% to 85% yield). Step 3:

To a solution of the compound from step 2 (0.86 mmol, 1 eq) in acetic acid (4 mL) was added 2-ethoxymethylene-malonic acid diethyl ester (190 L, 0.94 mmol, 1.1 eq). The mixture was heated at 120 °C for 20 h in a sealed tube. After cooling, the precipitate was filtered and washed with ethanoi to afford the expected compound (from 25% to 65% yield). Example 58

2-Benzylamino-7-oxo-4,7-dihydro-[1 ,2,4}triazolo[1 ,5-a]pyrimidine-6-carboxylic acid ethy! ester

The expected compound was obtained according to general procedure A using benzyiamine. The expected compound was isolated as white powder.

MS: 314.1

Mp: 275°C - 278°C

Example 59

2-(4-Bromobenzylamino)-7-oxo-4,7-dihydro-[1,2,4]tria20lo[1 ,5-aJpyrirnidine-6-carboxyl acid ethyi ester

The expected compound was obtained according to general procedure A using 4-bromo- benzyiamine. The expected compound was isolated as white powder.

MS: 392.2

Mp: 286°C - 287°C

Example 60

2-[(Naphthafen-1 -yimethyi)-amino]-7-oxo-4,7-dihydro-[1 ,2,4]triazolo[1 ,5a]pyrimidine-6- earboxyfie acid ethyl ester

The expected compound was obtained according to general procedure A using C-(2,3- dihydro-naphthalen-1 -y!)-methyiamine. The expected compound was isolated as white powder.

MS: 364.2

Mp: 273°C - 275°C Example 61

2-{4-l sopropoxy-phenyl ami no)-7-oxo-4,7^

carboxylic acid ethyl ester

The expected compound was obtained according to general procedure A using 4- isopropoxy-phenylamine. The expected compound was isolated as pale ye!low powder.

MS: 358.2

Mp: decomposes at 325°C - 330°C

Example 62

2-(4-Acetylamino-phenylamino)-7-oxo-4,7-dihydro-[1₎2,4]triazolo[1,5-a]pyrimi

carboxylic acid ethyl ester

The expected compound was obtained according to general procedure A using N-(4- phenyl)-acetamide. The expected compound was isolated as off-white powder.

MS: 357.2

Mp > 330°C

Example 63

2-{3-Chloro-4-methyl-phenylamino}-7-oxo-4,7-dihydro-[1 ,2,4]trIazoio[1 ,5-a]pyrimidine-6- carboxylic acid ethyl ester

The expected compound was obtained according to general procedure A using 3-chloro~4- methyl-phenylamine. The expected compound was isolated as white powder.

MS: 348.1

Mp > 340°C General Procedure B

step 3

Step 1 :

To a solution of the appropriate amine (4.3 mmoi, 1 eq) in ethanoi (10 mL) were added dimethyl N~cyanodithioiminocarbonate (1.0 g, 6.8 mmol, 1.6 eq). The mixture was stirred at 80 ^DC for 20 h. After cooling, the precipitate was filtered and rinsed with ethanoi to afford the expected compound (from 25% to 70% yield). Step 2:

To a solution of the compound from step 1 (1.1 mmol, 1 eq) in ethanoi (10 mL) were added a 1 solution of hydrazine in tetrahydrofuran (2.3 mL, 2.3 mmol, 2 eq). The mixture was heated at 80 °C for 20 h and was evaporated to dryness. The product was then triturated with diethyl ether, filtered and washed with diethyl ether to afford the expected compound (from 75% to 85% yield).

Step 3:

To a solution of the compound from step 2 (1.2 mmol, 1 eq) in acetic acid (6 mL) were added 2-formyl-succinic acid diethyl ester (Key Intermediate I) (277 mg, 1.37 mmol, 1.1 eq). The mixture was heated in a sealed tube at 120 °C for 20 h. After cooling, the mixture was evaporated to dryness. The residue was diluted in ethyl acetate (10 mL) and washed with a saturated solution of sodium bicarbonate (2 x 10 mL). The organic layers were dried over magnesium sulfate, filtered and evaporated in vacuo. If necessary, the crude compound was purified by flash chromatography using dichloromethane and methanol to afford the expected compound (from 35% to 45% yield).

Exarnpie 64

(7-Oxo-2-phenylamino-4,7-dihydro-[1 ,2,4]triazolo[1 ,5-a]pyrimidin-6-yl)-acetic acid ethyl ester

The expected compound was obtained according to genera! procedure B using aniiine. The expected compound was isolated as white powder.

MS: 314.2

Mp: 255°C - 257°C

Example 65

[2-{4-!sopropoxy-phenylamtno)-7-oxo-4 ~di ydro-[1,2,4jtriazolo[1 ,5-a]pyrirnidin-6-yi]- acetic acid ethyl ester

The expected compound was obtained according to general procedure B using 4- isopropoxy-phenylamine. The expected compound was isolated as pale yellow powder.

MS: 372.2

Mp: 235°C - 240°C General Procedure C

step 1

Step 1 :

To a solution of 2H-pyrazol-3-yiamine (2.3 mmol, 1 eq) in acetic acid (3 mL) was added 2- ethoxymethyiene-malonic acid diethyl ester (500 μΐ_, 2.5 mmol, 1.1 eq). The mixture was heated' at 120 °C for 20 h in a sealed tube. After cooling, the precipitate was filtered and washed with ethanol to afford the expected compound (from 30% to 80% yield). Step 2:

To a solution of the compound from step 1 (1.7 mmol, 1 eq) in ethanol (2 mL) was added sodium hydroxide (170 mg, 4.24 mmol, 2.5 eq) and water (2 mL). The mixture was heated in a sealed tube at 100 °C for 4 h. After cooling, the mixture was evaporated to dryness and water (30 mL) and citric acid (980 mg, 5.1 mmo!, 3 eq) were added. The precipitate obtained was filtered, washed with water and dried under vacuum to afford the expected compound (50% to quant, yield).

Example 66

3-Bromo-2-methyl-7-oxo-4,7-dihydro-pyrazolo[1 ,5-a]pyrimidine-6-carboxyJic acid ethyl ester

The expected compound was obtained according to general procedure C step 1 using 4- bromo-5-methyl-2H~pyrazoi-3-ylamine. The expected compound was isolated as pale yeliow powder.

MS: 300.0

p: decomposes at 270°C - 275°C Example 67

3-Cyano-2-(3-methylamino-propyl)-7-oxo- ,7-dihydro-pyrazolo[1 ,5-a]pyrimidine-6- carboxylic acid ethyl ester

The expected compound was obtained according to general procedure C step 1 using 5- imino-3-(3-methylamino-propyl}-4,5-dihydro-1 H-pyrazole-4-carbonitrile. The expected compound was isolated as white powder.

