EP3423053A1 - Use of 2-oxo-2h-pyrrol-1(5h)-carboxamide derivatives as anti-hiv agents and process for the production thereof - Google Patents

Use of 2-oxo-2h-pyrrol-1(5h)-carboxamide derivatives as anti-hiv agents and process for the production thereof

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Publication number
EP3423053A1
EP3423053A1 EP17718413.2A EP17718413A EP3423053A1 EP 3423053 A1 EP3423053 A1 EP 3423053A1 EP 17718413 A EP17718413 A EP 17718413A EP 3423053 A1 EP3423053 A1 EP 3423053A1
Authority
EP
European Patent Office
Prior art keywords
formula
compound
pharmaceutically acceptable
acceptable salt
linear
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP17718413.2A
Other languages
German (de)
French (fr)
Inventor
Nicola DELLA CA'
Bartolo Gabriele
Beatrice Macchi
Antonio Mastino
Salvatore Vincenzo GIOFRE'
Roberto Romeo
Mirco Costa
Michele QUEIROLO
Raffaella MANCUSO
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Universita' Degli Studi Dl Messina
Universita degli Studi di Parma
Consiglio Nazionale delle Richerche CNR
Universita degli Studi di Roma Tor Vergata
Universita della Calabria
Original Assignee
Universita' Degli Studi Dl Messina
Universita degli Studi di Parma
Consiglio Nazionale delle Richerche CNR
Universita degli Studi di Roma Tor Vergata
Universita della Calabria
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Application filed by Universita' Degli Studi Dl Messina, Universita degli Studi di Parma, Consiglio Nazionale delle Richerche CNR, Universita degli Studi di Roma Tor Vergata, Universita della Calabria filed Critical Universita' Degli Studi Dl Messina
Publication of EP3423053A1 publication Critical patent/EP3423053A1/en
Withdrawn legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/4015—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil having oxo groups directly attached to the heterocyclic ring, e.g. piracetam, ethosuximide
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/402—1-aryl substituted, e.g. piretanide
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/18—Antivirals for RNA viruses for HIV

Definitions

  • the present invention relates to the use of 2-oxo-2 -/-pyrrol-1 (5)-carboxamide derivatives as anti-HIV agents, and to the process for the production thereof.
  • the present invention originates in the pharmaceutical industry, specifically in the field of drugs for the treatment of HIV infections.
  • the present invention relates to the use in the medical field of 2-oxo- 2 - -pyrrol-1 (5)-carboxamide derivatives in the treatment of HIV infections, pharmaceutical compositions containing these derivatives as active ingredients, and a new process for the preparation of such derivatives.
  • the goal of drug therapy is to prevent the viral replication in the body in order to reduce the damage to the immune system and allow the survival with an acceptable quality of life.
  • the anti-HIV drugs yet available are grouped into 5 major classes of compounds: integrase inhibitors; reverse transcriptase inhibitors (NRTIs, NtRTIs, NNRTIs); protease inhibitors (PI); fusion inhibitors; inhibitors of the co-receptor involved in the entry of the virus.
  • integrase inhibitors reverse transcriptase inhibitors
  • PI protease inhibitors
  • fusion inhibitors inhibitors of the co-receptor involved in the entry of the virus.
  • Another important aspect, which originates from the chronicization of HIV infections, relates to the so-called reservoirs of infection in which the virus remains in a latent form preventing eradication of the infection [Siliciano J.D., Siliciano R.F., Recent developments in the search for a cure for HIV-1 infection: targeting the latent reservoir for HIV-1 , J Allergy Clin Immunol. 2014 Jul, 134(1 ), 12-9]
  • An area of pharmacological research in the field of treatment of HIV infection is directed to find new molecules that targets both the resistant forms and the reservoirs.
  • the lack of a prophylactic therapy and the lack of vaccines capable of treating or preventing the development of AIDS generates a high demand for new drug therapies for the treatment of HIV that are effective and do not contribute to determine the onset of resistant forms.
  • One of the objects of the present invention is to provide compounds with anti-HIV activity having a low degree of toxicity, and thus enabling to carry out prolonged chronic treatments of subjects who have contracted the infection.
  • a further object of the present invention is to provide molecules with anti-HIV activity that are effective, and whose the preparation does not involve neither high production costs nor complex procedures.
  • the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof
  • Ri , R2, R3, and R4 are as defined in the appended claims, for use in the prevention and/or treatment of HIV.
  • the present invention provides for a pharmaceutical composition
  • a pharmaceutical composition comprising one or more compounds of formula (I) as defined above, and/or pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable excipient, carrier, or diluent for use in the prevention and/or treatment of HIV.
  • the present invention provides for a method for the treatment of HIV and/or diseases or disorders associated with HIV infection in a human being, said method comprising the administration of a compound of formula (I) as defined above, or a pharmaceutically acceptable salt thereof.
  • the present invention provides for original methods for preparing compounds of formula (I) as defined above, through a process that provides for appropriate synthetic transformations, as an alternative to more cumbersome procedures [Pifferi, G.; Pinza, M.; Ger. Offen. (1977), DE 2635854 A1 19770224].
  • Figure 1 schematically illustrates a first chemical synthesis route of compounds of formula (I) of the invention.
  • Figure 2 schematically illustrates a second chemical synthesis route of compounds of formula (I). DETAILED DESCRIPTION OF THE INVENTION
  • the present invention consists, in a general aspect, in having identified that 2-oxo- 2 -/-pyrrol-1 (5H)-carboxamide derivatives find application in the medical field in the treatment and/or prevention of HIV infection.
  • the present invention provides compounds of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of HIV infection
  • Ri represents H, (C1-C10) linear or branched alkyl, an allyl group, -Si(Ci-C6 alkyl)3, aryl, heteroaryl, -CH2COOR5, -CH2CONR6R7, -C(S)NR 6 R7, -CORe, -SO2R6, -SORe;
  • R2 represents H, (C1-C10) linear or branched alkyl, aryl, heteroaryl, COOR5;
  • R3, R4 represent, each one independently, H, (C1-C6) linear or branched alkyl, -(CH 2 )n-;
  • R5 represents H, (C1-C6) linear or branched alkyl
  • R6, R7 represent, each one independently, H, (C1-C6) linear or branched alkyl, aryl, heteroaryl.
  • Ri is a (Ci-C6)alkyl, preferably (Ci-C4)alkyl, for example tert-butyl.
  • the R2 group is an aryl, typically phenyl, for example substituted by a (Ci-C6)alkyl or preferably is H.
  • R3 and R4 are H, and R2 has the meaning of any of the previously described substituent groups, and, in particular, is a phenyl optionally substituted by a (Ci-C6)alkyl.
  • the heteroaryl is a furyl, pyrrolyl, thiophenyl, or pyridyl, each optionally substituted, for example, by a (Ci-C6)alkyl group.
  • the Ri and R2 substituent groups are H.
  • 2-oxo-2 - -pyrrol-1 (5H)-carboxamide derivatives of formula (I) combine a high anti-HIV power associated with a low degree of toxicity.
  • the criterion that defines the cytotoxic activity in relation to the antiviral one, SI indicated that the activity of the compounds of formula (I) is high, and close to that of the latest generation of drugs that fall within the class of non-nucleoside inhibitors of HIV RT, while cytotoxicity is lower.
  • the cytotoxic activity of the compounds 1 , 2, and 3 having the formula (I) has proved to be 10-20 times lower than the activity of Ripilvirine and Etravirine molecules, known antiretroviral drugs widely used.
  • the present invention provides a process for the production of compounds of formula (I) or a pharmaceutically acceptable salt thereof.
  • Ri, R2, R3, and R4 have the meaning previously referred to, or in accordance with any one of the preceding embodiments, said method consists in reacting a substituted urea of formula
  • Ri , R2, R3, and R4 are as previously defined, in the presence of a catalyst typically based on Pd, such as Pdl2, a iodide, for example Kl, and an organic solvent, typically DME, and adding carbon oxide (CO).
  • a catalyst typically based on Pd, such as Pdl2, a iodide, for example Kl, and an organic solvent, typically DME, and adding carbon oxide (CO).
  • the carbonylation reaction is carried out by heating the reaction mixture, for example at a temperature from 30 to 150°C, preferably from 70 to 100°C.
  • the production of compounds of formula (I) can be made through the reaction between a) an isocyanate of formula
  • R2, R3, and R4 represents substituents as previously referred to, added to an organic solvent, typically DME, in the presence of carbon oxide and a suitable catalyst, typically palladium based.
  • reaction between the isocyanate and the propargylamine previously illustrated provides, in situ an urea of formula, as previously defined,
  • the inventions also relates to the intermediate compound
  • the palladium based catalyst is Pdl2.
  • the process for the production of compounds of formula (I) comprises the addition of Kl to the starting materials a) and b).
  • the carbonylation reaction is carried out in the presence of a suitable organic solvent, conveniently dimethoxyethane (DME).
  • a suitable organic solvent conveniently dimethoxyethane (DME).
  • the carbonylation reaction is carried out at pressures of carbon oxide from 1 to 100 bar, typically from 10 to 40 bar.
  • the catalyst concentration is comprised in the range of 0,1 - 0.001 molar (moles/liter of solution), preferably of 0.02-0.005 molar (moles/liter of solution).
  • palladium iodide is employed as catalyst in amounts comprised between 0.01 % and 5% by moles, based on the starting material propargylamine, preferably 0.5-2%.
