ES2358550T3 - BICYCLIC PYRIMIDINONES AND ITS USES. - Google Patents

Abstract

A compound of Formula I or a hydrate, salt or prodrug thereof in which: A is a monocyclic or bicyclic aromatic or heteroaromatic moiety fused with the nitrogen containing ring; X is selected from the group consisting of O and S; And it is selected from the group consisting of O and S; R1 has 0-3 substituents each of which is independently selected from the group consisting of CN, C1-10 alkyl, C2-10 alkenyl, C1-10PO3H2 alkyl, -O-C1-10 alkyl, C1-10NR3R4 alkyl, -O-C1-10NR3R4 alkyl, halo, NR3R4, alkylaryl, alkylheteroaryl, aryl, heteroaryl, -O-alkylaryl, SO2NR3R4; R3 and R4 are each independently selected from the group consisting of hydrogen, CO2 C1-4 alkyl, C (O) C1-4 alkyl C1-10 alkyl, C1-10NR5R6, -NH (CO) (CO) NH C1 alkyl -4, or R3 and R4 taken together with the bound nitrogen form a 5-7 membered heterocyclic ring containing from zero to two additional heteroatoms selected from N, O or S, in which S may be in the oxidation state S , S (O) or S (O) 2 and wherein said heterocyclic ring is optionally substituted on the carbon or nitrogen atoms with one or more substituents selected from halo, aryl, C (O) C1-4 alkyl, SO2 alkyl C1-4, SO2H, C1-4 alkyl, CO2H, CO2 C1-4 alkyl, NR5R8; C1-4NR5R6 alkyl; R5 and R6 are each independently selected from the group consisting of H, C1-4 or R5 alkyl and R6 and together with the attached nitrogen form a 5-7 membered heterocyclic ring containing from zero to two additional heteroatoms selected from N , O or S, in which S may be in the oxidation state S, S (O) or S (O) 2 and in which said heterocyclic ring is optionally substituted at the carbon or nitrogen atoms with one or more substituents selected from halo and C1-4 alkyl; when R1 is alkylaryl or -O-alkylaryl, the aryl group of said alkylaryl substituent is optionally substituted with a substituent selected from C1-10 alkyl, -O-C1-10 alkyl, C1-10NR3R4 alkyl, -O-C1-10NR3R4 alkyl , halo, NR3R4, alkylaryl, -O-alkylaryl, SO2NR3R4 B is absent or is -C (O) -; C is absent or is selected from the group consisting of -O-, -NH- and -NH-NH-C (O) -; R2 is selected from the group consisting of heteroaryl and heterocyclyl; and wherein said prodrug comprises (i) a compound in which an amino acid residue, or a polypeptide chain or two or more amino acid residues covalently binds to an amino, hydroxyl or carboxylic acid group of the compound of formula I, amino acid residues include the 20 natural amino acids usually designated by a three-letter symbol and also includes, 4-hydroxyproline, hydroxylysine, demosin, isodemosin, 3-methylhistidine, norvlina, betaalanine, gamma-aminobutyric acid, citrulline, homocysteine, homoserine, ornithine and methionine sulfone (ii) a compound in which an alkyl carbonate, carbamate, amide or ester is covalently linked to an amino, hydroxyl, or carboxylic acid group of the compound of formula I by the secondary chain of the carbon-containing prodrug carbonyl, or (iii) a compound comprising a phosphate derivative of a compound of formula I linked by a phosphorus-oxygen bond to a hydroxyl 1 I will be of the compounds of formula I; and wherein in addition each alkyl, cycloalkyl, alkylaryl, aryl, heterocyclyl, or heteroaryl group may optionally be substituted with one or more of C1-C3 alkyl, C3-C6 cycloalkyl, C6 aryl, heterocyclyl, heteroaryl, C1-3OH alkyl, alkylaryl, OH, O C1-3O alkyl, halo, CN, NO2, CO2H, CO2 C1-3 alkyl, CONH2, CONH (C1-C3 alkyl), CON (C1-C3 alkyl) 2, trifluoromethyl, NH2, NH (alkyl C1-C3) or N (C1-C3 alkyl) 2 and each substituent alkyl, cycloalkyl, alkylaryl, aryl heterocyclyl, or optional heteroaryl may also be optionally substituted.

Description

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FIELD OF THE INVENTION

The present invention relates to novel bicyclic pyrimidinone compounds and their analogs for the treatment of viral infections, particularly HIV infections.

BACKGROUND OF THE INVENTION

The retrovirus called "human immunodeficiency virus" or "HIV" is the etiologic agent of a complex disease that progressively destroys the immune system. This disease is known as acquired immunodeficiency syndrome or AIDS. In December 2005, it was estimated that 40 million people living with HIV lived in the world and there were approximately 3 million deaths per year.

A feature of retrovirus replication includes the reverse transcription of the viral genome in the proviral DNA and its integration into the genome of the host cell. These stages are necessary for HIV replication and are mediated by enzymes encoded by the virus, reverse transcriptase and integrase, respectively.

HIV infection follows a route of binding of viral particles to cell surface receptors and simultaneous receptors that results in the fusion of viral particles with the cell. Virus contents are released inside the cytoplasm, in which reverse transcription of the HIV genome occurs. Through a series of stages, a double-stranded Proviral DNA copy is produced. Proviral DNA is transported to the nucleus in a complex known as a preintegration complex (PIC) that contains integrase and other viral and possibly cellular proteins. Once inside the nucleus, proviric DNA is integrated into the genome of the host cell through the action of integrase. Once integrated, transcription and translation of the viral genome can occur resulting in the production of viral proteins and a new genome of viral RNA. These proteins and the genome are assembled on the cell surface and, depending on the type of cell, possibly in other intracellular membranous compartments. These infectious particles then sprout out of the cell and during, or shortly after, this process matures into infectious HIV particles through the action of the viral protease.

The integration of the proviric genome into the genome of the host cell requires the action of an integrase that carries out this process in at least three stages, possibly four. The first stage involves the assembly of the viral genome in a stable nucleoprotein complex, secondly, the processing of two nucleotides from the 3 'ends of the genome to give staggered ends with free OH residues in 3' and third, the transfer of these extremes in the genome of the host cell. The final stage involves filling holes and repairing the insertion site in the host genome. There are still some conjectures about whether integrase carries out this final stage or if it is carried out by cellular repair enzymes.

Currently, HIV infection can be treated with numerous inhibitors on the market that target reverse transcriptase, protease or cell entry. The treatment of HIV infection with these, or with a combination of these drugs, is known to be an effective treatment for AIDS and similar diseases. The drawbacks of current inhibitors include the rapid onset and increase in the incidence of resistance and the numerous side effects and, therefore, there is a need for new types of inhibitors that target proteins such as integrase.

WO 2006103399 refers to inhibitors of FHV integrase and inhibitors of FHV replication.

SUMMARY OF THE INVENTION

In a first aspect, the present invention provides a compound of Formula I or a hydrate, salt or prodrug thereof in which:

image 1

A is a monocyclic or bicyclic aromatic or heteroaromatic moiety fused with the nitrogen containing ring;

X is selected from the group consisting of O and S;

And it is selected from the group consisting of O and S;

R1 has 0-3 substituents each of which is independently selected from the group consisting of CN, C1-10 alkyl, C2-10 alkenyl, C1-10PO3H2 alkyl, -O-C1-10 alkyl, C1-10NR3R4 alkyl, -O-C1-10NR3R4 alkyl, halo, NR3R4, alkylaryl, alkylheteroaryl, aryl, heteroaryl, -O-alkylaryl, SO2NR3R4;

R3 and R4 are each independently selected from the group consisting of hydrogen, CO2 C1-4 alkyl, C (O) C1-4 alkyl C1-10 alkyl, C1-10NR5R6, -NH (CO) (CO) NH C1 alkyl -4, or R3 and R4 taken together with the bound nitrogen form a 5-7 membered heterocyclic ring containing from zero to two additional heteroatoms selected from N, O or S, in which S may be in the oxidation state S , S (O) or S (O) 2 and wherein said heterocyclic ring is optionally substituted on the carbon or nitrogen atoms with one or more substituents selected from halo, aryl, C (O) C1-4 alkyl, SO2 alkyl C1-4, SO2H, C1-4 alkyl, CO2H, CO2 C1-4 alkyl, NR5R8; C1-4NR5R6 alkyl;

R5 and R6 are each independently selected from the group consisting of H, C1-4 or R5 alkyl and R6 and together with the attached nitrogen form a 5-7 membered heterocyclic ring containing from zero to two additional heteroatoms selected from N , O or S, in which S may be in the oxidation state S, S (O) or S (O) 2 and in which said heterocyclic ring is optionally substituted at the carbon or nitrogen atoms with one or more substituents selected from halo and C1-4 alkyl;

when R1 is alkylaryl or -O-alkylaryl, the aryl group of said alkylaryl substituent is optionally substituted with a substituent selected from C1-10 alkyl, -O-C1-10 alkyl, C1-10NR3R4 alkyl, -O-C1-10NR3R4 alkyl , halo, NR3R4, alkylaryl, -O-alkylaryl, SO2NR3R4

B is absent or is -C (O) -;

C is absent or selected from the group consisting of -O-, -NH- and -NH-NH-C (O) -;

R2 is selected from the group consisting of heteroaryl and heterocyclyl;

and wherein said prodrug comprises (i) a compound in which an amino acid residue, or a polypeptide chain or two

or more amino acid residues covalently bind to an amino, hydroxyl or carboxylic acid group of the compound of formula I, (ii) a compound in which an alkyl carbonate, carbamate, amide or ester covalently binds to an amino, hydroxyl group , or of carboxylic acid of the compound of formula I by the secondary chain of the prodrug containing carbonyl carbon, or (iii) a compound comprising a phosphate derivative of a compound of formula I linked by a phosphorus-oxygen bond to a hydroxyl free of the compounds of formula I;

and wherein each alkyl, cycloalkyl, alkylaryl, aryl, heterocyclyl, or heteroaryl group may be optionally substituted with one or more of C1-C3 alkyl, C3-C6 cycloalkyl, C6 aryl, heterocyclyl, heteroaryl, C1-3OH alkyl, alkylaryl , OH, O C1-3O alkyl, halo, CN, NO2, CO2H, CO2 C1-3 alkyl, CONH, CONH (C1-C3 alkyl), CON (C1-C3 alkyl) 2, trifluoromethyl, NH2, NH (C1 alkyl -C3) or N (C1-C3 alkyl) 2 and each substituent alkyl, cycloalkyl, alkylaryl, aryl heterocyclyl, or optional heteroaryl may also be optionally substituted. In a second aspect, the present invention provides a compound of the first aspect or a salt or prodrug thereof for use in the treatment or prophylaxis of a viral infection in a subject.

In a fourth aspect, the present invention provides a pharmaceutical composition comprising a compound according to the first aspect and a pharmaceutically acceptable carrier, diluent or excipient.

BRIEF DESCRIPTION OF THE FIGURES

Figure 1 shows a table of activity of the compounds, which includes the compounds according to the present invention, in chain transfer assays using the natural and mutant HIV integrase enzyme. The mutant enzyme includes the Q148k mutation and is resistant to published integrase inhibitors such as S-1360, Raltegravir (Merck MK-0518) and GS9137 (Gilead GS-9137).

DETAILED DESCRIPTION OF THE INVENTION

In a first aspect, the present invention provides a compound of formula I or a hydrate, salt or prodrug thereof as defined above in a preferred form, R2 is substituted with aryl or alkylaryl.

In a preferred form, C2-10 alkenyl is allyl.

In a preferred form, the compound of formula I is selected from the group consisting of the compounds of formula II, III, IV, V and VI:

image2

In the compounds of formula III, IV and V, Z is O, S or NR8 in which R8 is H, C1-10 alkyl, C1-10 alkyl3R4 alkyl, alkylheteroaryl, aryl and heteroaryl.

In a preferred form, NR3R4 is morpholine.

Preferably, heteroaryl is selected from the group consisting of tetrazole, triazole, pyrazole, imidazole, oxazole, oxadiazole, thiazole, thiadiazole.

Preferably, the compound is selected from:

7- (3-Chloro-2-fluoro-benzyl) -3-hydroxy-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid; (1H-tetrazol-5-yl) -amide; 7- (3

Chloro-2-fluoro-benzyl) -3-hydroxy-2- (2H-tetrazol-5-yl) -pyrido [1,2-a] pyrimidin-4-one; 7- (3-Chloro-2-fluoro-benzyl) -3-hydroxy-2 (2H- [1,2,4] triazol-3-carbonyl) -pyrido [1,2-a] pyrimidin-4-one; 7- (3-Chloro-2-fluoro-benzyl) -3-hydroxy-2- (1H-tetrazol-5-carbonyl) pyrido [1,2-a] pyrimidin-4-one and 2- [5- (4 -fluoro-benzyl) - [1,3,4] oxadiazol-2-yl] -3-hydroxy-pyrido [1,2-a] pyrimidin-4-one.

In a second aspect, the use of a compound of Formula I or a pharmaceutically acceptable salt or prodrug derivative thereof is provided for the preparation of a medicament for the treatment or prophylaxis of a

30 viral infection in a subject.

Preferably, the viral infection of the second aspect is an HIV or VIS infection.

More preferably, HIV or VIS infection comprises a viral strain resistant to other integrase inhibitors. Even more preferably, the viral strain comprises the HIV integrase enzyme with the Q148K mutation. Some of the compounds of the present invention have been shown to have surprisingly greater activity against the enzyme

35 HIV integrase (Q148K) which is resistant to published integrase inhibitors such as S-1360, Raltegravir (Merck MK-0518) and GS9137 (Gilead GS-9137). A compound that shows a particularly high activity against Q148K is the compound of Example 6.13 (see Figure 1).

An example of a bibliographic reference of the resistance profiles of Q148K can be found in the 14th CROI (Conference on Retroviral and Opportunistic Infections), Los Angeles, February 27, 2007, by John Wai, Merck

40 Research Labs, ’Next generation inhibitors of HIV-1 Integrase Strand Transfer: Structural diversity and resistance profiles.

As used herein, the term "halo" or "halogen" refers to fluorine (fluoro), chlorine (chlorine), bromine (bromine) or iodine (iodine).

As used herein, the term "alkyl" used both alone and in terms of compounds such as NH (alkyl) or N (alkyl) 2, refers as appropriate to monovalent straight or branched chain hydrocarbon groups, which have 1 to 3, 1 to 6, or 1 to 10 carbon atoms. For example, suitable alkyl groups include, but are not limited to methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 2-, 3 - or 4-methylpentyl, 2-ethylbutyl, n-hexyl, or 2, 3-, 4-or 5-methylpentyl.

As used herein, the term "alkenyl" refers to straight or branched chain hydrocarbon groups having one or more double bonds between carbon atoms. Suitable alkenyl groups include, but are not limited to, ethenyl, allyl, propenyl, iso-propenyl, butenyl, pentenyl and hexenyl.

The term "cycloalkyl" as used herein refers to cyclic hydrocarbon groups. Suitable cycloalkyl groups include, but are not limited to cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

The term "aryl" as used herein refers to a C6-C10 aromatic hydrocarbon group, for example, phenyl or naphthyl.

The term "alkylaryl" includes, for example, benzyl.

The term "heterocycle", when used alone or in compound words includes monocyclic, polycyclic, fused or conjugated hydrocarbon moieties, preferably C3-6, in which one or more carbon atoms (and when appropriate, the attached hydrogen atoms to the above) are substituted by a heteroatom in such a way that they provide a non-aromatic moiety. The bonds between atoms can be saturated or unsaturated. Suitable heteroatoms include, O, N and S. When two or more carbon atoms are substituted, this may be by two or more of the same heteroatom or by different heteroatoms. Suitable examples of heterocyclic groups may include pyrrolidinyl, piperidyl, piperazinyl, morpholino, quinolinyl, isoquinolinyl, thiomorpholine, dioxanyl, 2,2'-dimethyl- [1,3] -dioxolanyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydropyrrolyl, etc.

The term "heteroaryl" includes a 5- or 6-membered heteroaromatic ring containing one or more heteroatoms selected from O, N and S. Suitable examples of heteroaryl groups include furanyl, thiophenyl, tetrazolyl, 1,2,3-triazolyl, 1, 2 , 4-triazolyl, imidazolyl, pyrazolyl, pyridinyl, pyrimidinyl, oxazolyl, oxadiazolyl, thioazolyl, thiodiazolyl, etc. The heteroaromatic ring may be fused to a 5- or 6-membered aromatic or heteroaromatic ring to form a bicyclic aromatic ring system, for example, benzofuran.

Unless otherwise defined, each alkyl, cycloalkyl, alkylaryl, aryl, heterocyclyl, or heteroaryl group may be optionally substituted with one or more of C1-C3 alkyl, C3-6 cycloalkyl, C6 aryl, heterocyclyl, heteroaryl, C13OH alkyl, alkylaryl, OH, O C1-C3 alkyl, halo, CN, NO2, CO2H, CO2 C1-3 alkyl, CONH2, CONH (C1-C3 alkyl), CON (C1C3 alkyl) 2, trifluoromethyl, NH2, NH ( C1-C3 alkyl) or N (C1-C3 alkyl) 2. For example, an aryl group may optionally be a 4-methylphenyl or 4-hydroxyphenyl group, and an optionally substituted alkyl group may be 2-hydroxyethyl, trifluoromethyl, or difluoromethyl.

Each substituent alkyl, cycloalkyl, alkylaryl, aryl, heterocyclyl, or optional heteroaryl may also be optionally substituted.

Examples of optional substituents also include suitable nitrogen protecting groups (see "Protective Groups in Organic Synthesis" Theodora Greene and Peter Wuts, third edition, Wiley, Interscience, 1999).

The salts of the compound of formula I are preferably pharmaceutically acceptable, but it will be appreciated that non-pharmaceutically acceptable salts are within the scope of the present invention, because they are useful as intermediates in the preparation of pharmaceutically acceptable salts.

The term "pharmaceutically acceptable derivative" may include any pharmaceutically acceptable salt, hydrate or prodrug, or any other compound that, upon administration to a subject, is capable of providing (directly or indirectly) a compound of formula I or a metabolite or residue of the same antibacterially active.

Suitable pharmaceutically acceptable salts include, but are not limited to, pharmaceutically acceptable inorganic acid salts such as hydrochloric, sulfuric, phosphoric, nitric, carbonic, boric, sulfamic, and hydrobromic acids, or pharmaceutically acceptable organic acid salts such as acetic, propionic, butyric, tartaric, maleic, hydroximaleic, fumaric, malic, citric, lactic, mucic, gluconic, benzoic, succinic, oxalic, phenylacetic, methanesulfonic, toluenesulfonic, benzenesulfonic, salicylic, sulfanyl, aspartic, aspartic, glutamic, aspartic, aspartic , palmitic, oleic, lauric, pantothenic, tannic, ascorbic and valeric.

Base salts include, but are not limited to, those formed with pharmaceutically acceptable cations, such as sodium, potassium, lithium, calcium, magnesium, zinc, ammonium, alkylammonium such as salts formed from triethyl amine, alkoxy ammonium such as those formed with ethanolamine and the salts formed from ethylenediamine, choline or amino acids such as arginine, lysine or histidine. Those skilled in the art know the general information about the types of pharmaceutically acceptable salts and it is as described in general texts such as "Handbook of Pharmaceutical salts" P.H. Stahl, C.G. Wermuth, 1st edition, 2002, Wiley-VCH.

The groups containing basic nitrogen can be quaternized with agents such as lower alkyl halide, such as methyl, ethyl, propyl and butyl chlorides, bromides and iodides; dialkyl sulfates of dimethyl and diethyl sulfate type; and others.

hydroxyl groups can be esterified with groups that include lower alkyl carboxylic acids such as acetic acid and 2,2-dimethylpropionic acid, or sulfonated with groups that include alkylsulfonic acids, such as methylsulfonic acid (see, for example, the compound of Example 15.10 ).

This invention also encompasses pharmaceutical compositions containing prodrugs of the compounds of formula I. This invention also encompasses the methods of treating or preventing a viral infection in a subject by administering prodrugs of the compounds of the formula I. The compounds of Formula I having free amino, amido, hydroxy or carboxylic groups can be converted into prodrugs.

Prodrugs include compounds in which the amino acid residue or a polypeptide chain of two or more (eg, two, three or four) amino acid residues that covalently bind to the free amino, hydroxy and carboxylic groups of the compounds of formula I. The amino acid residues include the 20 naturally occurring amino acids designated by three-letter symbols and also include, 4-hydroxyproline, hydroxylysine, demosin, isodemosin, 3-methylhistidine, norvaline, beta-alanine, gamma-aminobutyric acid, citrulline, homocysteine, homoserine, ornithine and methionine sulfone. Prodrugs also include compounds in which the carbonates, carbamates, amides and alkyl esters covalently bind to the above substituents of formula I by the secondary chain of the prodrug containing carbonyl carbon. Prodrugs also include phosphate derivatives of compounds of formula I (such as acids, acid salts, or esters) linked by a phosphorus-oxygen bond to a free hydroxyl of the compounds of formula I.

It will also be recognized that the compounds of formula I may have asymmetric centers and are therefore capable of existing in more than one stereoisomeric form. The invention also therefore relates to compounds in substantially pure isomeric form in one or more asymmetric centers, for example, more than about 90%, for example, such as about 95% or 97%, for example, or more than 99% for example, as well as the mixtures, which include the racemic mixtures thereof. Such isomers can be prepared by asymmetric synthesis, using for example chiral intermediates, or by chiral resolution.

In a fourth aspect, the present invention provides a pharmaceutical composition comprising a compound according to the first aspect and a pharmaceutically acceptable carrier, diluent or excipient.

The compositions of the present invention may contain other therapeutic agents as described below, and may be formulated, for example, using conventional solid or liquid carriers or diluents, as well as pharmaceutical additives of a type suitable to the desired mode of administration (by for example, excipients, binders, preservatives, stabilizers, aromas, etc.) according to techniques such as those known in the field of pharmaceutical formulation.

The compounds of the present invention can be administered by any suitable means, for example, parenterally, such as by subcutaneous, intravenous, intramuscular, or intracisternal injection or infusion techniques (for example, as sterile injectable aqueous or non-aqueous solutions or suspensions) .

Pharmaceutical formulations include those for oral, rectal, nasal, topical (including buccal and sublingual), vaginal or parenteral (which includes intramuscular, subcutaneous and intravenous) administration or in a form suitable for administration by inhalation or insufflation. The compounds of the invention, together with a conventional adjuvant, vehicle or diluent, can thus be placed in the form of pharmaceutical compositions and unit dosages thereof, and can be used in such form as solids, such as tablets or capsules. filled, or liquids such as solutions, suspensions, emulsions, elixirs or capsules filled therewith, all for oral use, in the form of suppositories for rectal administration; or in the form of sterile injectable solutions for parenteral use (including subcutaneous).

In addition to primates, such as humans, a variety of other mammals can be treated according to the method of the present invention. For example, mammals that include, but are not limited to, cows, sheep, goats, horses, dogs, cats, guinea pigs, rats or other species of cattle, sheep, equines, canines, felines, rodents or murines can be treated. However, the procedure can also be practiced in other species, such as bird species (eg, chickens).

The subjects treated in the previous procedure are mammals, which include, but are not limited to, cows, sheep, goats, horses, dogs, cats, guinea pigs, rats or other species of cattle, sheep, horses, dogs, cats, rodents or murine and preferably a human being, male or female.

The term "effective amount" means the amount of the subject composition that will stimulate the biological or medical response of a tissue, system, animal or human, which is being sought by the researcher, veterinarian, medical doctor or other specialist.

As those skilled in the art of treating viral infections, and particularly HIV infections, will understand, the term "treatment" does not necessarily mean that the viral infection is completely cured. The term "treatment" encompasses any reduction in viral load and / or inhibition of replication in the subject being treated.

It is intended that the term "composition" as used herein encompasses a product comprising the specified ingredients in the specified amounts, as well as any product that is the result, directly or indirectly, of the combination of the ingredients specified in the specified quantities. By "pharmaceutically acceptable" is meant that the vehicle, diluent or excipient must be compatible with the rest of the ingredients of the formulation and not detrimental to the recipient thereof.

It should be understood that the terms "administration of" and / or "administering a" compound means providing a compound of the invention to the individual in need of treatment.

Pharmaceutical compositions for the administration of the compounds of this invention may conveniently be presented in unit dosage form and may be prepared by any of the procedures well known in the pharmacopoeia technique. All procedures include the stage of carrying the active ingredient in association with the vehicle constituting one or more additional ingredients. In general, pharmaceutical compositions are prepared uniformly and intimately by carrying the active ingredient in association with a liquid vehicle or a finely divided solid vehicle or both, and then, if necessary, forming the product in the desired formulation. In the pharmaceutical composition, the object active ingredient is included in an amount sufficient to produce the desired effect on the disease process or condition. As used herein, the term "composition" is intended to encompass a product comprising the specified ingredients in the specified amounts, as well as any product that results, directly or indirectly, from the combination of the specified ingredients. in the specified quantities.

The pharmaceutical compositions may be in the form of a sterile injectable aqueous or oleaginous suspension. This suspension can be formulated according to the known technique using those suitable dispersing agents or humectants and the suspending agents mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be used are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile fixed oils are conventionally used as solvent or suspension medium. For this purpose, soft fixed oil that includes synthetic mono or diglycerides can be used. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

The pharmaceutical composition and the process of the present invention may further comprise other therapeutically active compounds that are normally applied in the treatment of the aforementioned pathological conditions. A technician normally skilled in the art can make the selection of appropriate agents for use in combination therapy, in accordance with conventional pharmaceutical principles. The combination of therapeutic agents can act synergistically to carry out the treatment or prevention of the various disorders described above. Using this approach, one may be able to achieve therapeutic efficacy with lower dosages of each agent, thereby reducing the potential for adverse side effects.

When other therapeutic agents are used in combination with the compounds of the present invention, they can be used, for example, in the amounts indicated in the Physician Desk Reference (PDR) or as otherwise determined by a technician normally skilled in the art .

In the treatment or prevention of ailments that require HIV inhibition or inhibition of the HIV integrase enzyme, an appropriate dosage level will generally be from about 0.01 to 500 mg per kg of the patient's body weight per day, which They can be administered in single or multiple doses. Preferably, the dosage level will be about 0.1 to about 250 mg / kg per day, more preferably about 0.5 to about 100 mg / kg per day. A suitable dosage level may be approximately 0.01 to 250 mg / kg per day, approximately 0.05 to 100 mg / kg per day, or approximately 0.1 to 50 mg / kg per day. Within this range, the dosage may be 0.05 to 0.5, 0.5 a5 or 5 to 50 mg / kg per day. For oral administration, the compositions are preferably provided in the form of tablets containing 1.0 to 1000 milligrams of the active ingredient, particularly 1.0, 5.0, 10.0, 15.0, 20.0, 25.0 , 50.0, 75.0, 100.0, 150.0, 200.0, 250.0, 300.0, 400.0, 500.0, 600.0, 750.0, 800.0, 900 , 0, and 1000.0 milligrams of the active substance for the symptomatic adjustment of the dosage to the patient being treated. The compounds may be administered on a schedule of 1 to 4 times per day, preferably once or twice per day.

It will be understood, however, that the specific dose level and dosage frequency of any particular patient can be varied and this will depend on a variety of factors including the activity of the specific compound employed, metabolic stability and duration. of the action of this compound, age, body weight, general health, sex, diet, mode and time of administration, excretion rate, drug combination, severity of the specific condition, and The therapy the host experiences.

In order that the nature of the present invention can be more clearly understood, preferred forms thereof will now be described by reference to the following non-limiting Examples.

EXAMPLES

Procedures HPLC Conditions: All HPLC measurements were performed on a Waters 2690 Alliance System instrument.

Procedure 1

Column: Waters Exterra C18 column (part number 186000410) at 30˚ C, flow rate 0.4 ml / min, spectrum measured at 254 Nm Buffers: Buffer A: 100% water, Buffer B: 100% acetonitrile, Buffer C: 2% aqueous TFA Gradient: (linear gradient curve 6)

  5 min 2 min 0.25 min 2.75 min 85% A10% B5% C → 0% A95% B5% C → 0% A95% B5% C → 85% A10% B5% C → 85% A10% B5% C

Procedure 2

Column: Merck C18 Chromolith column (part number 1.02129.0001) at 30˚ C, flow rate 4 ml / min, spectrum measured at 254 nM Buffers: Buffer A: 100% water, Buffer B: 100% acetonitrile, Buffer C: 2% aqueous TFA Gradient: (linear gradient curve 6)

2 min 1 min 0.15 min 0.85 min 92% A3% B5% C → 80% A15% B5% C → 80% A15% B5% C → 92% A3% B5% C → 92% A3% B5% C

Procedure 3

Column: Merck C18 Chromolith column (part number 1.02129.0001) at 30˚ C, flow rate 4 ml / min, spectrum measured at 254 nM Buffers: Buffer A: 100% water, Buffer B: 100% acetonitrile, Buffer C: 2% aqueous TFA Gradient: (linear gradient curve 6)

 2.3 min 0.7min 0.15 min 0.85 min 85% A10% B5% C → 45% A50% B5% C → 45% A50% B5% C → 85% A10% B5% C → 85% A10 % B5% C

Procedure 4

Column: Merck C18 Chromolith column (part number 1.02129.0001) at 30˚ C, flow rate 4 ml / min, spectrum measured at 254 nM Buffers: Buffer A: 100% water, Buffer B: 100% acetonitrile, Buffer C: 2% aqueous TFA Gradient: (linear gradient curve 6)

 2.3 min 0.7min 0.15 min 0.85 min 70% A25% B5% C → 20% A75% B5% C → 70% A25% B5% C → 70% A25% B5% C → 70% A25 % B5% C

Procedure 5

Column: Phenomenex Gemini C18 column (part number 344382-3) at 30˚C, flow rate 0.4 ml / min, spectrum measured at 254 nM Buffers: Buffer A: 100% water, Buffer B: 100% acetonitrile, Buffer C: 2% aqueous TFA Gradient: (linear gradient curve 6)

  5 min 1min 0.25 min 3.75 min 49% A40% B1% C → 4% A95% B1% C → 4% A95% B1% C → 49% A50% B1% C → 49% A50% B1% C

Procedure 6

Column: Phenomenex Gemini C18 column (part number 344382-3) at 30˚C, flow rate 0.4 ml / min, spectrum measured at 254 nM Buffers: Buffer A: 100% water, Buffer B: 100% acetonitrile, Buffer C: 2% aqueous TFA Gradient: (linear gradient curve 6)

  4 min 1min 0.25 min 4.75 min 69% A30% B1% C → 39% A60% B1% C → 39% A60% B1% C → 69% A30% B1% C → 69% A30% B1% C Procedure 7 Column: Waters Symmetry® C18 column (part number WAT045905) at 25˚ C, flow rate 1 ml / min, spectrum measured at 254 nM Buffers: Buffer A: 100% acetonitrile, Buffer B: 0.1% aqueous TFA Gradient: (linear gradient curve 6)

5 min 10min 10 min 10% A90% B → 10% A90% B → 100% A0% B → 100% A0% B General scheme 1: Synthesis Although the following example is not included in the scope of the claims, it is introduced as Useful background information for the reader.

image 1

The amino compound is reacted as in Scheme 1 with the fumarate derivative or suitable fumarate analogs, in which, for example, acetyl groups can be substituted with other leaving groups such as tosyl or mesyl. The reaction can be carried out in a suitable solvent such as methanol, DME, DMA, DMSO, chloroform, THF or dioxane. The reaction can be heated or subjected to microwave radiation (see for example B. R. Roberts and C.

R. Strauss, Acc. Chem. Res. 2005, 38, 653-661, "Toward Rapid,’ Green ’Predictable Microwave-assisted Synthesis"). The reaction can be carried out in the absence or presence of catalytic amounts of acid or base.

General scheme 2: Alternative synthesis 1

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image2

General scheme 3: Alternative synthesis 2

Although the following example is not included in the scope of the claims, it is introduced as background information useful to the reader.

image 1

Compounds whose substituent is halogen can be further reacted by methods known to the person skilled in the art as shown above, in which 'Pd coupling' includes reactions such as those of Suzuki, Buchwald, Heck or Sonogashira and substitution aromatic nucleophilic (see, for example, the reactions described in LS Hegedus, "Transition Metals in the Synthesis of Complex Organic Molecules", University Science books, 1994, first edition, or in M. Smith "Organic synthesis", 2001, McGraw- Hill Science, 2nd edition) which optionally use a metal catalyst such as a suitable form of palladium and reagent 1 is a derivative of R1 (for example derivatives of R1 including halogen or boronate).

image 1

OH can be protected by methods known to the person skilled in the art as shown in scheme (5), for example in which 'P' can be benzyl (see I. Stansfield et al., 'Active site inhibitors of HCV NS5B polymerase 'Bio-Org. Med. Chem. Lett, 2004, 14, 5085-5088) or a suitable protective group such as those described in "Protective Groups in Organic Synthesis" Theodora Greene and Peter Wuts, third edition, Wiley , Interscience, 1999).

image 1

Example 1: Preparation of dimethyl diacetoxybumarate

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The procedure described in OPPI, 22 (4), 1990, 532-534 was followed.