MS: 304.2

Mp: 285°C - 287°C Example 68

7-Oxo-2-phenyl-4,7-dihydro-pyrazo -a}pyrimidine-6-carboxylic acid

The expected compound was obtained according to general procedure C using 5-phenyl-2H- MS: 256.0

Mp: decomposes at 325°C

Example 69

2-(4-Ethoxy-phenyl)-7-oxo-4₅7-dihydro-pyrazolo[1 ,5-a]pyrimidine-6-carboxylic acid

The expected compound was obtained according to genera! procedure C using 5-{4-ethoxy- phenyi)-2H~pyrazol-3-ylamine. The expected compound was isoiated as white powder.

MS: 300.1

Mp: decomposes at 310°C - 315°C

Exampie 70

2-Cyclopropyl-7-oxo-4,7-dihydro-pyrazolo[1 ,5-a]pyrimidine-6-carboxylic acid

The expected compound was obtained according to general procedure C using cyc!opropyl-2H-pyrazoi-3-ylamine. The expected compound was isolated as white powder. MS: 220.0

Mp: 275°C - 278°C. Example 71

2-lsopropyl-7-oxo-4,7-dihydro-pyrazolo[1 ,5-a]pyrimidine-6-carboxylic acid

The expected compound was obtained according to general procedure C using 5-isopropyl 2H-pyrazoi-3-ylamine. The expected compound was isolated as white powder.

MS: 222.0

Mp: decomposes at 280°C - 285°C

Example 72

2-Cyclopentyl-7-oxo^4,7-dihydro-pyrazolo[1,5-a]pyrimidine-6-carboxylic acid

The expected compound was obtained according to general procedure C using 5- cyclopentyl-2H-pyrazol-3-ylamine. The expected compound was isolated as white powder, MS: 248.1

Mp: decomposes at 300°C

Example 73

nrnidine-6-carboxy[ic acid

The expected compound was obtained according to general procedure C using 5- trifiuoromethyi-2H-pyrazol-3-yiamine. The expected compound was isolated as white powder. MS: 248,0

Mp > 340°C General Procedu

To a solution of 7-oxo-4,7-dihydro-pyrazolo[1 ,5-a]pyrimidine-6-carboxylic acid ethyl ester (0.81 mmoi, 1 eq) in dimethylformamide (5 mL) were added potassium carbonate (560 mg, 4 mmol, 5 eq) and the appropriate bromide (3.2 mmol, 4 eq). The mixture was heated at 50 °C for 4 h. After cooling, the mixture was poured on brine (15 mL) and extracted with ethyl acetate (3 x 20 mL). The organic layers were dried over magnesium sulfate, fi!tered and evaporated in vacuo. The crude residue was purified by flash chromatography using dichloromethane and methanol (100/0 to 95/5) to afford the expected compound (13% to 97% yieid).

Example 74

4-Benzyl-2-cyclopropyl-7-oxo-4,7-dihydro-pyrazolo[1,5-a]pyrimidine-6-carboxylic acid ethyl ester

The expected compound was obtained according to genera! procedure D using Key Intermediate II and benzyl bromide. The expected compound was isolated as white powder. MS: 338.2

Mp: 160°C - 165°C

2-Cyclopropyl-7-oxo-4-phenethyl-4,7-dihydro-pyra2olo[1 ,5-a]pyrimidine-6-carboxyiic acid ethyl ester

The expected compound was obtained according to general procedure D using Key Intermediate If and phenethyl bromide. The expected compound was isolated as white powder.

MS: 352.2

Mp: 155°C ~ 160°C

Example 76

2-Cyclopropyl-4-[2-(4-hydroxy-phenyl)-ethyl]-7-oxo-4,7-dihydro-pyrazolo[1,5- a]pyrimidine-6-carboxyiic acid ethyl ester

The expected compound was obtained according to general procedure D using Key

Intermediate II and 4-(2-bromo-ethyl)-phenol. The expected compound was isolated as white powder. MS: 368.2

Mp: 95°C - 100°C

Example 77

4-[2-(4-Chloro-phenyl)-ethyl]-2-cyclopropyl-7-oxo-4,7-dihydro-pyrazolo[1,5- a]pyrimidine-6-carboxylic acid ethyl ester

The expected compound was obtained according to genera! procedure D using Key Intermediate II and 1-(2-bromo-ethyf)-4-chioro-benzene. The expected compound was isolated as white powder,

MS: 386.2

Mp: 190°C - 195°C

Example 78

2-Cyclopropyi-4-[2-(4-methoxy-phenyl)-ethyl]-7-oxo-4,7-dihydro-pyrazolo[1 ,5- a]pyrimidine-6-carboxylic acid ethyl

The expected compound was obtained according to general procedure D using Key

Intermediate il and 1-(2-bromo-ethyl)-4-methoxy-benzene. The expected compound was isolated as white powder.

MS: 382.2

Mp: 160°C - 165°C

Example 79

4-[2-(3-Chloro-phenyl)-ethyl]-2-cyclopropyl-7-oxo-4,7-dihydro-pyrazolo[1 ,5- a]pyrimidine-6-carboxylic acid ethyl ester

The expected compound was obtained according to general procedure D using Key Intermediate li and 1-(2-bromo-ethyl)-3-chloro-benzene. The expected compound was isolated as white powder.

MS: 386.2

p: 160°C - 165°C

Example 80

2-CyciopropyS-4-[2-{3-fluoro-phenyl)-ethyl]-7-oxo-4,7-dihydro-pyra∑olo[1,5-a]pyrimidine 6-carboxylic acid ethyl ester

The expected compound was obtained according to general procedure D using Key Intermediate II and 1-(2-bromo-ethyl)-3-fluoro-benzene. The expected compound was isolated as white powder.

MS: 370.2

Mp: 160X - 165°C

Example 81

2-Cyclopropyi-7-oxo-4-[2-(3-trifluoromethyl-phenyl)-ethyl}-4,7-dihydro-pyrazolo[ a] py ri m i d i ne-6-ca rboxy i i c acid ethyl ester

The expected compound was obtained according to general procedure 0 using Key intermediate II and 1-(2-bromo-ethyl)-3 rifiuoromethyl-benzene. The expected compound was isolated as white powder.

MS: 420.2

Mp: 140°C - 145°C

Example 82

2-Cyclopropyl-7-oxo-4-(3-phenyl-propyl)-4,7-dihydro-pyrazolo[1 ,5-a]pyrimidine-6- carboxyiic acid ethyl ester

The expected compound was obtained according to general procedure E using Key

Intermediate !E and (3-bromo-propyl)-benzene. The expected compound was isolated as white powder.