  • potassium iodide is employed as co-catalyst in amounts comprised between 0% and 50% by moles, based on the starting material propargylamine, preferably 5-20%.
  • the carbonylation reaction is performed by heating the reaction mixture, for example at a temperature comprised in the range from 30 to 150°C, preferably from 70 to 100°C.
  • the process for the production of compounds of formula (I) comprises adding propargylamine, isocyanate, and an organic solvent into a reactor in the presence of a catalyst of Pdl2 and Kl, and charge carbon oxide at a pressure of 25 bar, while stirring the reaction mixture at a temperature from 70 to 100°C.
  • the carbonylation reaction by which the compounds of formula (I) are obtained is carried out in the absence of air and other oxidants.
  • the process of the invention has the advantage to be highly convenient, as it uses a production plant of easy realization and low operating costs.
  • the process of the invention in both embodiments, has the advantage to take place with high production yields.
  • alkyl indicates a saturated aliphatic hydrocarbon radical, including straight chain and branched chain radicals of 1 to 10 carbon atoms.
  • Non limiting examples of alkyl are ( ⁇ - ⁇ ) alkyl, for example, methyl, ethyl, propyl, isopropyl, n-butyl, iso-butyl, tert-butyl, n-amyl, iso-amyl, n-hexyl, and the like.
  • aryl indicates a hydrocarbon consisting of a mono-, bi- or tricyclic ring system wherein the rings are fused together or linked covalently to one another, and at least one of the carbocyclic rings is aromatic.
  • aryl groups comprise phenyl, alpha- or beta-naphthyl, 9,10-dihydroanthryl, indanyl, fluorenyl, biphenyl, and the like.
  • heteroaryl indicates a mono-, bi- or tricyclic ring system containing from one to four heteroatoms selected from nitrogen, oxygen, and sulfur, wherein the rings are fused together or linked covalently to one another, and at least one of the rings is aromatic.
  • heteroaryl groups comprise furyl, pyrrolyl, thiophenyl, pyridyl.
  • alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heterocyclyl, or heterocyclic cyclic group may be unsubstituted or substituted by one or more substituents.
  • substituted or substituted group indicates that one or more hydrogen atoms of the previously mentioned groups are substituted by another atom or group including, as example, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, alkoxy, aryloxy, heteroaryl, trifluoromethyl, trifluoromethoxy, carboxyl, acyl, aroyl, heteroaryl, halogen, nitro, cyano, alkoxycarbonyl, aryloxycarbonyl, alkylthio, arylthio, alkylsulfonyl, arylsulfonyl.
  • Examples of compounds of the invention are:
  • the present invention further reports processes for producing compounds of formula (I), as previously defined, through a carbonylation reaction in the presence of a catalyst, as previously reported.
  • a compound of formula (I) may be obtained by applying the chemical reaction shown in the synthetic scheme depicted in Figure 1 and referred to as Method A.
  • this scheme illustrates the preparation of 2-oxo-2H-pyrrol-1 (5)- carboxamide derivatives through a single stage which comprises the carbonylation of the illustrated urea catalyzed by palladium salts.
  • a steel autoclave with a capacity of 50 imL is charged with N-propargylurea, Pdl2 catalyst, and Kl, and then dimethoxyethane (DME) is added.
  • the autoclave is purged four times with carbon oxide, pressurized to about 15 bar, to eliminate any residual air, then carbon oxide (25 bar measured at room temperature) is charged.
  • the autoclave immersed in an oil bath, is maintained under magnetic stirring at 80°C for 24 hours. At the end, the autoclave is cooled down to 0°C and vented slowly.
  • the reaction mixture is taken up with dichloromethane and filtered to remove any inorganic solid residue.
  • the reaction mixture obtained after the previously reported treatment is subjected to silica gel column chromatography, eluting with a hexane/ethyl mixture, thereby obtaining the pure final product.
  • a compound of formula (I) may be obtained by applying the chemical reaction shown in the synthetic scheme illustrated in Figure 2.
  • a steel autoclave with a capacity of 50 imL is charged with propargylamine and isocyanate, in an equimolar ratio, Pdl2 catalyst, and Kl, then dimethoxyethane (DME) is added.
  • the autoclave is purged four times with carbon oxide, pressurized to about 15 bar, to eliminate any residual air, then carbon oxide (25 bar measured at room temperature) is charged.
  • the autoclave immersed in an oil bath, is maintained under magnetic stirring at 100°C for 24 hours. At the end, the autoclave is cooled down to 0°C and vented slowly.
  • the reaction mixture is taken up with dichloromethane and filtered to remove any inorganic solid residue.
  • the raw reaction product is subjected to silica gel column chromatography, eluting with a hexane/ethyl mixture, thereby obtaining the pure product with a yield based on the starting substrate.
  • compounds of formula (I) may form an acid addition salt or a salt with a base, depending on the type of substituents, and these salts are included in the present invention, provided that they are pharmaceutically acceptable salts.
  • salt refers to any salt of a compound according to the present invention prepared from and inorganic or organic acid or base, and internally formed salts. Typically, such salts have a physiologically acceptable anion or cation.
  • physiologically or pharmaceutically acceptable salts of compounds of the present invention comprise hydrochloride, acetate, citrate, gluconate, lactate, tartrate, phosphate, borate, maleate, sulfate, and nitrate.
  • Physiologically and pharmaceutically acceptable salts may also be suitable for medical uses because of their greater aqueous solubility compared to the parent compound.
  • Pharmaceutically acceptable salts may also be prepared from other salts, including other pharmaceutically acceptable salts of compounds of formula (I), using conventional methods.
  • the compounds may only be described in one stereoisomeric form, but the present invention comprise all possible stereoisomers, whether in a pure state or in a mixture. And, thus, in the object of the present invention comprises all possible stereoisomers, including racemates and enantiomerically pure compounds.
  • compounds or salts of the invention should be interpreted as excluding those compounds (if any) that are chemically unstable, either per se or in water, which are clearly unsuitable for pharmaceutical use via any route of administration, whether oral, parenteral, or otherwise.
  • Such compounds are known to an expert chemist.
  • Prodrugs or compounds which are stable ex vivo, and which are convertible in the body of a mammal (for example, a human being) in the compounds of the invention are, however, included.
  • the present invention also comprises active metabolites of compounds of formula (I).
  • Another aspect of the present invention relates to pharmaceutical compositions containing a compound of formula (I).
  • compositions of the present invention comprise all compositions produced by mixing a compound of the present invention and a pharmaceutically acceptable carrier. Such compositions are suitable for pharmaceutical use in an animal or in a human being.
  • compositions of the present invention comprise a therapeutically effective amount of one or more compounds of formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
  • a pharmaceutical composition may optionally contain other active ingredients.
  • carrier refers to a transport agent, excipient, diluent or adjuvant with which the therapeutic or active ingredient is administered. Any carrier and/or excipient suitable for the form of preparation desired for administration is considered for use with the compounds described herein.
  • the carrier may take many forms, depending on the form of preparation desired for administration, for example, oral or parenteral (including intravenous).
  • any of the usual pharmaceutical means may be used such as, for example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents, and the like, in the case of oral liquid preparations such as, for example, suspensions, elixirs, and solutions; or carriers such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents, and the like, in the case of oral solid preparations such as, for example, powders, hard and soft capsules, and tablets, with solid oral preparations being preferable compared to liquid preparations.
  • the compounds of the present invention may be combined as the active ingredient in intimate admixture with a carrier and/or pharmaceutically suitable excipient according to conventional pharmaceutical compounding techniques.
  • compositions suitable for parenteral administration including subcutaneous, intramuscular, and intravenous, pulmonary, nasal, rectal, topical, or oral.
  • suitable route of administration in any given case will depend in part on the nature and severity of the conditions being treated, and on the nature of the active ingredient.
  • An exemplary route of administration is the oral route.
  • the compositions may advantageously be presented in unit dosage pharmaceutical forms prepared by any of the methods well known in the pharmacy art.
  • the preferred compositions comprise compositions suitable for oral, parenteral, topical, subcutaneous or pulmonary administration, in the form of nasal or buccal inhalation.
  • the compositions may be prepared by any of the methods well known in the pharmacy art.
  • compositions may be in the form of tablets, pills, capsules, solutions, suspensions, emulsions, powders, suppositories, and sustained-release formulations.
  • the tablets may be coated by means of standard aqueous or nonaqueous techniques.
  • such compositions and preparations may contain at least 0.1 percent of active compound.
  • the percentage of active compound in these compositions may, of course, vary and may advantageously be from 1 percent to about 60 percent of the weight of the unit.
  • the amount of active compound in such therapeutically useful compositions is such that the therapeutically active dosage will be obtained.
  • the active compounds may also be administered intranasally, for example, as liquid drops or spray.
  • the tablets, pills, capsules, and the like may also contain a binder such as gum tragacanth, acacia, corn starch, or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as corn starch, potato starch, alginic acid; a lubricant such as magnesium stearate; and a sweetening agent such as sucrose, lactose, or saccharin.
  • a liquid carrier such as a fatty oil.
  • Various other materials may be present as coatings or to modify the physical form of the dosage unit. For example, tablets may be coated with shellac, sugar, or both.
  • a syrup or elixir may contain, in addition to the active ingredient, sucrose as a sweetening agent, methyl- and propylparaben as preservatives, a dye and a flavoring agent, such as cherry flavor or orange.