A slightly cloudy stirred solution of dihydroxyfumic acid (10.7 g, 72.5 mmol) in anhydrous methanol (50 ml) under nitrogen was cooled (ice / water bath). Thionyl chloride (10.5 ml, 144 mmol) was added for 20 min under the surface of the methanolic solution with a syringe. After the addition the cooling bath was removed and the mixture was stirred at room temperature for 3 d. The resulting precipitate was collected by filtration and washed with cold methanol (10 ml) and water (80 ml). Dimethyl dihydroxybumarate was obtained as a white solid (11.8 g). This material (10.8 g) and isopropenyl acetate (36 ml) were combined and heated under reflux with stirring under nitrogen for 8 h. The reaction was cooled to room temperature and stored at 0 ° C overnight. The resulting precipitate was collected by filtration and washed with cold methanol (5 ml) and water (80 ml) to give dimethyl diacetoxybumarate as a white solid. (6.4 g).

Example 2: Preparation of the 7-bromo-3-hydroxy-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid methyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

2-Amino-5-bromo-pyridine (664 mg, 3.85 mmol), dimethyl diacetoxybumarate (1.00 g, 3.84 mmol) and glacial acetic acid (10 drops) in dry methanol (10 ml) were heated and combined at reflux. After 48 hours, the reaction was cooled to room temperature and concentrated in vacuo. Ethyl acetate (2 ml) was added to the residue and sonicated for 2 min and the resulting precipitate was collected by filtration and washed with cold ethyl acetate (1 ml) and dried by pump to give the acid methyl ester 7 -bromo-3-hydroxy-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid as a yellow solid (24 mg, 2%): 1H NMR (300 MHz, CDCl3) δ 4.19 (3H, s, OCH3), 7.53 (2H, d, J = 1.4 Hz, H8 and H9), 9.00 (1H, d, J = 1.4 Hz, H6), 10.53 ( 1H, br s, OH). MS (ESI +) m / z 299 (M [Br79] +1), 301 (M [Br81] +1)

HPLC procedure 1 96.0% / 4.30 min.

Example 2.1: Preparation of the 3-hydroxy-7-methyl-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid methyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

1H NMR (300 MHz, CDCl3) δ 2.40 (3H, s, CH3), 4.11 (3H, s, OCH3), 7.39 (1H, dd, J = 9.1 Hz, 1.8 Hz , H8), 7.60 (1H, d J = 9.1 Hz, H9), 8.68 (1H, m, H6). MS (ESI +) m / z 235 (M + 1), (ESI-) m / z 233 (M-1) HPLC procedure 1 100% / 3.74 min.

Example 2.2: Preparation of the 7-chloro-3-hydroxy-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid methyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image3

MS (ESI +) m / z 255 (M + 1), (ESI-) m / z 253 (M-1) HPLC procedure 1 96% / 14.14 min.

Example 2.3: Preparation of the 3-hydroxy-7-morpholin-4-yl-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid methyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

5-Morpholin-4-yl-pyridin-2-ylamine was prepared by adapting the procedure described in J. Med. Chem., 2005, 48 (7), 23882406. In summary, 5-bromo-2-nitro-pyridine was reacted with morpholine and potassium carbonate in DMSO at 60-70 ° C to give 4- (6-nitro-pyridin-3-yl) -morpholine in a yield of 84%. Reduction with palladium on carbon under nitrogen atmosphere provided 5-morpholin-4-yl-pyridin-2-ylamine in 70% yield. This was converted into 3-hydroxy-7-morpholin-4-yl-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid methyl ester in 25% yield by adaptation of the described process in Example 2, although glacial acetic acid was used instead of p-toluenesulfonic acid.

1H NMR (300 MHz, D6-DMSO): δ 3.22 (4H, t, J = 5.0 Hz, -NCH2CH2O-), 3.89 (4H, t, J = 5.0 Hz, -NCH2CH2O- ), 4.10 (3H, s, OCH3), 7.45 (1H, dd, J = 9.9, 2.7 Hz, H8), 7.63 (1H, d, J = 9.9 Hz, H9), 8.17 (1H, d, J = 2.7 Hz, H6), 10.32 (1H, s, OH)

MS (ESI +) m / z 305 (M + 1) HPLC procedure 7 97.4% / 11.7 min.

Example 2.4: Preparation of the 3-hydroxy-8-morpholin-4-yl-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid methyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The 2-picolinic acid was reacted with thionyl chloride and methanol to give methyl 4-chloro-2-picolinate which was hydrolyzed and the hydrochloride salt was subjected to Curtius rearrangement. Excision of the Boc protecting group resulted in 2-amino-4-chloropyridine. The procedure described in WO2006040520 was adapted to introduce morpholine into position 4. This was cycled to the ester intermediate using an adaptation of the procedure described in Example 2 in which the reaction was carried out at 60 ° C.

1H NMR (300 MHz, DMSO-d6) δ 3.43 (t, J = 4.7 Hz, 4H), 3.73 (t, J = 4.7 Hz, 4H), 3.85 (s, 3H ), 6.67 (d, J = 2.4 Hz, 1H), 7.27 (dd, J = 8.2 Hz, 2.5 Hz, 1H), 8.59 (d, J = 8.2 Hz, 1H), 9.29-9.63 (brs, 1H) MS (ESI +) m / z 328 (M + 23)

Example 2.5: Preparation of the 3-hydroxy-9-morpholin-4-yl-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid methyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The procedure described in Example 2.3 was adapted to 3-bromo-2-nitro-pyridine to give 2-amino-3-morpholinopyridine which became the desired ester by adaptation of the procedure described in Example 2.

1H NMR (300 MHz, CDCl3) δ 3.45-3.56 (m, 4H), 4.00-4.11 (m, 7H), 6.97 (t, J = 7.3 Hz, 1 H ), 7.22 (d, J = 7.3 Hz, 1H), 8.63 (dd, J = 7.2, 1.1 Hz, 1H), 10.31 (s, 1H)

MS (ESI +) m / z 328 (M + 23)

Example 2.6: Preparation of the 3-hydroxy-4-oxo-7-piperidin-1-yl-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid methyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The procedure described in Example 2.3 was adapted except that piperidine was used. The resulting 2-amino-5-piperidinopyridine was converted into the desired ester by adaptation of the procedure described in Example 2. The crude product was reacted by adapting the procedure of Example 17.1 (Step 1) except that pivaloyl chloride was used for Give the desired product.

1H NMR (300 MHz, CDCl3) δ 1.43 (s, 9H), 1.58-1.82 (m, 6H), 3.24 (t, J = 4.8 Hz, 4H), 3.96 (s, 3H), 7.64-7.78 (m, 2H) 8.30 (d, J = 1.8 Hz, 1H).

Example 3: Preparation of the 3-cyano-6-hydroxy-7-oxo-1,7-dihydro-pyrazolo [1,5-a] pyrimidine-5-carboxylic acid methyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

3-amino-4-pyrazole carbonitrile (400 mg, 3.7 mmol) and dihydroxybumarate (978 mg, 5.5 mmol) were dissolved in glacial acetic acid (5 ml) and the mixture was heated to 100 ° C. After 2 d, the reaction was considered complete by HPLC analysis and cooled to room temperature. Ethyl acetate (10 ml) was added to start precipitation of the product and the resulting precipitate was collected by filtration. The product was isolated with a yield of 62% (534 mg).

1H NMR (300 MHz, D6-DMSO): δ 3.90 (3H, s, OCH3) and 8.39 (1H, s, CHC [CN]).

MS (ESI +) m / z 235 (M + 1)

HPLC procedure 2 89.3% / 1.07 min.

Example 3.1: Preparation of the 4-fluoro-benzylamide of 3-cyano-6-hydroxy-7-oxo-1,7-dihydro-pyrazolo [1,5a] pyrimidine-5-carboxylic acid compound with 4-fluorobenzylamine.

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image4

The 3-cyano-6-hydroxy-7-oxo-1,7-dihydro-pyrazolo [1,5-a] pyrimidine-5-carboxylic acid methyl ester (100 mg, 0.427 mmol) was suspended in MeOH (10 ml ) and 4-fluorobenzylamine (122 µL, 1.07 mmol) was added thereto and the reaction was heated at reflux for 2 days before cooling to room temperature and filtered. The filtrate was concentrated and recrystallized from hot methanol to give the product as a yellow solid (25 mg, 18%).

1H NMR (300 MHz, D6-DMSO): δ 4.01 (2H, s, H3N + CH2Ph), 4.48 (2H, d, J = 6.6 Hz, [C = O] NHCH2), 7, 49 (8H, m, ArCH), 8.08 (1H, s, CHC [CN]), 9.13 (1H, t, J = 6.6 Hz, [C = O] NHCH2). MS (ESI +) m / z 328 [M + H] +

Example 3.2: Preparation of the 3-cyano-6-hydroxy-7-oxo-1,7-dihydro-pyrazolo [1,5a] pyrimidine-5-carboxylic acid 3,4-dichloro-benzylamide; compound with 3,4-dichloro-benzylamine

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Compound prepared by adapting the procedure of Example 3.1

1H NMR (300 MHz, D6-DMSO): δ 8.45 (1H, s, Ar-CH), 7.79-7.57 (4H, m, Ar-CH), 7.39-7.32 ( 2H, m, Ar-CH), 4.78 (2H, s, H3N + CH2), 4.49 (2H, d, J = 6.3 Hz, NHCH2). MS (ESI +) m / z 379 [M + H] +

Example 3.3: Preparation of 6-hydroxy-7-oxo-2-phenyl-1,7-dihydro-pyrazolo [1,5-a] pyrimidine-5-carboxylic acid methyl ether

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Compound prepared by adapting the procedure of Example 3.

1H NMR (300 MHz, D6-DMSO): δ 3.94 (3H, s, OCH3), 6.47 (1H, s, CHC [Ph]), 7.42-7.51 (3H, m, Ar -CH), 7.95 (2H, d J = 6.6 Hz, Ar-CH). MS (ESI +) m / z 286 [M + 1] +.

HPLC procedure 2 95.6% / 1.24 min.

Example 3.4: Preparation of 6-hydroxy-7-oxo-2-phenyl-1,7-dihydro-pyrazolo [1,5a] pyrimidine-5-carboxylic acid 4-fluoro-benzylamide; composed with 4-fluoro-benzylamine

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Compound prepared by adapting the procedure of Example 3.1.

1H NMR (300 MHz, D6-DMSO): δ 4.01 (2H, s, NH3CH2Ph), 4.47 (2H, d, J = 6.0 Hz, [C = O] NHCH2), 6.03 ( 1H, s, CHC [Ph]), 7.11-7.51 (11H, m, Ar-CH), 7.91 (2H, d, J = 7.2 Hz, Ar-CH). MS (ESI +) m / z 379 [M + H] +

Example 3.5: Preparation of 6-hydroxy-7-oxo-2-phenyl-1,7-dihydro-pyrazolo [1,5a] pyrimidine-5-carboxylic acid 3,4-dichloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Compound prepared by adapting the procedure of Example 3.1.

1H NMR (300 MHz, D6-DMSO): δ 4.58 (2H, d, J = 6.0 Hz, NCH2), 6.49 (1H, s, CHC [Ph]), 7.29-7, 67 (6H, m, Ar-CH) 7.92 (2H, d, J = 7.2 Hz, Ar-CH), 9.20 (1H, t, J = 6.0 Hz, [C = O] NHCH2). MS (ESI +) m / z 429 [M + H] +

Example 3.6: Preparation of the 6-hydroxy-7-oxo-1,7-dihydro-pyrazolo [1,5-a] pyrimidine-5-carboxylic acid methyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Compound prepared by adapting the procedure of Example 3. 1H NMR (300 MHz, D6-DMSO): δ 7.96 (1H, s, CHCHNH), 6.05 (1H, d, J = 1.8 Hz, CHCHNH), 3.87 (3H, s, OCH3 ). MS (ESI +) m / z 210 (M + 1)

HPLC procedure 3 97% / 1.18 min

Example 3.7: Preparation of 6-hydroxy-7-oxo-1,7-dihydro-pyrazolo [1,5-a] pyrimidine-5-carboxylic acid 4-chloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Compound prepared by adapting the procedure of Example 3.1.

1H NMR (300 MHz, D6-DMSO): δ 10.0 (1H, s, OH), 9.35 (1H, t, J = 6.3 Hz, NHCH2), 7.93 (1H, d, J = 8.4 Hz, Ar-CH) 7.68 (1H, d, J = 8.4 Hz, Ar-CH), 7.38-7.26 (4H, m, Ar-CH), 4.30 (2H, d, J = 6.3 Hz, NHCH2). MS (ESI +) m / z Without ionization.

Example 3.8: Preparation of 1- (4-fluoro-benzyl) -6-hydroxy-7-oxo-1,7-dihydro-pyrazolo [1,5a] pyrimidine-5-carboxylic acid methyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Compound prepared by adapting the procedure of Example 3.

1H NMR (300 MHz, D6-DMSO): δ 8.48 (1H, d, J = 3.6 Hz, Ar-CH), 7.23-7.08 (4H, m, Ar-CH), 6 , 47 (1H, d, J = 3.6 Hz Ar-CH), 5.76 (2H, s, CH2Ar), 3.81 (3H, s, OCH3). MS (ESI +) m / z 318 [M + H] +

Example 3.9: Preparation of 1- (4-fluoro-benzyl) -6-hydroxy-7-oxo-1,7-dihydropyrazolo [1,5-a] pyrimidine-5-carboxylic acid 4-fluoro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Compound prepared by adapting the procedure of Example 3.1.

1H NMR (300 MHz, D6-DMSO): δ 11.96 (1H, s, OH), 9.53 (1H, s, NH), 8.48 (1H, d, J = 3.3 Hz, Ar -CH), 7.36 (1H, d, J = 5.7 Hz, Ar-CH), 7.33 (1H, d, J = 5.4 Hz, Ar-CH), 7.23 (1H, d, J = 5.4 Hz, Ar-CH), 7.20 (1H, d, J = 5.4 Hz, Ar-CH), 7.18-7.0 (4H, m, Ar-CH) , 6.40 (2H, d, J = 3.6 Hz, CH2NH), 5.73 (2H, s, CH2Ar). MS (ESI +) m / z 411 [M + H] +.

Example 3.10: Preparation of 1- (4-fluoro-benzyl) -6-hydroxy-7-oxo-1,7-dihydropyrazolo [1,5-a] pyrimidine-carboxylic acid 3,4-dichloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Compound prepared by adapting the procedure of Example 3.1.

1H NMR (300 MHz, D6-DMSO): δ 11.83 (1H, s, OH), 9.57 (1H, t, J = 6.9 Hz, NHCH2), 8.50 (1H, d, J = 3.6 Hz, Ar-CH) 7.56 (1H, m, Ar-CH), 7.31-7.04 (5H, m, Ar-CH), 6.41 (1H, d, J = 3.6 Hz, Ar-CH), 5.74 (3H, m, CH2Ar and Ar-CH), 4.41 (2H, d, J = 6.9 Hz, CH2NH).

MS (ESI +) m / z 461 [M] +.

Example 3.11: Preparation of the 6-hydroxy-1-methyl-7-oxo-1,7-dihydro-pyrazolo [1,5-a] pyrimidine-5-carboxylic acid methyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The compound prepared by adapting the procedure of Example 3 was used in the next Stage without further purification.

Example 3.12: Preparation of 6-hydroxy-1-methyl-7-oxo-1,7-dihydro-pyrazolo [1,5a] pyrimidine-5-carboxylic acid 3,4-dichloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Compound prepared by adapting the procedure of Example 3.1.

1H NMR (300 MHz, D6-DMSO): δ 9.77 (1H, s, NH), 8.12 (1H, d, J = 3.3 Hz, Ar-CH), 7.60-7.52 (2H, m, Ar-CH), 7.34-7.30, (1H, m, Ar-CH), 6.32 (1H, d, J = 3.6 Hz, Ar-CH), 4.46 (2H, d, J = 5.7 Hz, CH2NH), 4.02 (3H , s, CH3). MS (ESI +) m / z 367 and 369 [M + H] +.

Example 3.13: Preparation of 6-hydroxy-1-methyl-7-oxo-1,7-dihydro-pyrazolo [1,5a] pyrimidine-5-carboxylic acid 4-fluoro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Compound prepared by adapting the procedure of Example 3.1.

1H NMR (300 MHz, D6-DMSO): δ 11.8 (1H, s, OH), 9.54 (1H, s, NH), 8.12 (1H, d, J = 3.3 Hz, Ar -CH), 7.38 (1H, d, J = 9.3 Hz, Ar-CH), 7.35 (1H, d, J = 8.7 Hz, Ar-CH), 7.15 (1H, d, J = 9.3 Hz, Ar-CH), 7.12 (1H, d, J = 8.7 Hz, Ar-CH), 6.33 (1H, d, J = 3.6 Hz, Ar -CH), 4.45 (2H, d, J = 6.6 Hz, CH2NH), 4.05 (3H, s, CH3).

MS (ESI +) m / z 317 [M + H] +.

Example 3.14: Preparation of the 3-bromo-6-hydroxy-7-oxo-1,7-dihydro-pyrazolo [1,5-a] pyrimidine-5-carboxylic acid methyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The compound prepared by adapting the procedure of Example 3 was used in the next Stage without further purification.

Example 3.15: Preparation of 3,4-dichloro-benzylamide of 3-bromo-6-hydroxy-7-oxo-1,7-dihydro-pyrazolo [1,5a] pyrimidine-5-carboxylic acid

Although the following example is not included in the scope of the claims, it is entered as information from

image5

Compound prepared by adapting the procedure of Example 3.1.

1H NMR (300 MHz, D6-DMSO): δ 4.78 (2H, s, CH2NH), 7.35 (1H, d, J = 8.1 Hz, CHCHC [Cl]), 7.60 (1H, d, J = 8.1 Hz, CHCHC [Cl]), 7.61 (1H, d, J = 1.8 Hz, NHCHC [Br]), 7.75 (1H, s, [C] CHC [Cl]), 8.00 (1H, s, CH2NH), 8.50 (1H, s, OH). MS (ESI +) m / z 473 [M + MeCN] +.

Example 3.16: Preparation of 3-Bromo-6-hydroxy-7-oxo-1,7-dihydro-pyrazolo [1,5a] pyrimidine-5-carboxylic acid 2-fluoro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Compound prepared by adapting the procedure of Example 3.1.

1H NMR (300 MHz, D6-DMSO): δ 4.89 (2H, s, CH2NH), 7.21-7.45 (5H, m, 4 x Ar-CH and 1 x NH), 8.04 ( 1H, dd, J = 8.1, 7.4 Hz, Ar-CH), 8.74 (1H, s, CH2NH). MS (ESI +) m / z Without ionization.

Example 3.17: Preparation of the 6-hydroxy-7-oxo-3-phenyl-1,7-dihydro-pyrazolo [1,5-a] pyrimidine-5-carboxylic acid methyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Phenylpyrazolamine (100 mg, 0.63 mmol), diacetoxybumarate (180 mg, 0.69 mmol) and p-toluenesulfonic acid (5 mg) were heated at 100 duranteC for 20 minutes before cooling to room temperature and sonicated with ethanol : i-propanol for 10 min. The resulting precipitate was collected and washed with ethanol. The product was isolated as a yellow solid (47 mg, 26%).

1H NMR (300 MHz, D6-DMSO): δ 8.53 (1H, s, Ar-CH), 7.82 (2H, d, J = 6.9 Hz, Ar-CH), 7.47 (2H , m, Ar-CH), 7.29 (1H m, Ar-CH), 3.89 (3H, s, OCH3).

MS (ESI +) m / z 286 [M + H] +.

Example 3.18: Preparation of 6-hydroxy-7-oxo-3-phenyl-1,7-dihydro-pyrazolo [1,5a] pyrimidine-5-carboxylic acid 3,4-dichloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The compound was prepared by adapting the procedure of Example 3.1.

1H NMR (300 MHz, D6-DMSO): δ 9.29 (1H, m, NH), 8.48 (1H, bs, CHNH), 7.63 (1H, d, J = 1.8 Hz, Ar -CH), 7.61 (2H, m, Ar- CH), 7.59 (1H, s, Ar-CH), 7.47-7.35 (4H, m, Ar-CH), 7.24 (1H, m, Ar-CH), 4.59 ( 2H, d, J = 6.3 Hz, NHCH2). MS (ESI +) m / z 429 [M] +.

Example 3.19: Preparation of 3-cyano-1- (4-fluoro-benzyl) -6-hydroxy-7-oxo-1,7-dihydro-pyrazolo [1,5-a] pyrimidine-5 acid 4-fluoro-benzylamide -carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Sodium hydride (11.0 mg, 0.367 mmol) was added in one portion to a stirred suspension of 3-cyano-6-hydroxy-7-oxo1,7-dihydro-pyrazolo [1,5-fluoro-benzylamide [1,5] -a] pyrimidine-5-carboxylic compound composed of 4-fluorobenzylamine (Example 3.1) (100 mg, 0.306 mmol) in DMF (2 ml) under nitrogen at room temperature. The mixture was stirred for 30 min before adding p-fluorobenzyl chloride (40 µL, 0.366 mmol) and the mixture was heated at 90 ° C for 2 days. After that period, the reaction was cooled to room temperature and partitioned between ethyl acetate (10 ml) and aqueous hydrochloric acid (M, 10 ml). The organic layer was separated, and the aqueous layer was extracted with ethyl acetate (3 x 10 ml). The combined organic layers were washed with water (3 x 10 ml), brine (10 ml) and then concentrated. The residue was purified by column chromatography (95: 5: 1 dichloromethane: methanol: aqueous ammonia to give the desired product (50 mg, 36%).

1H NMR (300 MHz, D6-DMSO): δ 8.11 (1H, s, CHC [CN]), 7.45 (1H, d, J = 6.0 Hz, Ar-H), 7.42 ( 1H, d, J = 6.0 Hz, Ar-H), 7.36 (1H, d, J = 5.4 Hz, Ar-H), 7.34 (1H, d, J = 5.4 Hz , Ar-H), 7.15 (1H, d, J = 9.3 Hz, Ar-H), 7.09 (1H, d, J = 8.7 Hz, ArH), 7.05 (1H, d, 9.3 Hz, Ar-H), 7.03 (1H, d, J = 8.7 Hz, Ar-H), 4.96 (2H, s, CH2N), 4.38 (2H, d , J = 6.3 Hz, NHCH2).

MS (ESI +) m / z 434 [M + H] +.

Example 4: Preparation of the 6-hydroxy-2-methyl-7-oxo-7H-isoxazolo [2,3-a] pyrimidine-5-carboxylic acid methyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image6

(10 mg) were combined in a capped vial and heated to 100 ° C. After 5 h the reaction was cooled to room temperature and ethanol (2.5 ml) and iso-propi ether (2.5 ml) were added to the residue, which was sonicated for 15 min. The resulting precipitate was collected by filtration and washed with cold ethanol (5 ml) and dried by pump to give the 6-hydroxy-2-methyl-7-oxo-7H-isoxazolo [2,3-a] pyrimidine acid methyl ester -5-carboxylic (331 mg, 37%): 1 H NMR (300 MHz, D6DMSO) δ 2.50 (3H, s, CH3), 3.84 (3H, s, OCH3), 6.67 (1H, s , H3), 10.31 (1H, br s, OH).

MS (ESI +) m / z 225 (M + 1

HPLC procedure 4 99.3% / 0.52 min.

Example 5: Preparation of the 3-hydroxy-4-oxo-4,10-dihydro-benzo [4,5] imidazo [1,2a] pyrimidine-2-carboxylic acid methyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

2-Aminobenzimidazole (200 mg, 1.50 mmol), dimethoxy diacetoxybumarate (430 mg, 1.65 mmol) and p-toluenesulfonic acid (5 mg) were heated in a closed tube at 100 duranteC for 2 h. The residue was triturated with ethanol / propyl ether (5 ml) and sonicated for 10 min. The resulting precipitate was collected and recrystallized from hot acetonitrile to give the 3-hydroxy-4-oxo-4,10-dihydro-benzo [4,5] imidazo [1,2a] pyrimidine-2-carboxylic acid methyl ester (153 mg, 39%)

1H NMR (300 MHz, D6-DMSO): δ 3.90 (3H, s, OCH3), 7.10 (1H, m, Ar-CH), 7.25-7.51 (3H, m, 2 3 Ar-CH and NH) and 8.43 (1H, d, J = 7.8 Hz, Ar-CH).

MS (ESI +) m / z 260 (M + 1)

Example 6: Preparation of 7-bromo-3-hydroxy-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid 4-fluorobenzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

7-Bromo-3-hydroxy-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid methyl ester (20 mg, 0.07 mmol) and pfluorobenzylamine (19 uL, 0.17 mmol) in dry methanol (4 ml) were heated to reflux with stirring. The progress of the reaction was monitored by HPLC. After 6 h, the reaction was cooled to room temperature and concentrated in vacuo. The residue was triturated with diethyl ether (2 ml) and the precipitate was collected by filtration and washed with diethyl ether (10 ml) and dried by pump to give 7-bromo-3-hydroxy acid 4-fluoro-benzylamide. -4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid (24 mg, 92%) as a yellow-brown solid.

1H NMR (300 MHz, D6DMSO) δ 4.51 (2H, s, NCH2), 7.33 (6H), 8.57 (1H, m, H6), 11.21 (1H, br s, NH).

MS (ESI +) m / z 392 (M [Br79] +1), 394 (M [Br81) +1), HPLC procedure 1 99.6% / 6.5 min.

The following compounds (6.1-6.3) were obtained by adaptation of the procedure described in Example 6:

Example 6.1: Preparation of 3-hydroxy-7-methyl-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid 4-fluorobenzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image7

HPLC procedure 1 94.8% / 6.20 min.

Example 6.2: Preparation of 3-hydroxy-7-methyl-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid 3,4-dichloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

MS (ESI +) m / z 378 (M [Cl35] +1). HPLC procedure 1 100% / 6.74 min.

Example 6.3: Preparation of the 7-chloro-3-hydroxy-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid 4-fluorobenzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

1H NMR (300 MHz, CDCl3) δ 9.95 (1H, bs, NHa), 8.93 1H, app. t, NHb), 8.62 (1H, s, H6a), 7.58 (1H, d, J =, 4 Hz, H8), 7.457.27 (2H, m, ArH), 7.26-7, 21 (2H, m, ArH), 7.07-6.99 (2H, m, ArH), 6.99-6.90 (2H, m, ArH), 4.52 (0.32H, d, = , 4 Hz, CH2a), 4.31 (0.68H, d, J 6.6 Hz, CH2b).

MS (ESI +) m / z 348 (M + 1), (ESI-) m / z 346 (M-1)

HPLC procedure 1 93% / 6.35 min.

Example 6.4: Preparation of the 7-chloro-3-hydroxy-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid 3,4-dichloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

1H NMR (300 MHz, CDCl3) δ 9.95 (1H, bs, NHa), 9.13 (1H, app. T, NHb), 8.65 (1H, s, H6a), 7.57 (1H, d, J 8.4 Hz, H8) 7.407.33 (2H, m, ArH), 7.26-7.24 (1H, m, ArH), 7.153 (1H, dd, J = 8.1 Hz, 1 , 8 Hz, ArH), 7.05 (1H, d, J = 7.8 Hz, ArH), 4.51 (0.41H, d, J = 6.3 Hz, CH2a), 4.30 (0 , 59H, d, J = 6.3 Hz, CH2b) MS (ESI +) m / z 400 (M + 1), (ESI-) m / z 396 (M-1)

HPLC procedure 1 91% / 6.89 min.

Example 6.5: Preparation of 3-hydroxy-4-oxo-4,10-dihydro-benzo [4,5] imidazo [1,2a] pyrimidine-2-carboxylic acid 4-fluoro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

1H NMR (300 MHz, D6-DMSO): δ 4.51 (2H, d, J = 6.3 Hz, NHCH2), 7.14-7.50 (7H, m, Ar-CH), 8.44 (1H, d, J = 8.1 Hz, Ar-CH) and 9.27 (1H, t, J = 6.3 Hz, NHCH2).

MS (ESI +) m / z 353 (M + 1) HPLC procedure 5 92% / 3.10 min.

Example 6.6: Preparation of 6-hydroxy-2-methyl-7-oxo-7H-isoxazolo [2,3-a] pyrimidine-5-carboxylic acid 4-fluoro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

1H NMR (300 MHz, D6DMSO) δ 2.49 (3H, s, CH3), 4.48 (2H, d, J = 5.9 Hz, NCH2), 6.50 (s, 1H, H3), 6 , 95 (2H, m, ArH) 7.30 (2H, m, ArH), 9.14 (1H, br s, NH).

MS (ESI +) m / z 318 (M + 1), (ESI-) m / z 316 (M-1)

HPLC procedure 1 90% / 5.50 min.

Example 6.7: Preparation of the 6-hydroxy-2-methyl-7-oxo-7H-isoxazolo [2,3a] pyrimidine-5-carboxylic acid 3,4-dichloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

1H NMR (300 MHz, D6DMSO) δ 2.49 (3H, s, CH3), 4.48 (2H, d, J = 5.9 Hz, NCH2), 7.30 (1H, dd, J = 8, 2, 2.3 Hz, ArH), 7.52 (2H, m, ArH), 10.45 (1H, br s, NH).

MS (ESI +) m / z 368 (M [CI35, CL35] +1), (ESI-) m / z 366 (M [CI35, CL35] -1)

HPLC procedure 4 92% / 1.83 min.

Example 6.8: Preparation of 3-hydroxy-7-morpholin-4-yl-4-oxo-4H-pyrido [1,2a] pyrimidine-2-carboxylic acid 4-fluoro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

1H NMR (300 MHz, D6-DMSO): δ 12.13 (1H, s, OH), 9.67 (1H, t, J = 6.9 Hz, NHCH2), 8.00 (1H, s, CHC [morpholine]), 7.85 (1H, d, J = 9.6 Hz, CHCHC [morpholine]), 7.50 (1H, d, J = 9.6 Hz, CHCHC [morpholine]), 7.39 (2H, m, Ar-CH), 7.16 (2H, m, Ar-CH), 4.50 (2H, d, J = 6.9 Hz, NHCH2), 3.76 (4H, m, CH2OCH2 ) and 3.16 (4H, m, CH2NCH2).

MS (ESI +) m / z 397 (M + 1)

HPLC procedure 6 98% / 6.40 min

Example 6.9: Preparation of 3-hydroxy-7-morpholin-4-yl-4-oxo-4H-pyrido [1,2a] pyrimidine-2-carboxylic acid 4-methoxy-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

1H NMR (300 MHz, D6-DMSO): δ 12.25 (1H, s, OH), 9.55 (1H, t, J = 6.9 Hz, CH2NH), 7.99 (1H, s, CHC [morpholine]) 7.84 (1H, d, J = 9.6 Hz, CHCHC [morpholine]), 7.50 (1H, d, J = 9.6 Hz, CHCHC [morpholine]), 7.29 ( 2H, d, J = 8.4 Hz, ArCH), 6.91 (2H, d, J = 8.4 Hz, Ar-CH), 4.44 (2H, d, J = 6.9 Hz, CH2NH ), 3.79-3.72 (4H, m, CH2OCH2), 3.70 (3H, s, OCH3) and 3.18 (4H, m, CH2NCH2).

MS (ESI +) m / z 411 (M + 1)

HPLC procedure 6 99% / 6.21 min

Example 6.10: Preparation of 3-hydroxy-7-morpholin-4-yl-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

1H NMR (300 MHz, D6-DMSO): δ 12.17 (1H, s, OH), 9.66 (1H, t, J = 6.3 Hz, CH2NH), 7.99 (1H, s, CHC [morpholine]), 7.85 (1H, d, J = 9.6 Hz, CHCHC [morpholine]), 7.50 (1H, d, J = 9.6 Hz, CHCHC [morpholine]), 7.38 -7.25 (5H, m, Ar-CH) 4.52 (2H, d, J = 6.3 Hz, CH2NH), 3.77 (4H, m, CH2OCH2) and 3.18 (4H, m, CH2NCH2).

MS (ESI +) m / z 381 (M + 1)

HPLC procedure 6 97% / 6.32 min

Example 6.11: Preparation of 3-hydroxy-7-morpholin-4-yl-4-oxo-4H-pyrido [1,2a] pyrimidine-2-carboxylic acid 4-chloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

1H NMR (300 MHz, D6-DMSO): δ 12.10 (1H, s, OH), 9.69 (1H, t, J = 6.9 Hz, CH2NH), 7.99 (1H, s, CHC [morpholine]) 7.85 (1H, d, J = 9.9 Hz, CHCHC [morpholine]), 7.50 (1H, d, J = 9.9 Hz, CHCHC [morpholine]), 7.52- 7.36 (4H, m, Ar-CH) 4.50 (2H, d, J = 6.9 Hz, CH2NH), 3.76 (4H, m, CH2OCH2) and 3.18 (4H, m, CH2NCH2 ).

MS (ESI +) m / z 415 (M + 1)

HPLC procedure 6 95% / 7.22 min

Example 6.12: Preparation of 3-hydroxy-7-morpholin-4-yl-4-oxo-4H-pyrido [1,2a] pyrimidine-2-carboxylic acid 2-chloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

 1H NMR (300 MHz, D6-DMSO): δ 12.00 (1H, s, OH), 9.63 (1H, t, J = 6.3 Hz, NHCH2), 8.00 (1H, s, CHC [morpholine]), 7.85 (1H, d, J = 9.6 Hz, CHCHC [morpholine]), 7.53 (1H, d, J = 9.6 Hz, CHCHC [morpholine]), 7.48 (1H, m, Ar-CH), 7.37-7.31 (3H, m, Ar-CH), 4.61 (2H, d, J = 6.3 Hz, NHCH2), 3.78 (4H , m, CH2OCH2) and 3.19 (4H, m, CH2NCH2).