MS: 366.2

Mp: 150°C - 155°C

Example 83

4-Ben2yl-2-isopropyl-7-oxo-4,7-dihydro-pyrazolo[1 ,5-a]pyrimidine-6-carboxylic acid ethyl ester

The expected compound was obtained according to general procedure D using Key Intermediate III and benzyl bromide. The expected compound was isolated as white powder. MS: 340.2

Mp: 135X - 140°C

Example 84

2-lsopropyl-7-oxo-4-phenethyl-4,7-dihydro-pyrazolo[1,5-a]pyrimidine-6-carboxylic acid ethyl ester

The expecied compound was obtained according to general procedure D using Key Intermediate III and phenethyl bromide. The expected compound was isolated as white powder.

MS: 354.2

p: 130°C - 135°C

Example 85

2-lsopropyl-7-oxo-4-(3-phenyl-propyl)-4₎7-dihydro-pyrazolo[1,5-a]pyrimidine-6- carboxylic acid ethyl ester

The expected compound was obtained according to genera! procedure D using Key Intermediate III and (3-bromo-propyi)-benzene. The expected compound was isolated as colorless oil

MS: 368.3

Example 86

4-Benzyl-2-cyclopentyl-7-oxo-4,7-dihydro-pyrazolo[1,5-a]pyrimidine-6-carboxylic acid ethyl ester

The expected compound was obtained according to general procedure D using Key Intermediate IV and benzyl bromide. The expected compound was isolated as white powder. MS: 366.2

Mp: 148°C - 150°C Example 87

2-Cyclopentyl-7-oxo-4-phenethy!-4J-dihydro-pyra2olo[1 ,5-a]pyrimidine-6-carboxyIic acid ethyl ester

The expected compound was obtained according to general procedure D using Key Intermediate !V and phenethyl bromide. The expected compound was isolated as white powder.

MS: 380.3

Mp: 162X - 164°C

General Procedure E

sealed tube, 120°C To a solution of 2H-pyrazol-3-ylamine {1.3 mmol, 1 eq) in acetic acid (8 mL) was added 2- formyl-succinic acid diethyl ester (Key Intermediate I) (286 mg, 1.4 mmol, 1.1 eq). The mixture was heated in a sealed tube at 120 °C for 20 h. After cooling, the precipitate was filtered, rinsed with ethanol and dried under vacuum to afford the expected compound (from 18% to 86% yield).

Example 88

(7-Oxo-2-phenyl-4,7-dihydro-pyra -a3pyrimidin-6-yl)-acetic acid ethyl

The expected compound was obtained according to general procedure E using 5-phenyl-2H- pyrazol-3-ylamine. The expected compound was isolated as white powder.

MS: 298.1 p: 245°C - 250

Example 89

(7-Oxo-2-trifluoromethyl-4,7-dihydropyrazolo[1 ,5-a]pyrimidin-6-yl)-acetic acid ethyl ester

The expected compound was obtained according to general procedure E using 5 trifluoromethyl-2H-pyrazol-3-y[amine. The expected compound was isolated as white powder. MS: 290.0

Mp: 290°C - 293°C

Example 90

(2-Cyclopropyl-7-oxo-4,7-dihydro-pyra2olo[1,5-a]pyrimidin-6-yl)-acetic acid ethyl ester

The expected compound was obtained according to general procedure E using 5- cyclopropyl-2H-pyrazol-3-yiamine. The expected compound was isolated as white powder. MS: 262.1

Mp: 280°C - 283°C

Example 91

(2-Cyclopropyl-4-methyl-7-oxo-4,7-dihydro-pyrazolo[1 ,5-a]pyrimidin-6-yl)-acetic acid ethyl ester

To a suspension of (2-cyclopropyi-7-oxo-4,7-dihydro-pyrazolo[1,5-a]pyrimidin-6-yl)-acetic acid ethy! ester (80 mg, 0.3 mmo!, 1 eq) described in example 90 in tetrahydrofuran (2 mL) was added sodium hydride (16 mg, 3.9 mmol, 1.3 eq). The mixture was stirred during 30 min at room temperature and methyl iodide (30 pL, 0.5 mmol, 1.5 eq) was added. The mixture was stirred at room temperature for 5 h. The mixture was then diluted with ethy! acetate (5 mL) and water (5 mL) was added. The aqueous layer was extracted with ethy! acetate (2 x 10 mL) and the aqueous phases were dried over magnesium sulfate, filtered and evaporated in vacuo. The crude residue was purified by flash chromatography using cyclohexane and ethy! acetate (100/0 to 0/100) to afford the expected compound as white powder (16 mg, 59% yield).

MS: 276.1

Mp: 147°C - 150°C

Example 92

(3-Bromo-2-methyl-7-oxo-4,7-dihydro-pyrazolo[1 ,5-a]pyrimidin-6-yl)-acetic acid ethyl ester

The expected compound was obtained according to general procedure E using 4-bromo-5- methyi-2H-pyrazol-3-ylarnine. The expected compound was isolated as pale pink powder. MS: 316.0

Mp: decomposes at 245°C - 250°C

Example 93

2-[2-(4-Chloro-phenyl)-ethylcarbamoyl]-7-oxo-4,7-dihydro-pyrazolo[1 ,5-a]pyrtmidine-6- carboxylic acid ethyl ester

Step 1 :

To a solution of 5-nitro-1 H-pyrazole-3-carboxylic acid (200 mg, 1.3 mmol, 1 eq) in tetrahydrofuran (5 mL) were added triethylamine (350 μΙ_, 1.9 mmol, 1.5 eq), hydroxybenzoiriazole (HOBT) (257 mg, 1.27 mmo!, 1 eq), 2-(4-chloro-phenyl)-ethylamine (180 uL, 1.27 mmol, 1 eq) and EDCI (364 mg, 1.9 mmoi, 1.5 eq). The mixture was stirred at room temperature during 20 h. Water (10 mL) was then added and the aqueous phase was extracted with ethyl acetate (2 x 15 mL). The organic layers were dried over magnesium sulfate, filtered and evaporated in vacuo. The crude residue was purified by flash chromatography using cyclohexane and ethyl acetate (100/0) to (50/50) to afford 5-nitro-1 H- pyrazole-3-carboxylic acid [2-(4-chloro-phenyl)-ethyl]-amide as white solid (160 mg, 43% yield). Step 2:

To a solution of 5-nitro- H-pyrazole-3-carboxylic acid [2-(4-chloro-phenyl)-ethyl]amide (160 mg, 5.42 mmol, 1 eq) in tetrahydrofuran and ethanol (1 mL / 3 mL) was added a saturated solution of ammonium chloride (1 mL) and iron (97 mg, 1.73 mmol, 3.2 eq). The mixture was stirred at 105 °C for 16 h. After cooling, the mixture was filtrated on a short pad of celite and washed with ethanol (10 mL), tetrahydrofuran (10 mL) and water (10 mL). The filtrate was evaporated, water (10 mL) was added and the aqueous phase was extracted with dichloromethane (2 x 15 mL). The organic layers were dried over magnesium sulfate, filtered and evaporated in vacuo to afford 5-amino-1 H-pyrazole-3-carboxylic acid [2-(4-chioro-phenyt)- ethyl]-amide as beige powder (100 mg, 70% yield).