  • sucrose as a sweetening agent
  • methyl- and propylparaben as preservatives
  • a dye and a flavoring agent such as cherry flavor or orange.
  • the composition will be an enteric coated formulation.
  • compositions for topical administration comprise, but are not limited to, ointments, creams, lotions, solutions, pastes, gels, sticks, liposomes, nanoparticles, patches, bandages, and dressings for wounds.
  • the topical formulation comprises a penetration enhancer.
  • compositions for pulmonary administration comprise, but are not limited to, dry powder compositions consisting of the powder of a compound of formula (I), or a salt thereof, and the powder of a suitable carrier and/or lubricant.
  • the compositions for pulmonary administration may be inhaled from any suitable dry powder inhaler device known to one skilled in the art.
  • compositions of the present invention are carried out according to a protocol, and at a sufficient dosage to reduce inflammation and pain in the subject.
  • the active ingredient or the active ingredients are generally formulated in dosage units.
  • the dosage units may contain from 0.01 to 1 ,000 mg of a compound of formula (I), per dosage unit for daily administration.
  • the effective amounts for topical formulations will depend on the severity of the disease, disorder or condition, on prior therapy, on the individual's health status, and on the response to the drug. In some embodiments, the dose is in the range from 0.001 % by weight to about 60% by weight of the formulation.
  • the compound of the present invention and the other active ingredient may be used in lower doses compared to when each one is used alone.
  • the present invention provides for compounds of formula (I) for use in the treatment of diseases or disorders associated with HIV infection.
  • the compounds of formula (I), and pharmaceutical compositions thereof, and methods for administering them are useful in the treatment of HIV complicated infections or infection where also a tumoral form is present.
  • the subject to be treated may be an animal (for example, a mouse, a rat, a non- human primate, and a non-human mammal) or a human being.
  • the present invention provides for a method for the treatment or prevention of an HIV infection, comprising administering a therapeutically effective amount of an active compound of formula (I), according to one or more of the embodiments previously described, in a subject in need of treatment.
  • the compounds of formula (I), and pharmaceutical compositions thereof, and methods for administering them are useful in the treatment or prevention of a disease or disorder when administered in combination with other treatments.
  • the present invention further relates to combined therapies or treatments with a compound of formula (I), or a pharmaceutical composition that contains them.
  • the compounds of formula (I), and pharmaceutical compositions thereof, and methods for administering them are useful in the treatment of viral infections, when administered in combination with other pharmacological agents or active ingredients.
  • the compounds of formula (I) find medical application in the prevention, treatment or in maintenance therapy of AIDS.
  • these pharmacological agents are chemotherapeutic agents including, for example, the compounds 1 , 2, and 3.
  • a steel autoclave with a capacity of 50 imL is charged with 1 -ter-butyl-3-(prop-2-yn- 1 -yl)urea (0.308 g, 2.0 mmol), Pdl2 (0.014 g, 0.04 mmol) 2% by moles based on the substrate, and Kl (0.066 g, 0.4 mmol) 20% by moles based on the substrate, then dimethoxyethane (DME) (4 imL) is added.
  • DME dimethoxyethane
  • reaction mixture obtained after the previously reported treatment is subjected to silica gel column chromatography, eluting with a 8/2 hexane/ethyl acetate mixture, affording the pure product in a 73% yield, based on the starting substrate (0.266 g, 1 .46 mmol).
  • a steel autoclave with a capacity of 50 mL is charged with 1 -ter-butyl-3-(prop-2-yn- 1 -yl)urea (0.308 g, 2.0 mmol), Pdl2 (0.0018 g, 0.005 mmol) 0.25% by moles based on the substrate, and Kl (0.0083 g, 0.05 mmol) 2.5% by moles based on the substrate, then dimethoxyethane (DME) (4 mL) is added. After it was purged four times with carbon oxide, the autoclave is pressurized with carbon oxide (2.5 MPa) and submitted to magnetic stirring at 80°C, in an oil bath, for 24 hours.
  • DME dimethoxyethane
  • a steel autoclave with a capacity of 50 mL is charged with 1 -ter-butyl-3-(3- phenylprop-2-yn-1 -yl)urea (0.460 g, 2.0 mmol), Pdl2 (0.014 g, 0.04 mmol) 2% by moles based on the substrate, and Kl (0.066 g, 0.4 mmol) 20% by moles based on the substrate, and Kl, then dimethoxyethane (DME) (4 mL) is added. After it was purged four times with carbon oxide, the autoclave is pressurized with carbon oxide (2.5 MPa) and submitted to magnetic stirring at 80°C, in an oil bath, for 24 hours.
  • DME dimethoxyethane
  • the reaction mixture is recovered and submitted to the described treatment (Method A).
  • the pure title compound is obtained by separation through silica gel column chromatography, eluting with a 8/2 hexane/ethyl acetate mixture, with a yield of 82%, based on the starting substrate (0.423 g, 1 .64 mmol).
  • a steel autoclave with a capacity of 50 imL is charged with 1 -allyl-3-(prop-2-yn-1 - yl)urea (0.276 g, 2.0 mmol), Pdl2 (0.014 g, 0.04 mmol) 2% by moles based on the substrate, and Kl (0.066 g, 0.4 mmol) 20% by moles based on the substrate, then dimethoxyethane (DME) (4 imL) is added.
  • DME dimethoxyethane
  • the autoclave is pressurized with carbon oxide (2.5 MPa) and submitted to magnetic stirring at 80°C, in an oil bath, for 24 hours.
  • the reaction mixture is recovered and submitted to the described treatment (Method A).
  • the pure title compound is obtained by separation through silica gel column chromatography, eluting with a 8/2 hexane/ethyl acetate mixture, with a yield of 70%, based on the starting substrate (0.232 g, 1 .40 mmol).
  • a steel autoclave with a capacity of 50 mL is charged with 1 -carboethoxymethyl-3- (prop-2-yn-1 -yl)urea (0.368 g, 2.0 mmol), Pdl2 (0.014 g, 0.04 mmol) 2% by moles based on the substrate, and Kl (0.066 g, 0.4 mmol) 20% by moles based on the substrate, then dimethoxyethane (DME) (4 mL) is added.
  • DME dimethoxyethane
  • the pure title compound is obtained by separation through silica gel column chromatography, eluting with a 7/3 hexane/ethyl acetate mixture, with a yield of 73%, based on the starting substrate (0.309 g, 1 .46 mmol).
  • a steel autoclave with a capacity of 50 mL is charged with propargylamine (0.1 12 g, 2.0 mmol), tert-butylisocyanate (0.199 g, 2.0 mmol), Pdl2 (0.014 g, 0.04 mmol) 2% by moles based on the substrate, and Kl (0.066 g, 0.4 mmol) 20% by moles based on the substrate, then dimethoxyethane (DME) (4 mL) is added. After it was purged four times with carbon oxide, the autoclave is pressurized with carbon oxide (2.5 MPa) and submitted to magnetic stirring at 100°C, in an oil bath, for 24 hours. The reaction mixture is recovered and submitted to the described treatment (Method B).
  • reaction mixture is submitted to silica gel column chromatography, eluting with a 8/2 hexane/ethyl acetate mixture, affording the pure title compound with a yield of 74%, based on the starting substrate (0.269 g, 1 .48 mmol).
  • a steel autoclave with a capacity of 50 mL is charged with propargylamine (0.1 12 g, 2.0 mmol), carboethoxymethylisocyanate (0.286 g, 2.0 mmol), Pdl2 (0.014 g, 0.04 mmol) 2% by moles based on the substrate, and Kl (0.066 g, 0.4 mmol) 20% by moles based on the substrate, then dimethoxyethane (DME) (4 imL) is added.
  • DME dimethoxyethane
  • the pure title compound is obtained by separation through silica gel column chromatography, eluting with a 7/3 hexane/ethyl acetate mixture, with a yield of 68%, based on the starting substrate (0.288 g, 1 .36 mmol).
  • a steel autoclave with a capacity of 50 imL is charged with 1 -(2-methylbut-3-yn-2- yl)urea (0.252 g, 2.0 mmol), Pdl2 (0.014 g, 0.04 mmol) 2% by moles based on the substrate, and Kl (0.066 g, 0.4 mmol) 20% by moles based on the substrate, then dimethoxyethane (DME) (4 imL) is added.
  • DME dimethoxyethane
  • the autoclave is pressurized with carbon oxide (2.5 MPa) and submitted to magnetic stirring at 80°C, in an oil bath, for 24 hours.
  • the reaction mixture is recovered and submitted to the described treatment (Method A).
  • the pure title compound is obtained by separation through silica gel column chromatography, eluting with a 7/3 hexane/ethyl acetate mixture, with a yield of 84%, based on the starting substrate (0.259 g, 1 .68 mmol).
  • N-benzoyl-2,2-dimethyl-5-oxo-2,5-dihydro-1 H-pyrrolo-1-carboxamide to a suspension of NaH (1 .1 mmol) in dry THF (50 imL), 2,2-dimethyl-5-oxo-2,5- dihydro-1 -/-pyrrol-1 -carboxamide (77 mg, 0.5 mmol) was added at room temperature under a nitrogen atmosphere. The mixture was maintained under stirring for 15 minutes at room temperature and at reflux for 2 hours. The solution was then cooled down to 0°C, and benzoyl chloride (140 mg, 1 mmol) was added dropwise. The resulting mixture was maintained under stirring at room temperature for 2 hours and at reflux for 15 hours.