MS (ESI +) m / z 415 (M + 1) +

HPLC procedure 5 90% / 3.85 min

Example 6.13: Preparation of 3-hydroxy-7-morpholin-4-yl-4-oxo-4H-pyrido [1,2a] pyrimidine-2-carboxylic acid 3,4-dichloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

1H NMR (300 MHz, D6-DMSO): δ 12.00 (1H, s, OH), 9.73 (1H, bs, CH2NH), 7.99 (1H, s, CHC [morpholine]), 7, 85 (1H, d, J = 9.9 Hz, CHCHC [morpholine]), 7.61 (2H, m, CHCHC [morpholine] and CHC [CI] C [CI]), 7.50 (2H, d, J = 8.1 Hz, ArCH), 7.35 (2H, d, J = 8.1 Hz, Ar-CH), 4.51 (2H, d, J = 6.6 Hz, CH2NH), 3, 77 (4H, m, CH2OCH2) and 3.18 (4H, m, CH2NCH2).

MS (ESI +) m / z 449 (M [CI35, CL35] +1), (ESI-) m / z 447 (M [CI35, CL35] -1)

HPLC procedure 5 94% / 4.84 min

Example 6.14: Preparation of 3-hydroxy-8-morpholin-4-yl-4-oxo-4H-pyrido [1,2a] pyrimidine-2-carboxylic acid 4-fluoro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Using the product of Example 2.4, the procedure described in Example 6 was adapted (except that only 1.3 eq. Of 4-fluorobenzylamine was used) to give the desired compound.

1H NMR (300 MHz, D6-DMSO): δ 11.60 (1H, s, OH), 9.50 (1H, t, J = 6.3 Hz, NH), 8.55 (1H, d, J = 8.4 Hz, Ar-CH), 7.41-7.36 (2H, m, Ar-CH), 7.22-7.12 (3H, m, Ar-CH), 6.51 (1H , s, Ar-CH), 4.47 (2H, d, J = 6.3 Hz, NHCH2), 3.72 (4H, m, CH2OCH2), 3.34 (4H, m, CH2NCH2).

(ESI-) m / z 397 (M-1)

HPLC procedure 7 94.4% / 9.0 min

Example 6.15: Preparation of 3-hydroxy-8-morpholin-4-yl-4-oxo-4H-pyrido [1,2a] pyrimidine-2-carboxylic acid 3,4-dichloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Using the product of Example 2.4, the procedure described in Example 6 was adapted (except that only 1.3 eq. Of 3,4-dichlorobenzylamine was used) to give the desired compound.

1H NMR (300 MHz, D6-DMSO): δ 11.50 (1H, s, OH), 9.61 (1H, t, J = 6.3 Hz, NH), 8.57 (1H, d, J = 8.4 Hz, Ar-CH), 7.63-7.60 (2H, m, Ar-CH), 7.34 (1H, d, J = 8.4 Hz, Ar-CH), 7, 22 (1H, d, J = 8.4 Hz, Ar-CH), 6.53 (1H, s, Ar-CH), 4.49 (2H, d, J = 6.3 Hz, NHCH2), 3 , 75 (4H, m, CH2OCH2), 3.37 (4H, m, CH2NCH2).

(ESI-) m / z 447 (M [Cl35] -1)

HPLC procedure 7 93.2% / 10.2 min

Example 6.16: Preparation of 3-hydroxy-9-morpholin-4-yl-4-oxo-4H-pyrido [1,2a] pyrimidine-2-carboxylic acid 4-fluoro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Using the product of Example 2.5, the procedure described in Example 6 was adapted (except that only 1.3 eq of 4-fluorobenzylamine was used) to give the desired compound.

1H NMR (300 MHz, CDCl3) δ 3.23 (4H, s, -NCH2CH2O-), 3.76 (4H, s, -NCH2CH2O-), 4.61 (2H, d, J = 5.7 Hz, - (O = C) NHCH2-), 6.91 (2H, m, ArH), 7.09 (2H, t, J = 8.4 Hz, ArH), 7.34 (2H, bt, ArH), 7.98 (1H, s, (O = C) NHCH2-), 8.61 (1H, d, J = 7.2 Hz, ArH), 11.80 (1H, s, OH).

(ESI +) m / z 399 (M + 1)

HPLC procedure 7 97.0% / 11.6 min

Example 6.17: Preparation of 3-hydroxy-9-morpholin-4-yl-4-oxo-4H-pyrido [1,2a] pyrimidine-2-carboxylic acid 3,4-dichloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Using the product of Example 2.5, the procedure described in Example 6 was adapted (except that only 1.3 eq of 3,4-dichlorobenzylamine was used) and the reaction was carried out in a 1: 1 mixture of methanol / tetrahydrofuran) to give the desired compound.

1H NMR (300 MHz, CDCl3) δ 3.35 (4H, s, -NCH2CH2O-), 3.97 (4H, s, -NCH2CH2O-), 4.64 (2H, d, J = 6.0 Hz, - (O = C) NHCH2-), 6.99 (2H, m, ArH), 7.24 (1H, m, ArH), 7.48 (2H, m, ArH), 8.50 (1H, bs , - (O = C) NHCH2-), 8.69 (1H, d, J = 7.8 Hz, ArH), 11.84 (1H, s, OH). (ESI +) m / z 471 (M + Na)

HPLC procedure 7 91.0% / 13.1 min

Example 6.18: Preparation of 3-hydroxy-4-oxo-7-piperidin-1-yl-4H-pyrido [1,2a] pyrimidine-2-carboxylic acid 3,4-dichloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Using the product of Example 2.6, the procedure described in Example 6 was adapted (except that only 1.3 eq of 3,4-dichlorobenzylamine was used) to give the desired compound.

1H NMR (300 MHz, CDCl3) δ 1.58 (2H, bm, cyclic-N (CH2) 3CH2NCH2-), 1.73 (4H, bs, cyclic-N (CH2) 3CH2NCH2-), 3.20 (4H , bm, cyclic-N (CH2) 2CH2NCH2-), 4.62 (2H, d, J = 6.0 Hz, - (O = C) NHCH2-), 7.50 (4H, m, ArH), 8 , 24 (1H, dd, J = 1.8 Hz ArH), 8.51 (1H, bs, - (O = C) NHCH2-), 11.86 (1H, s, OH). (ESI-) m / z 445 (M [Cl35] -1)

HPLC procedure 7 91.0% / 14.9 min

Example 7: Preparation of 3-cyano-6-hydroxy-7-oxo-1,7-dihydro-pyrazolo [1,5-a] pyrimidine-5-carboxylic acid 4-fluorobenzylamide acid

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

5- (4-Fluoro-benzylcarbamoyl) -7-oxo-2-phenyl-1,7-dihydro-pyrazolo [1,5-a] pyrimidin-6-olate; 4-Fluorobenzyl-ammonium (25 mg) was suspended in water (1 ml) and aqueous hydrochloric acid (1.0 M, 1 ml) was added. The mixture was sonicated for 5 min and the resulting precipitate was collected by filtration and washed with water (2 ml) and dried by pump to give the 3-cyano-6-hydroxy-7-oxo-1 acid 4-fluorobenzylamide. , 7-dihydro-pyrazolo [1,5-a] pyrimidine-5-carboxylic acid (13 mg) as a colorless solid.

1H NMR (300 MHz, D6-DMSO): δ 4.53 (2H, d, J = 5.9 Hz, NHCH2), 7.15 (2H, m, ArH), 7.42 (2H, m, ArH ), 8.36 (1H, s H2), 7.92 (2H, d, J = 7.2 Hz, Ar-CH), 9.14 (1H, t, J = 5.9 Hz, NHCH2), 11.25 (1H, br s, OH) MS (ESI-) m / z 326 (M-1)

HPLC procedure 5 95.4% / 4.14 min

By adaptation of the procedure described in Example 7, the following compounds were obtained.

Example 7.1: Preparation of 6-hydroxy-7-oxo-2-phenyl-1,7-dihydro-pyrazolo [1,5a] pyrimidine-5-carboxylic acid 4-fluoro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

1H NMR (300 MHz, D6-DMSO): δ 4.58 (2H, d, J = 5.9 Hz, NHCH2), 6.50 (1H, s, H3), 7.15 (2H, m, ArH ), 7.42 (5H, m, ArH), 7.93 (2H, m, ArH), 9.10 (1H, t, J = 5.9 Hz, NHCH2), 10.80 (1H, br s , OH), 11.84 (1H, br s, NH)

MS (ESI +) m / z 379 (M + 1)

Example 8: Preparation of 3-hydroxy-7-methyl-2- (5-m-tolyl- [1,3,4] oxadiazol-2-yl) -pyrido [1,2-a] pyrimidin-4-one

image8

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The product of Example 2.1 (1.5 g, 6.4 mmol) and potassium carbonate (2.7 g, 19.6 mmol) were mixed with acetone (30 ml) under an N2 atmosphere. The mixture was stirred at 70 ° C for 25 min, after which benzyl bromide (2.0 g, 11.7 mmol) was added and the mixture was refluxed for 10 h. After cooling to room temperature, the mixture was poured onto water, (100 ml), extracted with dichloromethane. The organic phase was washed with water, dried and concentrated in vacuo. Purification by flash column chromatography (dichloromethane) achieved the desired compound (1.5 g, 70%).

1H NMR (300 MHz, CDCl3): δ 2.44 (d, J = 0.9 Hz, 3H), 3.92 (s, 3H), 5.32 (s, 2H), 7.27-7, 41 (m, 3H), 7.47-7.57 (m, 3H) 7.65 (d, J = 9.1 Hz, 1H), 8.76-8.85 (m, 1H).

MS (ESI +) m / z 325 (M + 1), 347 (M + 23).

Example 8.2: Preparation of 3-benzyloxy-7-methyl-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

To a stirred solution of the product from Example 8.1 (400 mg, 1.23 mmol) in methanol (20 ml) was added a 1N aqueous solution of aluminum hydroxide and lithium (2.46 ml) at room temperature. After 3 h, 1N aqueous hydrochloric acid (20 ml) was added. The mixture was extracted with ethyl acetate and the organic phase was washed with brine, dried (Na2SO4) and concentrated in vacuo. The product was used directly in Example 8.3.

Example 8.3: Preparation of 3-Methyl-benzoic acid N ’- (3-benzyloxy-7-methyl-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carbonyl) hydrazide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

To a solution of the product of Example 8.2 (200 mg, 0.644 mmol) in tetrahydrofuran (10 ml), 3-methylbenzoyl hydrazine (94.8 mg, 0.632 mmol), 1-hydroxybenzotriazole (6.98 mg, 0.0576) mmol) and 1- (3-dimethylaminopropol) -3-ethylcarbodiimide hydrochloride (98 mg, 0.632 mmol) successively at room temperature. After 12 h the reaction solution was rapidly stopped with water (20 ml) and extracted with ethyl acetate. The extract was washed with 2N aqueous hydrochloric acid (20 ml), 2N aqueous sodium hydroxide (20 ml), dried (Na2SO4) and concentrated in vacuo to give the desired compound (53%).

1H NMR (300 MHz, DMSO-d6): δ 2.39 (s, 3H), 2.44 (s, 3H), 5.21 (s, 2H), 7.30-7.45 (m, 5H ), 7.56-7.63 (m, 2H), 7.69-7.79 (m, 3H), 7.84 (dd, J = 9.4, 2.1 Hz, 1H), 8, 78-8.85 (m, 1H), 10.56 (d, J = 11.1 Hz, 2H).

MS (ESI +) m / z 443 (M + 1), 465 (M + 23).

Example 8.4: Preparation of 3-benzyloxy-7-methyl-2- (5-m-tolyl- [1,3,4] oxadiazol-2-yl) -pyrido [1,2-a] pyrimidin4-one

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The product of Example 8.3 (202 mg, 0.457 mmol), carbon tetrachloride (0.221 ml, 2.28 mmol) and triethylamine (0.165 ml, 1.19 mmol) were mixed with acetonitrile (10 ml). To this mixture was added triphenylphosphine (291 mg, 1.11 mmol) at room temperature. After stirring at room temperature overnight, the reaction solution was diluted with ethyl acetate (100 ml), washed with saturated aqueous sodium bicarbonate (50 ml), water (50 ml) and brine (50 ml) successively , and then dried (Na2SO4). The crude product was subjected to flash chromatography (hexane-ethyl acetate 1: 1) to give the desired compound.

1H NMR (300 MHz, DMSO-d6): δ 2.41 (s, 3H), 2.46 (d, J = 1.1 Hz, 3H), 5.32 (s, 2H), 7.28- 7.33 (m, 3H), 7.46-7.52 (m 4H), 7.75-7.90 (m, 4H), 8.82-8.86 (m, 1H).

MS (ESI +) m / z 425 (M + 1), 447 (M + 23).

Example 8.5: Preparation of 3-hydroxy-7-methyl-2- (5-m-tolyl- [1,3,4] oxadiazol-2-yl) -pyrido [1,2-a] pyrimidin-4-one

image 1

To a stirred solution of the product from Example 8.4 (20 mg, 0.047 mmol) in acetonitrile (5 ml), trimethylsilyl iodide (54 uL, 0.38 mmol) was added dropwise under N2 at room temperature. After 2 h, methanol (5 ml) was added and the solution was stirred for 10 min. Water (10 ml) was added and the reaction was extracted with dichloromethane. The organic phase was washed with aqueous sodium bisulfite solution, dried (Na2SO4) and concentrated in vacuo to give the desired compound (88.6%).

1H NMR (300 MHz, CDCl3): δ 2.43 (s, 3H), 2.49 (s, 3H), 7.38-7.50 (m, 3H), 7.65 (d, J = 8 , 8 Hz, 1H), 8.03-8.09 (m, 2H) 8.78 (s, 1H), 9.92 (brs, 1H).

MS (ESI +) m / z 335 (M + 1), 357 (M + 23).

Example 8.6: Preparation of 3-hydroxy-7-methyl-2- (5-phenyl- [1,3,4] oxadiazol-2-yl) -pyrido [1,2-a] pyrimidin-4-one

image 1

The procedure described in Example 8.1-8.5 was adapted to prepare 3-hydroxy-7-methyl-2- (5-phenyl- [1,3,4] oxadiazol2-yl) pyrido [1,2-a] pyrimidin- 4-one 1H NMR (300 MHz, CDCl3) δ 2.43 (s, 3H), 7.45 (d, J = 8.5 Hz, 1H), 7.54-7.68 (m, 4H), 8.26 (d, J = 6.6 Hz, 2H), 8.77 (s, 1H) MS (ESI +) m / z 321 (M + 1) HPLC procedure 7 82.8% / 14.3 min

Example 8.7: Preparation of 2- [5- (2-Chloro-phenyl) - [1,3,4] oxadiazol-2-yl] -3-hydroxy-7-methyl-pyrido [1,2-a] pyrimidin -4-one

image 1

The procedure described in Example 8.1-8.5 was adapted to prepare 2- [5- (2-chloro-phenyl) - [1,3,4] oxadiazol-2-yl] -3-hydroxy7-methyl-pyrido [1 , 2-a] pyrimidin-4-one.

1H NMR (300 MHz, CDCl3) δ 2.43 (d, J = 1.1 Hz, 3H), 7.44 (dd, J = 9.2, 2.2 Hz, 1H), 7.49 (dd , J = 7.7, 1.5 Hz, 1H), 7.55 (dt, J = 7.5, 1.8 Hz, 1H), 7.60-7.66 (m, 2H), 8, 13 (dd, J = 7.7, 1.8 Hz, 1H), 8.76-8.79 (m, 1H), 9.71-9.91 (brs, 1H)

MS (ESI +) m / z 377 (M + Na +)

HPLC procedure 7 92.2% / 15.4 min

Example 8.8: Preparation of 2- [5- (4-methoxy-phenyl) - [1,3,4] oxadiazol-2-yl] -3-hydroxy-7-methyl-pyrido [1,2-a] pyrimidin -4ona

image 1

The procedure described in Example 8.1-8.5 was adapted to prepare 2- [5- (4-methoxy-phenyl) - [1,3,4] oxadiazol-2-yl] -3-hydroxy-7-methyl-pyrido [1 , 2-a] pyrimidin-4-one.

1H NMR (300 MHz, CDCl3) δ 2.43 (s, 3H), 3.92 (s, 3H), 7.07 (d, J = 8.9 Hz, 2H), 7.44 (dd, J = 9.5 Hz, 1.9 Hz, 1H), 7.64 (d, J = 9.4 Hz, 1H), 8.21 (d, J = 8.9 Hz, 2H), 8.78 ( d, J = 1.9 Hz, 1H), 9.88-10.10 (brs, 1H)

MS (ESI +) m / z 373 (M + Na +)

HPLC procedure 7 92.4% / 15.3 min

Example 8.9: Preparation of 2- [5- (4-fluoro-phenyl) - [1,3,4] oxadiazol-2-yl] -3-hydroxy-7-morpholin-4-yl-pyrido [1,2a ] pyrimidin-4-one

image 1

Using the product of Example 2.3 as a starting material, the procedure described in Example 8.1-8.5 was adapted to prepare 2- [5- (4-fluoro-phenyl) - [1,3,4] oxadiazol-2 -yl] -3-hydroxy-7-morpholin-4-yl-pyrido [1,2-a] pyrimidin-4-one.

1H NMR (300 MHz, DMSO-d6) δ 3.23 (t, J = 4.8 Hz, 4H), 3.80 (t, J = 4.8 Hz, 4H), 7.52 (t, J = 9.0 Hz, 2H), 7.67 (d, J = 10.0 Hz, 1H), 7.87 (dd, J = 10.0 Hz, 2.4 Hz, 1H), 8.03 ( d, J = 2.3 Hz, 1H), 8.16 (dd, J = 8.8 Hz, 5.1 Hz, 2H), 10.46-10.60 (brs, 1H)

HPLC procedure 7 98.4% / 8.5 min

Example 9: Preparation of 2- [5- (4-fluoro-benzyl) - [1,3,4] oxadiazol-2-yl] -3-hydroxy-7-methyl-pyrido [1,2-a] pyrimidin -4-one

image9

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

To a stirred solution of the product from Example 8.1 (800 mg, 2.56 mmol) in methanol (30 ml), hydrazine (6.0 eq) was added at room temperature. The mixture was then heated at 45 ° C for 4 h and then partially concentrated in vacuo (not to dryness) and then cooled to room temperature. The resulting solid was filtered, washed with water and dried in vacuo to give the desired compound (650 mg, in a yield of 78%).

1H NMR (300 MHz, DMSO-d6): δ 2.42 (s, 3H), 5.15 (s, 2H), 7.28-7.45 (m, 3H), 7.48-7.53 (m, 2H), 7.66 (d, J = 8.8Hz, 1H) 7.80 (dd, J = 8.8, 2.1 Hz, 1H), 8.78 (s, 1H), 8 , 93 (brs, 2H), 9.7 (brs, 1H).

MS (ESI +) m / z 325 (M + 1), 347 (M + 23).

Example 9.2: Preparation of the 3-benzyloxy-7-methyl-4-oxo-4H-pyrido [1,2a] pyrimidine-2- N '- [2- (4-fluoro-phenyl) -acetyl] -hydrazide carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The product of Example 9.1 (160 mg, 0.524 mmol) and sodium carbonate (106 mg, 1 mmol) were mixed with anhydrous tetrahydrofuran (25 ml) and then cooled in an ice bath. To this stirred solution was added 4fluorophenylacetyl chloride (90 mg, 0.55 mmol) dropwise. The mixture was stirred at room temperature for 2 h and then concentrated in vacuo. The residue was partitioned between ethyl acetate and water and the organic phase was washed with water, dried (Na2SO4) and concentrated in vacuo. Short column chromatography gave the desired compound (210 mg, in 86% yield)

1H NMR (300 MHz, CDCl3): δ 2.45 (s, 3H), 3.67 (s, 2H), 5.40 (s, 2H), 6.95-7.10 (m, 2H), 7.30-7.42 (m, 5H), 7.50-7.60 (m, 3H), 7.64 (d, J = 9.4 Hz, 1H), 8.70-8.80 ( m, 2H), 10.42 (brs, 1H).

MS (ESI +) m / z 461 (M + 1), 483 (M + 23).

Example 9.3: Preparation of 3-benzyloxy-2- [5- (4-fluoro-benzyl) - [1,3,4] oxadiazol-2-yl] -7-methyl-pyrido [1,2-a] pyrimidin -4ona

image10

The procedure described in Example 8.4 was adapted to the product obtained in Example 9.2 to give the desired product (70%).

1H NMR (300 MHz, CDCl3): δ 2.46 (d, J = 0.9 Hz, 3H), 4.25 (s, 2H), 5.39 (s, 2H), 6.96-7, 05 (m, 2H), 7.27-7.34 (m, 5H) 7.387.45 (m, 2H), 7.56 (dd, J = 9.1 Hz, 2.0Hz, 1H), 7, 70 (d, J = 9.1 Hz, 1H), 8.78-8.83 (m, 1H).

MS (ESI +) m / z 443 (M + 1), 465 (M + 23).

Example 9.4: Preparation of 2- [5- (4-fluoro-benzyl) - [1,3,4] oxadiazol-2-yl] -3-hydroxy-7-methyl-pyrido [1,2-a] pyrimidin -4ona

image 1

The procedure described in Example 8.5 was adapted to the product obtained in Example 9.3 to give the desired product (52%).

1H NMR (300 MHz, CDCl3): δ 2.42 (s, 3H), 4.37 (s, 2H), 7.02-7.11 (m, 2H), 7.32-7.48 (m , 3H), 7.61 (d, J = 9.6 Hz, 1H) 8.76 (s, 1H), 9.79 (brs, 1H).

MS (ESI-) m / z 351 (M-1) HPLC procedure 7 97.3% / 8.5 min

Example 9.5: Preparation of 2- [5- (3,4-dichloro-benzyl) - [1,3,4] oxadiazol-2-yl] -3-hydroxy-7-methyl-pyrido [1,2-a ] pyrimidin-4ona

image 1

The procedure described in Example 9.1-9.4 was adapted to prepare 2- [5- (3,4-dichloro-benzyl) - [1,3,4] oxadiazol-2-yl] -3-hydroxy-7-methyl -pyrido [1,2-a] pyrimidin-4-one.

1H NMR (300 MHz, CDCl3) δ 2.43 (s, 3H), 4.35 (s, 2H), 7.26 (1H, overlap), 7.41-7.55 (m, 3H), 7 , 64 (d, J = 9.2 Hz 1H), 8.76 (s, 1H), 9.55-9.85 (brs, 1H)

MS (ESI-) m / z 401 (M-1)

HPLC procedure 7 97.6% / 18.0 min

Example 9.6: Preparation of 2- [5- (3,4-dichloro-benzyl) - [1,3,4] oxadiazol-2-yl] -3-hydroxy-7-morpholin-4-yl-pyrido [1 , 2a] pyrimidin-4-one

image 1

Using the starting material prepared in Example 2.3, the procedure described in Example 9.1-9.4 was adapted to prepare 2- [5- (3,4-dichloro-benzyl) - [1,3,4] oxadiazole -2-yl] -3-hydroxy-7-methyl-pyrido [1,2-a] pyrimidin-4-one.

1H NMR (300 MHz, DMSO-d6) δ 3.18-3.24 (m, 4H), 3.75-3.83 (m, 4H), 4.47 (s, 2H), 7.40 ( dd, J = 8.3, 2.0 Hz, 1H), 7.61 (d, J = 9.8 Hz, 1H), 7.65 (d, J = 8.5 Hz, 1H), 7, 72 (d, J = 2.0 Hz, 1H), 7.85 (dd, J = 9.9, 2.5 Hz, 1H), 8.01 (d, J = 2.5 Hz, 1H), 10.41 (s, 1 H)

HPLC procedure 7 94.1% / 17.2 min

Example 9.7: Preparation of 2- [5- (4-fluoro-benzyl) - [1,3,4] oxadiazol-2-yl] -3-hydroxy-7-morpholin-4-yl-pyrido [1,2a ] pyrimidin-4-one

image 1

Using the starting material prepared in Example 2.3, the procedure described in Example 9.1-9.4 was adapted to

prepare 2- [5- (4-fluoro-benzyl) - [1,3,4] oxadiazol-2-yl] -3-hydroxy-7-methyl-pyrido [1,2-a] pyrimidin-4-one . 1H NMR (300 MHz, DMSO-d6) δ 3.20 (t, J = 4.8 Hz, 4H), 3.79 (t, J = 4.8 Hz, 4H), 4.42 (s, 2H ), 7.21 (t, J = 9.0 Hz, 2H), 7.43 (dd, J = 8.8, 5.5 Hz, 2H), 7.61 (d, J = 9.9 Hz, 1H), 7.85 (dd, J = 9.8, 2.5 Hz, 1H), 8.00 (d, J = 2.5 Hz, 1H), 10.39 (s, 1H).

MS (ESI-) m / z 422 (M-1) HPLC procedure 7 94.1% / 14.7 min

Example 10: Preparation of 2- [5- (4-fluoro-benzyl) - [1,3,4] thiadiazol-2-yl] -3-hydroxy-7-methyl-pyrido [1,2-a] pyri- midin-4-one

image11

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image12

. The product of Example 9.2 (80 mg, 0.173 mmol) and Lawensson's reagent (200 mg, 0.5 mmol) were mixed with toluene (15 ml) and refluxed for 10 h. The reaction mixture was concentrated in vacuo and flash chromatography (ethyl acetate / dichloromethane / diethyl ether 2: 6: 1) gave the desired compound (60 mg, 75.3%).

1H NMR (300 MHz, DMSO-d6): δ 2.42 (s, 3H), 4.53 (s, 2H), 5.26 (s, 2H), 7.17-7.26 (m, 2H ), 7.29-7.35 (m, 3H), 7.40-7.49 (m, 4H), 7.67 (d, J = 9.5 Hz, 1H), 7.80 (dd, J = 9.4Hz, 1.9 Hz, 1H), 8.77 (d, J = 1.1 Hz, 1H).

MS (ESI +) m / z 459 (M + 1), 481 (M + 23).

Example 10.2: Preparation of 2- [5- (4-fluoro-benzyl) - [1,3,4] thiadiazol-2-yl] -3-hydroxy-7-methyl-pyrido [1,2-a] pyrimidin -4ona

image 1

The procedure described in Example 8.5 was adapted to the product obtained in Example 10.1 to give the desired product (34%)

1H NMR (300 MHz, CDCl3): δ 2.39 (d, J = 0.8 Hz, 3H), 4.48 (s, 2H), 7.02-7.12 (m, 2H), 7, 30-7.45 (m, 4H), 8.71-8.77 (m, 1H), 10.80 (brs, 1H).

MS (ESI +) m / z 369 (M + 1), 391 (M + 23).

HPLC procedure 7 96.7% / 15.8 min

Example 10.3: Preparation of 2- [5- (3,4-dichloro-benzyl) - [1,3,4] thiadiazol-2-yl] -3-hydroxy-7-methyl-pyrido [1,2-a ] pyrimidin-4ona

image 1

The procedure described in Example 10.1-10.2 was adapted to prepare 2- [5- (3,4-dichloro-benzyl) - [1,3,4] thiadiazol-2-yl] -3-hydroxy-7-methyl-pyrido [1,2-a] pyrimidin-4-one

1H NMR (300 MHz, CDCl3) δ 3.16-3.25 (m, 4H), 3.73-3.83 (m, 4H), 4.29 (s, 2H), 7.37 (d, J = 8.6 Hz, 1H), 7.63 (d, J = 8.3 Hz, 2H), 7.69 (s, 1H), 7.86 (d, J = 8.8 Hz, 1H) , 7.98 (s, 1H), 10.71 (s, 1H)

MS (ESI-) m / z 417 (M [Cl35] -1)

HPLC procedure 7 97.8% / 19.8 min

Example 10.4: Preparation of 2- [5- (3,4-dichloro-benzyl) - [1,3,4] thiadiazol-2-yl] -3-hydroxy-7-morpholin-4-yl-pyrido [1 , 2a] pyrimidin-4-one

image 1

Using the starting material prepared in Example 2.3, the procedure described in Example 10.1-10.2 was adapted to prepare 2- [5- (3,4-dichloro-benzyl) - [1,3,4] thiadiazole -2-yl] -3-hydroxy-7-morpholin-4-yl-pyrido [1,2-a] pyrimidin-4-one

1H NMR (300 MHz, DMSO-d6) δ 3.15-3.25 (m, 4H), 3.70-3.85 (m, 4H), 4.60 (s, 2H), 7.42 ( dd, J = 8.2 Hz, 2.1 Hz, 1H) 7.60 (d, J = 9.8 Hz, 1H), 7.65 (d, J = 8.2 Hz, 1H), 7, 74 (d, J = 2.1 Hz, 1H), 7.86 (dd, J = 9.8 Hz, 2.5 Hz, 1H), 8.01 (d, J = 2.3 Hz, 1H) , 10.50-11.10 (brs, 1H)

MS (ESI-) m / z 488 (M-1)

HPLC procedure 7 97.6% / 19.3 min

Example 10.5: Preparation of 2- [5- (4-fluoro-benzyl) - [1,3,4] thiadiazol-2-yl] -3-hydroxy-7-morpholin-4-yl-pyrido [1,2a ] pyrimidin-4-one

image 1

Using the starting material prepared in Example 2.3, the procedure described in Example 9.1-9.4 was adapted to prepare 2- [5- (4-fluoro-benzyl) - [1,3,4] thiadiazol-2 -yl] -3-hydroxy-7-morpholin-4-yl-pyrido [1,2-a] pyrimidin-4-one

1H NMR (300 MHz, DMSO-d6) δ 3.20 (t, J = 4.8 Hz, 4H), 3.78 (t, J = 4.8 Hz, 4H), 4.56 (s, 2H ), 7.21 (t, J = 8.8 Hz, 2H), 7.47 (dd, J = 8.8 Hz, 5.5 Hz, 2H), 7.59 (d, J = 9.8 Hz, 1H), 7.85 (dd, J = 9.9 Hz, 2.7 Hz, 1H), 8.01 (d, J = 2.6 Hz, 1H), 10.80 (s, 1H)

MS (ESI-) m / z 438 (M-1)

HPLC procedure 7 94.1% / 14.2 min

Example 11: Preparation of 2- [5- (4-fluoro-benzyl) - [1,2,4] oxadiazol-3-yl] -3-hydroxy-7-methyl-pyrido [1,2-a] pyrimidin- 4-one

image13

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The product of Example 8.1 (3.1 g, 10 mmol) was dissolved in anhydrous tetrahydrofuran (50 ml) and cooled to -78 ° C. To this stirred solution was added dropwise diisobutylaluminum hydride (13 ml, 1 N in tetrahydrofuran). After 4 h, TLC showed that the starting material had been consumed and the reaction solution was rapidly stopped with aqueous sodium sulfate solution. The insoluble material was removed by filtration and the filtrate was concentrated in vacuo. The residue was dissolved in a combined ethyl acetate / dichloromethane solvent (1: 1 5 ml) and washed with brine, dried (Na2SO4) and filtered. To a solution of hydroxylamine hydrochloride (760 mg, 11 mmol) in water (120 ml), the above aldehyde solution was added followed by the addition of sodium bicarbonate (900 mg, 10.7 mmol). The mixture was stirred at room temperature for 2 h and the resulting precipitate was collected by filtration and washed with water and dried in vacuo to give the desired product (2.77g, 90% overall two-stage yield).

Example 11.2: Preparation of 3-benzyloxy-7-methyl-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carbonitrile

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Trichloro-1,3,5-triazine (576 mg, 3.15 mmol) was dissolved in anhydrous N, N-dimethylformamide (DMF) (1 ml) and stirred at room temperature for 30 min. To this solution was added dropwise a solution of the product of Example 11.1 (927 mg, 3 mmol) in DMF (5 ml). The mixture was kept at room temperature for 2 h, then ethyl acetate (50 ml) was added and the organic phase was separated and washed with brine, dried (Na2SO4) and concentrated in vacuo. Purification by short flash chromatography gave the desired compound (530 mg, 60.7%).

1H NMR (300 MHz, CDCl3): δ 2.45 (d, J = 1.2 Hz, 3H), 5.54 (s, 2H), 7.30-7.40 (m, 3H), 7, 48-7.54 (m, 2H), 7.55-7.58 (m 2H), 8.77 (dd, J = 2.7, 1.2 Hz, 1H).

Example 11.3: Preparation of 3-benzyloxy-N-hydroxy-7-methyl-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxamidine

Although the following example is not included in the scope of the claims, it is entered as information from

image5

The product of Example 11.2 (530 mg, 1.82 mmol) and hydroxylamine hydrochloride (0.549 g, 7.9 mmol) were mixed with ethanol (50 ml). To this stirred solution was added sodium bicarbonate (663 mg, 7.9 mmol) and the mixture was heated at 70 ° C for 3 h. The solvent was removed in vacuo and the residue was dissolved in a combined solvent (dichloromethane / ethanol 200 ml: 10 ml), washed with water, dried (Na2SO4) and concentrated in vacuo to give the desired compound (472 mg, 80%)

MS (ESI +) m / z 325 (M + 1), 347 (M + 23), 379 (M + 55).