Step 3:

To a solution of 5-amino-1 H-pyrazoie-3-carboxy(ic acid [2-(4-chloro-phenyl)-ethyl]-amide (100 mg, 0.4 mmol, 1 eq) in acetic acid (2 mL) was added 2-ethoxymethylene-malonic acid diethyl ester (80 pL, 0.44 mmol, 1.1 eq). The mixture was heated at 120 °C for 16 h in a sealed tube. After cooling, the precipitate was filtered and washed with ethanol (2 x 10 mL) to afford the expected compound as white powder (55 mg, 38% yield).

MS: 389.2

Jvlp > 300°C General Procedure F

Key Intermediate V To a solution of 7-oxo-4,7-dihydro-pyrazoSo[1 ,5-a]pyrimidine-2,6-dicarboxylic acid 6-ethyl ester (Key intermediate V) (176 mg, 0.7 mmol, 1 eq) in dichloromethane (5 mL) were added triethyiamine (195 μΐ, 1.4 mmoi, 2 eq), HOBT (142 mg, 1.05 mmoi, 1.5 eq), the appropriate amine (0.8 mmoi, 1.1 eq) and EDCI (201 mg, 1.05 mmoi, 1.5 eq). The mixture was stirred at room temperature during 20 h. Water (10 mL) was then added and the aqueous phase was extracted with dichloromethane (2 x 15 mL). The organic layers were dried over magnesium sulfate, filtered and evaporated in vacuo. The crude residue was purified by flash chromatography using dichioromethane and methanol (100/0) to (80/20). The compound obtained was taken up in methanol and filtered to afford the expected compound as white powder (145 mg, 49% yield).

Example 94

2-(1 -Benzylpiperidin-4-ylcarbamoyf)-7-oxo-4,7-dihydropyrazolo 1,5-a3pyrimidine-6- carboxylic acid ethyl ester

The expected compound was obtained according to general procedure F using Key

Intermediate V and 1-benzyS-piperidin-4-ylamine. The expected compound was isolated as white powder.

MS: 424.3

Mp: 264°C - 266X

Example 95

2-Benzylcarbamoyl-7-oxo-4,7-dihydropyrazolo[1 ,5-a]pyrimidine-6-carboxyHc acid ethy! ester

The expected compound was obtained according to general procedure F using Key Intermediate V and benzylamine. The expected compound was isolated as pate grey powder. MS: 341.2

Mp: 290°C - 292°C

General Procedure G

To a solution of the ester (0,32 mmol, 1 eq) in ethanol {6 mL) was added a 5N solution of sodium hydroxide (0.5 mL). The mixture was heated in a sealed tube at 80 °C for 20 h to 48 h. After cooling, the mixture was evaporated to dryness. Then water (5 mL) and citric acid (3 mL) were added. The precipitate obtained was filtered and washed with water to afford the expected compound (65% to quant, yield).

Example 96

2-(4-lsopropoxy-phenyIamino)-7^xo-4J-dihydro-[1,2,4ltriazolo[1,5-a]pyrimiciine-6- carboxy!ic acid

The expected compound was obtained according to genera! procedure G using 2-(4~ isopropoxy-phenylamino)-7-oxo-4,7-dihydro-[1 ,2,4]triazolo[1 ,5-a]pyrimidSne-6-carboxylic acid ethyl ester described in example 61. The expected compound was isolated as yeliow powder. MS: 330.1

Mp: decomposes at 260°C - 265X

Example 97

2-Benzylamino-7-oxo-4,7-dihydr -a]pyrimidine-6-carboxylic acid

The expected compound was obtained according to general procedure G using 2- benzyiamino- -oxo^.y-dihydro-fl ^^jtriazolofl^-ajpyrimidine-e-carboxyiic acid ethyl ester described in example 58. The expected compound was isolated as pale yellow powder.

MS: 286.1

Mp: 240°C - 245°C.

Example 98 2-[(Naphfhalen-1 -yimethyl)-amino]-7-oxo-4,7-dihydro-[1 ,2,4]triazolo[1 ,5a]pyrimidine-6- carboxylic acid

The expected compound was obtained according to general procedure G using 2- [(naphthaien-l-ylmethylJ-aminoj- -oxo^ -dihydrofl ^.^triazoioCl ^ajpyrimidine-B-carboxyiic acid ethyl ester described in example 60. The expected compound was isolated as pate orange powder.

MS: 336.1

Mp: 245°C - 250°C. Example 99

2-[(Benzo[1 ,3]dioxol-5-ylmethyl)-amino]-7-oxo-4,7-dihydro-[1 ,2,4]triazolo[1 ,5- a]pyrimidine-6-carboxylic acid sodium salt

The expected compound was obtained according to general procedure G using 2- [(benzo[1 ,3]dioxol-5-ylmethyl)-amino]-7-oxo-4,7-dihydro-[1 ,2,4]triazolo[1.5-a]pyrimidine-6- carboxyiic acid ethyl ester. This starting material was obtained according to general procedure A using C-benzo[1 ,3]dioxol-5-yl-methylamine. The expected acid was isolated without treatment as sodium salt and as yellow powder.

MS: 330.1

Mp decomposes at 300°C.

Example 100

(7-Oxo-2-phenylamino-4,7-dihydro- 1 ,2,4]triazolo[1,5-a]pyrimidin-6-yl)-acetic acid

The expected compound was obtained according to generai procedure G using {7-oxo-2- phenylamino-4,7-dihydro-[1 ,2,4]iriazoio[1 ,5-a]pyrimidin-6~y])-aCetic acid ethyl ester described in example 64. The expected compound was isolated as white powder.

MS: 286.1

Mp: 279°C - 281 °C.

Example 101

[2-(4-lsopropoxy-phenylamino)-7-oxo-4,7-dihydro-[1,2,4}triazolo[1,5-a3pyrimidin-6-yl]- acetic acid

The expected compound was obtained according to general procedure G using [2-(4- isopropoxy-phenylamino)-7-oxo-4,7-dihydro-[1 ,2,4]triazoio[1 ,5-a3pyrimidin-6-yl]-acetic acid ethyl ester described in example 65.

Example 102

4-Benzyl-2-cyclopropyl-7-oxo-4,7-dihydro-pyra2olo[1,5-a]pyrimidine-6-carboxylic acid

The expected compound was obtained according to general procedure G using 4-benzyl-2- cyciopropyl-7-oxo-4,7-dihydro-pyrazolo[1 ,5-a]pyrimidine-6-carboxylic acid ethyl ester described in example 74. The expected compound was isolated as beige powder.