  • 2,2-dimethyl-5-oxo-N-tosyl-2,5-dihydro-1 H-pyrrol-1-carboxamide to a suspension of NaH (1 .1 mmol) in dry THF (50 imL), 2,2-dimethyl-5-oxo-2,5- dihydro-1 -/-pyrrol-1 -carboxamide (77 mg, 0.5 mmol) was added at room temperature under a nitrogen atmosphere. The mixture was maintained under stirring for 15 minutes at room temperature and at reflux for 2 hours. The solution was then cooled down to 0°C, and tosyl chloride (140 mg, 1 mmol) was added. The resulting mixture was maintained under stirring at room temperature for 2 hours and at reflux for 15 hours.
  • 2-oxo-2 - -pyrrol-1 (5H)carboxamide derivatives have shown antiretroviral activity in in vitro experimental infections with HIV virus whose description will be reported in details below in the experimental section.
  • an infection with HIV virus was used. The assay was performed on three compounds identified as 1 , 2, and 3.
  • a cytotoxicity test after 72 hours of culture was set up and assessed as inhibition of the cellular metabolism of U937 human monocytoid cells in the presence of concentrations of compounds 1 , 2 and 3 of 1000, 100, 10 and 1 ⁇ , and calculating the CC50 (concentration able to inhibit the oxidative cellular metabolism in 50% of cells).
  • a MTT assay was employed that uses an oxidizing chromogen system (MTT bromide) having a tetrazole ring which may be reduced by mitochondrial dehydrogenase to form a nitrogenous chromogenic compound, designated as formazan, that forms insoluble crystals that remain within the cell because the cell membranes are waterproof.
  • MTT bromide oxidizing chromogen system having a tetrazole ring which may be reduced by mitochondrial dehydrogenase to form a nitrogenous chromogenic compound, designated as formazan, that forms insoluble crystals that remain within the cell because the cell membranes are waterproof.
  • the formazan has been properly metabolized by the mitochondrial enzymes, it accumulates in the cell, it does not come out.
  • the test molecules inhibited the production of formazan and, in particular, the compound 1 showed a CC50 equal to 324 ⁇ , the compound 2 of 848 ⁇ , and the compound 3 of 529 ⁇ , respectively.
  • the cytotoxicity assays were also performed on drugs already used in therapy such as Etravirian and Ripilvirine. In this case, Etravirine showed a CC50 of 55 ⁇ while Ripilvirine of 34 ⁇ .
  • the cytotoxicity experiments were performed in duplicate and repeated twice. Assessment of the Antiviral Activity of Compounds 1 , 2 e 3
  • HIV human immunodeficiency virus 1
  • HIV is the etiological agent of acquired immunodeficiency.
  • the virus infects T lymphocytes, in particular CD4, and causes a cytopathic effect.
  • lymphomonocytes were isolated from peripheral blood of healthy individuals and exposed to infection with HIV.
  • the HIV NL4-3 virus was prepared after transfection of a cell line of human epithelial origin, HEK293T, with a purified plasmid DNA containing the infectious clone of HIV NL4-3 (pNL4-3, AIDS reagent, Programm NIH, Bethesda, MD, Adachi et al., 1986). Subsequently, the HIV-1 virus was purified from 20-50 imL of the culture supernatant of transfected cells by clarification (3,000 revolutions per minute), and ultracentrifugation on sucrose continuous gradient (40,000 revolutions per minute). The purified virus fractions were obtained by further centrifugation on a discontinuous gradient of iodixanol.
  • the fractions separated on gradient were recovered and the amount of virus was assayed by assessing the presence of HIV p24 viral nucleocapsid protein.
  • the lymphomonocytes were pre- treated and not with molecules 1 , 2 and 3 at concentrations of 1 , 10, and 100 ⁇ , respectively, for 2 hours at 37°C.
  • microcultures were set up in which the cells were exposed to infection by a "cell free" method consisting in the presentation of 1 x10 5 lymphomonocytes to 250 ng of purified HIV NL4-3 virus. To promote virus adsorption, the cells and the virus were centrifuged for 1 hour at 400 rpm at room temperature.
  • the adsorption phase was extended for another 3 hours in a humidified atmosphere at 37°C, in the presence of 5% carbon oxide. After this time, the excess virus, i.e. the one not adsorbed on the cells, was removed by washings and it was replaced by fresh culture medium containing the compounds 1 , 2 and 3 at the same concentrations used in the pre-treatment. After 72 hours culture, the infection was assessed by an ELISA method measuring the production of the p24 protein.
  • NNRTIs i.e. HIV non-nucleoside reverse transcriptase inhibitors
  • ICso concentration of compound that causes inhibition of infection in 50% of the cells
  • the compound 1 was the most active in inhibiting the infection
  • the compounds 2 and 3 showed an ICso, always in the same assay, of 169 ⁇ and 105 ⁇ , respectively.
  • the reference compounds Etravirine Ripilvirine showed an ICso of 9 and 10, respectively.
  • the global activity of the compounds was expressed as "selectivity index" (SI), which defines as a whole the in vitro activity of a molecule as the ratio of the toxic concentration to the concentration at which the molecule produces an effect.

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Abstract

The present invention relates to the use in the medical field of 2-oxo-2H-pyrrol- 1 (5)-carboxamide derivatives in the treatment of HIV infections, pharmaceutical compositions containing these derivatives as active ingredients, and a process for the preparation of such derivatives.

Description

"USE OF 2-OXO-2H-PYRROL-1 (5H)-CARBOXAMIDE DERIVATIVES AS ANTI- HIV AGENTS AND PROCESS FOR THE PRODUCTION THEREOF"
** ** ** ** **
DESCRIPTION FIELD OF THE INVENTION
The present invention relates to the use of 2-oxo-2 -/-pyrrol-1 (5)-carboxamide derivatives as anti-HIV agents, and to the process for the production thereof. The present invention originates in the pharmaceutical industry, specifically in the field of drugs for the treatment of HIV infections.
In particular, the present invention relates to the use in the medical field of 2-oxo- 2 - -pyrrol-1 (5)-carboxamide derivatives in the treatment of HIV infections, pharmaceutical compositions containing these derivatives as active ingredients, and a new process for the preparation of such derivatives.
PRIOR ART
It is estimated that at least 30 million people in the world are affected by HIV infection, and that every year there are between 2.5 to 3 million new cases.
In recent years, the pharmacological research in the field of HIV infection has made significant progress, and the discovery of new therapies has enabled us to limit the number of fatal cases. The goal of drug therapy is to prevent the viral replication in the body in order to reduce the damage to the immune system and allow the survival with an acceptable quality of life.
The anti-HIV drugs yet available are grouped into 5 major classes of compounds: integrase inhibitors; reverse transcriptase inhibitors (NRTIs, NtRTIs, NNRTIs); protease inhibitors (PI); fusion inhibitors; inhibitors of the co-receptor involved in the entry of the virus.
The combined use of these classes of drugs has substantially changed the clinical course of HIV infections from a short course pathological form with a fatal outcome into a chronic disease with an increased life expectancy.
The use of cocktails of HIV drugs belonging to different categories allows to increase the therapeutic response, while limiting the risks of the onset of mutant viral strains that are resistant to such drugs [lyidogan P., Anderson K.S., Current perspectives on HIV-1 antiretroviral drug resistance, Viruses, 2014 Oct 24, 6(10), 4095-4139. doi: 10.3390/v610409]. In fact, one of the major issues associated with the therapeutic control of the viral load in HIV-positive patients is in fact connected to the phenomena of resistance to chemotherapy.
Consequently, a need currently exists to develop new molecules that act in a selective manner on known viral targets or phylogenetically conserved viral targets but different from the targets, that can reduce the risks to originate, as a result of the treatment, resistant forms of the virus [Mori M., Kovalenko L, Lyonnais S., Antaki D., Torbett B.E., Botta M., Mirambeau G., Mely Y.. Nucleocapsid Protein: A Desirable Target for Future Therapies Against HIV-1 , Curr Top Microbiol Immunol. 2015, 389, 53-92. doi: 10.1007/82_2015_433].
Another important aspect, which originates from the chronicization of HIV infections, relates to the so-called reservoirs of infection in which the virus remains in a latent form preventing eradication of the infection [Siliciano J.D., Siliciano R.F., Recent developments in the search for a cure for HIV-1 infection: targeting the latent reservoir for HIV-1 , J Allergy Clin Immunol. 2014 Jul, 134(1 ), 12-9]
An area of pharmacological research in the field of treatment of HIV infection is directed to find new molecules that targets both the resistant forms and the reservoirs.
At present, the lack of a prophylactic therapy and the lack of vaccines capable of treating or preventing the development of AIDS, generates a high demand for new drug therapies for the treatment of HIV that are effective and do not contribute to determine the onset of resistant forms.
SUMMARY OF THE INVENTION
One of the objects of the present invention is to provide compounds with anti-HIV activity having a low degree of toxicity, and thus enabling to carry out prolonged chronic treatments of subjects who have contracted the infection.
A further object of the present invention is to provide molecules with anti-HIV activity that are effective, and whose the preparation does not involve neither high production costs nor complex procedures.
The Inventors have now found that certain pyrrol-2-(5H)one derivatives, in particular 2-oxo-2 -/-pyrrol-1 (5)-carboxamide derivatives, are provided with a specific anti-HIV activity, and have a low in vitro toxicity. According to a first aspect, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof
wherein Ri , R2, R3, and R4 are as defined in the appended claims, for use in the prevention and/or treatment of HIV.