Example 11.4: Preparation of 3-benzyloxy-2- [5- (4-fluoro-benzyl) - [1,2,4] oxadiazol-3-yl] -7-methyl-pyrido [1,2-a] pyrimidin4 -ona

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The product of Example 11.3 (472 mg, 1.46 mmol) was dissolved in a combined dichloromethane / tetrahydrofuran solvent (120 ml: 120 ml) with stirring. Triethylamine (155 mg, 1.53 mmol) was then added followed by the dropwise addition of 4-fluorophenyl acetyl colide (263 mg, 1.53 mmol). The mixture was stirred at room temperature for 2 h and then concentrated in vacuo and the resulting residue was dissolved in ethyl acetate and washed with water, dried (Na2SO4) and concentrated in vacuo. The resulting solid was used without purification.

The previous solid (668 mg) was suspended in toluene (25 ml) and the mixture was refluxed for 24 h. The solvent was concentrated in vacuo to give the desired compound quantitatively.

1H NMR (300 MHz, CDCl3): δ 2.46 (d, J = 0.7 Hz, 3H), 4.32 (s, 2H), 5.38 (s, 2H), 7.00-7, 08 (m, 2H), 7.20-7.30 (m, 3H?), 7.327.37 (m, 2H), 7.44-7.52 (m, 2H), 7.55 (dd, J = 9.2, 1.9 Hz, 1H), 7.74 (d, J = 9.1 Hz, 1H), 8.81-8.85 (m, 1H).

MS (ESI +) m / z 443 (M + 1), 465 (M + 23).

Example 11.5: Preparation of 2- [5- (4-fluoro-benzyl) - [1,2,4] oxadiazol-3-yl] -3-hydroxy-7-methyl-pyrido [1,2-a] pyrimidin -4ona

image 1

The procedure described in Example 8.5 was adapted to the product obtained in Example 11.4 to give the desired product (68%).

1H NMR (300 MHz, CDCl3): δ 2.41 (d, J = 0.7 Hz, 3H), 4.39 (s, 2H), 7.01-7.12 (m, 2H), 7, 32-7.44 (m, 3H), 7.68 (d, J = 9.2 Hz, 1H), 8.70 (s, 1H), 8.72-8.90 (brs, 1H).

MS (ESI +) m / z 353 (M + 1), 375 (M + 23).

HPLC procedure 7 94.5% / 14.4 min

Example 12: Preparation of 3-hydroxy-7-methyl-2- (5-phenyl-oxazol-2-yl) -pyrido [1,2-a] pyrimidin-4-one

image14

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

α-Bromoacetophenone (7.0 g, 0.035 mol), urotropin (5.4 g, 0.0385 mol) and sodium iodide (5.8 g, 0.0385 mol) were mixed in ethanol (425 ml) and stirred at room temperature for 24 h. The reaction mixture was filtered and the filter cake was washed with cold ethanol and the resulting solid was dissolved in ethanol (100 ml) and 6N aqueous hydrochloric acid (20 ml) was added. The mixture was refluxed for 5 h and then cooled to room temperature. The mixture was filtered and the filtrate was concentrated in vacuo. The resulting residue was recrystallized from diisopropyl ether / concentrated hydrochloric acid (100/1) to give the desired product (4.1 g, 69%).

 1H NMR (300 MHz, CDCl3): δ 4.57 (s, 2H), 7.56-7.691 (m, 2H), 7.71-7.76 (m, 1H), 8.00-8.03 (m, 2H), 8.52 (br s, 3H).

Example 12.2: Preparation of 3-benzyloxy-7-methyl-4-oxo-4H-pyrido [1,2a] pyrimidine-2-carboxylic acid (2-oxo-2-phenyl-ethyl) -amide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

To a stirred solution of the product from Example 8.2 (324 mg, 1 mmol) in tetrahydrofuran (15 ml) at room temperature, the product of Example 12.1 (162 mg, 1.2 mmol), 1-hydroxybenzotriazole hydrochloride ( 162 mg, 1.2 mmol), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide (191 mg, 1 mmol) and triethylamine (112 mg, 1.1 mmol) successively. After 3 h, the reaction solution was stopped quickly with saturated sodium bicarbonate solution (5 ml). The mixture was extracted with ethyl acetate and the organic phase was washed with water, brine, then dried with (Na2SO4) and concentrated in vacuo. Flash chromatography of the residue provided the desired product (215 mg, 45%).

1H NMR (300 MHz, CDCl3): δ 2.45 (s, 3H), 4.93 (d, J = 4.5Hz, 2H), 5.44 (s, 2H), 7.26-7.32 (m, 3H), 7.51-7.72 (m, 7H) 8.03 (d, J = 7.5Hz, 2H), 8.65 (s, 1H, NH), 8.79 (s, 1 HOUR).

MS (ESI +) m / z 428 (M + 1), 450 (M + Na +), 482 (M + MeOH + Na +).

Example 12.3: Preparation of 3-benzyloxy-7-methyl-2- (5-phenyl-oxazol-2-yl) -pyrido [1,2-a] pyrimidin-4-one

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

To a stirred solution of the product from Example 12.2 (170 mg, 0.4 mmol) in acetonitrile (5 ml) at room temperature was added carbon tetrachloride (360 mg, 2.4 mmol), triethylamine (130 mg, 1, 28 mmol) and triphenylphosphine (320 mg, 1.2 mmol) successively. After 2 h, saturated sodium bicarbonate solution (5 ml) was added and the products were extracted with ethyl acetate. The organic phase was washed with water, brine, then dried (Na2SO4) and concentrated in vacuo. Flash chromatography of the residue gave the desired compound (142 mg, 86%).

1H NMR (300 MHz, CDCl3) δ2.45 (s, 3H), 5.46 (s, 2H), 7.29-7.37 (m, 6H), 7.54-7.61 (m, 6H ), 7.75 (d, 1H), 8.80 (s, 1H).

MS (ESI-) m / z 380 (M-1); MS (ESI +) m / z 410 (M + 1), 432 (M + Na +), 464 (M + MeOH + Na +), 841

(2M + Na +)

Example 12.4: Preparation of 3-hydroxy-7-methyl-2- (5-phenyl-oxazol-2-yl) -pyrido [1,2-a] pyrimidin-4-one

image 1

The product of Example 12.3 (62 mg, 0.5 mmol) and sodium iodide (440 mg, 2.9 mmol) were mixed with acetonitrile (5 ml). To this stirred solution was added trimethylsilyl chloride (316 mg, 2.9 mmol) dropwise. The mixture was stirred for 1 h, then quickly stopped by the addition of methanol (5 ml) followed by water (20 ml) and then extracted with ethyl acetate. The combined organic layers were washed with brine, dried (Na2SO4) and concentrated in vacuo to a volume of 1 ml. Hexane (15 ml) was added dropwise and the resulting solid was collected by filtration and dried in vacuo to give the desired product (38 mg, 79%).

1H NMR (300 MHz, CDCl3) δ 2.40 (s, 3H), 7.40-7.57 (m, 6H), 7.84 (s, 2H), 8.74 (s, 1H). MS (ESI +) m / z 320 (M + 1), 342 (M + Na +), 374 (M + MeOH + Na +), 661 (2M + Na +). HPLC procedure 7 95.0% / 15.6 min

Example 12.5: Preparation of 2- [5- (4-fluoro-phenyl) -oxazol-2-yl] -3-hydroxy-7-methyl-pyrido [1,2-a] pyrimidin4-one

image 1

The procedure described in Example 12.1-12.4 was adapted to prepare 2- [5- (4-fluoro-phenyl) -oxazol-2-yl] -3-hydroxy-7-methyl-pyrido [1,2-a] pyrimidine- 4-one 1H NMR (300 MHz, CDCl3) δ 2.43 (s, 3H), 7.45 (d, J = 8.5 Hz, 1H), 7.54-7.68 (m, 4H), 8.26 (d, J = 6.6 Hz, 2H), 8.77 (s, 1H) MS (ESI +) m / z 321 (M + 1) HPLC procedure 7 82.8% / 15.5 min

Example 12.6: Preparation of 2- [5- (4-methoxy-phenyl) -oxazol-2-yl] -3-hydroxy-7-methyl-pyrido [1,2-a] pyrimidin-4-one

image 1

The procedure described in Example 12.1-12.4 was adapted to prepare 2- [5- (4-methoxy-phenyl) -oxazol-2-yl] -3-hydroxy7methyl-pyrido [1,2-a] pyrimidin-4- ona.

1H NMR (300 MHz, CDCl3) δ 2.40 (s, 3H), 3.87 (s, 3H), 6.99 (d, J = 8.3 Hz, 2H), 7.39 (d, J = 9.2 Hz, 1H), 7.45 (s, 1H), 7.62 (d, J = 9.2 Hz, 1H), 7.77 (d, J = 8.5 Hz, 2H), 8.75 (s, 1H), 10.75-11.35 (brs, 1H)

MS (ESI +) m / z 372 (M + Na +) HPLC procedure 7 94.1% / 16.1 min

Example 12.7: Preparation of 1-amino-3- (4-fluoro-phenyl) -propan-2-one hydrochloride

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image15

The procedures described in Tetrahedron were adapted. 1994, 50 (21), 6287-6298 and Chem. Pharm. Bull. 1984, 32 (7), 2536-2543 to give the 1-amino-3- (4-fluoro-phenyl) -propan-2-one hydrochloride.

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The procedures described in Tetrahedron were adapted. 1994, 50 (21), 6287-6298 and Chem. Pharm. Bull. 1984, 32 (7), 2536-2543 to give the 1-amino-3- (3,4-dichloro-phenyl) -propan-2-one hydrochloride.

Example 12.9: Preparation of 2- [5- (4-fluoro-benzyl) -oxazol-2-yl] -3-hydroxy-7-methyl-pyrido [1,2-a] pyrimidin4-one

image 1

Using the material from Example 12.7 and adapting the procedures of Examples 12.2-4, 2 [5- (4-fluorobenzyl) -oxazol-2-yl] -3-hydroxy-7-methyl-pyrido [1,2] was achieved -a] pyrimidin-4-one.

1H NMR (300 MHz, CDCl3) δ 2.41 (s, 3H), 4.17 (s, 2H), 6.81-7.18 (m, 3H), 7.26-7.60 (m, 4H), 8.77 (s, 1H), 10.40-11.80 (brs, 1H)

MS (ESI +) m / z 352 (M + 1) HPLC procedure 7 89.6% / 15.5 min

Example 13: Preparation of 3-hydroxy-7-methyl-2- (5-phenyl-thiazol-2-yl) -pyrido [1,2a] pyrimidin-4-one

image16

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The product of Example 12.2 (100 mg, 0.23 mmol) and Lawensson reagent (120 mg, 0.3 mmol) were mixed with toluene (10 ml) and refluxed for 12 h. The reaction mixture was concentrated in vacuo and flash chromatography gave the

desired compound (27 mg, with a 27% yield).

1H NMR (300 MHz, CDCl3): δ 2.44 (s, 3H), 5.55 (s, 2H), 7.30-7.70 (m, 11H), 7.80 (d, J = 9 , 2 Hz, 1H), 8.28 (s, 1H), 8.80 (s, 1H).

Example 13.2: Preparation of 3-hydroxy-7-methyl-2- (5-phenyl-thiazol-2-yl) -pyrido [1,2a] pyrimidin-4-one

image 1

The procedure described in Example 8.5 was adapted to the product obtained in Example 13.1 to give the desired product (80%).

 1H NMR (300 MHz, CDCl3): δ 2.39 (s, 3H), 7.32-7.56 (m, 5H), 7.62-7.70 (m, 2H), 8.12 (s , 1H), 8.75 (s, 1H), 11.65 (brs, 1H). MS (ESI +) m / z 336 (M + 1) HPLC procedure 7 98.7% / 17.5 min

Example 13.3: Preparation of 2- [5- (4-fluoro-benzyl) -thiazol-2-yl] -3-hydroxy-7-methyl-pyrido [1,2-a] pyrimidin-4-one

image 1

Using the material from Example 12.7 and adapting the procedures of Examples 13.1 to 13.2, 2- [5- (4-fluorobenzyl) -thiazol-2-yl] -3-hydroxy-7-methyl-pyrido [1, 2-a] pyrimidin-4-one.

1H NMR (300 MHz, DMSO-d6) δ 2.35 (d, J = 1.2 Hz, 3H), 4.31 (s, 2H), 7.18 (t, J = 9.9 Hz, 2H ), 7.39 (dd, J = 8.9 Hz, 5.5 Hz 2H), 7.45-7.60 (m, 2H), 7.95 (s, 1H), 8.58-8, 64 (m, 1H), 11.31 (s, 1H) MS (ESI +) m / z 390 (M + Na +)

HPLC procedure 7 96.7% / 18.5 min

Example 13.4: Preparation of 2- [5- (3,4-Dichloro-benzyl) -thiazol-2-yl] -3-hydroxy-7-methyl-pyrido [1,2-a] pyrimidin-4-one

image 1

Using the material from Example 12.8 and adapting the procedures of Examples 13.1 to 13.2, 2- [5- (3,4-dichloro-benzyl) -thiazol-2-yl] -3-hydroxy-7-methyl- pyrido [1,2-a] pyrimidin-4-one.

1H NMR (300 MHz, DMSO-d6) δ 2.36 (d, J = 1.2 Hz, 3H), 4.34 (s, 2H), 7.37 (dd, J = 8.3 Hz, 2 , 0 Hz, 1H), 7.47-7.60 (m 2H), 7.62 (d, J = 8.2 Hz, 1H), 7.67 (d, J = 2.1 Hz, 1H) , 7.98 (s, 1H), 8.60-8.65 (m, 1H), 11.28 (s, 1H)

MS (ESI +) m / z 418 (M + 1)

HPLC procedure 7 98.8% / 19.8 min

Example 13.5: Preparation of 2- [5- (4-fluoro-benzyl) -thiazol-2-yl] -3-hydroxy-7-morpholin-4-yl-pyrido [1,2-a] pyrimidin-4- ona

image 1

Using the materials from Example 2.3 and Example 12.7 and adapting the procedures of Examples 13.1 to

13.2 2- [5- (4-Fluoro-benzyl) -thiazol-2-yl] -3-hydroxy-7-morpholin-4-yl-pyrido [1,2-a] pyrimidin-4-one was obtained

1H NMR (300 MHz, DMSO-d6) δ 3.14-3.21 (m, 4H), 3.74-3.81 (m, 4H), 4.31 (s, 2H), 7.18 ( t, J = 8.9 Hz, 2H), 7.39 (dd, J = 8.8 Hz, 5.5 Hz, 2H), 7.53 (d, J = 9.9 Hz, 1H), 7 , 83 (dd, J = 9.9 Hz, 2.6 Hz, 1H), 7.95 (s, 1H), 8.04 (d, J = 2.5 Hz, 1H) 11.25 (s, 1H) MS (ESI +) m / z 461 (M + Na +)

HPLC procedure 7 86.3% / 19.6 min

image17

Using the materials from Example 2.3 and Example 12.8 and adapting the procedures of Examples 13.1 to

13.2 2- [5- (3,4-Dichloro-benzyl) -thiazol-2-yl] -3-hydroxy-7-morpholin-4-yl-pyrido [1,2-a] pyrimidin-4- was obtained ona.

1H NMR (300 MHz, DMSO-d6) δ 3.14-3.21 (m, 4H), 3.74-3.82 (m, 4H), 4.33 (s, 2H), 7.36 ( dd, J = 8.2 Hz, 2.1 Hz, 1H) 7.53 (d, J = 10.0 Hz, 1H), 7.62 (d, J = 8.2 Hz, 1H), 7, 67 (d, J = 2.1 Hz, 1H), 7.74-7.86 (m, 1H), 7.96 (s, 1H), 8.01-8.06 (m, 1H), 11 , 1811.28 (brs, 1H)

MS (ESI-) m / z 487 (M-1)

HPLC procedure 7 97.1% / 19.7 min

Example 13.7: Preparation of 2- [5- (4-fluoro-benzyl) -thiazol-2-yl] -3-hydroxy-7-morpholin-4-ylmethyl-pyrido [1,2-a] pyrimidin-4one

image 1

Stage 1:

The product of Example 2.1 (3.66 g, 15.6 mmol), t-butyldimethylsilyl chloride (3.52 g) and imidazole (2.66 g) were added to dichloromethane / DMF (30 mU10 ml) and The mixture was stirred at room temperature for 2 h. The mixture was diluted with dichloromethane (30 ml) and the organic phase was washed with water, dried, filtered and concentrated in vacuo. The residue was subjected to column chromatography (hexane / ethyl acetate 4: 1) to give the desired compound (5.02 g, 92%).

1H NMR (300 MHz, CDCl3): δ 0.32 (s, 6H), 0.99 (s, 9H), 2.39 (s, 3H), 3.97 (s, 3H), 7.42 ( dd, J = 9.1, 1.8 Hz, 1H), 7.62 (d, J = 9.2 Hz, 1H), 8.68 (bs, 1H).

Stage 2:

To a stirred solution of the product from Step 1 (5 g, 14 mmol) in carbon tetrachloride (80 ml) was added Nbromosuccinimide (4.1 g) and t-butyl peroxide (0.348 g) under a nitrogen atmosphere. The reaction mixture was refluxed for 5 h and then cooled to room temperature. The solution was diluted with dichloromethane (200 ml), washed with water, dried, filtered and concentrated in vacuo. The residue was subjected to column chromatography (hexane / ethyl acetate 8: 1) to give the desired compound (3.0 g, 48%) as a yellow solid.

1H NMR (300 MHz, DMSO-d6): δ 0.26 (s, 6H), 0.94 (s, 9H), 3.86 (s, 3H), 4.88 (s, 2H), 7, 66 (d, J = 9.2 Hz, 1H), 7.79 (dd J = 9.3, 2.0 Hz, 1H), 9.03 (d, J = 1.8 Hz, 1H)

Stage 3:

The product of Step 2 (1.1 g, 2.6 mmol) and morpholine (672 mg, 7.73 mmol) were dissolved in a combined dichloromethane / methanol solvent (1: 1, 20 ml). The solution was stirred at room temperature for 4 h and then partially concentrated in vacuo and diluted with dichloromethane (40 ml) which was washed with brine, dried, filtered and evaporated under reduced pressure. Purification by silica gel column chromatography (hexane / ethyl acetate 1: 1) obtained the desired product (1.03 g, 92%).

1H NMR (300 MHz, DMSO-d6): δ 0.26 (s, 6H), 0.93 (s, 9H), 2.43 (t, J = 4.5 Hz, 4H), 3.53- 3.62 (m, 6H), 3.86 (s, 3H), 7.64 (dd, J = 9.1, 0.6 Hz, 1H), 7.76 (dd, J = 9.2, 1.9 Hz, 1H), 8.74 (dd, J = 1.8, 0.6 Hz, 1H)

Stage 4:

The product of Step 4 (100 mg, 0.23 mmol) was added to a stirred mixture of solvents glacial acetic acid / water / tetrahydrofuran (1: 1: 3, 5 ml) and the mixture was stirred overnight at temperature ambient. Water (10 ml) was added and then solid sodium carbonate was added to adjust pH 7. The mixture was extracted twice with dichloromethane and the combined organic layers were washed, dried and concentrated in vacuo to give the desired compound (65 mg, 88%).

1H NMR (300 MHz, DMSO-d6): δ 2.42 (t, J = 4.5 Hz, 4H), 3.53-3.63 (m, 6H), 3.88 (s, 3H), 7.58 (d, J = 9.2 Hz, 1H), 7.64 (dd, J = 9.4, 1.7 Hz, 1H), 8.62-8.67 (m, 1H), 10 , 24 (s, 1H)

Stages 5-9:

The procedures described in Example 8.1 (except that the reaction was carried out at 70 ° C using DMF as solvent), Example 8.2, Example 12.2, Example 13.1 and Example 12.4 were adapted to give 2- [5- (4-fluoro -benzyl) -thiazol-2-yl] -3-hydroxy-7-morpholin-4-ylmethyl-pyrido [1,2-a] pyrimidin-4-one.

1H NMR (300 MHz, DMSO-d6) δ 2.40 (m, 4H, N-CH2-CH2-O), 3.53 (s, 2H, Ar-CH2-N), 3.57 (t, J = 4.7 Hz, 4H, NCH2-CH2-O), 4.30 (s, 2H, CH2-thiazole), 7.17 (t, J = 8.9 Hz, 2H, ArH), 7.39 ( dd, J = 8.9 Hz, 5.4 Hz, 2H, ArH), 7.52 (d, J = 8.9 Hz, 1H, H9), 7.65 (dd, J = 8.9, 2 , 4 Hz, 1H, H8), 7.95 (s, 1H, CH (thiazole)), 8.66 (m, 1H, H6), 11.33 (s, 1H, OH).

MS (ESI +) m / z 453 (M + 1)

Example 14: Preparation of 3-hydroxy-4-oxo-4,10-dlihydro-benzo [4,5] imidazo [1,2a] pyrimidine2-carboxylic acid substituted benzylamides: general route

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The 2-aminobenzimidazole was rented by adaptation of the procedure described in WO2005 / 058869. The procedure described in Example 5 was adapted to prepare the methyl esters that were converted to the amido derivatives by adaptation of the procedure described in Example 6. The final products were purified by recrystallization or with preparative HPLC (obtaining the salt form). The following Examples (14.1-14.17) were prepared by adapting the above procedure.

Example 14.1: Preparation of 3-hydroxy-4-oxo-4,10-dihydro-benzo [4,5] imidazo [1,2-α] pyrimidine-2-carboxylic acid 3,4-dichloro-benzylamide acid

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

1H NMR (300 MHz, D6 DSMO): δ 9.38 (1H, m, NHCH2), 8.43 (1H, d, J = 8.1 Hz, Ar-CH), 7.72-7.25 ( 6H, m, Ar-CH), 4.52 (2H, d, J = 6.3 Hz, NHCH2).

MS (ESI-) m / z 403 (M-1)

Example 14.2: Preparation of 3-hydroxy-4-oxo-4,10-dihydro-benzo [4,5] imidazo [1,2α] pyrimidine-2-carboxylic acid 4-fluoro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

1H NMR (300 MHz, D6 DMSO): δ 9.27 (1H, m, NHCH2), 8.44 (1H, d, J = 8.1 Hz, Ar-CH), 7.50-7.14 ( 7H, m, Ar-CH), 4.51 (2H, d, J = 6.3 Hz, NHCH2).

MS (ESI +) m / z 353 (M + 1).

Example 14.3: Preparation of 3,4-difluoro-benzylamide of 3-hydroxy-4-oxo-4,10-dihydro-benzo [4,5] imidazo [1,2α] pyrimidine-2-carboxylic acid

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

1H NMR (300 MHz, D6-DMSO): δ 9.31 (1H, t, J = 6.3 Hz, NHCH2), 8.44 (1H, d, J = 8.1 Hz, Ar-CH), 7.51-7.18 (6H, m, Ar-CH), 4.51 (2H, d, J = 6.3 Hz, NHCH2).

MS (ESI-) m / z 370 (M-1).

Example 14.4: Preparation of 3-hydroxy-10-methyl-4-oxo-4,10-dihydro-benzo [4,5] imidazo [1,2-α] pyrimidine2-carboxylic acid 4-fluoro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

MN 1H (300 MHz, D6 DMSO): δ 11.89 (OH), 9.61 (1H, t, J = 6.6 Hz, NHCH2), 8.44 (1H, d, J = 7.8 Hz , Ar-CH), 7.54 (2H, m, Ar-CH), 7.41 (2H, dd, J = 9.0, 5.7 Hz, Ar-CH), 7.35-7.30 (1H, m, Ar-CH), 7.17 (2H, m, Ar-CH), 4.54 (2H, d, J = 6.6 Hz, NHCH2), 3.78 (3H, s, CH3 ).

MS (ESI +) m / z 367 (M + 1).

Example 14.5: Preparation of 3-hydroxy-10-methyl-4-oxo-4,10-dihydro-benzo [4,5] imidazo [1,2-α] pyrimidine2-carboxylic acid 4-fluoro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

1H NMR (300 MHz, D6 DMSO): δ 11.71 (OH), 9.61 (1H, t, J = 6.6 Hz, NHCH2), 8.44 (1H, d, J = 8.1 Hz , Ar-CH), 7.63-7.52 (4H, m, Ar-CH), 7.38-7.34 (2H, m, Ar-CH), 4.54 (2H, d, J = 6.6 Hz, NHCH2), 3.79 (3H, s, CH3).

MS (ESI +) m / z 417 (M + 1).

Example 14.6: Preparation of 3,4- dichloro-benzylamide of 10- (4-fluoro-benzyl) -3-hydroxy-4-oxo-4,10-dihydrobenzo [4,5] imidazo [1,2-α ] pyrimidine-2-carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

1H NMR (300 MHz, D6 DMSO): δ 11.8 (1H, s, OH), 9.75 (1H, bs, NH), 8.47 (1H, d, J = 4.8 Hz, Ar- CH), 7.64 (1H, d, J =, 1 Hz, Ar-CH), 7.63 (1H, s, Ar-CH), 7.55 (1H, d, J = 5.1 Hz, Ar-CH), 7.54 (1H, d, J = 4.8 Hz, Ar-CH), 7.48 (1H, dd, J = 4.8, 4.5 Hz, Ar-CH), 7 , 45 (1H, dd, J = 4.8, 4.2 Hz, Ar-CH), 7.38-7.32 (2H, m, Ar-CH), 7.17 (1H, d, J = 5.4 Hz, Ar-CH) 7.15 (1H, d, J = 5.1 Hz, Ar-CH), 5.61 (2H, s, Ar-CH2), 4.57 (2H, d, J = 3.6 Hz, CH2NH).

MS (ESI +) m / z 511 (M + 1).

MS (ESI +) m / z 516 (M) +.

Example 14.8: Preparation of 3-hydroxy-4-oxo-10- (2-pyrrolidin-1-yl-ethyl) -4,10-dihydro-benzo [4,5] imidazo [1-3,4-dichloro-benzylamide , 2-α] pyrimidine-2-carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image18

image 1

1H NMR (300 MHz, D6 DMSO): δ 8.09 (1H, s, NH), 7.70-7.65 (3H, m, Ar-CH), 7.37-7.29 (4H, m , Ar-CH), 5.30 (0.7H, s, Tautomer B NCH2), 4.62 (1.3H, d, J = 6.3 Hz, Tautomer A NCH2), 4.31 (2H, t , J = 6.9 Hz, NCH2CH2N), 2.98-2.92 (2H, m, NCH2CH2N), 2.65 (2H, m, CH2NCH2), 2.59 (2H, m, CH2NCH2), 1, 82-1.77 (4H, m, NCH2CH2CH2).

MS (ESI +) m / z 500 (M) +.

Example 14.9: Preparation of 3-hydroxy-4-oxo-10- (2-piperidin-1-yl-ethyl) -4,10-dihydro-benzo [4,5] imidazo [1-3,4-dichloro-benzylamide , 2-α] pyrimidine-2-carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

1H NMR (300 MHz, D6 DMSO): δ 11.58 (1H, bs, OH), 8.69 (1H, d, J = 7.8 Hz, Ar-CH), 8.12 (1H, bs, NH), 7.48 (2H, d, J = 7.0 Hz, Ar-CH), 7.45 (1H, d, J = 8.4 Hz, Ar-CH), 7.35-7.30 (2H, m, Ar-CH), 7.23 (1H, dd, J = 8.1, 1.8 Hz, Ar-CH) 4.62 (2H, d, J = 6.0 Hz, CH2NH) , 4.31 (2H, t, J = 6.9 Hz, NCH2CH2N), 2.72 (2H, t, J = 6.9 Hz, NCH2CH2N), 2.46 (4H, t, J = 5.4 Hz, CH2NCH2), 1.62-1.41 (6H, m, NCH2CH2CH2CH2).

MS (ESI +) m / z 514 and 516 (M) +.

Example 14.10: Preparation of 3,4-Dichloro-benzylamide of 3-hydroxy-10- (2-methoxy-ethyl) -4-oxo-4,10-dihydrobenzo [4,5] imidazo [1,2-α ] pyrimidine-2-carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

1H NMR (300 MHz, D6 DMSO): δ 11.78 (1H, s, OH), 9.66 (1H, bs, NH), 8.45 (1H, d, J = 8.1 Hz, Ar- CH), 7.65-7.60 (3H, m, Ar- image 1 CH), 7.53 (1H, m, Ar-CH), 7.37-7.30 (2H, m, Ar-CH), 4.55 (2H, d, J = 6.3 Hz, CH2NH) , 4.52 (2H, t, J = 5.4 Hz, OCH2CH2N), 3.72 (2H, t, J = 5.4 Hz, OCH2CH2N), 3.27 (3H, s, OCH3).

MS (ESI +) m / z 461 and 463 (M + 1).

Example 14.11: Preparation of 3,4-Dichloro-benzylamide of 3-hydroxy-4-oxo-10- (3-piperidin-1-yl-propyl) -4,10-dihydro-benzo [4,5] imidazo [1] , 2-α] pyrimidine-2-carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

1H NMR (300 MHz, D6 DMSO): δ 11.70 (1H, bs, OH), 9.39 (1H, t, J = 6.3 Hz, NHCH2), 8.47 (1H, d, J = 7.8 Hz, Ar-CH) 7.657.51 (4H, m, Ar-CH), 7.37-7.24 (2H, m, Ar-CH), 4.42 (2H, t, J = 6 , 0 Hz, CH2N), 4.31 (2H, d, J = 6.3 Hz, NHCH2), 2.25 (2H, t, J = 6.0 Hz, CH2N), 1.99-1.92 (6H, m, CH2NCH2 and NCH2CH2CH2N), 1.17-1.11 (6H, m, NCH2CH2CH2CH2).

MS (ESI +) m / z 528 and 530 (M + 1).

Example 14.12: Preparation of 3-hydroxy-7,8-dimethyl-4-oxo-4,10-dihydro-benzo [4,5] imidazo [1,2-a] pyrimidine acid 3,4-dichloro-benzylamide -2-carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

1H NMR (300 MHz, D6 DMSO): δ 9.30 (1H, t, J = 6.6 Hz, NH), 8.22 (1H, s, Ar-CH), 7.60 (2H, m, Ar-CH), 7.35 (1H, d, J = 8.1 image 1 Hz, Ar-CH), 7.21 (1H, s, Ar-CH), 4.51 (2H, d, J = 6.6 Hz, CH2NH), 2.34 (6H, s, 2 xCH3).

MS (ESI +) m / z 431 (M) +.

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

2.38 (3H, s, CH3), 2.35 (3H, s, CH3). MS (ESI +) m / z 544 and 546 (M + 1).

Example 14.14: Preparation of 3-hydroxy-10- (2-morpholin-4-yl-ethyl) -4-oxo-4,10-dihydro-benzo [4,5] imidazo [3-chloro-4-fluoro-benzylamide 1,2-a] pyrimidine-2-carboxylic

Useful background for the reader.

image19

image20

1H NMR (300 MHz, CDCl3) δ 2.52 (4H, bdd, J = 4.5 Hz, 23-NCH2CH2O-), 2.79 (2H, t, J = 6.3, 7.2 Hz, - NCH2CH2NAr-) 3.59 (4H, bdd, J = 4.2 Hz, 23-NCH2CH2O-), 4.31 (t, J = 6.6 Hz, -NCH2CH2NAr-), 4.61 (2H, dd, J = 6.0 Hz, -NHCH2-), 7.15 (1H, t, J = 8.4, 8.7 Hz, ArCH), 7.25 (1H, dd, J = 2.4, 4, 5 Hz ArCH), 7.32 (2H, m, ArCH), 7.42 (1H, dd, J = 2.4, 6.9 Hz ArCH) 7.50 (2H, dt, J = 1.5, 7.8 Hz, ArCH), 8.00 (1H, s, NH), 8.72 (1H, s, NH), 8.68 (1H, d, J = 8.1 Hz, ArCH), 11, 62 (1H, s, OH).

MS (ESI +) m / z 500 (M [Cl35] +1)

Example 14.15: Preparation of 3-hydroxy-10- (2-morpholin-4-yl-ethyl) -4oxo-4,10-dihydro-benzo [4,5] imidazo 4-chloro-3-trifluoromethyl-benzylamide [1,2-a] pyrimidine-2-carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

1H NMR (300 MHz, CDCl3) δ 2.51 (4H, bdd, image 1 J = 4.5 Hz, 23-NCH2CH2O-), 2.77 (2H, t, J = 6.6 Hz, -NCH2CH2NAr-), 3.59 (4H, bdd, J = 4.5 Hz, 23- NCH2CH2O-), 4.31 (t, J = 6.6 Hz, -NCH2CH2NAr-), 4.68 (2H, dd, J = 6.0 Hz, -NHCH2-), 7.30 (2H, m, ArCH), 7.47 (2H, dt, J = 0.9, 8.0 Hz, ArCH), 7.50 (2H, bdd, J = 0.9, ArCH), 7.68 (1H, bs, ArCH), 8.07 (1H, d, J = 7.8 Hz ArCH), 11.55 (1H, s, OH).