Mp: 210°C - 215

Example 103

2-Cyclopropyi-7-oxo-4-phenethyl-4,7-dihydro-pyrazolo[1,5-a]pyrimidine-6-carboxylic acid

The expected compound was obtained according to general procedure G using 2- cydopropyl-7-oxo-4-phenethyl-4,7-dihydro-pyrazolo[1 ,5-a]pyrimidine-6-carboxyfic acid ethyi ester described in example 75. The expected compound was isolated as beige powder.

MS: 324.1

Mp: 185°C - 190°C

Example 104

2-Cyclopropyl-4-[2-(4-hydroxy-phenyl)-ethyl]-7-oxo-4,7-dihydro-pyrazolo[1,5-a]-

The expected compound was obtained according to general procedure G using 2- cyciopropyl-4-[2-{4-hydroxy-phenyl)-ethyi]-7-oxo-4,7-dihydro-pyrazoio[1 ,5-a]pyrimidine-6- carboxylic acid ethyl ester described in example 76. The expected compound was isolated as white powder.

MS: 340.1

Mp: 265°C - 270°C

Example 105

4-[2-(4-Chloro-phenyl)-ethyl]-2-cyclopropyl-7-oxo-4,7-dihydro-pyrazolo[1,5- a] py r i m i d i n e-6-car boxy I i c acid

The expected compound was obtained according to genera! procedure G using 4-[2-(4- chloro-phenyl)-ethyl]-2-cyclopropyl-7-oxo-4,7-dihydro-pyrazolo[1 ,5-a]pyrimidine-6-carboxylic acid ethyl ester described in example 77, The expected compound was isolated as white powder.

MS: 358.1

Mp: 220°C ~ 225°C

Example 106

2-Cyclopropyl-4-[2-(4-methoxy-phenyl)-ethyl]-7-oxo-4,7-dihydro-pyrazolo[1,5-a]- pyrimidine-6-carboxylic acid

The expected compound was obtained according to general procedure G using 2- cyclopropyl-4-[2-(4-methoxy-phenyl)-ethy[ -7-oxo-4,7-dihydro-pyrazolo[1 ,5-a]pyrimidine-6- carboxylic acid ethyl ester described in example 78. The expected compound was isolated as white powder.

MS: 354.2

Mp: 145°C - 150°C

Example 107

2-Cyclopropyl-7-oxo-4-[2-(4-trifluoromethyl-phenyl)-ethyl]-4,7-dihydro-pyra20lo[1 ,5-a] pyrimidine-6-carboxyiic acid

The expected compound was obtained according to general procedure G using 2- cyclopropyl-7-oxo-4-[2-(4-trifluoromethyl-phenyl)-ethyl]-4,7-dihydro-pyrazoio[ _l5-a]pyrimidine- 6-carboxylic acid ethyl ester. The starting materia! was obtained according to general procedure D using Key intermediate i! and 1 -(2-bromo-ethyl)-4-irifiuoromethy!-benzene. The expected compound was isolated as white powder.

MS: 392.2

Mp: 225°C - 230°C

Example 108

4-[2-{3-Ch!oro-phenyi)-ethyI]-2-cycfopropyl-7-oxo-4,7-dihydro-pyrazolo[1_I5-aJ- pyrimidine-6-ca r boxy i i c acid

The expected compound was obtained according to general procedure G using 4-[2-(3- chloro-pheny -ethyI]-2-cyclopropyl-7-oxo-4,7-dihydro-pyrazo!o[1 ,5-a]pyrimidine-6-carboxylic acid ethyl ester described in example 79. The expected compound was isolated as white powder.

MS: 358.1

Mp: 230°C - 235°C

Example 109

2-Cyclopropyl-4-[2-{3-fiuoro-phenyl)-ethyl]-7-oxo-4,7-dihydro-pyrazolo[1 ,5-a]pyrimidine- 6 -carboxylic acid

The expected compound was obtained according to general procedure G using 2- cycSopropy!-4-[2-(3-fluoro-pheny[}-ethyl]~7-oxo-4,7-d!hydro-pyrazolo[1 ,5-a]pyrimidine-6- carboxylic acid ethyi ester described in example 80. The expected compound was isolated as white powder.

MS: 342.1

Mp: 220°C - 225°C Example 110

2-Cyciopropyl-7-oxo-4-[2-(3-trifluoromethyl-pheny!)-ethyl]-4,7-dihydro-pyra2olo[1 ,5-a]- pyrimidine-6-carboxylic acid

The expected compound was obtained according to general procedure G using 2- cyclopropyl-7-oxo-4-[2-{3-trifiuoromethyl-phenyl)-ethyi]-4,7-dihydro-pyrazolo[1 ,5-a]pyrimidine~ 6-carboxylic acid ethyl ester described in example 81. The expected compound was isolated as white powder.

MS: 392.2

Mp: 200°C - 205°C

Example 111

2-Cyclopropyi-7-oxo-4-(3-phenyl-propyl)-4,7-dihydro-pyrazolo[1,5-a]pyrimidine-6- carboxyiic acid

The expected compound was obtained according to general procedure G using 2- cyclopropyi-7-oxo-4-(3-phenyl-propyl)-4,7-dihydro-pyrazolo[1 ,5-a]pyrimidine-6-carboxylic acid ethyl ester described in example 82. The expected compound was isolated as beige powder. MS: 338.2

Mp: 95°C - 100°C

Example 112

4-Benzyl-2-isopropyl-7-oxo-4,7-dihydro-pyrazolo[1,5-a]pyrimidine-6-carboxylic acid

The expected compound was obtained according to genera! procedure G using 4-benzyS-2- isopropyf-7-oxo-4,7-dihydro-pyrazolo[1 ,5-a]pyrimidine-6-carboxylic acid ethyl ester described in example 83. The expected compound was isolated as beige powder.

MS: 312.1

Mp: 180°C ~ 185°C

Example 113

2-lsopropyl-7-oxo-4-phenethyf-4,7-dihydro-pyrazolo[1,5-a}pyrimidine-6-carboxylic acid

The expected compound was obtained according to genera! procedure G using 2-isopropyl- 7-oxo-4-phenethyl-4,7-dihydro-pyrazolo[1 ,5-a]pyrimidine-6-carboxylic acid ethyl ester described in example 84. The expected compound was isolated as white powder.

MS: 326.2

Mp: 220°C - 225°C

2-lsopropyl-7-oxo-4-(3-phenyl-propyl)-4,7-dihydro-pyrazolo[1 ,5-a]pyrimidine-6- carboxylic acid

The expected compound was obtained according to genera! procedure G using 2-isopropyl- 7-oxo-4-(3-phenyi-propyl)-4,7-dihydro-pyrazo!o[1 ,5-a]pyrtmidine-6-carboxylic acid ethyl ester described in example 85. The expected compound was isolated as orange oil.