Further embodiments of the compounds of the invention are the subject matter of claims 2-6.
In a second aspect, the present invention provides for a pharmaceutical composition comprising one or more compounds of formula (I) as defined above, and/or pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable excipient, carrier, or diluent for use in the prevention and/or treatment of HIV.
In a third aspect, the present invention provides for a method for the treatment of HIV and/or diseases or disorders associated with HIV infection in a human being, said method comprising the administration of a compound of formula (I) as defined above, or a pharmaceutically acceptable salt thereof.
In a fourth aspect, the present invention provides for original methods for preparing compounds of formula (I) as defined above, through a process that provides for appropriate synthetic transformations, as an alternative to more cumbersome procedures [Pifferi, G.; Pinza, M.; Ger. Offen. (1977), DE 2635854 A1 19770224].
BRIEF DESCRIPTION OF THE DRAWINGS
Figure 1 schematically illustrates a first chemical synthesis route of compounds of formula (I) of the invention.
Figure 2 schematically illustrates a second chemical synthesis route of compounds of formula (I). DETAILED DESCRIPTION OF THE INVENTION
The present invention consists, in a general aspect, in having identified that 2-oxo- 2 -/-pyrrol-1 (5H)-carboxamide derivatives find application in the medical field in the treatment and/or prevention of HIV infection.
Accordingly, in a first aspect, the present invention provides compounds of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of HIV infection
(I)
wherein
Ri represents H, (C1-C10) linear or branched alkyl, an allyl group, -Si(Ci-C6 alkyl)3, aryl, heteroaryl, -CH2COOR5, -CH2CONR6R7, -C(S)NR6R7, -CORe, -SO2R6, -SORe; R2 represents H, (C1-C10) linear or branched alkyl, aryl, heteroaryl, COOR5; R3, R4 represent, each one independently, H, (C1-C6) linear or branched alkyl, -(CH2)n-;
R5 represents H, (C1-C6) linear or branched alkyl;
R6, R7 represent, each one independently, H, (C1-C6) linear or branched alkyl, aryl, heteroaryl.
According to some embodiments, Ri is a (Ci-C6)alkyl, preferably (Ci-C4)alkyl, for example tert-butyl.
According to some embodiments, Ri is an allyl CH2=CH-CH2- or CH2COOR5, wherein R5 is (Ci-C6)alkyl, for example CH3-CH2-.
In some embodiments, for example according to any one of those previously described, the R2 group is an aryl, typically phenyl, for example substituted by a (Ci-C6)alkyl or preferably is H.
According to certain embodiments of compounds of formula (I), R3 and R4 are H, and R2 has the meaning of any of the previously described substituent groups, and, in particular, is a phenyl optionally substituted by a (Ci-C6)alkyl.
Preferably, the heteroaryl is a furyl, pyrrolyl, thiophenyl, or pyridyl, each optionally substituted, for example, by a (Ci-C6)alkyl group.
According to some embodiments, the Ri and R2 substituent groups are H.
Compounds of formula (I) preferred as antivirals are 2-oxo-2 -/-pyrrol-1 (5H)- carboxamides having the following formulae 1 , 2, 3:
wherein Et is ethyl and Ph is phenyl.
The inventors have found that 2-oxo-2 - -pyrrol-1 (5H)-carboxamide derivatives of formula (I) combine a high anti-HIV power associated with a low degree of toxicity. In particular, it has been observed that the criterion that defines the cytotoxic activity in relation to the antiviral one, SI, indicated that the activity of the compounds of formula (I) is high, and close to that of the latest generation of drugs that fall within the class of non-nucleoside inhibitors of HIV RT, while cytotoxicity is lower.
In particular, the cytotoxic activity of the compounds 1 , 2, and 3 having the formula (I) has proved to be 10-20 times lower than the activity of Ripilvirine and Etravirine molecules, known antiretroviral drugs widely used.
According to another aspect, the present invention provides a process for the production of compounds of formula (I) or a pharmaceutically acceptable salt thereof.
wherein
Ri, R2, R3, and R4 have the meaning previously referred to, or in accordance with any one of the preceding embodiments, said method consists in reacting a substituted urea of formula
wherein Ri , R2, R3, and R4 are as previously defined, in the presence of a catalyst typically based on Pd, such as Pdl2, a iodide, for example Kl, and an organic solvent, typically DME, and adding carbon oxide (CO). Typically, the carbonylation reaction is carried out by heating the reaction mixture, for example at a temperature from 30 to 150°C, preferably from 70 to 100°C.
According to another embodiment, the production of compounds of formula (I) can be made through the reaction between a) an isocyanate of formula
N=C=0 wherein Ri represents a substituent as previously defined, with b) a propargylamine of formula
wherein R2, R3, and R4 represents substituents as previously referred to, added to an organic solvent, typically DME, in the presence of carbon oxide and a suitable catalyst, typically palladium based.
Typically, the reaction between the isocyanate and the propargylamine previously illustrated provides, in situ an urea of formula, as previously defined,
that further reacts through a carbonylation reaction catalyzed by palladium salts, to give the compounds of formula (I).
According to certain aspects, the inventions also relates to the intermediate compound
as previously defined.
The realization of the process of the invention has proved to be very simple to produce compounds with high anti-HIV activity according to any one of claims 9- 14.
According to certain embodiments, the palladium based catalyst is Pdl2.
According to some embodiments, the process for the production of compounds of formula (I) comprises the addition of Kl to the starting materials a) and b).
Typically, the carbonylation reaction is carried out in the presence of a suitable organic solvent, conveniently dimethoxyethane (DME).
According to some embodiments, the carbonylation reaction is carried out at pressures of carbon oxide from 1 to 100 bar, typically from 10 to 40 bar. According to some embodiments, the catalyst concentration is comprised in the range of 0,1 - 0.001 molar (moles/liter of solution), preferably of 0.02-0.005 molar (moles/liter of solution). Generally, palladium iodide is employed as catalyst in amounts comprised between 0.01 % and 5% by moles, based on the starting material propargylamine, preferably 0.5-2%. Generally, potassium iodide is employed as co-catalyst in amounts comprised between 0% and 50% by moles, based on the starting material propargylamine, preferably 5-20%. According to some embodiments, the carbonylation reaction is performed by heating the reaction mixture, for example at a temperature comprised in the range from 30 to 150°C, preferably from 70 to 100°C.
According to some embodiments, the process for the production of compounds of formula (I) comprises adding propargylamine, isocyanate, and an organic solvent into a reactor in the presence of a catalyst of Pdl2 and Kl, and charge carbon oxide at a pressure of 25 bar, while stirring the reaction mixture at a temperature from 70 to 100°C.
Typically, the carbonylation reaction by which the compounds of formula (I) are obtained, is carried out in the absence of air and other oxidants.
The process of the invention has the advantage to be highly convenient, as it uses a production plant of easy realization and low operating costs.
In addition, the process of the invention, in both embodiments, has the advantage to take place with high production yields.
All the scientific and technical terms used herein have the same meanings commonly known by one of ordinary skill in the art, unless otherwise defined. The following terms used in the description and in the claims of the present application have the meanings specified below, unless otherwise defined.
I. Definitions
As used herein, the term "alkyl" indicates a saturated aliphatic hydrocarbon radical, including straight chain and branched chain radicals of 1 to 10 carbon atoms.
Non limiting examples of alkyl are (Οι-Οβ) alkyl, for example, methyl, ethyl, propyl, isopropyl, n-butyl, iso-butyl, tert-butyl, n-amyl, iso-amyl, n-hexyl, and the like.
As used herein, the term "aryl" indicates a hydrocarbon consisting of a mono-, bi- or tricyclic ring system wherein the rings are fused together or linked covalently to one another, and at least one of the carbocyclic rings is aromatic. Examples of aryl groups comprise phenyl, alpha- or beta-naphthyl, 9,10-dihydroanthryl, indanyl, fluorenyl, biphenyl, and the like.
As used herein, the term "heteroaryl" indicates a mono-, bi- or tricyclic ring system containing from one to four heteroatoms selected from nitrogen, oxygen, and sulfur, wherein the rings are fused together or linked covalently to one another, and at least one of the rings is aromatic.
Examples of heteroaryl groups comprise furyl, pyrrolyl, thiophenyl, pyridyl.
Any previously mentioned alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heterocyclyl, or heterocyclic cyclic group may be unsubstituted or substituted by one or more substituents.
Unless otherwise indicated, as used herein, the term "substituent" or "substituent group" indicates that one or more hydrogen atoms of the previously mentioned groups are substituted by another atom or group including, as example, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, alkoxy, aryloxy, heteroaryl, trifluoromethyl, trifluoromethoxy, carboxyl, acyl, aroyl, heteroaryl, halogen, nitro, cyano, alkoxycarbonyl, aryloxycarbonyl, alkylthio, arylthio, alkylsulfonyl, arylsulfonyl. Examples of compounds of the invention are:
II. Processes for Preparing Compounds of Formula (I)
In another aspect, the present invention further reports processes for producing compounds of formula (I), as previously defined, through a carbonylation reaction in the presence of a catalyst, as previously reported.
In one embodiment, a compound of formula (I) may be obtained by applying the chemical reaction shown in the synthetic scheme depicted in Figure 1 and referred to as Method A.