MS (ESI +) m / z 550 (M [Cl35] +1)

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

(4H, bdd, J = 4.2 Hz, 23-NCH2CH2O-), 4.31 (t, J = 6.6 Hz, -NCH2CH2NAr-), 4.61 (2H, dd, J = 6.0 Hz , -NHCH2-), 7.15 (1H, t, J = 8.4, 8.7 Hz, ArCH), 7.25 (1H, dd, J = 2.4, 4.5 Hz ArCH), 7 , 32 (2H, m, ArCH), 7.42 (1H, dd, J = 2.4, 6.9 Hz ArCH), 7.50 (2H, dt, J = 1.5, 7.8 Hz, ArCH), 8.00 (1H, s, NH), 8.72 (1H, s, NH), 8.68 (1H, d, J = 8.1 Hz, ArCH), 11.62 (1H, s , OH).

MS (ESI +) m / z 530 (M [Cl35] + 1-salt)

Example 14.17: Preparation of 2- [2- (3,4-Dichloro-phenyl) -pyrrolidine-1-carbonyl] -3-hydroxy-10- (2-morpholin-4-yl-ethyl) -10Hbenzo [4,5] imidazo [1,2-a] pyrimidin-4-one

image21

image 1

1H NMR (300 MHz, CDCl3) δ 2.52 (4H, bdd, J = 4.5 Hz, 23-NCH2CH2O-), 2.79 (2H, t, J = 6.3, 7.2 Hz, - NCH2CH2NAr-) 3.59 (4H, bdd, J = 4.2 Hz, 23-NCH2CH2O-), 4.31 (t, J = 6.6 Hz, -NCH2CH2NAr-), 4.61 (2H, dd, J = 6.0 Hz, -NHCH2-), 7.15 (1H, t, J = 8.4, 8.7 Hz, ArCH), 7.25 (1H, dd, J = 2.4, 4, 5 Hz ArCH), 7.32 (2H, m, ArCH), 7.42 (1H, dd, J = 2.4, 6.9 Hz ArCH) 7.50 (2H, dt, J = 1.5, 7.8 Hz, ArCH), 8.00 (1H, s, NH), 8.72 (1H, s, NH), 8.68 (1H, d, J = 8.1 Hz, ArCH), 11, 62 (1H, s, OH) MS (ESI +) m / z 556 (M [Cl35] +1)

Example 15: Preparation of benzylamides of 3-hydroxy-4-oxo-10- (2-piperazin-1-yl-ethyl) -4,10-dihydrobenzo [4,5] imidazo [1,2-a] pyrimidine- 2-carboxylic: general procedure

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image22

Example 15.1.1: Preparation of 4- (2-Chloro-ethyl) -piperazine-1-carboxylic acid tert-butyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

twenty

image 1

4- (2-Chloro-ethyl) -piperazine-1-carboxylic acid tert-butyl ester was prepared according to a patented process described in WO2002 / 44141.

1 H NMR (300 MHz, CDCl3): δ 3.45 (2H, t, J = 6.9 Hz, NCH2), 3.40 (4H, t, J = 4.8 Hz, CH2NCH2), 2.52 (2H, t, J = 6.9 Hz, CH2Cl), 2.42 (4H, t, J = 4.8 Hz, CH2NCH2), 1.49 (9H, s, C [CH3] 3.

MS (ESI +) m / z 249 (M + 1).

Example 15.1.2: Preparation of 4- [2- (2-Amino-benzoimidazol-1-yl) -ethyl] -piperazine-1-carboxylic acid tert-butyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

35

4- [2- (2-Amino-benzoimidazol-1-yl) -ethyl] -piperazine-1-carboxylic acid tert-butyl ester was prepared according to the image 1 procedure described in WO2005 / 058869.

1H NMR (300 MHz, CDCl3): δ 7.43 (1H, d, J = 7.5 Hz, Ar-CH), 7.17-7.06 (3H, m, Ar-CH), 5.78 (2H, s, NH2), 3.64 (2H, t J = 5.7 Hz, NCH2CH2N), 3.46 (4H, m, CH2NCH2), 2.56 (4H, m, CH2NCH2), 2.43 (2H, t, J = 5.7 Hz, NCH2CH2N), 1.46 (9H, s, C [CH3] 3).

MS (ESI +) m / z 346 (M + 1).

Example 15.2: Preparation of the 3-acetoxy-10- [2- (4-tert-butoxycarbonyl-piperazin-1-yl) -ethyl] -4oxo-4,10-dihydrobenzo [4,5] imidazo [1] methyl ester , 2-a] pyrimidine-2-carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

3-Acetoxy-10- [2- (4-tert-butoxycarbonyl-piperazin-1-yl) -ethyl] -4-oxo-4,10-dihydrobenzo [4,5] imidazo [1,2a ] pyrimidine-2-carboxylic acid was prepared by adaptation of the procedure shown in Example 5.

1H NMR (300 MHz, D6 DMSO): δ 8.66 (1H, d, J = 8.7 Hz, Ar-CH), 7.58 (1H, m, Ar-CH), 7.42 (2H, m, Ar-CH), 4.29 (2H, t, J = 5.9 Hz, NCH2CH2N) 3.96 (3H, s, OCH3), 3.55 (4H, t, J = 5.4 Hz, CH2NCH2), 2.88 (2H, t, J = 5.9 Hz, NCH2CH2N), 2.70 (4H, t, J = 5.4 Hz CH2NCH2), 2.20 (3H, s, O = CCH3) , 1.47 (9H, s, C [CH3] 3).

MS (ESI +) m / z 514 (M + 1).

Example 15.3: Preparation of the 3-acetoxy-4-oxo-10- (2-piperazin-1-yl-ethyl) -4,10-dihydrobenzo [4,5] imidazo [1,2-a] pyrimidine acid methyl ester -2-carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

3-Acetoxy-10- [2- (4-tert-butoxycarbonyl-piperazin-1-yl) -ethyl] -4-oxo-4,10-dihydrobenzo [4,5] imidazo [1,2a ] pyrimidine2-carboxylic acid (33 mg, 0.064 mmol) was treated with trifluoroacetic acid (0.20 ml) and stirred for one hour at room temperature. After that period, the mixture was concentrated and the crude residue was the desired product (27 mg, 100%).

1H NMR (300 MHz, D6-DMSO): δ 8.76 (1H, bs, NH), 8.47 (1H, d, J = 8.1 Hz, Ar-CH), 7.85 (1H, d , J = 8.1 Hz, Ar-CH) 7.65 (1H, dd, J = 8.1, 7.8 Hz, Ar-CH), 7.47 (1H, dd, J = 8.1, 7.8 Hz, Ar-CH), 4.57 (2H, t, J = 5.4 Hz, NCH2CH2N), 3.88 (3H, s, OCH3), 3.27-3.07 (10H, m , 2 x NCH2CH2N and 8 x NHCH2CH2NCH2CH2), 2.31 (3H, s, [C = O] CH3).

MS (ESI +) m / z 414 (M + 1).

Example 15.4: Preparation of the tert-butyl ester of 4- {2- [2- (3,4-dichloro-benzylcarbamoyl) -3-hydroxy-4-oxo-4Hbenzo [4,5] imidazo [1,2-a] pyrimidin-10-yl] -ethyl} -piperazine-1-carboxylic acid

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The product of Example 15.2 was converted to the 4- {2- [2- (3,4-dichloro-benzylcarbamoyl) -3-hydroxy-4oxo-4H-benzo [4,5] imidazo [1] tert-butyl ester , 2-a] pyrimidin-10-yl] -ethyl} -piperazine-1-carboxylic acid following a procedure adapted from Example 6.

1H NMR (300 MHz, D6 DMSO): δ 8.68 (1H, d, J = 7.5 Hz, Ar-CH), 8.02 (1H, t, J = 6.3 Hz, NHCH2), 7 , 49-7.14 (6H, m Ar-CH), 4.63 (2H, d, J = 6.3 Hz, NHCH2), 4.31 (2H, t, J = 6.9 Hz, NCH2CH2N) , 3.34 (4H, m, CH2NCH2), 2.79 (2H, t, J = 6.9 Hz, NCH2CH2N), 2.45 (4H, m, CH2NCH2), 1.47 (9H, s, C [CH3] 3).

 MS (ESI +) m / z 615 and 617 (M + 1).

Example 15.5: Preparation of 3-hydroxy-4-oxo-10- (2-piperazin-1-yl-ethyl) -4,10-dihydrobenzo [4,5] imidazo [1,2-3,4-dichloro-benzylamide -a] pyrimidine-2-carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The product of Example 15.3 was converted to 3-hydroxy-4-oxo-10- (2-piperazin-1-yl-ethyl) -4,10-dihydrobenzo [4,5] imidazo [3,4-dichloro-benzylamide. 1,2-a] pyrimidine-2-carboxylic acid following a procedure adapted from Example 6.

1H NMR (300 MHz, D6 DMSO): δ 9.65 (1H, t, J = 6.6 Hz, NHCH2), 8.46 (1H, d, J = 8.1 Hz, Ar-CH), 7 , 68-7.60 (3H, m Ar-CH), 7.58-7.50 (1H, m, Ar-CH), 7.38-7.31 (2H, m, Ar-CH), 4 , 54 (2H, d, J = 6.6 Hz, NHCH2), 4.47 (2H, t, J = 6.0 Hz NCH2CH2N), 3.16 (4H, m, CH2NCH2), 2.66 (2H , t, J = 6.0 Hz, NCH2CH2N), 2.41 (4H, m, CH2NCH2).

MS (ESI +) m / z 515 and 517 (M + 1).

Example 15.6: Preparation of 3,4-dichloro-benzylamide of 3-hydroxy-10- [2- (4-methanesulfonyl-piperazin-1-yl) ethyl] -4-oxo-4,10-dihydrobenzo [4, 5] imidazo [1,2-a] pyrimidine-2-carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

3-Acetoxy-4-oxo-10- (2-piperazin-1-yl-ethyl) -4,10-dihydrobenzo [4,5] imidazo [1,2-a] pyrimidine-2-carboxylic acid methyl ester (Example 15.3) (97 mg, 0.235 mmol) was dissolved in dichloromethane (1 ml) and triethylamine (98 µL, 0.71) was added

mmol) to the previous one, followed by mesyl chloride (21 µL, 0.26 mol). The reaction was stirred at room temperature before evaporating the solvents and the residue was converted to 3-hydroxy-10- [2- (4-methanesulfonylpiperazin-1-yl) -ethyl] 4-3,4-dichloro-benzylamide. -oxo-4,10-dihydrobenzo [4,5] imidazo [1,2-a] pyrimidine-2-carboxylic acid by adaptation of the procedure described in Example 6.

1H NMR (300 MHz, D6 DMSO): δ 9.60 (1H, t, J = 6.3 Hz, NHCH2), 8.47 (1H, d, J = 8.1 Hz, Ar-CH), 7 , 68-7.52 (4H, m, Ar-CH), 7.36-7.31 (2H, m, Ar-CH), 4.55 (2H, d, J = 6.3 Hz, NHCH2) , 4.51 (2H, t, J = 5.7 Hz, NCH2CH2N), 2.87 (4H, m CH2NCH2), 2.79 (2H, t, J = 5.7 Hz, NCH2CH2N), 2.74 (3H, s, SCH3), 2.57 (4H, m, CH2NCH2).

MS (ESI +) m / z 593 and 595 (M + 1).

Example 15.7: Preparation of 3,4- dichloro-benzylamide of 10- [2- (4-Acetyl-piperazin-1-yl) -ethyl] -3-hydroxy-4-oxo4,10-dihydrobenzo [4,5 ] imidazo [1,2-a] pyrimidine-2-carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

3-Acetoxy-4-oxo-10- (2-piperazin-1-yl-ethyl) -4,10-dihydrobenzo [4,5] imidazo [1,2-a] pyrimidine-2-carboxylic acid methyl ester (Example 15.3) (100 mg, 0.242 mmol) was dissolved in dichloromethane (1 ml) and triethylamine (96 µL, 0.70 mmol) was added thereto, followed by acetyl chloride (21 µL, 0.266 mol). The reaction was stirred at room temperature for 3 hours before evaporating the solvents and the residue was converted to the 10- [2- (4-acetylpiperazin-1-yl) -ethyl] -3 3,4-dichloro-benzylamide. -hydroxy-4-oxo-4,10-dihydrobenzo [4,5] imidazo [1,2-a] pyrimidine-2-carboxylic acid by adaptation of the procedure described in Example 6.

1H NMR (300 MHz, D6 DMSO): δ 9.61 (1H, t, J = 6.3 Hz, NHCH2), 8.46 (1H, d, J = 8.4 Hz, Ar-CH), 7 , 70-7.62 (3H, m Ar-CH), 7.57 (1H, m, Ar-CH), 7.37 (2H, m, Ar-CH), 4.56 (2H, d, J = 6.3 Hz, NHCH2), 4.51 (2H, t, J = 5.7 Hz, NCH2CH2N) 3.20 (2H, m, CH2NCH2), 3.12 (2H, m, CH2NCH2), 2, 73 (2H, t, J = 5.7 Hz, NCH2CH2N), 2.45-2.38 (4H, m, CH2NCH2), 1.89 (3H, s, CH3).

MS (ESI +) m / z 557 (M) +.

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image23

[4,5] imidazo [1,2-a] pyrimidine-2 acid methyl ester

sodium (21 mg, 0.63 mmol) and sodium acetate (30 mg, 0.38 mmol), followed by formaldehyde (38 µL, 0.47 mmol). The reaction was stirred at room temperature for 2 hours before evaporating the solvents and the residue was converted to 3-hydroxy- [2- (4-methyl-piperazin-1-yl) -ethyl] 3,4-dichlorobenzylamide. -4-oxo-4,10-dihydrobenzo [4,5] imidazo [1,2-a] pyrimidine-2-carboxylic acid by adaptation of the procedure described in Example 6.

1H NMR (300 MHz, D6 DMSO): δ 9.58 (1H, t, J = 6.3 Hz, NHCH2), 8.46 (1H, d, J = 7.2 Hz, Ar-CH), 7 , 66-7.53 (4H, m Ar-CH), 7.36-7.31 (2H, m, Ar-CH), 4.54 (2H, d, J = 6.3 Hz, NHCH2), 4.47 (2H, t, J = 6.0 Hz, NCH2CH2N), 2.67 (2H, t, J = 6.0 Hz, NCH2CH2N), 2.54-2.26 (8H, m, CH2NCH2CH2NCH2) , 1.99 (3H, s, CH3).

MS (ESI +) m / z 527 (M) +.

Example 15.9.1: Preparation of the 3-acetoxy-10- [2- (4-methoxymethyl-piperazin-1-yl) -ethyl] -4-oxo-4,10-dihydrobenzo [4,5] imidazo [1] methyl ester , 2-a] pyrimidine-2-carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

3-Acetoxy-4-oxo-10- (2-piperazin-1-yl-ethyl) -4,10-dihydrobenzo [4,5] imidazo [1,2-a] pyrimidine-2-carboxylic acid methyl ester (Example 15.3) (97 mg, 0.235 mmol) was dissolved in dichloromethane (1 ml) and diisopropylethylamine (95 µL, 0.52 mmol) was added thereto, followed by methoxymethoxy chloride (20 µL, 0.258 mmol). The reaction was stirred at room temperature for 24 h before evaporating the solvents and the residue was purified by column chromatography (98: 1.5: 0.5 dichloromethane: methanol: aqueous ammonia) to give the desired product (30 mg, 28%)

1H NMR (300 MHz, D6-DMSO): δ 8.68 (1H, d, J = 8.4 Hz Ar-CH), 7.54 (1H, dd, J = 8.1, 7.8 Hz, Ar-CH), 7.41-7.35 (2H, m, Ar-CH), 4.42 (2H, t, J = 6.3 Hz, NCH2CH2N), 3.99 (3H, s, OCH3) , 3.60 (2H, s, CH2OCH3), 3.52-3.45 (2H, m, CH2N) 3.31 (2H, m, CH2N), 2.83 (2H, t, J = 6.3 Hz, NCH2CH2N), 2.55-2.46 (4H, m, CH2NCH2), 2.09 (3H, s, OCH3), 2.06 (3H, s, O = CCH3).

 MS (ESI +) m / z 458 (M + 1).

Example 15.9.2: Preparation of 3-hydroxy-10- [2- (4-methoxymethyl-piperazin-1-yl) -ethyl] 4-oxo-4,10-dihydrobenzo [-3,4-dichloro-benzylamide 4,5] imidazo [1,2-a] pyrimidine-2-carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The product of Example 15.9.1 was converted to 3-hydroxy-10- [2- (4-methoxymethyl-piperazin-1-yl) -ethyl] -4-oxo4,10-dihydrobenzo [3,4-dichloro-benzylamide. 4,5] imidazo [1,2-a] pyrimidine-2-carboxylic acid by adaptation of a method adapted from Example 6.

1H NMR (300 MHz, D6 DMSO): δ 8.70 (1H, d, J = 8.1 Hz, Ar-CH), 7.90 (1H, m, NH), 7.52-7.19 ( 6H, m, Ar-CH), 4.63 (2H, d, J = 6.0 Hz, CH2NH), 4.32 (2H, t, J = 6.6 Hz, NCH2CH2N), 3.85 (2H , s, CH2OCH3), 3.65 (2H, m, CH2N), 3.50 (2H, m, CH2N), 2.81 (2H, t, J = 6.6 Hz, NCH2CH2N), 2.51 ( 4H, m, CH2NCH2), 2.04 (3H, s, OCH3).

MS (ESI +) m / z 557 and 559 (M + 1).

Example 15.10: Preparation of 2- (3,4-dichloro-benzylcarbamoyl) -10- [2- (4-methanesulfonyl-piperazin-1-yl) -ethyl] -4-oxo-4,10-dihydrobenzo [4,5] imidazo [1,2-a] pyrimidin-3-yl methanesulfonic acid ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

3-Hydroxy-4-oxo-10- (2-piperazin-1-yl-ethyl) -4,10-dihydrobenzo [4,5] imidazo [1,2-a] 3,4-dichloro-benzylamide pyrimidine2-carboxylic acid, (Example 15.5) (75 mg, 0.146 mmol) and triethylamine (60 µL, 0.31 mmol) were dissolved in dichloromethane (1 mL) and to this was added mesyl chloride (18 µL, 0.31 mmol). The reaction was stirred at room temperature for 15 minutes after which the solvents were concentrated in vacuo and the residue was purified by column chromatography (95: 4.5: 0.5 dichloromethane: methanol: aqueous ammonia) to give the desired product. (65 mg, 76%).

1H NMR (300 MHz, D6 DMSO): δ 8.48 (1H, t, J = 6.6 Hz, NHCH2), 7.82 (1H, d, J = 8.1 Hz, Ar-CH), 7 , 62 (4H, m, Ar-CH), 7.35 (2H, m, Ar-CH), 4.57 (2H, m, CH2N), 4.49 (2H, t, J = 6.0 Hz , NCH2CH2N), 4.16 (2H, d, J = 6.6 Hz, NHCH2), 2.88 (4H, m, CH2NCH2), 2.78 (2H, m, NCH2CH2N), 2.55 (2H, m, CH2N), 2.06 (6H, s, 2 x S-CH3).

MS (ESI +) m / z 671 and 673 (M + 1).

Example 16: Preparation of 6-hydroxy-5-oxo-5H-thiazolo [3,2-a] pyrimidine-7-carboxylic acid substituted methyl esters

Example 16.1: Preparation of 6- (2,2-Dimethyl-propionyloxy) -5-oxo-5H-thiazolo [3,2-a] pyrimidine-7-carboxylic acid methyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Stage 1:

The procedure described in Example 5 was applied to 2-aminothiazole to give the desired ester. 1H NMR (300 MHz, DMSO-d6) δ 3.86 (s, 3H), 7.55 (d, J = 4.7 Hz, 1H)) 7.96 (d, J = 4.9 Hz, 1H ), 10.21 (s, 1 H)

Stage 2:

The above ester (620 mg, 2.7 mmol) and triethylamine (2.21 g, 21 mmol) were dissolved in dichloromethane (30 ml). To the above solution was added pivaloyl chloride (362 mg, 3.0 mmol) dropwise at room temperature. After completion of the addition, the mixture was stirred for 1 h then concentrated in vacuo. The residue was purified by column chromatography using (hexane / ethyl acetate 1: 1) to give the desired product (610 mg, 83%).

1H NMR (300 MHz, DMSO-d6) δ 1.30 (s, 9H), 3.84 (s, 3H), 7.73 (d, J = 5.0 Hz, 1H), 8.08 (d , J = 4.8 Hz, 1H)

MS (ESI +) m / z 333 (M + Na +)

Example 16.2: Preparation of the 6-hydroxy-2-methyl-5-oxo-5H-thiazolo [3,2-a] pyrimidine-7 carboxylic acid methyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The procedure described in Example 5 was applied to 2-amino-5-methylthiazole to give the desired ester. 1H NMR (300 MHz, DMSO-d6) δ 2.41 (d, J = 1.4 Hz, 3H), 3.85 (s, 3H), 7.81 (d, J = 1.5 Hz, 1H ), 10.21 (s, 1 H). MS (ESI +) m / z 263 (M + Na +)

Example 16.3: Preparation of the 6-hydroxy-3-methyl-5-oxo-5H-thiazolo [3,2-a] pyrimidine-7-carboxylic acid methyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

2-Amino-4-methylthiazole was converted to N-acetate by treatment with acetic anhydride at 90˚C in 80% yield. This was oxidized with mCPBA in a 45% yield and then hydrolyzed to give the 2-amino-4-methylthiazole N-oxide, which was subjected to the conditions described in J. Heterocyclic Chem., 1979, 16 to give the crude ester . This was acylated by the procedure described in Example 16.1 (Step 2) to give the desired product.

1H NMR (300 MHz, CDCl3) δ 1.41 (s, 9H), 2.82 (s, 3H), 3.93 (s, 3H), 6.55 (s, 1H)

Example 16.4: Preparation of the 6-hydroxy-2-isopropyl-5-oxo-5H-thiazolo [3,2-a] pyrimidine-7-carboxylic acid methyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The procedure described in Example 5 was applied to 2-amino-5-isopropylthiazole to give the desired ester.

1H NMR (300 MHz, DMSO-d6) δ 1.29 (d, J = 7.0 Hz, 6H), 3.12-3.23 (m, 1H), 3.85 (s, 3H), 7 , 75 (d, J = 1.2 Hz, 1H), 10.23 (s, 1 HOUR). MS (ESI-) m / z 267 (M-1)

Example 17: Preparation of substituted benzylamides of 6-hydroxy-5-oxo-5H-thiazolo [3,2-a] pyrimidine-7-carboxylic acid

Example 17.1: Preparation of 6-hydroxy-5-oxo-5H-thiazolo [3,2-a] pyrimidine-7-carboxylic acid 4-fluoro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Starting from the product of Example 16.1 and by adapting the procedure of Example 6, 6-hydroxy-5-oxo-5H-thiazolo [3,2-a] pyrimidine-7-carboxylic acid 4-fluoro-benzylamide was obtained.

1H NMR (300 MHz, D6-DMSO): δ 12.35 (1H, s, OH), 9.74 (1H, t, J = 6.3 Hz, CH2NH), 7.97 (1H, d, J = 5.1 Hz, Ar-CH) 7.52 (1H, d, J = 5.1 Hz, Ar-CH), 7.38 (2H, dd, J = 7.8, 8.0 Hz, Ar -CH), 7.58 (2H, dd, J = 7.8, 8.0 Hz, Ar-CH), 4.45 (2H, d J = 6.3 Hz, CH2NH).

 MS (ESI-) m / z 318 (M-H).

Example 17.2: Preparation of 6-hydroxy-5-oxo-5H-thiazolo [3,2-a] pyrimidine-7-carboxylic acid 3,4-dichlorobenzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Starting from the product of Example 16.1 and by adapting the procedure of Example 6, 6-hydroxy-5-oxo-5H-thiazolo [3,2-a] pyrimidine-7-carboxylic acid 3,4-dichlorobenzylamide was obtained.

1H NMR (300 MHz, D6-DMSO): δ 12.22 (1H, s, OH), 9.79 (1H, t, J = 5.7 Hz, CH2NH), 7.97 (1H, d, J = 5.4 Hz, Ar-CH) 7.60 (1H, d, J = 8.1 Hz, Ar-CH), 7.60 (1H, s, Ar-CH), 7.52 (1H, d , J = 5.4 Hz, Ar-CH), 7.34 (1H, d, J = 8.1 Hz, Ar-CH), 4.47 (2H, d, J = 5.7 Hz, CH2NH) .

 MS (ESI-) m / z 368 (M [Cl35] -1)

HPLC procedure 7 99.1% / 15.4 min

Example 17.3: Preparation of 6-hydroxy-2-methyl-5-oxo-5H-thiazolo [3,2-a] pyrimidine-7-carboxylic acid 4-fluoro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

(300 MHz, D6-DMSO) δ 2.38 (3H, s, -CHC (S) CH3), 4.42 (2H, d, J = 5.7 Hz, -NH-CH2-), 7.13 (2H, m, ArH), 7.37 (2H, m ArH), 7.79 (1H, s, -CHC (S) CH3), 9.72 (1H, t, J = 6.3 Hz, - NHCH2-).

MS (ESI-) m / z 332 (M [Cl35] -1).

HPLC procedure 7 98.4% / 10.6 min

Example 17.4: Preparation of 6-hydroxy-2-methyl-5-oxo-5H-thiazolo [3,2-a] pyrimidine7-carboxylic acid 3,4-dichloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image24

image 1

Starting from the product of Example 16.2 and by adapting the procedure of Example 6, 3,4-dichlorobenzylamide of 6-hydroxy-2-methyl-5-oxo-H-thiazolo [3,2-a] pyrimidine-7- acid was obtained carboxylic.

1H NMR (300 MHz, D6-DMSO): δ 12.21 (1H, s, OH), 9.76 (1H, t, J = 6.3 Hz, CH2NH), 7.81 (1H, s, Ar -CH), 7.59 (2H, d, J = 7.8 Hz, Ar-CH), 7.33 (1H, d J = 7.8 Hz, Ar-CH), 4.46 (2H, d , J = 6.3 Hz, CH2NH), 2.40 (3H, s, CH3).

MS (ESI-) m / z 382 (M [Cl35] -1). HPLC procedure 7 99.1 / 14.6 min

Example 17.5: Preparation of 6-hydroxy-3-methyl-5-oxo-5H-thiazolo [3,2-a] pyrimidine7-carboxylic acid 3,4-dichloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Starting from the product of Example 16.3 and by adapting the procedure of Example 6, 3,4-dichlorobenzylamide of 6-hydroxy-3-methyl-5-oxo-H-thiazolo [3,2-a] pyrimidine-7- acid was obtained carboxylic.

 1H NMR (300 MHz, CDCl3) δ 2.80 (3H, s, - (CH) C (N) CH3), 4.57 (2H, d, J = 6.6 Hz, -NH-CH2-), 6.37 (1H, s, - (CH3) C = CH-S-), 7.18 (1H, m, ArH), 7.43 (2H, m, ArH), 7.95 (1H, m, -NHCH2-).

MS (ESI-) m / z 332 (M [Cl35] -1).

HPLC procedure 7 99.6% / 10.6 min

Example 17.6: Preparation of 6-hydroxy-2-isopropyl-5-oxo-5H-thiazolo [3,2a] pyrimidine-7-carboxylic acid 3,4-dichloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Starting with the product of Example 16.4 and by adapting the procedure of Example 6, 6-hydroxy-2-isopropyl-5-oxo-H-thiazolo [3,2-a] pyrimidine-7- acid 3,4-dichlorobenzylamide was obtained carboxylic.

1H NMR (300 MHz, D6-DMSO) δ 1.27 (6H, d, 23 (CH3) 2CH-), 3.15 (1H, m, CH3) 2CH-), 4.44 (2H, d, J = 6.6 Hz, - (O = C) NHCH2), 7.31 (1H, dd, J = 1.8, 8.1 Hz, ArH), 7.58 (2H, m, ArH), 7, 74 (1H, d, J = 1.2 Hz, ArH), 9.78 (1H, t, J = 6.0 Hz, - (O = C) NHCH2), 12.22 (1H, s, OH) .

MS (ESI +) m / z 434 (M [Cl35] + Na).

HPLC procedure 7 99.0% / 15.2 min

Example 17.7: Preparation of substituted benzylamides of 2-aminomethyl-6-hydroxy-5-oxo-5H-thiazolo [3,2a] pyrimidine-7-carboxylic acid - General procedures

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image25

ice water). N, N-dimethylaminopyridine (500 mg, 4.13 mmol) was added and after 10 min, acetyl chloride (320 mg 4.13 mmol) was added dropwise. The mixture was heated to room temperature and stirred for 6 h. TLC indicated that all of the starting ester had been consumed and the mixture was washed twice with aqueous hydrochloric acid (4.0 M), brine, dried, (Na2SO4), filtered and concentrated in vacuo to give the compound. desired as a yellow solid (70 mg, 90%).

1H NMR (300 MHz, CDCl3) δ 2.38 (s, 3H), 2.49 (s, 3H), 3.96 (s, 3H), 7.70 (s, 1H)

MS (ESI +) m / z 305 (M + 23)

Stage 2

image 1

The intermediate of Step 1 (6 g, 21 mmol), N-bromosuccinimide (3.02 g) and t-butyl peroxide (1.03 g) in carbon tetrachloride (400 ml) were heated and combined at reflux. After 1 h, another portion of N-bromosuccinimide (1.14 g) was added. The reaction mixture was refluxed for another 4 h, then cooled to room temperature. The solids were collected by filtration and then dissolved in dichloromethane and the solution was washed with water, dried, and evaporated to dryness to give the crude product. Further purification by recrystallization from dichloromethane / hexane gave the desired compound (3.0 g, 39%) as a white solid.

1H NMR (300 MHz, DMSO-d6) δ 2.31 (s, 3H), 3.87 (s, 3H), 5.02 (d, J = 0.9 Hz, 2H), 8.33 (t , J = 0.8 Hz, 1H).

MS (ESI +) m / z 383 (M [Br79] +23), 385 (M [Br81] +23)

Stage 3: A

The product of Step 2 (0.33 mmol) and amine (1 mmol) in dichloromethane (6 ml) was stirred at room temperature for 20 h. The resulting precipitate was collected by filtration, washed with cold methanol and used directly in the next reaction step.

Stage 3: B

To a stirred solution of the product from Step 3: A (0.83 mmol) in dichloromethane (20 ml), amine (2.49 mmol) was added dropwise at room temperature. The mixture was kept at room temperature for 24 h. The mixture was diluted with dichloromethane (20 ml) and extracted with aqueous hydrochloric acid (1.0 M, 20 ml). The pH of the aqueous phase was adjusted to pH = 10 with aqueous sodium hydroxide (1.0 M) and then extracted with dichloromethane (2 x 30ml). The combined organic layers were dried (Na2SO4), filtered and concentrated in vacuo. The product was further purified by recrystallization or by preparative HPLC.

Stage 4: A

The procedure described in Example 6 was adapted. The product was further purified by recrystallization or by preparative HPLC.

Stage 4: B

To a stirred solution of the product of Step 3: B (0.14 mmol) in methanol (20 ml) was added 1.3 equivalents of benzylamine. The resulting mixture was refluxed for 24 h and then concentrated in vacuo. The residue was dissolved in dichloromethane (20 ml) and washed with aqueous sodium hydroxide (1.0 M, 10 ml) and to the organic layer was added aqueous hydrochloric acid (1.0 M, 15 ml). The resulting precipitate was collected by filtration, washed with cold water followed by hexane. The solid was dried in vacuo to give the desired compound as hydrochloride salt.

Example 17.7.1: Preparation of 6-hydroxy-2-morpholin-4-ylmethyl-5-oxo-5H-thiazolo [3,2a] pyrimidine-7-carboxylic acid 4-fluoro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

MS (ESI-) m / z 419 (M [Cl35] -1)

HPLC procedure 7 96.7% / 12.2 min

Example 17.7.2: Preparation of 6-hydroxy-2-morpholin-4-ylmethyl-5-oxo-5H-thiazolo [3,2-a] pyrimidine-7-carboxylic acid 3,4-dichloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

and prepared the following compound using Step 3: A and Step 4: A: (300 MHz, D6-DMSO) δ 2.49 (4H, s, N-CH2-CH2-O), 3.61 (4H, s, N-CH2-CH2-O), 4.49 (2H, d, J = 6.3 Hz, -NH-CH2-) 7.36 (1H, dd, J = 8.4, 2.1 Hz , ArH), 7.62 (2H, m, ArH), 8.03 (1H, s, SC = CH-N-), 9.82 (1H, bt, O = C-NH-CH2), 12, 25 (1H s, OH).

image26

image 1

 MS (ESI-) m / z 467 (M [Cl35] -1) HPLC procedure 7 99.1% / 13.9 min

Example 17.7.3: Preparation of 6-hydroxy-5-oxo-2-piperidin-1-ylmethyl-5H-thiazolo [3,2-a] pyrimidine-7-carboxylic acid 3,4-dichloro-benzylamide acid

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

The following compound was prepared using Stage 3: A and Stage 4: A:

image 1

1H NMR (300 MHz, D6-DMSO): δ 11.84 (1H, s, OH), 8.03 (1H, m, NHCH2), 7.77 (1H, s, Ar-CH), 7.45 (2H, m, Ar-CH) 7.18 (1H, d, J = 8.7 Hz, Ar-CH), 4.58 (2H, d, J = 6.3 Hz, NHCH2), 3.56 (2H, s, NCH2 [C]), 2.45 (4H, m, CH2NCH2), 1.59-1.45 (6H, m, NCH2CH2CH2CH2).