MS: 340.2

Example 115

4-Benzyl-2-cyclopentyl-7-oxo-4,7-dihydro-pyrazolo[1,5-a]pyrimidine-6-carboxylic acid

The expected compound was obtained according to general procedure G using 4-benzyi-2- cyclopentyl-7-oxo-4,7-dihydro-pyrazolo[1 ,5-a]pyrimidine-6-carboxylic acid ethyl ester described in example 86. The expected compound was isolated as white powder.

Mp: 213°C - 2.15°C

Example 116

2-Cyclopentyi-7-oxo-4-phenethyi-4,7-dihydro-pyrazolo[1,5-a]pyrimidine-6-carboxyHc acid

The expected compound was obtained according to general procedure G using 2- cyciopenty!-7-oxo-4-phenethy(-4,7-dihydro-pyrazolo[1 ,5-a]pyrimidine-6-carboxylic acid ethyi ester described in example 87. The expected compound was isolated as white powder.

MS: 352.2

Mp: 198°C - 200°C

Example 117

{7-Oxo-2-phenyl-4,7-dihydro-pyrazolo[1,5-a]pyrimidin-6-yl)-acetic acid

The expected compound was obtained according to genera! procedure G using (2-phenyi-7- oxo-4,7-dihydro-pyrazoio[1 ,5-a]pyrimidin-6-yl)-acetic acid ethyi ester described in example 88. The expected compound was isolated as beige powder.

MS: 270.1

Mp decomposes at 285°C - 290°C Example 118

[2-(4-Et oxy-phenyf}-7-oxo-4,7-dihydro-pyrazoio[1,5-a]pyrimidiri-6-yt3-acetic acid

The expected compound was obtained according to general procedure G using [2-(4-ethoxy- phenyl)-7-oxo-4,7-dihydro-pyrazoloE1 ,5-a3pyrimidin-6-yl]-acetic acid ethyi ester. The starting material was obtained according to general procedure E using 5-(4-ethoxy-phenyl)-2H- pyrazol-3-yiamine. The expected compound was isolated as white powder.

MS: 314.1

Mp: decomposes at 295°C - 300°C

Example 1 9

{7-Oxo-2-trifluoromethyl-4,7-dihydro-pyrazolo[1,5-a]pyrimidin-6-yl)-acetic acid

The expected compound was obtained according to general procedure G using (7-oxo-2- trifluoromethyl-4,7-dthydropyra20(o[1 ,5-a]pyrimidin-6-y!)-acetic acid ethyl ester described in example 89. The expected compound was isolated as pale salmon colored powder.

MS; 262.0

Mp: 320°C - 324°C

Example 120

(2-Cyclopropyl-7-oxo-4,7-dihydro-pyrazolo[1,5-a]pyrimidin-6-yl)-acetic acid

The expected compound was obtained according to general procedure G using (2- cyciopropyl-7-oxo-4,7-dihydro-pyrazolo[1 ,5-a]pynmidin-6-yl)-acetsc acid ethyl ester described in example 90. The expected compound was isolated as white powder.

MS: 234.1

Mp > 300°C

Example 121 2-[2-(4-Chioro^henyi}-ethylcarbamoy!]-7-oxo^J-dihydro-pyrazolo[1,5-a]pyrimidine-6- carboxylic acid

The expected compound was obtained according to general procedure G using [2-[2-{4- chloro-pheny[)-ethylcarbamoyi]-7-oxo-4,7-dihydro-pyrazoio[1 ,5-a]pyrtmidine-6-carboxylic acid ethyl ester described in example 93. The expected compound was isolated as white powder. MS: 361.1

Mp > 300°C Example 122

Sodium 2-(1 -benzyl-piperi din-4-ylcarbamoyl)-7-oxo-4,7-dihydro-pyrazoio[1 ,5-a]- pyri mi di ne-6-carboxyiate

The expected compound was obtained according to general procedure G using 2-(1~ benzylpiperidin-4-yicarbamoy!)-7-oxo-4,7-dihydropyrazolo[1 ^-ajpynmidine-e-carboxylic acid ethyl ester described in example 94. Instead of the described treatment, the precipitate obtained was filtered to isolate the expected compound as the sodium salt and as white powder.

MS: 396.2

Mp: decomposes at 300°C

General Procedure H

Key Intermediate VI Key Intermediate VII

Step 1:

1 H-1 ,2,4-Triazole-3,5-diamine (12.4 g, 0.125 mol) was dissolved in AcOH (50 ml), and diethyl 2-(ethoxymethylene) malonaie (32.5 g, 0.15 mol) was added. The solution was refluxed overnight, then cooled, filtered, and dried to give Key intermediate VI (22 g, 79%) as a white solid.

Step2:

To a mixture of VI (500 mg, 2.2 mmoi) in N-methylpyrroiidone (20 ml), K₂C0₃ (619 mg, 4.5 mmol) and RBr (3.4 mmol) were added. The solution was stirred at 50 °C over night. The solution was cooled, filtered, and concentrated. The solid was washed with eOH (20 ml), and dried to give Key Intermediate VII as a white solid.

A mixture of VII and NaOH (2.0 eq. (mmol)) in CH₃OH/THF/H₂0 (5/5/1 ) was stirred at r.t. for 2 h. The solvent was removed in vacuum. The residue was dissolved in water (20 ml), the pH value was adjusted to 6, then filtered, and dried to give desired compounds as a white solid.

Example 123

2 -Am i n o-4-benzyl -7-oxo-4, 7-d i hyd ro-{1 ,2,4Jtriazolo[1 , 5-a] yri midi ne-6-carboxyl ic acid

VI was treated with benzylbromide according to the general procedure H to obtain compound

66 as a white solid.

Yield: 10 %

MS (ESI): 286 (M+H)⁺

¹H NMR (d₆-DMSO, 300 MHz):

δ 12.87 (br, s, 1 H), 8.86 (s, 1 H),7.34-7.41 (m, 5H), 6.42 (s, 2H), 5.43 (s, 2H)

Example 124

2-Amino-7 )XO-4-phenethyl ,7-dihydro-[1,2,4)triazolo[1,5-alpyrimidine-6-carboxylic acid

VI was treated with phenethylbromide according to the general procedure H to obtain compound 67 as a white solid.

Yield: 11 %

MS (ESI): 300 ( +Hf

¹H NMR (d₆-DMSO, 300 MHz):

δ 12.84 (s, 1 H), 8.69 (s, 1 H),7.30-7.40 (m, 5H), 6.54 (s, 2H), 4.49 (t, J = 7.2 Hz,2H), 3.19 (t, J = 7.2 Hz,2H)

¹³C NMR (de-DMSO, 300 MHz):

Example 125

2-Amino^-(cyclohexylmethyl)-7-oxo^ -dihydro-[1 ,2,4]triazolo[1 ,5-a3pyrimidine-6- carboxylic acid

VI was treated with (bromomethyl)cyclohexane according to the genera! procedure H to obtain compound 68 as a white solid.