Specifically, this scheme illustrates the preparation of 2-oxo-2H-pyrrol-1 (5)- carboxamide derivatives through a single stage which comprises the carbonylation of the illustrated urea catalyzed by palladium salts.
By way of example, an embodiment of the method A is illustrated below. A steel autoclave with a capacity of 50 imL is charged with N-propargylurea, Pdl2 catalyst, and Kl, and then dimethoxyethane (DME) is added. The autoclave is purged four times with carbon oxide, pressurized to about 15 bar, to eliminate any residual air, then carbon oxide (25 bar measured at room temperature) is charged. The autoclave, immersed in an oil bath, is maintained under magnetic stirring at 80°C for 24 hours. At the end, the autoclave is cooled down to 0°C and vented slowly. The reaction mixture is taken up with dichloromethane and filtered to remove any inorganic solid residue. The reaction mixture obtained after the previously reported treatment is subjected to silica gel column chromatography, eluting with a hexane/ethyl mixture, thereby obtaining the pure final product.
In an alternative embodiment, a compound of formula (I) may be obtained by applying the chemical reaction shown in the synthetic scheme illustrated in Figure 2.
In this second scheme, the preparation of 2-oxo-2 -/-pyrrol-1 (5)-carboxamide derivative in a single stage is illustrated, comprising the in situ formation of N- propargylurea starting from isocyanate and propargylamine, and the carbonylation of the same urea in the presence of CO and a Pd salts based catalyst.
By way of example, an embodiment of the method B is illustrated below.
A steel autoclave with a capacity of 50 imL is charged with propargylamine and isocyanate, in an equimolar ratio, Pdl2 catalyst, and Kl, then dimethoxyethane (DME) is added. The autoclave is purged four times with carbon oxide, pressurized to about 15 bar, to eliminate any residual air, then carbon oxide (25 bar measured at room temperature) is charged. The autoclave, immersed in an oil bath, is maintained under magnetic stirring at 100°C for 24 hours. At the end, the autoclave is cooled down to 0°C and vented slowly. The reaction mixture is taken up with dichloromethane and filtered to remove any inorganic solid residue. The raw reaction product is subjected to silica gel column chromatography, eluting with a hexane/ethyl mixture, thereby obtaining the pure product with a yield based on the starting substrate.
Compounds of formula (I) obtained with both Method A and B are particularly stable.
III. Pharmaceutically Acceptable Salts
It should be understood that, as used herein, the references and the compounds of formula (I) are also intended to comprise pharmaceutically acceptable salts and/or derivatives thereof.
In addition, compounds of formula (I) may form an acid addition salt or a salt with a base, depending on the type of substituents, and these salts are included in the present invention, provided that they are pharmaceutically acceptable salts.
The expressions "compound of the invention", and "compounds of the present invention", and "compounds of formula (I)" refer to each of the compounds of formula (I), and are intended to include pharmaceutically acceptable salts, hydrates, solvates, and crystalline forms, polymorphs thereof, and also any suitable form illustrated herein below.
As used herein, the term "salt" refers to any salt of a compound according to the present invention prepared from and inorganic or organic acid or base, and internally formed salts. Typically, such salts have a physiologically acceptable anion or cation.
Suitably physiologically or pharmaceutically acceptable salts of compounds of the present invention comprise hydrochloride, acetate, citrate, gluconate, lactate, tartrate, phosphate, borate, maleate, sulfate, and nitrate.
Physiologically and pharmaceutically acceptable salts may also be suitable for medical uses because of their greater aqueous solubility compared to the parent compound.
Pharmaceutically acceptable salts may also be prepared from other salts, including other pharmaceutically acceptable salts of compounds of formula (I), using conventional methods.
In the present description, the compounds may only be described in one stereoisomeric form, but the present invention comprise all possible stereoisomers, whether in a pure state or in a mixture. And, thus, in the object of the present invention comprises all possible stereoisomers, including racemates and enantiomerically pure compounds.
In general, compounds or salts of the invention should be interpreted as excluding those compounds (if any) that are chemically unstable, either per se or in water, which are clearly unsuitable for pharmaceutical use via any route of administration, whether oral, parenteral, or otherwise. Such compounds are known to an expert chemist. Prodrugs or compounds which are stable ex vivo, and which are convertible in the body of a mammal (for example, a human being) in the compounds of the invention are, however, included.
The present invention also comprises active metabolites of compounds of formula (I).
IV. Pharmaceutical Compositions
Another aspect of the present invention relates to pharmaceutical compositions containing a compound of formula (I).
Pharmaceutical compositions of the present invention comprise all compositions produced by mixing a compound of the present invention and a pharmaceutically acceptable carrier. Such compositions are suitable for pharmaceutical use in an animal or in a human being.
Pharmaceutical compositions of the present invention comprise a therapeutically effective amount of one or more compounds of formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
A pharmaceutical composition may optionally contain other active ingredients. The term "carrier" refers to a transport agent, excipient, diluent or adjuvant with which the therapeutic or active ingredient is administered. Any carrier and/or excipient suitable for the form of preparation desired for administration is considered for use with the compounds described herein.
The carrier may take many forms, depending on the form of preparation desired for administration, for example, oral or parenteral (including intravenous). In preparing the compositions for an oral dosage form, any of the usual pharmaceutical means may be used such as, for example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents, and the like, in the case of oral liquid preparations such as, for example, suspensions, elixirs, and solutions; or carriers such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents, and the like, in the case of oral solid preparations such as, for example, powders, hard and soft capsules, and tablets, with solid oral preparations being preferable compared to liquid preparations.
In some embodiments, the compounds of the present invention may be combined as the active ingredient in intimate admixture with a carrier and/or pharmaceutically suitable excipient according to conventional pharmaceutical compounding techniques.
The compositions comprise compositions suitable for parenteral administration, including subcutaneous, intramuscular, and intravenous, pulmonary, nasal, rectal, topical, or oral. The suitable route of administration in any given case will depend in part on the nature and severity of the conditions being treated, and on the nature of the active ingredient. An exemplary route of administration is the oral route. The compositions may advantageously be presented in unit dosage pharmaceutical forms prepared by any of the methods well known in the pharmacy art. The preferred compositions comprise compositions suitable for oral, parenteral, topical, subcutaneous or pulmonary administration, in the form of nasal or buccal inhalation. The compositions may be prepared by any of the methods well known in the pharmacy art.
The pharmaceutical compositions may be in the form of tablets, pills, capsules, solutions, suspensions, emulsions, powders, suppositories, and sustained-release formulations.
If desired, the tablets may be coated by means of standard aqueous or nonaqueous techniques. In some embodiments, such compositions and preparations may contain at least 0.1 percent of active compound. The percentage of active compound in these compositions may, of course, vary and may advantageously be from 1 percent to about 60 percent of the weight of the unit. The amount of active compound in such therapeutically useful compositions is such that the therapeutically active dosage will be obtained. The active compounds may also be administered intranasally, for example, as liquid drops or spray.
The tablets, pills, capsules, and the like, may also contain a binder such as gum tragacanth, acacia, corn starch, or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as corn starch, potato starch, alginic acid; a lubricant such as magnesium stearate; and a sweetening agent such as sucrose, lactose, or saccharin. When the dosage unit form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier such as a fatty oil. Various other materials may be present as coatings or to modify the physical form of the dosage unit. For example, tablets may be coated with shellac, sugar, or both. A syrup or elixir may contain, in addition to the active ingredient, sucrose as a sweetening agent, methyl- and propylparaben as preservatives, a dye and a flavoring agent, such as cherry flavor or orange. To prevent decomposition during the transit through the upper portion of the gastrointestinal tract, the composition will be an enteric coated formulation.
The compositions for topical administration comprise, but are not limited to, ointments, creams, lotions, solutions, pastes, gels, sticks, liposomes, nanoparticles, patches, bandages, and dressings for wounds. In some embodiments, the topical formulation comprises a penetration enhancer.
The compositions for pulmonary administration comprise, but are not limited to, dry powder compositions consisting of the powder of a compound of formula (I), or a salt thereof, and the powder of a suitable carrier and/or lubricant. The compositions for pulmonary administration may be inhaled from any suitable dry powder inhaler device known to one skilled in the art.
The administration of the compositions is carried out according to a protocol, and at a sufficient dosage to reduce inflammation and pain in the subject. In some embodiments, in the pharmaceutical compositions of the present invention the active ingredient or the active ingredients are generally formulated in dosage units. The dosage units may contain from 0.01 to 1 ,000 mg of a compound of formula (I), per dosage unit for daily administration.
In some embodiments, the effective amounts for topical formulations will depend on the severity of the disease, disorder or condition, on prior therapy, on the individual's health status, and on the response to the drug. In some embodiments, the dose is in the range from 0.001 % by weight to about 60% by weight of the formulation.
When used in combination with one or more other active ingredients, the compound of the present invention and the other active ingredient may be used in lower doses compared to when each one is used alone.
With reference to the formulations regarding any variety of routes of administration, methods and formulations for the administration of drugs are described in Remington's Pharmaceutical Sciences, 17th Edition, Gennaro et al. Ed., Mack Publishing Co., 1985, and Remington's Pharmaceutical Sciences, Gennaro AR ed. 20th Edition, 2000, Williams & Wilkins PA, USA, and Remington: The Science and Practice of Pharmacy, 21st Edition, Lippincott Williams & Wilkins Ed., 2005; and in Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems, 8th Edition, Lippincott Williams & Wilkins Ed., 2005.