MS (ESI-) m / z 465 (M [Cl35] -1)

HPLC procedure 7 97.7% / 10.3 min

Example 17.7.4: Preparation of dibutyl- [7- (3,4-dichloro-benzylcarbamoyl) -6-hydroxy-5-oxo-5H-thiazolo [3,2-a] pyrimidin-2-ylmethyl] -ammonium chloride

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

The following compound was prepared using Stage 3: B and Stage 4: B:

image 1

1H NMR (300 MHz, D6-DMSO): δ 12.25 (1H, s, OH), 10.79 (1H, bs, N + HCl), 9.80 (1H, t, J = 6.0 Hz , NHCH2). 8.30 (1H, s, Ar-CH), 7.57 (1H, d, J = 6.6 Hz, Ar-CH), 4.56 (2H, bs, NCH2 [C]), 4.45 (2H, d, J = 6.0 Hz, CH2NH), 3.01 (4H, m, 2 x NCH2CH2), 1.66 (4H, m, 2 x NCH2CH2CH2), 1.31 (4H, m, 2 x NCH2CH2CH2CH3), 0.89 (6H, t, J = 7.2 Hz, 2 x NCH2CH2CH2CH3).

MS (ESI-) m / z 509 (M [Cl35] -1)

HPLC procedure 7 98.5% / 11.5 min

Example 17.7.5: Preparation of 6-hydroxy-5-oxo-2-piperazin-1-ylmethyl-5-thiazolo [3,2-a] pyrimidine-7-carboxylic acid 3,4-dichloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

The following compound was prepared using Step 3: A and Step 4: A: 1H NMR (300 MHz, D6-DMSO): δ 10.94 (1H, s, OH), 7.85 (1H, s, Ar -CH), 7.59 (1H, d, J = 8.4 Hz, Ar-CH), 7.55 (1H, s, Ar-CH), 7.30 (1H, d, J = 8.4 Hz, Ar-CH), 4.47 (2H, d, J = 6.0 Hz, CH2NH), 4.32 (2H, s, NCH2 [C]), 2.89 (4H, m, CH2NCH2) 2 , 50 (4H, m, CH2NCH2).

image 1

MS (ESI-) m / z 466 (M [Cl35] -1)

HPLC procedure 7 96.3% / 9.97 min

Example 17.7.6: Preparation of 6-hydroxy-2- (4-methyl-piperazin-1-ylmethyl) -5-oxo-5H-thiazolo [3,2-a] pyrimidine-7 acid 3,4-dichloro-benzylamide -carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

The following compound was prepared using Stage 3: A and Stage 4: A:

image 1

1H NMR (300 MHz, DMSO) δ 2.17 (3H, -N-CH3), 2.33 (4H, bs, N-CH2-CH2-O), 2.46 (4H, bs, N-CH2- CH2-O), 4.44 (2H, d, J = 6.3 Hz, -NH-CH2-), 7.30 (1H, dd, J = 2.1, 8.1 Hz, ArH), 7 , 57 (2H, m, ArH), 7.94 (1H, s, SC = CH-N-), 9.87 (1H, bt O = C-NHCH2).

MS (ESI +) m / z 482 (M [Cl35] +1)

HPLC procedure 7 95.9% / 10.3 min

Example 17.7.7: Preparation of 6-hydroxy-2- (3-methyl-piperazin-1-ylmethyl) -5-oxo-5H-thiazolo [3,2-a] pyrimidine-7 acid 3,4-dichloro-benzylamide -carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

The following compound was prepared using Stage 3: A and Stage 4: A:

image 1

1H NMR (300 MHz, D6-DMSO): δ 7.82 (1H, s, Ar-CH), 7.57 (1H, d, J = 8.1 Hz, Ar-CH), 7.54 (1H , s, Ar-CH), 7.29 (1H, d, J = 8.1 Hz, Ar-CH), 4.56 (2H, d, J = 5.7 Hz, CH2NH), 3.69 ( 2H, s, CH2N), 3.67-2.83 (5H, m, NCH [CH3] CH2NCH2), 2.14 (1H, t, J = 6.3 Hz, CH2N), 1.86 (1H, t, J = 6.3 Hz, CH2N), 1.02 (3H, d, J = 6.3 Hz, CH3CH).

MS (ESI-) m / z 480 (M [Cl35] -1) HPLC procedure 7 81.4% / 10.1 min

Example 17.7.8: Preparation of 2-diethylaminomethyl-6-hydroxy-5-oxo5H-thiazolo [3,2-a] pyrimidine-7-carboxylic acid 3,4-dichloro-benzylamide hydrochloride

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

The following compound was prepared using Stage 3: B and Stage 4: B:

image 1

1H NMR (300 MHz, D6-DMSO): δ 12.25 (1H, s, OH), 11.09 (1H, bs, NH + Et2), 9.81 (1H, t, J = 6.3 Hz , NHCH2), 8.32 (1H, s, Ar-CH), 7.59 (1H, d, J = 8.1 Hz, Ar-CH), 7.56 (1H, s, Ar-CH), 7.32 (1H, d, J = 8.1 Hz, Ar-CH), 4.54 (2H, s, CH2NH +) 4.46 (2H, d, J = 6.3 Hz, NHCH2), 3, 06 (4H, m, 2 x CH2CH3), 1.26 (6H, m, 2 x CH2CH3).

MS (ESI-) m / z 453 (M [Cl35] -1) HPLC procedure 7 98.5% / 10.2 min

Example 17.7.9: Preparation of 6-hydroxy-5-oxo-2-pyrrolidin-1-methyl-5H-thiazolo [3,2-a] pyrimidine-7-carboxylic acid 3,4-dichloro-benzylamide hydrochloride

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

 1H NMR (300 MHz, D6-DMSO): δ 12.26 (1H, s, OH), 11.01 (1H, bs, NH +), 9.80 (1H, t, J = 6.3 Hz, CH2NH ), 8.26 (1H, s, Ar-CH), 7.58 (1H, d, J = 8.2 Hz, Ar-CH), 7.57 (1H, s, Ar-CH), 7, 31 (1H, d, J = 8.1 Hz, Ar-CH), 4.57 (2H, bs, CH2NH +), 4.45 (2H, d, J = 6.3 Hz, CH2NH), 3.47 (2H, m, CH2N), 3.10 (2H, m, CH2N), 2.04-1.85 (4H, m, NCH2CH2CH2).

MS (ESI-) m / z 451 (M [Cl35] -1)

HPLC procedure 7 99.4% / 10.1 min

Example 17.7.10: Preparation of 2-dimethylaminomethyl-6-hydroxy-5oxo-5H-thiazolo [3,2-a] pyrimidine-7-carboxylic acid 3,4-dichloro-benzylamide hydrochloride

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

The following compound was prepared using Stage 3: B and Stage 4: B:

image 1

1H NMR (300 MHz, D6-DMSO): δ 12.26 (1H, s, OH), 11.18 (1H, bs, NH +), 9.81 (1H, t, J = 6.3 Hz, CH2NH ), 8.25 (1H, s, Ar-CH), 7.60 (1H, d, J = 8.1 Hz, Ar-CH), 7.58 (1H, s, Ar-CH), 7, 33 (1H, d, J = 8.1 Hz, Ar-CH), 4.51 (2H, bs, CH2NH +), 4.48 (2H, d, J = 6.3 Hz, CH2NH), 2.75 (6H, s, 2 x CH3).

 MS (ESI-) m / z 427 (M [Cl35] -1)

HPLC procedure 7 98.0% / 9.9 min

Example 17.7.1: Preparation of 4- [7- (3,4-dichloro-benzylcarbamoyl) -6-hydroxy-5-oxo-5H-thiazolo [3,2-a] pyrimidin-2-ylmethyl] tert-butyl ester -piperazine-1-carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

The following compound was prepared using Stage 3: A and Stage 4: A:

image 1

 1H NMR (300 MHz, D6-DMSO): δ 12.24 (1H, s, OH), 9.80 (1H, m, CH2NH), 8.00 (1H, s, Ar-CH), 7.61 (1H, d, J = 8.4 Hz, Ar-CH), 7.59 (1H, s, Ar-CH), 7.33 (1H, d, J = 8.4 Hz, Ar-CH), 4.46 (2H, d, J = 6.0 Hz, CH2NH), 3.71 (2H, s, CH2N), 3.33 (4H, m, CH2NCH2), 2.41 (4H, m, CH2NCH2) , 1.40 (9H, s, C [CH3] 3).

MS (ESI-) m / z 566 (M [Cl35] -1)

HPLC procedure 7 98.3% / 11.4 min

Example 17.7.12: Preparation of 3,4-Dichloro-benzylamide of 6-hydroxy-2- [4- (2-methoxy-phenyl) -piperazin-1-methyl] -5-oxo-5H-thiazolo [3,2- a] pyrimidine-7-carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

The following compound was prepared using Stage 3: A and Stage 4: A:

image 1

(300 MHz, D6-DMSO) δ 2.61 (4H, bs, 23-NCH2CH2N-CH2-), 2.96 (4H, bs, 23-NCH2CH2N-CH2-), 3.60 (2H, s -NCH2CH2NCH2 -), 3.75 (3H, s, -OCH3), 4.45 (2H, dd, J = 6.6 Hz, - (O = C) NHCH2-), 6.90 (4H, m, ArH) , 7.26 (1H, dd J = 1.5, 9.1 Hz, ArH), 7.53 (3H, m, ArH), 12.1 (1H, bs, OH).

MS (ESI-) m / z 572 (M [Cl35] -1) HPLC procedure 7 97.0% / 11.2 min

Example 17.7.13: Preparation of 3,4-dichloro-benzylamide hydrochloride of 6-hydroxy-5-oxo-2-propylaminomethyl-5H-thiazolo [3,2-a] pyrimidine-7-carboxylic acid

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

The following compound was prepared using Stage 3: B and Stage 4: B:

image 1

1H NMR (300 MHz, D6-DMSO) δ 0.91 (3H, t, J = 7.2 Hz, CH3CH2-), 1.65 (2H, q, J = 7.2 Hz, CH3CH2-), 2 , 87 (2H, t, J = 7.2 Hz, -CH2CH2-), 4.37 (2H, s, -CH2CH2NH2 + CH2-), 4.47 (2H, dd, J = 6.6 Hz, - (O = C) NHCH2-), 7.33 (1H, dd, J = 2.1, 8.4 Hz ArH), 7.59 (2H, m, ArH), 8.23 (1H, s, ArH ), 9.36 (1H, bs, - (O = C) NHCH2-), 9.79 (1H, t, J = 6.0 Hz, ArH), 12.3 (1H, bs, OH).

MS (ESI-) m / z 439 (M [Cl35] -1)

HPLC procedure 7 96.0% / 10.2 min

Example 17.7.14: Preparation of 6-hydroxy-5-oxo-2- (4-phenyl-piperazin-1-ylmethyl) 5H-thiazolo [3,2-a] pyrimidine- acid 3,4-dichloro-benzylamide 7-carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

The following compound was prepared using Stage 3: A and Stage 4: A:

image 1

1H NMR (300 MHz, D6-DMSO) δ 2.62 (4H, bs, PhNCH2CH2N-), 3.14 (4H, bs, PhNCH2CH2O-), 3.74 (2H, s, PhNCH2CH2NCH2-), 4.46 (2H, d, J = 6.0 Hz, - (O = C) NHCH2-), 6.77 (2H, t, J = 7.5 Hz, ArH), 6.93 (2H, d, J = 8.1 Hz, ArH) 7.20 (2H, t, J = 7.5 Hz, ArH), 7.32 (1H, dd, J = 1.8, 8.2 Hz, ArH), 7.60 (2H, m, ArH), 8.0 (1H, s, ArH) 9.88 (1H, t, J = 6.6 Hz (O = C) NHCH2-).

MS (ESI-) m / z 542 (M [Cl35] -1)

HPLC procedure 7 5.0% / 11.4 min

Example 17.7.15: Preparation of 6-hydroxy-2- (4-methanesulfonyl-piperazin-1-ylmethyl) -5-oxo-5H-thiazolo [3,2-a] pyrimidine7-carboxylic acid 3,4-dichloro-benzylamide of the acid

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

The following compound was prepared using Stage 3: A and Stage 4: A:

image 1

1H NMR (300 MHz, D6-DMSO) δ 2.54 (4H, bs, (SO2CH3) NCH2CH2N-), 2.88 (3H, bs, (SO2CH3) NCH2CH2N-), 3.14 (4H, bs, ( SO2CH3) NCH2CH2N-), 3.65 (2H, s, -NCH2CH2NCH2-), 4.43 (2H, bs, - (O = C) NHCH2-), 7.25 (1H, bs, ArH), 7, 51 (2H, bs, ArH), 7.71 (1H, bs, ArH).

MS (ESI-) m / z 544 (M [Cl35] -1) HPLC procedure 7 92.0% / 10.9 min

Example 18: Preparation of 6-hydroxy-5-oxo-1,5-dihydro-imidazo [1,2-a] pyrimidine7-carboxylic acid substituted benzylamides

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image27

Example 18.1.1: Route A

Example 18.1.1.1: Preparation of the 6-hydroxy-5-oxo-1,5-dihydro-imidazo [1,2-a] pyrimidine-7-carboxylic acid methyl ester (Step 1)

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The 2-aminoimidazole monosulfate (7.4 g, 56 mmol) was suspended in anhydrous methanol (20 ml) and cooled to -78˚C. To this mixture was slowly added a solution of sodium methoxide (3.0 g, 56 mmol) in anhydrous methanol (20 ml). After finishing the addition, the mixture was heated to room temperature and kept at room temperature for 4 h. The solid was collected by filtration and washed with anhydrous methanol and the washings were combined and concentrated to dryness in vacuo to give 2-amino imidazole (4.1 g, with a yield of 90%).

1H NMR (300 MHz, DMSO-d6) δ 4.95-5.30 (brs, 2H), 6.40 (s, 2H)

Dimethyl diacetoxybumarate (3.2 g, 12 mmol), 2-amino imidazole (1.03 g, 12 mmol) and p-toluenesulfonic acid (395 mg, 2.0 mmol were mixed in a 25 ml flask and immersed in a preheated oil bath (120 ° C) After 6 h, the reaction mixture was cooled to room temperature and ethyl acetate (10 ml) and methanol (0.6 ml) were added to the residue and the mixture was sonicated for 2 min The resulting precipitate was collected by filtration, washed with cold ethyl acetate (1 ml) and dried by pump to give the crude product Recrystallization from methanol gave the 6-hydroxy-5- acid methyl ester oxo-1,5-dihydro-imidazo [1,2-a] pyrimidine-7-carboxylic acid (1.03 g, 41%).

1H NMR (300 MHz, DMSO-d6) δ 3.85 (s, 3H), 7.56-7.64 (m, 2H), 9.00-9.40 (brs, 1H), 12.30- 12.70 (brs, 1H)

Example 18.1.1.2: Preparation of the 6-acetoxy-5-oxo-1,5-dihydro-imidazo [1,2-a] pyrimidine-7-carboxylic acid methyl ester (Step 2):

Although the following example is not included in the scope of the claims, it is entered as information from

image5

The product of Example 18.1.1.1 (300 mg, 1.43 mmol) and N, N-dimethylamino pyridine (262 mg, 2.15 mmol) were dissolved in dichloromethane (20 ml), and then cooled to 5 ° C. To this stirred mixture was added dropwise a solution of acetyl chloride (112 mg, 1.43 mmol) in dichloromethane (5 ml). After maintaining this temperature for 30 min, the mixture was heated to room temperature and stirred for another 12 h. The solvent was removed under reduced pressure and the resulting residue was subjected to silica gel column chromatography using dichloromethane / methanol (20: 1) as eluent. The 6-acetoxy-5-oxo-1,5-dihydro-imidazo [1,2-a] pyrimidine 7-carboxylic acid methyl ester (195 mg, 54.2%) was obtained as a yellow solid. 1H NMR (300 MHz, DMSO-d6) δ 2.26 (s, 3H), 3.84 (s, 3H), 7.72 (d, J = 2.6Hz, 1H), 7.75 (d, J = 2.6Hz, 1H), 13.20-13.26 (brs, 1H)

Example 18.1.1.3: Preparation of the 6-acetoxy-1- (2-morpholin-4-yl-ethyl) -5-oxo-1,5-dihydroimidazo [1,2-a] pyrimidine-7-carboxylic acid methyl ester (Stage 3)

[0252] Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The product of Example 18.1.1.2 (30 mg, 0.12 mmol) and 18-crown-6 (3 mg, 10% w / w) were mixed with acetonitrile (5 ml) at room temperature. To this stirred solution was added anhydrous potassium carbonate (83 mg, 0.60 mmol) and 4- (2-chloroethyl) -morpholine hydrochloride (25 mg, 0.132 mmol). The mixture was heated at reflux for 2 h and then cooled to room temperature. The reaction was concentrated to near dryness in vacuo and ethyl acetate (15 ml) was added and the mixture was washed with water (2 x 10 ml). The organic layer was separated, dried and concentrated in vacuo and the residue was purified by column chromatography using dichloromethane / methanol (20: 1) as eluent. 6-Acetoxy-1- (2morpholin-4-yl-ethyl) -5-oxo-1,5-dihydro-imidazo [1,2-a] pyrimidine-7-carboxylic acid methyl ester (15 mg 34%) It was obtained as a white solid.

Example 18.1.1.4: Preparation of the 6-acetoxy-1-allyl-5-oxo-1,5-dihydro-imidazo [1,2-a] pyrimidine7-carboxylic acid methyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The procedure described in Example 18.1.1.3 was adapted except that sodium hydride (1.1 eq.) Was added to the reaction mixture at room temperature 30 min before the addition of allyl bromide (1.1 eq. ) and the reaction mixture was heated to 70 ° C.

1H NMR (500 MHz, CDCl3): δ 7.65 (1H, d, J = 2.7 Hz, Ar-CH), 7.10 (1H, d, J = 2.7 Hz, Ar-CH), 5.98 (1H, ddt, J = 17.1, 10.5,

, 7 Hz, CH2CH = CH2), 5.39 (2H, m, CH2CH = CH2), 4.78 (2H, dt, J = 5.7, 1.8 Hz, CH2CH = CH2), 3.95 ( 3H, s, OCH3), 2.37 (3H, s, C [= O] CH3).

Example 18.1.2: Route B Example 18.1.2.1: Preparation of 1- (2-Piperidin-1-yl-ethyl) -1H-imidazol-2-ylamine (Step 1)

[0254] Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The 2-aminoimidazole monosulfate (500 mg, 1.89 mmol) and potassium carbonate (580 mg, 4.16 mmol) were suspended in DMF (2 ml) and chloroethyl piperidine monohydrochloride (766 mg, 4.16 was added mmol). The reaction was heated to 100 ° C for 2.5 hours before cooling to room temperature and filtered. The filtrate was concentrated and purified by column chromatography (95: 4.5: 0.5 dichloromethane: methanol: aqueous ammonia) and the product was isolated as a brown oil (83 mg, 11%).

1H NMR (300 MHz, D6-DMSO): δ 6.59 (1H, s, Ar-CH), 6.45 (1H, s, Ar-CH), 5.41 (2H, bs, NH2), 3 , 80 (2H, t, J = 5.0 Hz NCH2CH2N), 2.59 (2H, t, J = 5.0 Hz, NCH2CH2N), 2.46 (4H, m, CH2NCH2), 1.57 (4H , m, NCH2CH2CH2CH2), 1.45 (2H, m, NCH2CH2CH2CH2).

Example 18.1.2.2: Preparation of the 6-acetoxy-5-oxo-1- (2-piperidin-1-yl-ethyl) -1,5-dihydroimidazo [1,2-a] pyrimidine-7-carboxylic acid methyl ester (Stage 2)

[0255] Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The procedure described in Example 4 was adapted to give the 6-acetoxy-5-oxo-1- (2-piperidin-1-ylethyl) -1,5-dihydroimidazo [1,2-a] pyrimidine- acid methyl ester 7-carboxylic.

1H NMR (300 MHz, D6-DMSO): δ 7.60 (1H, d, J = 2.4 Hz, Ar-CH), 7.42 (1H, d, J = 2.4 Hz, Ar-CH ), 4.33 (2H, m, CH2N), 3.95 (3H, s, OCH3), 2.76 (2H, m, CH2N), 2.54 (4H, m, CH2NCH2), 2.37 ( 3H, s, O = CCH3), 1.63-1.43 (6H, m NCH2CH2CH2CH2).

MS (ESI +) m / z 363 (M + 1).

Example 18.2: Preparation of 6-hydroxy-1-methyl-5-oxo-1,5-dihydro-imidazo [1,2a] pyrimidine-7-carboxylic acid 4-fluoro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Starting from iodomethane and adapting the procedure described in Example 18.1.2, the 6-hydroxy-1-methyl-5-oxo1,5-dihydroimidazo [1,2-a] pyrimidine-7-carboxylic acid methyl ester was obtained. By adapting the procedure described in Example 6, 6-hydroxy-1-methyl-5-oxo-1,5-dihydro-imidazo [1,2-a] pyrimidine-7-carboxylic acid 4-fluoro-benzylamide was obtained.

Example 18.3: Preparation of 6-hydroxy-1-methyl-5-oxo-1,5-dihydroimidazo [1,2a] pyrimidine-7-carboxylic acid 3,4-dichloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The procedure described in Example 18.2 was adapted to give the 6-hydroxy-1-methyl-5-oxo1,5-dihydro-imidazo [1,2-a] pyrimidine-7-carboxylic acid 3,4-dichloro-benzylamide .

(300 MHz, DMSO) δ 3.67 (3H, s, N-CH3), 4.50 (2H, d, J = 6.3 Hz, -NH-CH2-), 7.32 (1H, dd, J = 2.1, 8.1 Hz, - (CH3) NCH-CH-N), 7.58 (4H, m, ArH), 9.56 (1H, t, J = 6.3 Hz, -NHCH2 -).

MS (ESI-) m / z 365 (M [Cl35] -1)

Example 18.4: Preparation of 6-hydroxy-1- (2-morpholin-4-yl-ethyl) -5-oxo-1,5-dihydroimidazo [1,2-a] pyrimidine acid 3,4-dichloro-benzylamide -7-carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Using the product of Example 18.1.1.3 and by adapting the procedure described in Example 6, the 6-hydroxy-1- (2-morpholin-4-yl-ethyl) -5-oxo 3,4-dichloro-benzylamide was prepared -1,5-dihydro-imidazo [1,2-a] pyrimidine-7-carboxylic.

1H NMR (300 MHz, D6-DMSO): δ 9.57 (1H, t, J = 6.6 Hz, NHCH2), 7.67 (1H, d, J = 2.7 Hz, Ar-CH), 7.60 (3H, m, ArCH), 7.33 (1H, d, J = 8.4 Hz, Ar-CH), 4.53 (2H, d, J = 6.6 Hz, NHCH2), 7 , 26 (2H, t, J = 5.4 Hz, NCH2CH2N), 3.43 (4H t, J = 4.8 Hz, CH2OCH2), 2.68 (2H, t, J = 5.4 Hz, NCH2CH2N ), 2.45 (4H, m, CH2NCH2).

MS (ESI-) m / z 464 (M [Cl35] -1)

Example 18.5: Preparation of 6-hydroxy-5-oxo-1- (2-piperidin-1-yl-ethyl) -1,5-dihydroimidazo [1,2-a] pyrimidine7-3,4-dichloro-benzylamide -carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image28

The procedure of Example 6 was adapted to the product of Example 18.1.2.2 to give the 6-hydroxy-5-oxo-1- (2-piperidin-1-yl-ethyl) -1,5-dihydro- 3,4-dichloro-benzylamide imidazo [1,2-a] pyrimidine-7-carboxylic.

1H NMR (300 MHz, D6-DMSO): δ 11.43 (1H, s, OH), 9.55 (1H, bs, NH), 7.64-7.59 (4H, m, Ar-CH) , 7.32 (1H, d, J = 8.1 Hz, Ar-CH), 4.51 (2H, d, J = 6.6 Hz, CH2NH), 4.23 (2H, t, J = 6 , 3 Hz, NCH2CH2N), 2.62 (2H, t, J = 6.3 Hz, NCH2CH2NH), 2.41-2.38 (4H, m, CH2NCH2), 1.23 (6H, m, NCH2CH2CH2CH2) .

MS (ESI +) m / z 464 (M [Cl35] +1).

Example 18.6: Preparation of 1-allyl-6-hydroxy-5-oxo-1,5-dihydro-imidazo [1,2a] pyrimidine-7-carboxylic acid 3,4-dichloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The procedure described in Example 6 was applied to the product of Example 18.1.1.4 to give the desired product.

1H NMR (500 MHz, D6-DMSO): δ 9.57 (1H, m, NHCH2), 7.66-7.57 (4H, m, Ar-CH), 7.33 (1H, d, J = 7.8 Hz, Ar-CH) 6.10-5.97 (1H, m, CH2CH = CH2), 5.27 (1H, m, CH2CH = CH2), 5.23 (1H, m, CH2CH = CH2 ), 4.77 (2H, d, J = 5.4 Hz, CH2CH = CH2), 4.51 (2H, d, J = 6.6 Hz, NHCH2).

MS (ESI +) m / z 393 (M [Cl35] +1).

Example 19: Preparation of substituted benzylamides of 2-hydroxy-1-oxo-1H-9-oxa-4,9a-diaza-fluorene-3-carboxylic acid

Example 19.1: Preparation of the 2-hydroxy-1-oxo-1H-9-oxa-4,9a-diaza-fluorene-3-carboxylic acid methyl ester

Although the following example is not included in the scope of the claims, it is entered as information from

Useful background for the reader. For Stage 1 and Stage 2, the procedure described in J. Heterocyclic Chem., 1989, 26, 1293 was adapted to give 3-aminobenzoisoxazole. The procedure described in Example 5 was adapted to give the desired ester.

1H NMR (300 MHz, D6-DMSO) δ 3.90 (s, 3H), 7.52-7.62 (m, 1H), 7.80-7.90 (m, 2H), 8.12 ( d, J = 7.8 Hz, 1H), 10.92 (s, 1H). MS (ESI +) m / z 283 (M + Na).

image29

Although the following example is not included in the scope of the claims, it is entered as information from

image5

By adapting the procedure described in Example 19.1 and using the 2-fluoro-6-morpholin-4-yl-benzonitrile as the starting material, the desired ester was prepared.

1H NMR (300 MHz, D6-DMSO) δ 3.24-3.45 (4H), 3.85 (t, J = 4.6 Hz, 4H), 3.90 (s, 3H), 6.89 (d, J = 8.3 Hz, 1H), 7.25 (d J = 8.1 Hz, 1H), 7.68 (t, J = 8.4 Hz, 1H), 10.78 (s, 1 HOUR)

Example 19.3: Preparation of the 2-benzyloxy-1-oxo-1H-9-oxa-4,9a-diaza-fluorene-3-carboxylic acid methyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

A solution of the product of Example 19.1 (50 mg, 0.19 mmol) and benzyl alcohol (46 mg, 0.42 mmol) in tetrahydrofuran (10 ml) was cooled (ice / water bath). To this solution was added triphenylphosphine (111 mg, 0.423 mmol) and diisopropyl azodicarboxylate (85 mg, 0.42 mmol). The mixture was heated to room temperature and after 2 h, the volatiles were concentrated in vacuo. The residue was purified by column chromatography (hexane / ethyl acetate 1: 1) to give the desired compound (53 mg, 79%) as a white solid.

 1H NMR (300 MHz, DMSO-d6) δ 3.82 (s, 3H), 5.23 (s, 2H), 7.31-7.52 (m, 5H), 7.57-7.67 ( m, 1H), 7.93 (d, J = 3.2 Hz, 2H) 8.18 (d, J = 7.7 Hz, 1H)

Example 19.4: Preparation of the 2-benzyloxy-5-morpholin-4-yl-1-oxo-1H-9-oxa-4,9a-diaza-fluorene-3-carboxylic acid methyl ester

[0264] Although the following example is not included in the scope of the claims, it is entered as information

image30

The procedure described in Example 8.1 was adapted, but the reaction was carried out at 70 ° C using DMF as solvent, to give the desired compound.

1H NMR (300 MHz, DMSO-d6) δ 3.33-3.40 (m, 4H), 3.77-3.86 (m, 7H), 5.21 (s, 2H), 6.93 ( d, J = 8.2 Hz, 1H), 7.29-7.50 (m, 6H), 7.75 (t, J = 8.4 Hz, 1H).

Although the following example is not included in the scope of the claims, it is entered as information from

image31

Using the product of Example 19.3 and adapting the procedure described in Example 8.2, the desired compound was obtained.

1H NMR (300 MHz, DMSO-d6) δ 5.21 (s, 2H), 7.30-7.45 (m, 3H), 7.50 (d, J = 6.6 Hz, 2H), 7 , 56-7.69 (m, 1H), 7.93 (d J = 3.6 Hz, 2H), 8.19 (d, J = 8.1 Hz, 1H), 13.78-13.98 (brs, 1H).

Example 19.6: Preparation of 2-benzyloxy-5-morpholin-4-yl-1-oxo-1H-9-oxa-4,9a-diaza-fluorene-3-carboxylic acid

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Using the product of Example 19.4 and adapting the procedure described in Example 8.2, the desired compound was obtained.

1H NMR (300 MHz, DMSO-d6) δ 3.34-3.41 (m, 4H), 3.78-3.86 (m, 4H), 5.20 (s, 2H), 6.93 ( d, J = 8.2 Hz, 1H), 7.30-7.44 (m, 4H), 7.46-7.54 (m, 2H), 7.75 (t, J = 8.2 Hz , 1H), 13.58-13.79 (brs, 1H)

MS (ESI-) m / z 420 (M-1)

Example 19.7: Preparation of 2-benzyloxy-5-morpholin-4-yl-1-oxo-1H-9-oxa-4,9adiaza-fluorene-3-carboxylic acid 3,4-dichloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

N, N’-dicyclohexylcarbodiimide (110 mg, 0.522 mmol) was added to a stirred solution of the product from Example

19.6 (200 mg, 0.475 mmol) in dichloromethane (100 ml). After 30 min, N, N-dimethylaminopyridine (6 mg, 0.05 mmol), 3,4-dichlorobenzylamine (92 mg, 0.52 mmol) and 1-hiroxybenzotriazole (70 mg, 0.52 mmol) were added successively . The mixture was stirred at room temperature overnight and after processing and aqueous extraction the crude product was further purified by column chromatography (hexane / ethyl acetate 4: 1) as eluent to give the desired product (120 mg 44%). As a yellow solid.

1H NMR (300 MHz, CDCl3) δ 3.37-3.45 (m, 4H), 3.85-3.95 (m, 4H), 4.51 (d, J = 5.8 Hz, 2H) , 5.43 (s, 2H), 6.84 (d, J = 8.3 Hz, 1H), 7.06-7.16 (m, 2H), 7.28-7.36 (m, 3H ), 7.37-7.49 (m, 4H), 7.55-7.71 (m, 2H).

Example 19.8: Preparation of 2-hydroxy-5-morpholin-4-yl-1-oxo-1H-9-oxa-4,9adiaza-fluorene-3-carboxylic acid 3,4-dichloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Iron (III) chloride (10 mg, 0.062 mmol) was added to a stirred solution of the product from Example 19.7 (12 mg, 0.021 mmol) in dichloromethane (5 ml). The mixture was stirred at room temperature for 1.5 h and then aqueous hydrochloric acid (1.0 M) was added dropwise until the solution became clear. The products were extracted with ethyl acetate and the organic phase was dried and concentrated in vacuo. The residue was recrystallized from a solvent combination (10/1 hexane / ethyl acetate) to give the desired compound (8 mg, 80%) as a gray solid.

1H NMR (300 MHz, D6-DMSO): δ 12.28 (1H, s, OH), 8.62 (1H, m, NHCH2), 7.28 (3H, m, Ar-CH), 7.40 (1H, d, J = 8.7 Hz, Ar-CH), 7.30 (1H, d, J = 8.4 Hz, Ar-CH), 6.95 (1H, d, J = 8.1 Hz, Ar-CH), 4.62 (2H, d, J = 6.6 Hz, NHCH2), 3.77 (4H, m CH2OCH2), 2.49 (4H, m, CH2NCH2).

MS (ESI-) m / z 487 (M [Cl35] -1)

Example 19.9: Preparation of 2-hydroxy-1-oxo-1H-9-oxa-4,9a-diaza-fluorene-3-carboxylic acid 3,4-dichlorobenzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Using the product of Example 19.5 and adapting the procedures described Example 19.7 and Example 19.8 provided the desired compound.

1H NMR (300 MHz, D6-DMSO) δ 4.55 (2H, d, J = 6.0 Hz, -NH-CH2-), 7.37 (1H, dd, J = 8.4, 2.1Hz , ArH), 7.61 (3H, m, ArH), 7.87 (2H, m, ArH), 8.07 (1H, d, J = 7.5 Hz, ArH), 9.77 (1H, t, J = 6.0 Hz, O = C-NH-CH2), 12.79 (1H, s, OH).