Yield: 10 %

MS (ESI): 292 (M+Hf

¹H NMR (de-DMSO, 300 MHz):

δ 12.86 (s, 1H), 8.69 (s, 1 H), 6.44 (s, 2H), 4.05 (d, J = 7.2 Hz,2H), 1.89-1.95 (m, 1 H), 1.56- 1.67 (m, 5H), 0.90-1.15 (m, 5H)

Example 126

2-Amino-4-isopropyl-7-oxo-4,7-dihydro-[ ,2,4]triazolo[1 ,5-a3pyrimidine-6-carboxylic acid

VI was treated with 2-bromopropane according to the general procedure H to obtain compound 69 as a white solid.

Yield: 1 1 %

MS (ESI): 238 (M+H)⁺

1H NMR (de-DMSO, 300 MHz):

δ 12.97 (s, 1 H), 8.71 (s, 1 H), 6.50 (s, 2H), 4.86-4.95 (m, 1 H), 1.58 (d, J = 6.6 Hz, 6H)

Example 127

2-Amino-4-(biphenyi-2-ylmethyl)-7^

carboxyiic acid

V! was treated with 2-(bromomeihy!)biphenyl according to the genera! procedure H to obtain compound 70 as a white solid.

Yield: 13 %

MS (ESI): 362 (M+H)⁺

H NMR (de-DMSO, 300 MHz):

6 12.76 (br, s, 1 H), 8.47 (s, 1 H), 7.34-7.47 (m, 7H), 7.20-7.29 (m, 2H), 6.32 (s, 2H), 5.39 (s, 2H) Examples 128 and 129

2-Amino-4-[1 -adamantyl]-7-oxo-4,7-dihydro-[1 ,2,4]triazolo[1 , 5-a] py rim i d i n e-6-carboxy I i c acid and 2~arnino~ *[1-adamaniy!]=[1 ,2,4]triazoio[1,5=ajpyrtmidin=7{4H}-one

128 129 VI was treated with 1-bromoadamantane according to the general procedure to obtain compounds 128 and 129 as a brown solid.

Yield: 5 %

MS (ESI): 330 (M+H)⁺ , 286

A19, ¹H NMR (CDCI₃, 300 MHz):

δ 8.46 (s, 1 H), 2.00-2.22 (m, 9H), 1.58-1.70 (m, 3H)

A19-0, Ή NMR (CDC , 300 MHz):

δ 7.69 (d, J = 6.6 Hz, 1 H), 5.73 (d, J - 6.6 Hz,1 H) 2.00-2.22 (m, 9H), 1.58-1.70 (m, 3H) Activity data for compounds having general formula (C)

Claims

A compound having the general formula (C), optionally in the form of a pharmaceutically acceptable salt, solvate, polymorph, codrug, cocrystai, prodrug, tautomer, racemate, enantiomer, or diastereomer or mixture thereof,

wherein

V is N, or CR^e;

X¹ is O, S, or NR⁸;

X² is NR⁵, N(R⁵)C(0), C(0)NR⁵, O, C(O), C(0)0, OC{0); N(R⁵)S0₂, S0₂N(R⁵), S, SO, S0₂;

R^* is -H, -Hal, -(optionally substituted C-i„₆ alkyl), -(optionally substituted mono- or po!ycyciic group containing 3 to 20 carbon atoms and optionally 1 to 4 heteroatoms selected from O, N and S),— Ci_4 a!kyl— (optionally substituted mono- or po!ycyclic group containing 3 to 20 carbon atoms and optionally 1 to 4 heteroatoms selected from O, N and S) or -X²-R¹;

R¹ is -H, -(optionally substituted alkyl), -(optionally substituted mono- or polycyclic group containing 3 to 20 carbon atoms and optionally 1 to 4 heteroatoms selected from O, N and S), -C_M aikyl-(optionally substituted mono- or polycyclic group containing 3 to 20 carbon atoms and optionally 1 to 4 heteroatoms selected from O, N and S);

R² is -H, -(optionally substituted Ci_₃ alkyl), -(optionally substituted C₃_₇ cyctoa!kyl), -(optionally substituted aryi), - _4 alkyl— (optionally substituted C₃^₇ cycloalkyl), or -d-4 alkyl-(optionally substituted aryl) or if X¹ is NR' then R² can also be -OH;

R³ is -H, -(optionally substituted C_1-6 alky!), -R⁷, or -X²-R⁷;

R⁴ is -H, -(optionally substituted d-e alkyl), -(optionally substituted C₃_ cycloa!kyl), -(optionally substituted aryl), -C_M alkyi— (optionaliy substituted C₃_₇ cycloalkyl), or -d-4 aikyl— (optionally substituted aryl);

R⁵ is -H, -(optionally substituted d_e alkyl), -(optionally substituted C₃_₇ cycloalkyl), -(optionally substituted aryl), -C_M alkyl— (optionally substituted C₃„₇ cycloalkyl), or -d-4 alkyl-(optionaliy substituted aryi);

R⁶ H, -Ci-6 alkyl, -aryl, halogen or CN;

R⁷ is -(optionally substituted hydrocarbon group which contains from 5 to 20 carbon atoms and optionally 1 to 4 heteroatoms selected from O, N and S and which contains at least one ring);

R⁸ is -H, or -d-e alkyl; and

n is 0 to 4; wherein the optional substituent of the alky! group is selected from the group consisting of halogen, -CN, -NR⁵R⁵, -OH, and -0-d_₆ alkyl;

wherein the optional substituent of the cycloalkyl group, the aryl group, the mono- or polycyclic group or the hydrocarbon group is selected from the group consisting of -d-s alkyl, halogen, -CF₃, -CN, -X²-R^s and -d-4 alkyl— aryl;

wherein the compound is for use in the treatment, amelioration or prevention of a viral disease.

The compound according to claim 1 , wherein R^" is -(optionally substituted C₃_₇ cycloalkyl).

The compound according to claim 1 or 2, wherein X¹ is 0.

The compound according to any of claims 1 to 3, wherein R² is -H or -(optionally substituted _6 alkyl) or if X¹ is NR' then R² can also be -OH.

The compound according to any of claims 1 to 4, wherein R³ is -H, -d~ alkyi— (optionally substituted aryl) or S0₂-R⁵.