V. Clinical Uses of Compounds of Formula (I) and/or Therapeutic Treatments
According to some embodiments, the present invention provides for compounds of formula (I) for use in the treatment of diseases or disorders associated with HIV infection.
In some embodiments, the compounds of formula (I), and pharmaceutical compositions thereof, and methods for administering them are useful in the treatment of HIV complicated infections or infection where also a tumoral form is present.
The subject to be treated may be an animal (for example, a mouse, a rat, a non- human primate, and a non-human mammal) or a human being.
According to some embodiments, the present invention provides for a method for the treatment or prevention of an HIV infection, comprising administering a therapeutically effective amount of an active compound of formula (I), according to one or more of the embodiments previously described, in a subject in need of treatment.
The inventors have discovered that the compounds of formula (I) play an antiviral function.
In some embodiments, the compounds of formula (I), and pharmaceutical compositions thereof, and methods for administering them are useful in the treatment or prevention of a disease or disorder when administered in combination with other treatments.
In an additional aspect, the present invention further relates to combined therapies or treatments with a compound of formula (I), or a pharmaceutical composition that contains them. In some embodiments, the compounds of formula (I), and pharmaceutical compositions thereof, and methods for administering them are useful in the treatment of viral infections, when administered in combination with other pharmacological agents or active ingredients.
In some embodiments, the compounds of formula (I) find medical application in the prevention, treatment or in maintenance therapy of AIDS.
In some embodiments, these pharmacological agents are chemotherapeutic agents including, for example, the compounds 1 , 2, and 3.
Example 1
N-(t-butyl)-2-oxo-2,5-dihydro-1 H-pyrrol-1 -carboxamide
Production Process Following Method A.
A steel autoclave with a capacity of 50 imL is charged with 1 -ter-butyl-3-(prop-2-yn- 1 -yl)urea (0.308 g, 2.0 mmol), Pdl2 (0.014 g, 0.04 mmol) 2% by moles based on the substrate, and Kl (0.066 g, 0.4 mmol) 20% by moles based on the substrate, then dimethoxyethane (DME) (4 imL) is added. After it was purged four times with carbon oxide, the autoclave is pressurized with carbon oxide (2.5 MPa) and maintained under magnetic stirring at 80°C, in an oil bath, for 24 hours. The reaction mixture is recovered and submitted to treatment according to Method A. The reaction mixture obtained after the previously reported treatment is subjected to silica gel column chromatography, eluting with a 8/2 hexane/ethyl acetate mixture, affording the pure product in a 73% yield, based on the starting substrate (0.266 g, 1 .46 mmol).
Melting Point 96-97°C
1H NMR (400 MHz, CDC ): δ 8.34 (s, 1 H), 7.25 (dt, J = 5.9, 1 .8 Hz, 1 H), 6.17 (dt, J = 5.9, 1 .5 Hz, 1 H), 4.42 (t, J = 1 .8 Hz, 2H), 1 .40 (s, 9H); 13C NMR (100 MHz, CDCI3): δ 171 .4, 150.6, 145.7, 127.0, 50.8, 50.5, 28.8; IR (ZnSe) v/cnr1 : 3403, 3282, 3072, 2965, 2924, 171 1 , 1547, 1438, 1354, 1279, 1 197, 1 1 15, 966, 936, 919, 805, 755, 688; MS m/z. 182 (M+,1 ), 167 (100), 124 (25), 1 10 (12), 84 (77), 56 (12), 41 (1 1 ). Example 2
N-(t-butyl)-2-oxo-2,5-dihydro-1 H-pyrrol-1 -carboxamide
Production Process Following Method A.
A steel autoclave with a capacity of 50 mL is charged with 1 -ter-butyl-3-(prop-2-yn- 1 -yl)urea (0.308 g, 2.0 mmol), Pdl2 (0.0018 g, 0.005 mmol) 0.25% by moles based on the substrate, and Kl (0.0083 g, 0.05 mmol) 2.5% by moles based on the substrate, then dimethoxyethane (DME) (4 mL) is added. After it was purged four times with carbon oxide, the autoclave is pressurized with carbon oxide (2.5 MPa) and submitted to magnetic stirring at 80°C, in an oil bath, for 24 hours. The reaction mixture is recovered and submitted to the described treatment (Method A). The pure title compound is obtained by separation through silica gel column chromatography, using a 8/2 hexane/ethyl acetate mixture, with a yield of 63%, based on the starting substrate (0.228 g, 1 .26 mmol). Example 3
N-ter-butyl-2-oxo-3-phenyl-2,5-dihydro-1 H-pyrrol-1 -carboxamide
Production Process Following Method A.
A steel autoclave with a capacity of 50 mL is charged with 1 -ter-butyl-3-(3- phenylprop-2-yn-1 -yl)urea (0.460 g, 2.0 mmol), Pdl2 (0.014 g, 0.04 mmol) 2% by moles based on the substrate, and Kl (0.066 g, 0.4 mmol) 20% by moles based on the substrate, and Kl, then dimethoxyethane (DME) (4 mL) is added. After it was purged four times with carbon oxide, the autoclave is pressurized with carbon oxide (2.5 MPa) and submitted to magnetic stirring at 80°C, in an oil bath, for 24 hours. The reaction mixture is recovered and submitted to the described treatment (Method A). The pure title compound is obtained by separation through silica gel column chromatography, eluting with a 8/2 hexane/ethyl acetate mixture, with a yield of 82%, based on the starting substrate (0.423 g, 1 .64 mmol).
Melting Point 104-105°C
1H NMR (400 MHz, DMSO-afe): δ 8.47 (s, 1 H), 7.88-7.85 (m, 1 H), 7.82 (t, J = 2.2 Hz, 1 H), 7.45-7.40 (m, 4H), 4.41 (d, J = 2.2 Hz, 2H), 1 .37 (s, 9H); 13C NMR (100 MHz, DMSO-afe): δ 170.6, 150.4, 141 .8, 134.5, 130.9, 128.9, 128.7, 126.9, 50.3, 48.8, 28.8; IR (ZnSe) v/cnr1 : 3281 , 3060, 2963, 2929, 1701 , 1541 , 1444, 1355, 1279, 1226, 1 180, 995, 916, 812, 789, 763, 738, 689; MS m/z: 258 (M+, 1 ), 215 (53), 200 (44), 172 (41 ), 159 (100), 130 (86), 1 15 (60), 103 (18), 84 (52), 77 (14), 56 (23), 41 (34). xample 4
N-allyl-2-oxo-2,5-dihydro-1 H-pyrrol-1 -carboxamide
Production Process Following Method A.
A steel autoclave with a capacity of 50 imL is charged with 1 -allyl-3-(prop-2-yn-1 - yl)urea (0.276 g, 2.0 mmol), Pdl2 (0.014 g, 0.04 mmol) 2% by moles based on the substrate, and Kl (0.066 g, 0.4 mmol) 20% by moles based on the substrate, then dimethoxyethane (DME) (4 imL) is added. After it was purged four times with carbon oxide, the autoclave is pressurized with carbon oxide (2.5 MPa) and submitted to magnetic stirring at 80°C, in an oil bath, for 24 hours. The reaction mixture is recovered and submitted to the described treatment (Method A). The pure title compound is obtained by separation through silica gel column chromatography, eluting with a 8/2 hexane/ethyl acetate mixture, with a yield of 70%, based on the starting substrate (0.232 g, 1 .40 mmol).
Melting Point 62-64°C
1H NMR (400 MHz, CDC ): δ 8.43 (s, 1 H), 7.29 (dt, J = 6.0, 1 .9 Hz, 1 H), 6.19 (dt, J = 6.0, 1 .8 Hz, 1 H), 5.93-5.83 (m, 1 H), 5.23 (dd, J = 10.3, 2.0 Hz, 1 H), 5.14 (dd, J = 17.0, 2.0 Hz, 1 H), 4.45 (t, J = 1 .9 Hz, 2H), 3.96 (dt, J = 5.6, 1 .6 Hz, 2H); 13C NMR (100 MHz, CDCb): δ 171 .5, 151 .9, 146.2, 133.8, 126.9, 1 15.8, 51 .0, 41 .8; IR (ZnSe) v/cnr1 : 3296, 3083, 2975, 2934, 2870, 1709, 1537, 1453, 1325, 1246, 1 191 , 1 1 16, 993, 918, 819, 764, 709, 665; MS m/z: 166 (M+, 10), 139 (3), 123 (5), 1 10 (35), 83 (72), 66 (17), 56 (100), 41 (53). xample 5
N-Carboethoxymethyl-5-oxo-2,5-dihydro-1 H-pyrrol-1 -carboxamide
Production Process Following Method A.
A steel autoclave with a capacity of 50 mL is charged with 1 -carboethoxymethyl-3- (prop-2-yn-1 -yl)urea (0.368 g, 2.0 mmol), Pdl2 (0.014 g, 0.04 mmol) 2% by moles based on the substrate, and Kl (0.066 g, 0.4 mmol) 20% by moles based on the substrate, then dimethoxyethane (DME) (4 mL) is added. After it was purged four times with carbon oxide, the autoclave is pressurized with carbon oxide (2.5 MPa) and submitted to magnetic stirring at 80°C, in an oil bath, for 24 hours. The reaction mixture is recovered and submitted to the described treatment (Method A). The pure title compound is obtained by separation through silica gel column chromatography, eluting with a 7/3 hexane/ethyl acetate mixture, with a yield of 73%, based on the starting substrate (0.309 g, 1 .46 mmol).