MS (ESI +) m / z 404 (M [Cl35] +1)

HPLC procedure 7 96.2% / 19.0 min

Example 19.10: Preparation of 2-hydroxy-1-oxo-1H-9-oxa-4,9a-diaza-fluorene-3-carboxylic acid 4-fluoro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Using the product of Example 19.5 and adapting the procedures described in Example 19.7 and Example 19.8, the desired compound was obtained.

1H NMR (300 MHz, D6-DMSO) δ 4.54 (2H, d, J = 6.9 Hz, -NH-CH2-), 7.17 (2H, t, J = 9.0, 2.4 Hz, ArH), 7.42 (2H, m, ArH) 7.60 (1H, m, ArH), 7.87 (1H, m, ArH), 8.06 (1H, d, J = 8.1 Hz, ArH), 9.72 (1H, t, J = 6.6 Hz O = C-NH-CH2), 12.93 (1H, s, OH).

MS (ESI-) m / z 352 (M-1)

HPLC procedure 7 93.1% / 12.5 min

Example 19.11: Preparation of 2-hydroxy-5-morpholin-4-yl-1-oxo-1H-9-oxa-4,9a-diazafluorene-3-carboxylic acid 4-fluoro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Using the product of Example 19.6 and adapting the procedures described in Example 19.7 and Example 19.8, the desired compound was obtained

1H NMR (300 MHz, D6-DMSO) δ 3.29 (4H, s, N-CH2-CH2-O), 3.71 (4H, s, N-CH2-CH2-O), 4.60 (2H , dd, J = 6.0 Hz, -NHCH2-), 6.94 (1H, dd, J = 7.8 Hz, ArH), 7.21 (2H, t, J = 8.2 Hz, ArH) , 7.29 (1H, dd, J = 8.4Hz, ArH), 7.45 (2H, m, ArH), 7.70 (1H, dd, J = 8.4Hz, ArH), 8.43 (1H, t, O = C-NH-CH2), 12.42 (1H, s, OH).

MS (ESI-) m / z 437 (M [Cl35] -1)

HPLC procedure 7 91.0% / 15.7 min

Example 20: Preparation of substituted benzylamides of 3-hydroxy-4-oxo-4H-9-thia-1,4a-diaza-fluorene-2-carboxylic acid

The following compounds were prepared by adapting the procedures described in Example 4 and Example 6:

Example 20.1: Preparation of 3-hydroxy-4-oxo-4H-9-thia-1,4a-diaza-fluorene-2-carboxylic acid 3,4-dichlorobenzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

1H NMR (300 MHz, D6-DMSO): δ 12.35 (1H, s, OH), 9.82 (1H, t, J = 6.9 Hz, NHCH2), 8.90 (1H, m, Ar -CH), 8.01 (1H, m, Ar-CH), 7.63-7.56 (4H, m, Ar-CH), 7.35 (1H, d, J = 7.8 Hz, Ar -CH), 4.48 (2H, d, J = 6.9 Hz, CH2NH).

MS (ESI-) m / z 418 (M [Cl35] -1)

HPLC procedure 7 91% / 18.8 min

Example 20.2: Preparation of 3-hydroxy-7-methoxy-4-oxo-4H-9-thia-1,4a-diazafluorene-2-carboxylic acid 3,4-dichloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

 1H NMR (300 MHz, D6-DMSO): δ 12.30 (1H, s, OH), 9.75 (1H, t, J = 6.9 Hz, NHCH2), 8.77 (1H, d, J = 9.3 Hz, Ar-CH) 7.637.59 (3H, m, Ar-CH), 7.33 (1H, d, J = 8.4 Hz, Ar-CH), 7.13 (1H, d , J = 9.3 Hz, Ar-CH), 4.46 (2H, d, J = 6.9 Hz, CH2NH) 3.83 (3H, s, CH3).

HPLC procedure 7 95.3% / 19.1 min

Example 21: Preparation of substituted benzylamides of 3-hydroxy-4-oxo-4,6-dihydro-pyrimido [1,2-b] indazol-2-carboxylic acid

Example 21.1: Preparation of the 3-hydroxy-4-oxo-4,6-dihydro-pyrimido [1,2-b] indazol-2-carboxylic acid methyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Stage 1:

2-Fluorobenzonitrile (605 mg, 5 mmol) and 85% aqueous hydrazine hydrate (352 mg, 6 mmol) were mixed with 1-butanol (3 ml). The mixture was heated at reflux with stirring for 5 h and then cooled to room temperature. The resulting precipitate was collected by filtration and washed with dichloromethane and the filter cake dried in vacuo to give 3-aminobenzopyrazole (293 mg, 44%).

1H NMR (300 MHz, D6-DMSO): δ 5.26-5.36 (brs, 2H), 6.84-6.93 (m, 1H), 7.18-7.24 (m, 2H) , 7.67 (dt, J = 8.1, 0.9 Hz 1H), 11.33 (s, 1H).

Stage 2:

The procedure described in Example 3 was adapted to the product of Step 1 to give the desired compound.

1H NMR (300 MHz, D6-DMSO): δ 3.91 (s, 3H), 7.33 (t, J = 7.7 Hz, 1H), 7.48 (dt, J = 8.0, 0 , 8 Hz, 1H), 7.70 (t, J = 7.7 Hz 1H), 8.09 (d, J = 8.0 Hz, 1H), 10.25 (s, 1H), 13.10-13.80 (brs, 1H). MS (ESI-) m / z 258 (M-1)

Example 21.2: Preparation of the 3-hydroxy-10-morpholin-4-yl-4-oxo-4,6-dihydro-pyrimido [1,2b] indazol-2-carboxylic acid methyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

By adapting the procedure described in Example 21.1 and using the 2-fluoro-6-morpholin-4-yl-benzonitrile as the starting material, the desired ester was prepared.

 1H NMR (300 MHz, D6-DMSO): δ 3.30 (m, 4H (obscured by water peak)), 3.88 (t, J = 4.5 Hz, 4H), 3.92 (s , 3H), 6.67 (d J = 7.9 Hz, 1H), 6.98 (d, J = 8.1 Hz, 1H), 7.55 (t, J = 8.0 Hz, 1H) , 10.15 (s, 1H), 13.20-13.55 (brs, 1H) MS (ESI-) m / z 343 (M-1)

Example 21.3: Preparation of 3-hydroxy-10-morpholin-4-yl-4-oxo-4,6-dihydro-pyrimido [1,2-b] indazol-2-carboxylic acid 4-fluoro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The product of Example 21.2 (172 mg, 0.5 mmol), sodium methoxide (54 mg, 1.0 mmol) and 4-fluorobenzylamine (1.87 mg, 1.5 mmol) in methanol (15 ml) were combined and heated with stirring to reflux overnight. The mixture was cooled to room temperature and the resulting solid was collected by filtration and dissolved in dichloromethane (30 ml). The solution was washed with aqueous hydrochloric acid (2.0 M), water, dried and concentrated in vacuo to give the desired compound (84 mg, 38.4).

1H NMR (300 MHz, D6-DMSO): δ 13.61 (1H, s, NH), 11.97 (1H, s, OH), 8.45 (1H, t, J = 6.0 Hz, NHCH2 ), 7.56 (1H, t J = 6.0 Hz, Ar-CH), 7.45 (2H, dd, J = 9.0, 8.0 Hz, Ar-CH), 7.20 (2H , dd, J = 9.0, 9.0, Hz, Ar-CH), 7.00 (1H, d, J = 8.1 Hz, Ar-CH), 6.71 (1H, d, J = 7.8 Hz, Ar-CH), 4.61 (2H, d, J = 6.0 Hz, NHCH2), 3.69 (4H, m, CH2OCH2), 3.15 (4H, m, CH2NCH2).

MS (ESI-) m / z 436 (M-1)

HPLC procedure 7 98.9% / 13.6 min

Example 21.4: Preparation of 3-hydroxy-4-oxo-4,6-dihydro-pyrimido [1,2-b] indazol-2-carboxylic acid 4-fluorobenzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

By adapting the procedure described in Example 21.3, the desired compound was prepared.

1H NMR (300 MHz, DMSO) δ 4.55 (2H, d, J = 6.6 Hz, -NH-CH2-), 7.18 (2H, m, ArH), 7.42 (4H, m, ArH), 7.70 (1H, t, J = 7.2.7.8 Hz, ArH), 8.09 (1H, d, J = 7.8 Hz, ArH), 9.67 (1H, t, J = 6.6 Hz, O = C-NH -CH2), 12.40 (1H, s, OH). MS (ESI-) m / z 351 (M-1)

HPLC procedure 7 96.4% / 13.9 min

Example 21.5: Preparation of 3-hydroxy-10-morpholin-4-yl-4-oxo-4,6-dihydro-pyrimido [1,2-b] indazol-2-carboxylic acid 3,4-dichloro-benzylamide acid

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

By adapting the procedure described in Example 21.3, the desired compound was prepared.

1H NMR (300 MHz, DMSO) δ 3.19 (4H, s, N-CH2-CH2-O), 3.75 (4H, s, N-CH2-CH2-O), 4.63 (2H, d , J = 6.3 Hz, -NHCH2-), 6.70 (1H, dd, J = 7.8 Hz, ArH), 7.00 (1H, dd, J = 8.4 Hz, ArH), 7 , 41 (1H, dd, J = 6.9, 2.1 Hz, ArH), 7.56 (1H, t, J = 8.2 Hz, ArH), 7.66 (2H, m, ArH), 8.59 (1H, t, J = 5.7 Hz, O = C-NH-CH2), 11.81 (1H, s, OH).

MS (ESI-) m / z 486 (M [Cl35] -1) HPLC procedure 7 94.3% / 15.8 min

Example 21.6: Preparation of 3-hydroxy-6-methyl-4-oxo-4,6-dihydro-pyrimido [1,2-b] indazol-2-carboxylic acid methyl ester

Although the following example is not included in the scope of the claims, it is entered as information from

image32

3-Aminobenzopyrazole (266 mg, 2 mmol) and phthalic anhydride (296 mg, 2 mmol) were mixed and heated at 170 ° C for 30 min. The mixture was cooled to room temperature after which methanol (10 ml) was added and then the mixture was sonicated for 2 min. The solid was collected by filtration and washed with methanol to give the desired product (352 mg, 67%).

1H NMR (300 MHz, D6-DMSO) δ 7.13-7.20 (m, 1H), 7.40-7.48 (m, 1H), 7.60-7.66 (m, 1H), 7.70 (dd, J = 8.2, 0.8 Hz, 1H), 7,948.07 (m, 4H), 13.44 (s, 1H).

Stage 2:

Iodomethane (1.41 g, 10 mmol) t room temperature was added dropwise to a stirred solution of the product from Step 1 (2.63 g, 10 mmol) and potassium carbonate (2.76 g, 20 mmol) in DMF (50 ml). After 3 h the reaction mixture was cooled to room temperature and poured into water / ice (300 ml) and extracted with dichloromethane (3 x 100 ml). The combined organic layers were washed with brine, dried and evaporated under reduced pressure and the resulting residue was recrystallized from ethyl acetate to give the desired compound (2.27 g, 82%)

1H NMR (300 MHz, D6-DMSO) δ 4.12 (s, 3H), 7.19 (ddd, J = 8.0, 7.0, 0.8 Hz, 1H), 7.49 (ddd, J = 8.6, 6.9, 1.1 Hz, 1H), 7.71 (dt, J = 8.0, 1.1 Hz, 1H), 7.73 (dt, J = 8.8, 0.9 Hz, 1H), 7.94-8.06 (m, 4H).

Stage 3:

The product from Step 2 (277 mg, 1 mmol) was suspended in a mixture of methanol (15 ml) and 85% aqueous hydrazine hydrate (588 mg, 10 mmol). The mixture was heated at reflux for 1 h and then cooled to room temperature. Water (40 ml) was added and the mixture was extracted with dichloromethane (3 x 20 ml). The combined organic layers were washed with brine, dried and concentrated in vacuo. The residue was purified by flash chromatography (dichloromethane / methanol 10: 1) to give the desired compound (105 mg, 72%).

1H NMR (300 MHz, DMSO) δ 3.71 (s, 3H), 5.39 (s, 2H) 6.85-6.93 (m, 1H), 7.21-7.33 (m, 2H ), 7.66 (dt, J = 8.0, 1.0 Hz, 1H).

Stage 4:

The procedure described in Example 3 was adapted to the product of Step 3 to give the desired compound.

1H NMR (300 MHz, D6-DMSO) δ 3.87 (s, 3H), 3.90 (s, 3H), 7.35-7.47 (m, 1H), 7.76 (d, J = 3.5 Hz, 2H), 8.04 (d, J = 8.0 Hz, 1H), 10.35 (s, 1H).

MS (ESI +) m / z 296 (M + 23)

Example 21.7: Preparation of the 3-hydroxy-6-methyl-10-morpholin-4-yl-4-oxo-4,6-dihydro-pyrimido [1,2-b] indazol-2-carboxylic acid methyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Using 4-morpholin-4-yl-1H-indazol-3-ylamine and adapting the procedure described in Example 21.6, the desired compound was prepared.

1H NMR (300 MHz, D6-DMSO) δ 3.30 (4H, obscured by water peak), 3.82 (s, 3H), 3.84-3.93 (m, 7H), 6.79 (d, J = 8.0 Hz, 1H), 7.23 (d, J = 8.1 Hz, 1H), 7.61 (t, J = 8.2 Hz, 1H), 10.24 (s , 1H) MS (ESI +) m / z 381 (M + 23)

Example 21.8: Preparation of 3-hydroxy-6-methyl-10-morpholin-4-yl-4-oxo-4,6-dihydropyrimido [1,2-b] indazol-2-carboxylic acid 4-fluoro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

By adapting the procedure described in Example 21.3, the desired compound was prepared.

1H NMR (300 MHz, D6-DMSO) δ 3.15 (4H, s, N-CH2-CH2-O), 3.66 (4H, s, N-CH2-CH2-O), 4.62 (2H , d, J = 6.0 Hz, -NHCH2-), 6.70 (1H, dd, J = 7.8 Hz, ArH), 7.21 (3H, m, ArH), 7.45 (2H, m, ArH), 7.62 (1H, t, J = 8.1 Hz, ArH), 8.39 (1H, t, J = 6.3 Hz, O = C-NH-CH2), 12.02 (1H, s, OH).

 MS (ESI-) m / z 450 (M-1)

HPLC procedure 7 99.7% / 12.6 min

Example 21.9: Preparation of 3,4-dichloro-benzylamide of 3-hydroxy-6-methyl-10-morpholin-4-yl-4-oxo-4,6-dihydropyrimido [1,2-b] indazol-2 acid -carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

By adapting the procedure described in Example 21.3, the desired compound was prepared.

1H NMR (300 MHz, D6-DMSO) δ 3.23 (4H, bs, -NCH2CH2O-), 3.73 (4H, bs, -NCH2CH2O-), 3.82 (3H, s, -NCH3), 4 , 63 (2H, d, J = 6.6 Hz, - (O = C) NHCH2-), 6.84 (2H, d, J = 8.1 Hz, ArH), 7.27 (2H, d, J = 8.1 Hz, ArH), 7.39 (2H, dd, J = 2.4, 8.0 Hz, ArH), 7.65 (3H, m, ArH), 9.73 (1H, t , J = 6.6 Hz, - (O = C) NHCH2-), 11.87 (1H, s, OH).

MS (ESI-) m / z 524 (M [Cl35] + Na)

HPLC procedure 7 96.0% / 14.2 min

Example 21.10: Preparation of 3-hydroxy-6-methyl-4-oxo-4,6-dihydro-pyrimido [1,2b] indazol-2-carboxylic acid 4-fluoro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

By adapting the procedure described in Example 21.3, the desired compound was prepared.

1H NMR (300 MHz, D6-DMSO) δ 3.84 (3H, s, -NCH3), 4.55 (2H, d, J = 6.0 Hz, - (O = C) NHCH2-), 7, 18 (2H, m, ArH), 7.47 (3H, m, ArH), 7.76 (2H, dd, J = 1.5, 9.1 Hz, ArH), 8.06 (1H, dd, J = 0.9, 8.5 Hz, ArH), 9.68 (1H, bt, - (O = C) NHCH2-), 12.47 (1H, bs, OH).

MS (ESI-) m / z 365 (M-1) HPLC procedure 7 85.0% / 12.8 min

Example 21.11: Preparation of 3-hydroxy-6-methyl-4-oxo-4,6-dihydro-pyrimido [1,2b] indazol-2-carboxylic acid 3,4-dichloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

By adapting the procedure described in Example 21.3, the desired compound was prepared.

 1H NMR (300 MHz, D6-DMSO) δ 3.84 (3H, s, -NCH3), 4.56 (2H, d, J = 6.6 Hz, - (O = C) NHCH2-), 7, 37 (2H, dd, J = 2.1, 8.4 Hz, ArH), 7.47 (1H, m, ArH), 7.62 (2H, m, ArH), 7.77 (2H, d, J = 3.6 Hz, ArH), 8.06 (1H, d, J = 8.1 Hz, ArH), 9.73 (1H, bs (O = C) NHCH2-), 12.34 (1H, bs, OH).

MS (ESI-) m / z 415 (M [Cl35] -1), 417 (M [CI37] -1)

HPLC procedure 7 88.0% / 14.4 min

Example 22: Preparation of 3-hydroxy-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid disubstituted benzylamides

Example 22.1: Preparation of 3-hydroxy-9-iodo-7-morpholin-4-yl-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid 3,4-dichloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Stage 1:

Sodium periodate (1.53 g, 7.16 mmol) was added to a stirred solution of 5-fluoro-2-aminopyridine (2.0 g, 17.9 mmol) in aqueous sulfuric acid (2.0 M, 30 ml) and the reaction was heated to 100 ° C. A solution of sodium iodide (2.68 g 17.9 mmol) in water (10 ml) was added dropwise to the reaction mixture. After completion of the addition, the mixture was refluxed for 1 h and then cooled to room temperature. A saturated sodium bicarbonate solution was added dropwise to adjust the pH -7.0 and the mixture was extracted with dichloromethane (3 x). The combined organic layers were washed with aqueous sodium bisulfite solution, brine, dried and concentrated in vacuo. The residue was subjected to column chromatography (hexane / ethyl acetate 4: 1) to give the desired compound (2.56 g, 60%).

1HNMR (300 MHz, CDCl3) δ 4.70-5.03 (brs, 2H), 7.69 (dd, J = 7.2, 2.1 Hz, 1H), 7.93 (d, J = 2 , 1 Hz, 1H)

MS (ESI +) m / z 239 (M + 1)

Stage 2:

The product of Step 1 (400 mg, 1.68 mmol) was dissolved in concentrated sulfide acid (2 ml) and cooled to -10 ° C. To this stirred solution was added dropwise a mixture of 30% hydrogen peroxide (2.3 g, 20.2 mmol) and concentrated sulfuric acid (4.2 ml). The mixture was kept at -10 ° C for 30 min, after which it was heated to 8 ° C and stirred at this temperature overnight. The mixture was poured onto water / ice (50 ml) and extracted with dichloromethane (3 x). The combined organic layers were washed with aqueous sodium bisulfite solution, brine, dried and concentrated in vacuo. The residue was subjected to column chromatography (hexane / ethyl acetate 5: 1) to give the desired product (36 mg, 8.0%).

1H NMR (300 MHz, CDCl3) δ 8.13 (dd, J = 6.9, 2.3 Hz, 1H), 8.35 (d, J = 2.4 Hz, 1H).

Stage 3:

By adaptation of the procedure described in Example 2.3, the desired compound was obtained

1H NMR (300 MHz, CDCl3) δ 3.36 (t, J = 4.9 Hz, 4H), 3.88 (t, J = 4.8 Hz, 4H), 7.69 (d, J = 2 , 6 Hz, 1H), 8.06 (d, J = 2.6 Hz, 1 HOUR). MS (ESI +) m / z 358 (M + 23)

Stage 4:

The product of Step 3 (607 mg, 1.8 mmol) was dissolved in anhydrous ethanol (50 ml) under N2 atmosphere. They were added

successively aqueous tin (iv) chloride (2.75 g, 14.5 mmol) and 2-3 drops of water. The mixture was refluxed overnight, after which it was concentrated in vacuo. The residue was mixed with water and sodium hydroxide solution was added.

11. The resulting mixture was extracted with dichloromethane (3 x) and organic layers
׽ aqueous (0.2 M) to adjust the pH

The combined were washed with brine, dried and concentrated in vacuo. The residue was subjected to column chromatography (hexane / ethyl acetate 1: 2) to give the desired product (489 mg, 89%).

1H NMR (300 MHz, CDCl3) δ 2.94-3.03 (m, 4H), 3.77-3.87 (m, 4H), 4.50-4.76 (brs, 2H), 7, 56 (d, J = 2.6 Hz, 1H), 7.78 (d J = 2.6 Hz, 1H)

MS (ESI +) m / z 306 (M + 1).

Stage 5:

The procedure described in Example 2 was adapted to the product of Step 4 to give the desired compound.

1H NMR (300 MHz, DMSO-d6) δ 3.14-3.22 (m, 4H), 3.70-3.81 (m, 4H), 3.89 (s, 3H), 8.00 ( d, J = 2.6 Hz, 1H), 8.47 (d, J = 2.6 Hz, 1H), 10.31 (s, 1H)

Stage 6:

The procedure described in Example 6 was adapted to the product of Step 5 to give the desired compound.

1H NMR (300 MHz, DMSO-d6) δ 3.14-3.21 (m, 4H), 3.72-3.81 (m, 4H), 4.62 (d, J = 6.6 Hz, 2H), 7.39 (dd, J = 8.3, 1.8 Hz 1H), 7.61-7.67 (m, 2H), 8.01 (d, J = 2.6 Hz, 1H) , 8.50 (d, J = 2.6 Hz, 1H), 8.95 (t, J = 6.5 Hz, 1H), 11.82 (s, 1H).

MS (ESI-) m / z 573 (M [Cl35] -1) HPLC procedure 7 92.7% / 12.4 min

Example 22.2: Preparation of [2- (4-fluoro-benzylcarbamoyl) -3-hydroxy-7-iodo-4-oxo-4H-pyrido [1,2-a] pyrimidin-9-yl] -carbamic acid ethyl ester

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image33

2,3-Diamino-5-iodopyridine (2.35 g, 10 mmol) in pyridine (15 ml) was cooled in an ice bath. To the above stirred solution was added ethyl chloroformate (1.08 g, 10 mmol) dropwise. The mixture was stirred at 0 ° C for 15 min and then at room temperature for 3 h, after which it was diluted with water (30 ml) and ethyl acetate (30 ml). The organic phase was washed with water, dried and concentrated in vacuo. The residue was subjected to column chromatography (dichloromethane) to give the desired compound (2.52 g, 82%).

1H NMR (300 MHz, DMSO-d6) δ 1.24 (t, J = 7.1 Hz, 3H), 4.12 (q, J = 7.1 Hz, 2H), 6.05 (s, 2H ), 7.87 (d, J = 2.1 Hz, 1H) 7.92 (s, 1H), 8.78 (s, 1H).

MS (ESI +) m / z 308 (M + 1)

Stage 2:

The procedure described in Example 2 was adapted to give the desired product.

 1H NMR (300 MHz, DMSO-d6) δ 1.28 (t, J = 7.1 Hz, 3H), 3.90 (s, 3H), 4.23 (q, J = 7.0 Hz, 2H ), 8.19 (d, J = 1.7 Hz, 1H) 8.56 (d, J = 1.8Hz, 1H), 8.70 (s, 1H) 10.66 (s, 1H) MS ( ESI +) m / z 456 (M + 23).

Stage 3:

The procedure described in Example 6 was adapted to give the desired product.

1H NMR (300 MHz, DMSO-d6) δ 1.30 (t, J = 7.1 Hz, 3H), 4.26 (q, J = 7.1 Hz, 2H), 4.62 (d, J = 6.1 Hz, 2H), 7.20 (t, J = 9.0 Hz, 2H), 7.38 (dd, J = 8.8, 5.4 Hz, 2H), 8.39 (d , J = 1.7 Hz, 1H), 8.56 (d, J = 1.8 Hz, 1H), 9.99 (s, 1H), 10.47 (t, J = 6.3 Hz 1H) , 12.66 (s, 1 H)

MS (ESI-) m / z 525 (M-1) HPLC procedure 7 90.4%

Example 23: Preparation of 7-benzyl-3-hydroxy-4-oxo-4H-pyrido [1,2a] pyrimidine-2-carboxylic acid substituted acids and amides

Example 23.1: Preparation of 3- (tert-Butyl-dimethyl-silanyloxy) -7-iodo-4-oxo-4H-pyrido [1,2a] pyrimidine-2-carboxylic acid methyl ester

[0303] Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Stage 1:

Starting from the 2-amino-5-iodopyridine and adapting the procedure described in Example 5, the desired ester was obtained.

1H NMR (300 MHz, DMSO-d6) δ 3.85 (s, 3H), 7.37 (d, J = 9.1 Hz, 1H, H9), 7.79 (dd, J = 9.3, 2.1 Hz, 1H, H8), 8.86 (d, J = 2.1 Hz, 1H, H6), 8.50 (d, J = 2.6 Hz, 1H), 10.46 (s, 1H, OH). MS (ESI +) m / z 347 (M + 1)

Stage 2:

By adapting the procedure described in Example 13.7 (Step 1) the desired silyl compound was obtained

 1H NMR (300 MHz, D6-DMSO): δ 0.25 (s, 6H), 0.93 (s, 9H), 3.85 (s, 3H), 7.44 (dd, J = 9.2 , 0.8 Hz, 1H), 7.94 (dd, J = 9.3, 1.9 Hz, 1H), 8.97 (dd, J = 1.9, 0.8 Hz, 1H)

Example 23.2: Preparation of 3- (tert-butyl-dimethyl-silanyloxy) -7- (3-chloro-2-fluoro-benzyl) -4oxo-4H-pyrido [1,2-a] pyrimidine-2 acid methyl ester -carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The desired compound was prepared by adaptation of the procedure described in WO2004046115, except that tri-o-tolyl phosphane was used instead of tri-furan-2-yl-phosphane.

1H NMR (300 MHz, CDCl3): δ 0.33 (s, 6H), 1.00 (s, 9H), 3.97 (s, 3H), 4.03 (s, 2H), 7.01- 7.14 (m, 2H), 7.29-7.43 (m, 2H) 7.61 (d, J = 9.1 Hz, 1H), 8.75 (d, J = 1.3 Hz, 1 HOUR).

Example 23.3: Preparation of 7- (3-Chloro-2-fluoro-benzyl) -3-hydroxy-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Aqueous sodium hydroxide (0.5 M, 1.1 ml) was added to a stirred solution of the product from Example 23.2 (22 mg 0.046 mmol) in methanol (5 ml). The mixture was stirred at 50 ° C for 24 h. Then acid was added dropwise

4. The methanol was evaporated under reduced pressure and the resulting solid
׽ aqueous hydrochloric (1.0 M) to adjust the pH to 3

collected by filtration and dried in vacuo to give the desired compound as a brown solid (13 mg, 81%).

1H NMR (300 MHz, D6-DMSO): δ 8.73 (1H, s, Ar-CH), 7.78 (1H, d, J = 9.3 Hz, Ar-CH), 7.72 (1H , d, J = 9.3 Hz, Ar-CH), 7.49 (1H, dd, J = 7.5, 6.6 Hz, Ar-CH), 7.35 (1H, dd, J = 7 , 5, 5.7 Hz, Ar-CH), 7.20 (1H, dd, J = 7.8, 7.2 Hz, Ar-CH), 4.18 (2H, s, CH2Ar).

MS (ESI-) m / z 347 (M [CL35] -1)

HPLC procedure 7 96.1% / 13.2 min

Example 23.4: Preparation of 7- (3-Chloro-2-fluoro-benzyl) -3-hydroxy-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid 4-fluoro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Using the product of Example 23.3, 4-fluorobenzylamine and by adapting the procedure described in Example 6, the desired compound was obtained.

 1H NMR (300 MHz, DMSO) δ 4.13 (2H, s, Cl, F-Ph-CH2-), 4.48 (2H, d, J = 6.3 Hz, - (O = C) NHCH2- ), 7.17 (4H, m, ArH) 7.39 (3H, m, ArH), 7.50 (3H, m, ArH), 9.64 (1H, s, ArH), 9.68 (1H , t, J = 6.0 Hz, (O = C) NHCH2-), 12.21 (1H, s, OH)

MS (ESI-) m / z 454 (M [Cl35] -1)

HPLC procedure 7 94.0% / 18.1 min

Example 23.5: Preparation of 7- (3-Chloro-2-fluoro-benzyl) -3-hydroxy-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid 3,4-dichloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Using the product of Example 23.3, 3,4-dichlorobenzylamine and by adapting the procedure described in Example 6, the desired compound was obtained.

1H NMR (300 MHz, D6-DMSO) δ 4.13 (2H, s, Cl, F-Ph-CH2-), 4.49 (2H, d, J = 6.0 Hz, - (O = C) NHCH2-), 7.17 (4H, t, J = 6.9 Hz, ArH), 7.32 (2H, m, ArH), 7.59 (7H, m, ArH), 8.65 (1H, s, ArH), 9.74 (1H, t, J = 6.6 Hz, (O = C) NHCH2-), 12.081H, s, OH).

MS (ESI +) m / z 506 (M [Cl35] +1)

HPLC procedure 7 99.0% / 18.0 min

Example 24.1: Preparation of 2- [3- (4-fluoro-benzyl) - [1,2,4] oxadiazol-5-yl] -3-hydroxy-7-morpholin-4-yl-pyrido [1,2a ] pyrimidin-4-one

image 1

5

10

fifteen

twenty

25

30

35

Stage 1:

The procedure described in J. Med. Chem. 1999, 42 (20), 4088-4098 was followed.

Stage 2:

Using the product of Example 2.3 and adapting the procedures of Examples 8.1 and 8.2, 3-benzyloxy-7-morpholin-4-yl-4-oxo-4H-pyrido [1,2-a] pyrimidine-2 acid was prepared -carboxylic. This compound (159 mg) was combined with the product of Step 1 (300 mg, 0.79 mmol), triphenylphosphane (619 mg) and triethylamine (0.3 ml) in acetonitrile (30 ml) under nitrogen atmosphere and stirred at room temperature for 10 min. Carbon tetrachloride (0.4 ml) was added dropwise and the mixture was stirred for 11 h. The solvent was removed in vacuo and the residue was dissolved in dichloromethane, washed with brine, dried and evaporated. The residue was subjected to column chromatography (hexane / ethyl acetate 1: 4) and the desired compound was obtained as a yellow solid (280 mg 67%).

1H NMR (300 MHz, D6-DMSO): δ 3.18-3.29 (m, 4H), 3.40 (s, 2H), 3.74-3.86 (m, 4H), 5.17 (s, 2H), 6.29-6.72 (m, 2H) 7.11 (t, J = 8.8 Hz, 2H), 7.24-7.46 (m, 7H), 7.71 (d, J = 9.7 Hz, 1H), 8.05 (dd, J = 9.8, 2.4 Hz 1H), 8.22 (d, J = 2.3 Hz, 1H)

Stage 3:

A solution of the product of Step 2 260 mg 0.49 mmol in toluene (30 ml) was heated to reflux for 12 h. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. Purification by column chromatography (hexane / ethyl acetate 1: 4) gave the desired compound as a yellow solid (183 mg, 73%)

1H NMR (300 MHz, D6-DMSO): δ 3.27 (t, J = 4.7 Hz, 4H), 3.80 (t, J = 4.7 Hz, 4H), 4.22 (s, 2H), 5.21 (s, 2H), 7.17 (t, J = 8.9 Hz, 2H), 7.25-7.44 (m, 7H), 7.76 (d, J = 9 , 6 Hz, 1H), 8.07 (dd, J = 9.8, 2.6 Hz, 1H), 8.20 (d, J = 2.4 Hz, 1H)

Stage 4:

The procedure described in Example 19.8 was adapted to give the desired compound.

1H NMR (300 MHz, D6-DMSO): δ 10.67 (1H, s, OH), 7.97 (1H, s, Ar-CH), 7.84 (1H, d, J = 7.8 Hz , Ar-CH), 7.62 (1H, d, J = 9.0 Hz, Ar-CH), 7.42 (1H, d, J = 8.4 Hz, Ar-CH), 7.39 ( 1H, d, J = 8.7 Hz, Ar-CH), 7.19 (1H, d, J = 8.4 Hz, Ar-CH), 7.16 (1H, d, J = 9.0 Hz , Ar-CH), 4.23 (2H, s, ArCH2), 3.78 (4H, m, CH2OCH2), 3.20 (4H, m, CH2NCH2).

MS (ESI +) m / z 424 (M + 1)

HPLC procedure 7 91.7% / 12.0 min

Example 24.2: Preparation of 2- [3- (3,4-dichloro-benzyl) - [1,2,4] oxadiazol-5-yl] -3-hydroxy-7-morpholin-4-yl-pyrido [1 , 2a] pyrimidin-4-one

image 1

Starting from 3,4-dichlorobenzylnitrile and adapting the procedure described in Example 24.1, the desired compound was obtained.