6. The compound according to any of ciaims 1 to 5, wherein R⁴ is -H, or -(optionally substituted Ci_6 alkyi). 7. A method of treating, ameliorating or preventing a viral disease, the method comprising administering to a patient in need thereof an effective amount of a compound having the general formula (C) as defined in any of claims 1 to 6, optionally in the form of a pharmaceutically acceptable salt, solvate, polymorph, codrug, cocrysta!, prodrug, tautomer, racemate, enantiomer, or diastereomer or mixture thereof.

8. The compound according to any of claims 1 to 6 or the method according to claim 7, wherein the viral disease is caused by Herpesviridae, Retroviridae, Filoviridae, Paramyxoviridae, Rhabdoviridae, Orthomyxoviridae, Bunyaviridae, Arenaviridae, Coronaviridae, Picornaviridae, Togaviridae, Flaviviridae.

9. The compound or method according to claim 8, wherein the viral disease is influenza.

10. The compound or method according to any of claims 1 to 9, wherein a further antiviral agent is to be administered concurrently or sequentially with the compound having the genera! formula (C).

1 1 . A pharmaceutical composition comprising:

(i) a compound having the genera! formula (C) as defined in claim 1 ; and

(ii) a compound having the genera! formula (A), optionally in the form of a pharmaceutically acceptable salt, solvate, polymorph, codrug, cocrystal, prodrug, tautomer, racemate, enantiomer, or diastereomer or mixture thereof,

wherein

R^* is -H, -Hal, -{optionally substituted a!kyl), -(optionally substituted CV₇ cycloalkyl), -(optionally substituted aryi), -C1- a!kyl-(optionaliy substituted C₃_₇ cycloalkyl), -C1. aikyl-(optionalfy substituted aryl) or -X¹-R¹;

X¹ is O, C(O), C(0)0, OC(O); S, SO, SQ₂, NR⁴, N(R⁵)C(0), C(0)NR^s;

X² is O, S, NR⁴;

X³ is O or S;

X⁴ is O or S;

R¹ is -H, -(optionally substituted Ci_6 alkyi), -(optionally substituted C₃_₇ cycloalkyl), -(optionally substituted aryi), -C_H alkyi— (optionally substituted C₃„₇ cycloalkyl), -C1-4 aikyl-(optionaliy substituted aryl);

R² is a hydrocarbon group which contains from 5 to 20 carbon atoms and optionally 1 to 4 heteroatoms selected from 0, N and S and which contains at least one ring, wherein the hydrocarbon group can be optionafly substituted;

R³ is -H, -(optionally substituted Ci_e alkyi), -(optionally substituted C^.₇ cycloalkyl), -(optionally substituted aryl), or -C_M alkyi— (optionally substituted aryl) or if X² is NR⁴, then R³ can also be -OH;

R⁴ is -H, -(optionally substituted alkyi), -(optionally substituted C₃_₇ cycloalkyl), -(optionally substituted aryl), -C_H alkyi— (optionally substituted C₃_₇ cycloalkyl), or -C _-JS alkyi— (optionally substituted aryl) or if X¹ is NR⁴, then R⁴ and R¹ can be joined together to form a 5- to 7-membered ring, which can optionally contain O, S or further N or if X² is NR⁴, then R⁴ and R³ can be joined together to form a 5- to 7-membered ring, which can optionally contain O, S or further N; and

R⁵ is -H, -(optionally substituted C-_₆ alkyi), -(optionally substituted C₃_₇ cycloalkyl), -(optionally substituted aryl), -C_1- alkyi— (optionally substituted C₃_₇ cycloalkyl), or -C^ alkyi— (optionally substituted aryl); and

R⁶ is -H, or -Ci_6 alkyi;

wherein the optional substituent of the alkyi group is selected from the group consisting of halogen, -CN, -NR⁶R⁶, -OH, and -0-d_₆ alkyi;

wherein the optional substituent of the cycloalkyl group, the aryl group or the hydrocarbon group is selected from the group consisting of -C^ alkyi, halogen, -CF₃, - CN, -X¹-R⁵ and -C_w aikyl-aryl; and optionally one or more pharmaceutically acceptable excipient(s) and/or carrier(s).

12. A pharmaceutical composition comprising:

(i) a compound having the genera! formula (C) as defined in claim 1 or a compound having the general formula (A) as defined in claim 1 1; and

(ii) at least one polymerase inhibitor which is different from the compound having the general formula (C); and optionally one or more pharmaceuticaliy acceptable excipient(s) and/or carrier(s).

13. A pharmaceutical composition comprising:

(i) a compound having the general formula (C) as defined in claim 1 or a compound . having the general formula (A) as defined in ciaim 11; and

(ii) at least one neuramidase inhibitor; and optionally one or more pharmaceutically acceptable excipieni(s) and/or carrier(s).

14. A pharmaceutical composition comprising:

(i) a compound having the general formula (C) as defined in claim 1 or a compound having the general formula (A) as defined in claim 11 ; and

(ii) at least one M2 channel inhibitor; and optionally one or more pharmaceutically acceptable excipient(s) and/or carrier(s).

15. A pharmaceutical composition comprising:

(ii) at ieast one alpha glucosidase inhibitor; and optionally one or more pharmaceutically acceptable excipient(s) and/or carrier(s). 16. A pharmaceutical composition comprising: (i) a compound having the general formula (C) as defined in claim 1 or a compound having the general formula (A) as defined in claim 11; and

(ii) at least one ligand of another influenza target; and optionally one or more pharmaceutically acceptable excipient(s) and/or carher(s).

17. A pharmaceutical composition comprising: a compound having the general formula (C) as defined in claim 1 or a compound having the general formula (A) as defined in claim 1 1 ; and

at least one medicament selected from antibiotics, anti-inflammatory agents, lipoxygenase inhibitors, EP ligands, bradykinin ligands, and cannabinoid Sigands; and optionally one or more pharmaceutically acceptable excipient(s) and/or carrier(s).

The pharmaceutical composition as defined in any of claims 11 to 17 for use in the treatment, amelioration or prevention of a viral disease.

A method of treating, ameliorating or preventing a viral disease, the method comprising administering to a patient in need thereof an effective amount of a pharmaceutical composition as defined in any of claims 11 to 17.

The pharmaceutical composition according to claim 18 or the method according to claim 19, wherein the viral disease is caused by Herpesviridae, Retroviridae, Filoviridae, Paramyxoviridae, Rhabdoviridae, Orthomyxoviridae, Bunyaviridae, Arenaviridae, Coronaviridae, Picornaviridae, Togaviridae, Flaviviridae; more specifically wherein the virai disease is influenza. 21 The compound, pharmaceutical composition or method according to any of the preceding claims, wherein the compound having the general formula (C) exhibits a % reduction of at least about 30 % at 50 μΜ in the CPE assay disclosed herein. The compound, pharmaceutical composition or method according to any of the preceding claims, wherein the compound having the general formula (C) exhibits an IC₅₀ of at least about 40 μ in the FRET endonuclease activity assay disclosed herein.