1H NMR (300 MHz, D6-DMSO): δ 10.71 (1H, s, OH), 7.98 (1H, s, Ar-CH), 7.85 (1H, d, J = 9.6 Hz , Ar-CH), 7.66 (3H, m, Ar-CH), 7.36 (1H, d, J = 8.1 Hz, Ar-CH), 4.24 (2H, s, ArCH2), 3.78 (4H, m, CH2OCH2), 3.22 (4H, m, CH2NCH2).

MS (ESI +) m / z 474 (M [CI35 + 1)

Example 25.1: Preparation of 2- [4- (4-fluoro-benzyl) -4,5-dihydro-oxazol-2-yl] -3-hydroxy-7-morpholin-4-yl-pyrido [1,2a] pyrimidin-4-one; sodium salt

image 1

Stage 1:

A stirred mixture of the product of Example 2.3 (305 mg, 1 mmol) and 2-amino-3- (4-fluoro-phenyl) -propan1-ol (169 mg, 1 mmol) in ethanol (15 ml) was heated at reflux for 2 d. The solvent was evaporated in vacuo to give a crude product that was used directly in the next Step.

Stage 2:

Methanesulfonyl chloride (228 mg, 2.0 mmol) and triethylamine (0.5 ml, 3.59 mmol) were added to a stirred mixture of the product from Step 2 in dichloromethane (50 ml). After 2 h, the reaction mixture was diluted with ethyl acetate (50 ml) and the organic phase was washed with saturated aqueous sodium bicarbonate (50 ml), water (50 ml) and brine (50 ml), dried, It was filtered and concentrated in vacuo. The residue was purified by flash chromatography (hexane / ethyl acetate 1: 5) to give the desired compound (250 mg, 50% two steps).

1H NMR (300 MHz, DMSO-d6) δ 2.82 (dd, J = 13.8, 6.7 Hz, 1H), 2.97 (dd, J = 14.0 Hz, 6.8Hz, 1H) , 3.25-3.31 (m, 4H), 3.50 (s, 3H), 3.76-3.85 (m, 4H), 4.01-4.13 (m, 1H), 4 , 40-4.48 (m, 1H), 4.50-4.63 (m, 1H), 7.12 (t, J = 9.0 Hz, 2H), 7.37 (dd, J = 8 , 6, 5.7 Hz, 2H), 7.82 (d, J = 9.5 Hz, 1H), 8.18 (dd, J = 9.7, 2.6 Hz, 1H), 8.23 (d, J = 2.4 Hz, 1H)

MS (ESI +) m / z 503 (M + 1)

Stage 3:

The product of Step 3 (228 mg) and solid sodium hydroxide (40 mg, 1 mmol) were mixed in methanol (25 ml). The resulting mixture was stirred at room temperature for 1 h and water / ice (100 ml) was added. The resulting precipitate was collected by filtration and washed with cold water to give the desired compound as a sodium salt (170 mg, 76%).

1H NMR (300 MHz, D6-DMSO) δ 3.07 (2H, m, -CH2-Ph-F), 3.24 (4H, m, -OCH2CH2N-), 3.85 (4H, m, -OCH2CH2N -) 4.54 (1H, m, cyclic-NCHCH2O-), 7.00 (2H, t, J = 9.0 Hz, ArH), 7.30 (2H, m, ArH), 7.64 (1H , d, J = 10.0 Hz, ArH), 7.80 (1H dd, J = 2.7, 9.9 Hz, ArH), 8.13 (1H, d, J = 2.4 Hz, ArH ).

 MS (ESI-) m / z 423 (M-Na-1) HPLC procedure 7 87.0% / 17.7 min

Example 25.2: Preparation of 2- [4- (4-fluoro-benzyl) -4,5-dihydro-oxazol-2-yl] -3-hydroxy-7-morpholin-4-ylmethyl-pyrido [1,2a] pyrimidin -4-one; sodium salt

image34

Using the product of Example 13.7 (Step 4) and adapting the procedure described in Example 25.1, the desired compound was obtained.

MS (ESI +) m / z 461 (M-Na + 1)

HPLC procedure 7 85.4% / 11.4 min

 1H NMR (300 MHz, D6-DMSO): δ 8.55 (1H, s, Ar-CH), 7.42 (3H, m, Ar-CH), 6.89 (2H, m, Ar-CH) , 6.75 (1H, m, Ar-CH) 4.27 (2H, m, OCH2CH [N]), 3.72 (4H, m, CH2OCH2), 3.59 (2H, s, Ar-CH2) , 3.25 (1H, m, OCH2CH [N]), 2.51 (4H, m, CH2NCH2)

Example 26.1: Preparation of 7- (1,1-dioxo-isothiazolidin-2-yl) -3-hydroxy-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid 4-fluoro-benzylamide

Example 26.1.1: Preparation of 3-hydroxy-7-iodo-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid 4-fluorobenzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The procedure described in Example 6 was adapted to the product of Example 23.1 (Step 1) to give the desired compound.

1H NMR (300 MHz, D6-DMSO): δ 4.59 (2H, d, J = 6.9 Hz, NHCH2), 7.15 (2H, m, ArH), 7.29 (1H, d, J = 9.4 Hz, H9), 7.38 (2H, dd, J = 8.3, 5.9 Hz, ArH), 7.81 (1H, dd, J = 9.4, 1.7 Hz, H8), 9.71 (1H, t, J = 6, 9 Hz, NHCH2), 12.33 (1H, s, OH), MS (ESI +) m / z 440 (M + 1).

HPLC procedure 7 97.5% / 15.5 min

Example 26.1.2: Preparation of 3-benzyloxy-7-iodo-4-oxo-4H-pyrido [1,2a] pyrimidine-2-carboxylic acid 4-fluoro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The procedure described in Example 18.1 was adapted to give the desired product.

1H NMR (300 MHz, D6-DMSO) 4.41 (2H, d, J = 6.2 Hz, NHCH2), 5.12 (2H, s, ArCH2O), 7.04 (2H, t, J = 9 , 1 Hz, ArH) 7.32-7.39 (7H, m, ArH), 7.50 (1H, dd, J = 0.6, 9.3 Hz, H9), 8.06 (1H, dd, J = 2.1, 9.3 Hz, H8 ), 9.01-9.13 (2H, m, H6 and NHCH2) MS (ESI +) m / z 530 (M + 1)

Example 26.1.3: Preparation of 3-benzyloxy-7- (1,1-dioxo-isothiazolidin-2-yl) -4-oxo-4H-pyrido [1,2-a] pyrimidine-2 acid 4-fluoro-benzylamide -carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The product of Example 26.1.2 (100 mg, 0.189 mmol), isothiazolidine 1,1-dioxide (46 mg, 0.378 mmol), copper (I) iodide (4 mg, 0.019 mmol), N, N-dimethylethyl diamine ( 3 mg, 0.039 mmol) and potassium carbonate (55 mg, 0.378 mmol) were mixed in DMF (4.0 ml) and heated to 80 ° C. After 2 h, TLC indicated that the reaction was complete. The reaction mixture was cooled to room temperature and poured onto aqueous hydrochloric acid (1.0 M, 40 ml. The resulting solid was collected by filtration and washed with water, dried and subjected to column chromatography (dichloromethane / 50: 1 methanol) to give the desired product (93 mg, 95%).

1H NMR (300 MHz, D6-DMSO) δ 3.64 (2H, t, J = 7.3 Hz, cyclic- (SO2) -CH2CH2CH2N), 3.89 (2H, t, J = 6.5 Hz, cyclic- (SO2) CH2CH2CH2N-), 4.43 (2H, d, J = 5.9 Hz, NHCH2), 5.14 (2H, s, CH2O) 7.06 (2H, t, J = 9.0 Hz, ArH), 7.32-7.48 (7H, m, ArH), 7.84 (1H, d, J = 9.9 Hz, ArH), 8.00 (1H, dd, J = 2, 8, 9.7 Hz, ArH), 8.61 (1H, d, J = 2.6 Hz, ArH), 9.07 (1H, t J = 6.2 Hz, NHCH2).

Example 26.1.4: Preparation of 7- (1,1-Dioxo-isothiazolidin-2-yl) -3-hydroxy-4-oxo-4H-pyrido [1,2-a] pyrimidine2-carboxylic acid 4-fluoro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The procedure described in Example 19.8 was adapted to give the desired compound

1H NMR (300 MHz, D6-DMSO) δ 3.62 (2H, t, J = 7.2 Hz, cyclic- (SO2) -CH2CH2CH2N-), 3.84 (2H, t, J = 6.6 Hz , cyclic- (SO2) CH2CH2CH2N-), 4.50 (2H, d, J = 6.3 Hz, - (O = C) NHCH2-), 7.16 (2H, t, J = 8.7 Hz, ArH), 7.41 (2H, m, ArH), 7.64 (1H, d, J = 9.6 Hz, ArH), 7.83 (1H, dd, J = 2.7, 9.9 Hz , ArH), 8.35 (1H, d, J = 1.8 Hz, ArH), 9.74 (1H, bt, - (O = C) NHCH2-) 12.28 (1H, s, OH).

MS (API +) m / z 455 (M + Na)

Example 26.2: Preparation of 7- (1,1-dioxo-isothiazolidin-2-yl) -3-hydroxy-4-oxo-4H-pyrido [1,2-a] pyrimidine2-carboxylic acid 3,4-dichloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The procedure described in Example 26.1 was adapted to give the desired compound.

1H NMR (300 MHz, D6-DMSO) δ 3.62 (2H, t, J = 7.8 Hz, cyclic- (SO2) -CH2CH2CH2N-), 3.84 (2H, t, J = 6.3 Hz , cyclic- (SO2) CH2CH2CH2N-), 4.52 (2H, d, J = 6.3 Hz, - (O = C) NHCH2-), 7.36 (1H, dd, J = 2.1, 8 , 1 Hz, ArH), 7.62 (3H, m, ArH) 7.84 (1H, dd, J = 2.1, 9.9 Hz, ArH), 8.36 (1H, d, J = 2 , 4 Hz, ArH), 9.78 (1H, bt, - (O = C) NHCH2-), 12.14 (1H, s, OH).

 MS (ESI-) m / z 481 (M [Cl35] -1)

Example 26.3: Preparation of 7- (1,1-dioxo- [1,2] thiazinan-2-yl) -3-hydroxy-4-oxo-4H-pyrido [1,2-a] acid 4-fluoro-benzylamide pyrimidine2-carboxylic

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The procedure described in Example 26.1 was adapted to give the desired compound.

1H NMR (300 MHz, D6-DMSO): δ 12.32 (1H, s, OH), 9.72 (1H, t, J = 6.3 Hz, NH), 8.60 (1H, s, Ar -CH), 7.71 (1H, d, J = 9.6 Hz, Ar-CH), 7.56 (1H, d, J = 9.6 Hz, Ar-CH), 7.43 (2H, m, Ar-CH), 7.18 (1H, d, J = 8.7 Hz, Ar-CH), 7.15 (1H, d, J = 8.7 Hz, Ar-CH), 4.51 (2H, d, J = 6.3 Hz, NHCH2), 3.77 (2H, m, CH2N), 3.42 (2H, m, CH2S), 2.18 (2H, m, CH2CH2CH2S), 1, 86 (2H, m, CH2CH2CH2S).

MS (ESI +) m / z 447 (M + 1)

HPLC procedure 7 96.1% / 12.0 min

Example 26.4: Preparation of 7- (1,1-Dioxo- [1,2] thiazinan-2-yl) -3-hydroxy-4-oxo-4H-acid [1,2-] 3,4-dichloro-benzylamide a] pyrimidine2-carboxylic acid

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The procedure described in Example 26.1 was followed to give the desired compound.

1H NMR (300 MHz, D6-DMSO) δ 1.84 (2H, bm, cyclic- (SO2) -CH2CH2CH2CH2N-), 2.18 (2H, bm, cyclic- (SO2) CH2CH2CH2CH2N-), 3.40 ( 2H, bm, cyclic- (SO2) -CH2CH2CH2CH2N-), 3.74 (2H, bm, cyclic- (SO2) -CH2CH2CH2CH2N-), 4.50 (2H, d, J = 6.6 Hz, - (O = C) NHCH2-), 7.50 (1H, dd, J = 1.8, 8.2 Hz, ArH), 7.58 (3H, m ArH), 7.69 (1H, dd, J = 2 , 4, 9.9 Hz, ArH), 8.58 (1H, d, J = 1.8 Hz, ArH), 9.76 (1H, t, J = 6.9 Hz, - (O = C) NHCH2-), 12.16 (1H, s, OH).

MS (API +) m / z 497 (M [Cl35] +1)

HPLC procedure 7 92.0% / 13.2 min

Example 26.5: Preparation of 3-hydroxy-4-oxo-7- (2,2,2-trifluoro-acetylamino) -4H-pyrido [1,2-a] pyrimidine2-carboxylic acid 4-fluoro-benzylamide

Example 26.5.1: Preparation of 3-hydroxy-4-oxo-7- (2,2,2-trifluoro-acetylamino) -4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid 4-fluoro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The product of Example 6 was reacted under the conditions described in Example 8.1 to give the desired product.

 1H NMR (300 MHz, D6-DMSO) 4.43 (2H, d, J = 6.0 Hz, NHCH2), 5.15 (2H, s, CH2O), 7.06 (2H, t, J = 8 , 8 Hz, ArH), 7.28-7.51 (7H, m, ArH), 7.69 (1H, d, J = 9.5 Hz, H9), 8.02 (1H, dd, J = 1.7, 9.6 Hz, H8), 9.02 (1H, d, J = 1.5 Hz, H6), 9.09 (1H, t, J = 5.9 Hz, NHCH2).

MS (ESI +) m / z 482 (M [Br79] +1), 484 (M [Br81] +1)

Example 26.5.2: Preparation of 3-hydroxy-4-oxo-7- (2,2,2-trifluoro-acetylamino) -4H-pyrido [1,2-a] pyrimidine2-carboxylic acid 4-fluoro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The product of Example 26.5.1 was reacted under the conditions described in Example 26.1.3 using trifluoroacetamide to give the desired compound (22 mg, 31%)

1H NMR (300 MHz, D6-DMSO) δ 4.49 (2H, d, J = 6.0 Hz, -NH-CH2-), 7.15 (2H, m, ArH), 7.42 (2H, m, ArH), 7.57 (1H, d J = 9.6 Hz, ArH), 7.83 (1H, d, J = 9.6 Hz, ArH), 9.36 (1H, dd, J = 1.8 Hz, ArH), 9.72 (1H, bt, O = C-NH-CH2). 11.96 (1H, s, OH)

MS (ESI +) m / z 423 (M-1)

HPLC procedure 7 96.7% / 12.4 min

Example 26.6: Preparation of 3-hydroxy-4-oxo-7- (2,2,2-trifluoro-acetylamino) -4H-pyrido [1,2-a] pyrimidine2-carboxylic acid 3,4-dichloro-benzylamide

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The procedure described in Example 26.5 was adapted to give the desired compound.

1H NMR ((300 MHz, D6-DMSO) δ 4.49 (2H, bd, J = 6.6 Hz, - (C = O) NHCH2-), 7.34 (1H, m, ArH), 7, 60 (3H, m, ArH), 7.89 (1H, dd, J = 2.4, 9.9 Hz, ArH), 9.37 (1H, d, J = 2.1 Hz, ArH), 9.75 (1H, bt, - (O = C) NHCH2- ), 12.14 (1H, s, OH). MS (ESI +) m / z 423 (M-1) MS (ESI-) m / z 473 (M [Cl35] -1)

HPLC procedure 7 82.0% / 13.7 min

Example 27.1: Preparation of [2- (4-fluoro-benzylcarbamoyl) -3-hydroxy-4-oxo-4H-pyrido [1,2-a] pyrimidin-7-yl-methyl] phosphonic acid

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

Stage 1: (using the product of Example 13.7 (Stage 2))

To a stirred solution of the product from Example 13.7 (Step 2) (347 mg, 0.81 mmol) in toluene (10 ml) was added triethyl phosphite (268 mg, 1.62 mmol). The mixture was heated at reflux for 4 h, after which it was concentrated to dryness in vacuo. The residue was subjected to column chromatography (dichloromethane / methanol 30: 1) to give the desired compound (373 mg, 95%).

1H NMR (300 MHz, CDCl3) δ 0.33 (s, 6H), 0.99 (s, 9H), 1.30 (t, J = 7.0 Hz, 6H), 3.15 (d, J = 21.4 Hz, 2H), 3.98 (s, 3H), 4,044.16 (m, 4H), 7.55-7.68 (m, 2H), 8.75 (d, J = 3, 0 Hz, 1H) MS (ESI +) m / z 507 (M + 23)

Stage 2:

A mixture of the product from Step 1 (115 mg, 0.24 mmol) and p-toluenesulfonic acid (5 mg, 0.024 mmol) in methanol (5 ml) was stirred at room temperature overnight. The solution was concentrated in vacuo to quantitatively give the crude product that was used directly in the next step.

Stage 3:

The crude product from Step 2 was dissolved in acetonitrile (4 ml) and the stirred solution was cooled (ice / water bath). Trimethylsilyl iodide (191 mg, 0.97 mmol) was added dropwise and after 2 h, the solution was heated to room temperature and stirred overnight. The reaction mixture was rapidly stopped with methanol and then concentrated in vacuo. To the resulting residue was added acetonitrile (4 ml) and the mixture was sonicated for 5 min. The resulting solid was collected by filtration, washed with acetonitrile and dried in vacuo to give the desired product (62 mg, 87%).

1H NMR (300 MHz, CD3OD) δ 3.36 (d, overlap, 2H), 4.08 (s, 3H), 7.94 (d, J = 9.4 Hz, 1H), 8.13 (d , J = 9.2 Hz, 1H) 8.95 (s, 1H) MS (ESI-) m / z 313 (M-1)

Stage 4:

The procedure described in Example 6 was adapted to give the objective compound.

 1H NMR (300 MHz, D6-DMSO) δ 3.07 (2H, d, J = 20.7 Hz, -PCH2Ph-), 4.49 (2H, d, J = 5.7 Hz, - (O = C) NHCH2-), 7.15 (2H, m, ArH), 7.50 (4H, m, H7, H8 and 2 x ArH), 8.58 (1H, s, H6), 9.74 (1H , bs, - (O = C) NHCH2-), 12.15 (1H, bs, OH) MS (ESI-) m / z 406 (M-1)

Example 27.2: Preparation of [2- (3,4-dichloro-benzylcarbamoyl) -3-hydroxy-4-oxo-4H-pyrido [1,2-a] pyrimidin-7ylmethyl] -phosphonic acid

Although the following example is not included in the scope of the claims, it is introduced as useful background information for the reader.

image 1

The procedure described in Example 27.1 was adapted to give the desired compound.

1H NMR (300 MHz, D6-DMSO) δ 2.97 (2H, d, J = 21.3 Hz, PCH2), 4.49 (1H, d, J = 5.4 Hz, CH2NH), 7.49 (5H, m, H8, H9 and 2 x ArH), 8.58 (1H, bs, H6), 9.75 (1H, bs, CH2NH), 11.8 (1H, bs, OH) MS (ESI- ) m / z 456 (M [Cl35] -1)

Example 28 Biological tests

The biological activity of the compounds of the present invention can be tested using the following techniques:

Example 28.1 Combined test of 3 ’processing / chain transfer:

A combined 3 'processing / chain transfer test procedure similar to the one published could be used (Ovenden et al. Phytochemistry. 2004 Dec; 65 (24): 3255-9.) The test can be adapted to a plate format of 96 wells. In summary, 400ng of the compound to be tested are incubated with 30nM substrate DNA, which consists of a hybridized LTR U5 oligonucleotide sequence labeled with digoxigenin (DIG; 5'ACTGCTAGAGATTTTCCACACTGACTAAAAGGGTC-DIG-3 ') or biotin (5'-BioGACCCTTTTAGTAGTCCTATTTAGTAGTCCTTTAGTAGTCCGTTGTAGTCCTA ) so that each substrate has a DIG or Bio label on the opposite chains. The reactions are carried out for 2 h at 37 ° C, the products generated as a result of the 3 'processing and the chain transfer activity were bound to streptavidin plates and detected by DIG antibody conjugated to alkaline phosphatase and phosphate substrate of p-nitrophenyl.

Example 28.2 Specific chain transfer test:

The specific chain transfer test has a format similar to that of the combined 3 ’processing / chain transfer test except that it uses a biotinylated substrate that represents a preprocessed LTR end (5’Bio-GACCCTTTTAGTCAGTGTGGAAAATCTCTAGCA-3’).

The 5’-biotin-GACCCTTTTAGTCAGTGTGGAAAATCTCTAGCA-3 ’and 5’ACTGCTAGAGATTTTCCACACTGACTAAAAGGGTC-Dig-3’ oligonucleotides were hybridized in 10 mM Tris-Cl pH 8.0, 100 mM NaCl, a 30 mM EDTA concentration of 2 mM.

Each reaction (40ul) contains 30nm DNA substrate and 400ng of integrase in a reaction buffer containing 20mM Tris-Cl pH 7.5, 25mM NaCl, 5mM MnCl2, 5mM MgCl2, 5mM B-ME , 50 µg / ml BSA, 0.05% Tween-20 (v / v).

The compounds were added to DMSO at 1/10 of the final reaction volume.

The reactions were incubated at 37 ° C for 2 h followed by the addition of 60 µl of adjustment buffer containing 33 mM Tris-Cl pH 7.5, 664 mM NaCl, 16.6 mM EDTA, 0.166 mg / ml DNA of sonic salmon sperm.

Then, the samples were transferred to streptavidin coated plates and the products of the enzymatic reactions were allowed to bind for 1 h at room temperature.

The plates were then washed 3 x 5 min with 30 mM NaOH, 200 mM NaCl, 1 mM EDTA and then 3 x 5 min with wash 2: 10 mM Tris-Cl pH 8.0, 6 mM EDTA, BSA free 0.1 mg / ml nuclease.

Fab anti-digoxigenin phosphatase (Roche, 0.1 U / ml)) was added, diluted 1/2000 in wash buffer 2 and then added to each well and the plates were incubated for 1 h at 37˚C.

The plates were then washed 3 times with TBS-Tween-20 (0.1%) and then twice with TBS and 100ul of substrate (1 mg / ml of 0.1M Tris p-nitrophenyl phosphate, pH 9 was added , 8) and the plates were incubated until sufficient color appeared.

Example 28.3 Inhibition of HIV replication

The cells were seeded in 96-well microtiter plates at 50,000 cells per 50 µl per well in RF-10 containing 2 µg / ml of polybrene (RF-10/2). The compounds were prepared at a final 4 x concentration in RF-10/2, and 30 µl were added to the wells. Viruses (40 µl in RF-10/2 containing 1600 pfu) were added to each well or 40 µl of RF-10/2 for the negative controls and to test the cytotoxicity of the compound. After 24 h, an additional 90 µl of medium or medium containing 1 x compound was added to each well. 4 days after infection, 100 µl of medium was removed from each well and replaced with 100 µl of fresh medium with or without compound. Forty-eight hours later, supernatants were harvested and extracellular p24 levels were determined. The supernatants were diluted 1 in

10,000 and p24 levels were tested using the Vironostika p24 test kit. EC50 was calculated as the concentration necessary to inhibit the production of HIV p24 up to 50% of the non-drug controls.

image 1

X

Y R1 R2 IC50  EC50 CC50 (µM)





7-Br CONHCH2 (4F-Ph) ++  +++ + *





7-Me CONHCH2 (4F-Ph) ++ NT *





7-Me CONHCH2 (3,4-ClPh) +++  +++ + *





7-Me CONHCH2 (3,4-ClPh) NT +++  + *





7-Me CONHCH2 (4F-Ph) NT +++  + *





7-Me  + NT NT





7-Me image 1 +++ NT NT





7-Me + NT NT





7-Me +++ NT NT





7-Me +++ NT NT

N / A not applicable NT not tested. +++ indicates value between 0.001 mM and 1 mM ++ indicates value between 1 mM and 10mM + indicates value greater than 1mM Examples marked with "*" are not part of the invention

Example 28.4 Comparison of activity against natural and mutant HIV integrase (Q148K)

Example 28.4.1 Chain transfer test:

5 A combined 3 'processing / chain transfer test procedure similar to the one published could be used (Ovenden et al. Phytochemistry. 2004 Dec; 65 (24): 3255-9.) The test can be adapted to a plate format 96 wells In summary, 400 ng of the compound to be tested are incubated with 30 nM substrate DNA. The substrate DNA is designed to mimic the ends of HIV DNA that have undergone 3 ’processing and consist of a sequence of hybridized LTR U5 oligonucleotides labeled with digoxigenin (DIG; 5’

10 ACTGCTAGAGATTTTCCACACTGACTAAAAGGGTC-DIG-3 ’) or biotin (5’-BioGACCCTTTTAGTCAGTGTGGAAAATCTCTAGCAGT-3’) so that each substrate has a DIG or Bio label on the opposite chains. The reactions are carried out for 2 h at 37 ° C, the products generated as a result of the 3 'processing and the chain transfer activity were bound to streptavidin plates and detected by DIG antibody conjugated to alkaline phosphatase and phosphate substrate of p-nitrophenyl.

15 Figure 1 represents as an example, the results of the chain transfer test from selected compounds against natural integrase and the integrase containing the Q148K mutation.

Example 28.4.2 Mutant enzymes:

The HIV integrase was mutated into a shuttle vector (pGEM) containing most of the gag and pol sequences of HIV-1 using site-directed mutagenesis to generate the published integrase sequences indicating that they conferred resistance to the published integrase inhibitors. These include but are not limited to mutations such as Q148K. The integrase coding region was subsequently subjected to PCR and cloned into a bacterial expression vector. The

Specific introduction of the desired mutation or mutations was confirmed by sequence analysis. Proteins were expressed, purified and used in chain transfer assays.

Throughout this specification, it will be understood that the word "comprises", or variations such as "comprise" or "comprising", implies the inclusion of an element, integer or stage, or group of elements, integers or 5 stages specified , but not the exclusion of any other element, integer or stage, or group of elements, integers or stages. All publications mentioned in this specification are incorporated herein by reference. Any discussion of documents, acts, materials, Devices, articles or the like included herein are solely for the purpose of providing a context for the present invention.It should not be taken as an admission that any or all of these materials form part of the basis of the

The prior art or that these materials form part of the generalized knowledge in the relevant field of the present invention so that they existed in Australia or elsewhere before the priority date of each of the claims of this application.

Those skilled in the art will appreciate that numerous variations and / or modifications can be made in the invention as demonstrated in the specific embodiments without separating the scope of the invention as it has been

15 described broadly. The present embodiments, however, should be considered in all their aspects as illustrative and not restrictive.

References

Tisler, M. and Zupet, R., Organic Preparations and Procedures International, 22 (4), 1990, 532-534. Vompe A.F. and Turitsyna N.F., J. Gen. Chem. of the USSR, 1957, 27, 3318-3325.

20 Martinez-Barrasa V., Delgado F., Burgos C., Garcia-Navio J.L., Izquierdo M.L. and Alvarez-Builla J., Tetrahedron, 00, 56, 24812490.

Carceller R., Garcia-Navio J.L., Izquierdo M.L., Alvarez-Builla J., Fajardo M., Gomez-Sal P. and Gago F., Tetrahedron, 1994, 50 (17), 4995-5012.

Burgos C., Delgado F., Garcia-Navio J.L., Izquierdo M.L. and Alvarez-Builla J., Tetrahedron, 1995, 51 (31), 8649-8654. of the

25 Rosa R., Martinez-Barrasa V., Burgos C. and Alvarez-Builla J., Tet. Let., 2000, 41, 5837-5840. Behrman E.J., Kiser R.L., Garas W.F., Behrman E.C. and Pitt B.M., J. Chem. Res. (M), 1995, 1051-1063.

Claims (16)

1. A compound of Formula I or a hydrate, salt or prodrug thereof in which: image 1 A is a monocyclic or bicyclic aromatic or heteroaromatic moiety fused with the nitrogen containing ring; X is selected from the group consisting of O and S; And it is selected from the group consisting of O and S; R1 has 0-3 substituents each of which is independently selected from the group consisting of CN, C1-10 alkyl, C2-10 alkenyl, C1-10PO3H2 alkyl, -O-C1-10 alkyl, C1-10NR3R4 alkyl, -O-C1-10NR3R4 alkyl, halo, NR3R4, alkylaryl, alkylheteroaryl, aryl, heteroaryl, -O-alkylaryl, SO2NR3R4; R3 and R4 are each independently selected from the group consisting of hydrogen, CO2 C1-4 alkyl, C (O) C1-4 alkyl C1-10 alkyl, C1-10NR5R6, -NH (CO) (CO) NH C1 alkyl -4, or R3 and R4 taken together with the bound nitrogen form a 5-7 membered heterocyclic ring containing from zero to two additional heteroatoms selected from N, O or S, in which S may be in the oxidation state S , S (O) or S (O) 2 and wherein said heterocyclic ring is optionally substituted on the carbon or nitrogen atoms with one or more substituents selected from halo, aryl, C (O) C1-4 alkyl, SO2 alkyl C1-4, SO2H, C1-4 alkyl, CO2H, CO2 C1-4 alkyl, NR5R8; C1-4NR5R6 alkyl; R5 and R6 are each independently selected from the group consisting of H, C1-4 or R5 alkyl and R6 and together with the attached nitrogen form a 5-7 membered heterocyclic ring containing from zero to two additional heteroatoms selected from N , O or S, in which S may be in the oxidation state S, S (O) or S (O) 2 and in which said heterocyclic ring is optionally substituted at the carbon or nitrogen atoms with one or more substituents selected from halo and C1-4 alkyl; when R1 is alkylaryl or -O-alkylaryl, the aryl group of said alkylaryl substituent is optionally substituted with a substituent selected from C1-10 alkyl, -O-C1-10 alkyl, C1-10NR3R4 alkyl, -O-C1-10NR3R4 alkyl , halo, NR3R4, alkylaryl, -O-alkylaryl, SO2NR3R4 B is absent or is -C (O) -; C is absent or selected from the group consisting of -O-, -NH- and -NH-NH-C (O) -; R2 is selected from the group consisting of heteroaryl and heterocyclyl; and wherein said prodrug comprises (i) a compound in which an amino acid residue, or a polypeptide chain or two or more amino acid residues covalently bind to an amino, hydroxyl or carboxylic acid group of the compound of formula I, the amino acid residues include the 20 natural amino acids usually designated by a three-letter symbol and also includes, 4-hydroxyproline, hydroxylysine, demosin, isodemosin, 3-methylhistidine, norvlina, betaalanine, gamma-aminobutyric acid, citrulline, homocysteine, homoserine, ornithine and methionine sulfone (ii) a compound in which a carbonate, carbamate, amide or alkyl ester covalently binds to a amino, hydroxyl, or carboxylic acid group of the compound of formula I by the secondary chain of the prodrug containing carbonyl carbon, or (iii) a compound comprising a phosphate derivative of a compound of formula I linked by a phosphorus bond oxygen to a free hydroxyl of the compounds of formula I; and wherein in addition each alkyl, cycloalkyl, alkylaryl, aryl, heterocyclyl, or heteroaryl group may optionally be substituted with one or more of C1-C3 alkyl, C3-C6 cycloalkyl, C6 aryl, heterocyclyl, heteroaryl, C1-3OH alkyl, alkylaryl, OH, O C1-3O alkyl, halo, CN, NO2, CO2H, CO2 C1-3 alkyl, CONH2, CONH (C1-C3 alkyl), CON (C1-C3 alkyl) 2, trifluoromethyl, NH2, NH (alkyl C1-C3) or N (C1-C3 alkyl) 2 and each substituent alkyl, cycloalkyl, alkylaryl, aryl heterocyclyl, or optional heteroaryl may also be optionally substituted.

2.

A compound according to claim 1 wherein R2 is heteroaryl.

3.

A compound according to claim 2 wherein R2 is substituted with aryl or alkylaryl.

Four.

A compound according to claim 3 wherein R2 is substituted with alkylaryl.

5.

A compound according to claim 4 wherein said alkylaryl is benzyl.

6.

A compound according to claim 5 wherein said compound has the structure:

7.

A compound according to any one of claims 1 to 5 wherein the compound of formula I is selected from the group consisting of the compounds of formulas II, III, IV, V and VI:

wherein Z is O, S or NR8, where R8 is H, C1-10 alkyl, C1-10NR3R4 alkyl, alkylaryl, alkylheteroaryl, aryl and heteroaryl.

8.

A compound according to any one of claims 1 to 5 wherein NR3R4 is morpholine.

9.

A compound according to any one of claims 1 to 5 wherein the heteroaryl is selected from the group consisting of tetrazole, triazole, pyrazole, imidazole, oxazole, oxadiazole, thiazole, thiadiazole.

10.

 A compound according to claim 1 wherein B is absent.

eleven.

 A compound according to claim 1 wherein C is absent.

12.

 A compound according to claim 1 wherein said compound has the structure:

13.

 A compound according to any one of claims 1 to 12 or a salt or prodrug thereof for use in the treatment or prophylaxis of a viral infection in a subject.

14.

 A compound according to claim 13 for use in the treatment or prophylaxis of an HIV infection or VIS.

fifteen.

 A pharmaceutical composition comprising a compound according to any one of claims 1 to 12 and a pharmaceutically acceptable carrier, diluent or carrier.

16.

 The pharmaceutical composition of claim 15 further comprising another therapeutic active ingredient that is usually used in the treatment of the pathological conditions indicated in claim 13 or claim 14.