Melting Point 59-60°C
1H NMR (400 MHz, CDC ): δ 8.75 (s, 1 H), 7.32 (dt, J = 6.0, 1 .7 Hz, 1 H), 6.22 (dt, J = 6.0, 1 .6 Hz, 1 H), 4.47 (d, J = 1 .7 Hz, 2H), 4.22 (q, J = 7.2 Hz, 2H), 4.1 1 (d, J = 5.7 Hz, 2H), 1 .29 (t, J = 7.2 Hz, 3H); 13C NMR (100 MHz, CDCb): δ 171 .4, 169.3, 152.1 , 146.4, 126.8, 61 .2, 50.8, 41 .4, 13.9; IR (ZnSe) v/cnr1 : 3286, 3068, 2989, 2920, 2851 , 1736, 1701 , 1528, 1354, 1213, 1 166, 1071 , 1021 , 960, 803, 760, 694; MS m/z: 212 (M+, 3), 166 (20), 139 (100), 1 10 (80), 82 (15), 66 (9), 56 (20), 41 (4). xample 6
N-(t-butyl)-2-oxo-2,5-dihydro-1 H-pyrrol-1 -carboxamide
Production Process Following Method B.
A steel autoclave with a capacity of 50 mL is charged with propargylamine (0.1 12 g, 2.0 mmol), tert-butylisocyanate (0.199 g, 2.0 mmol), Pdl2 (0.014 g, 0.04 mmol) 2% by moles based on the substrate, and Kl (0.066 g, 0.4 mmol) 20% by moles based on the substrate, then dimethoxyethane (DME) (4 mL) is added. After it was purged four times with carbon oxide, the autoclave is pressurized with carbon oxide (2.5 MPa) and submitted to magnetic stirring at 100°C, in an oil bath, for 24 hours. The reaction mixture is recovered and submitted to the described treatment (Method B). The reaction mixture is submitted to silica gel column chromatography, eluting with a 8/2 hexane/ethyl acetate mixture, affording the pure title compound with a yield of 74%, based on the starting substrate (0.269 g, 1 .48 mmol).
Example 7
N-Carboethoxymethyl-5-oxo-2,5-dihydro-1 H-pyrrol-1 -carboxamide
Production Process Following Method B.
A steel autoclave with a capacity of 50 mL is charged with propargylamine (0.1 12 g, 2.0 mmol), carboethoxymethylisocyanate (0.286 g, 2.0 mmol), Pdl2 (0.014 g, 0.04 mmol) 2% by moles based on the substrate, and Kl (0.066 g, 0.4 mmol) 20% by moles based on the substrate, then dimethoxyethane (DME) (4 imL) is added. After it was purged four times with carbon oxide, the autoclave is pressurized with carbon oxide (2.5 MPa) and submitted to magnetic stirring at 80°C, in an oil bath, for 24 hours. The reaction mixture is recovered and submitted to the described treatment (Method B). The pure title compound is obtained by separation through silica gel column chromatography, eluting with a 7/3 hexane/ethyl acetate mixture, with a yield of 68%, based on the starting substrate (0.288 g, 1 .36 mmol).
Example 8
-dimethyl-5-oxo-2,5-dihydro-1 H-pyrrol-1 -carboxamide
Production Process Following Method A.
A steel autoclave with a capacity of 50 imL is charged with 1 -(2-methylbut-3-yn-2- yl)urea (0.252 g, 2.0 mmol), Pdl2 (0.014 g, 0.04 mmol) 2% by moles based on the substrate, and Kl (0.066 g, 0.4 mmol) 20% by moles based on the substrate, then dimethoxyethane (DME) (4 imL) is added. After it was purged four times with carbon oxide, the autoclave is pressurized with carbon oxide (2.5 MPa) and submitted to magnetic stirring at 80°C, in an oil bath, for 24 hours. The reaction mixture is recovered and submitted to the described treatment (Method A). The pure title compound is obtained by separation through silica gel column chromatography, eluting with a 7/3 hexane/ethyl acetate mixture, with a yield of 84%, based on the starting substrate (0.259 g, 1 .68 mmol).
1H NMR (400 MHz, CDC ): δ 8.20 (s, 2H), 7.32 (d, J = 6.0 Hz, 1 H), 6.23 (d, J = 6.0 Hz, 1 H), 1 .27 (s, 6H); 13C NMR (100 MHz, CDCb): δ 171 .3, 159.5, 152.6, 122.3, 66.4, 23.3; IR (ZnSe) v/cnr1 : 3367, 3202, 31 10, 3005, 2976, 2937, 1705, 1582, 1470, 1448, 1366, 131 1 , 1 190, 1 1 19, 1062, 1035, 938, 823, 786, 703; MS m/z: 154 (M+, 3), 139 (5), 1 10 (60), 96 (100), 83 (5), 68 (19), 53 (8), 42 (25). Example 9
N-benzoyl-2,2-dimethyl-5-oxo-2,5-dihydro-1 H-pyrrolo-1-carboxamide: to a suspension of NaH (1 .1 mmol) in dry THF (50 imL), 2,2-dimethyl-5-oxo-2,5- dihydro-1 -/-pyrrol-1 -carboxamide (77 mg, 0.5 mmol) was added at room temperature under a nitrogen atmosphere. The mixture was maintained under stirring for 15 minutes at room temperature and at reflux for 2 hours. The solution was then cooled down to 0°C, and benzoyl chloride (140 mg, 1 mmol) was added dropwise. The resulting mixture was maintained under stirring at room temperature for 2 hours and at reflux for 15 hours. The mixture was concentrated under reduced pressure, and the residue was dissolved in ethyl acetate/acidic water (HCI 0.1 M). The organic layer was recovered and the aqueous layer was extracted again with ethyl acetate (3 χ 100 imL). The organic layers were combined, washed with saturated NaCI solution (2 χ 100 imL), dried over MgSO4, and concentrated in vacuo. The residue was purified through flash column chromatography, using cyclohexane-ethyl acetate (7:3) as eluting mixture, to give the pure compound with a yield of 55%, based on the starting substrate (0.071 g, 0.27 mmol).
1H NMR (500 MHz, CDC ) δ = 8.42 (bs, 1 H), 8.12 (dd, J = 8.4, 1 .3 Hz, 2H), 7.64 - 7.58 (m, 1 H), 7.50 - 7.45 (m, 2H), 7.15 (d, J = 6.0 Hz, 1 H), 6.03 (d, J = 6.0 Hz, 1 H), 1 .63 (s, 6H). Example 10
2,2-dimethyl-5-oxo-N-tosyl-2,5-dihydro-1 H-pyrrol-1-carboxamide: to a suspension of NaH (1 .1 mmol) in dry THF (50 imL), 2,2-dimethyl-5-oxo-2,5- dihydro-1 -/-pyrrol-1 -carboxamide (77 mg, 0.5 mmol) was added at room temperature under a nitrogen atmosphere. The mixture was maintained under stirring for 15 minutes at room temperature and at reflux for 2 hours. The solution was then cooled down to 0°C, and tosyl chloride (140 mg, 1 mmol) was added. The resulting mixture was maintained under stirring at room temperature for 2 hours and at reflux for 15 hours. The mixture was concentrated under reduced pressure, and the residue was dissolved in ethyl acetate/acidic water (HCI 0.1 M). The organic layer was recovered and the aqueous layer was extracted again with ethyl acetate (3 χ 100 imL). The organic layers were combined, washed with saturated NaCI solution (2 χ 100 imL), dried over MgSO4, and concentrated in vacuo. The residue was purified through flash column chromatography, using cyclohexane-ethyl acetate (7:3) as eluting mixture, to give the pure compound with a yield of 65%, based on the starting substrate (0.100 g, 0.32 mmol).
1H NMR (500 MHz, CDC ) δ = 8.51 (bs, 1 H), 7.75 (d, J = 7.8 Hz, 2H), 7.40 (d, J = 7.8 Hz, 2H), 7.12 (d, J = 6.1 Hz, 1 H), 6.08 (d, J = 6.1 Hz, 1 H), 2.40 (s, 3H), 1 .59 (s, 6H).
Example 11
Toxicity an in vitro Activity of 2-oxo-2H-pyrrol-1 (5H)carboxamide Derivatives on HIV Infection
2-oxo-2 - -pyrrol-1 (5H)carboxamide derivatives have shown antiretroviral activity in in vitro experimental infections with HIV virus whose description will be reported in details below in the experimental section. To test the activity of 2-oxo-2 -/-pyrrol-1 (5H)carboxamide derivatives, an infection with HIV virus was used. The assay was performed on three compounds identified as 1 , 2, and 3.
Assessment of toxicity of compounds 1 , 2, and 3
For the three compounds above, a cytotoxicity test after 72 hours of culture was set up and assessed as inhibition of the cellular metabolism of U937 human monocytoid cells in the presence of concentrations of compounds 1 , 2 and 3 of 1000, 100, 10 and 1 μΜ, and calculating the CC50 (concentration able to inhibit the oxidative cellular metabolism in 50% of cells). To assess the effect of the test molecules on cell viability, a MTT assay was employed that uses an oxidizing chromogen system (MTT bromide) having a tetrazole ring which may be reduced by mitochondrial dehydrogenase to form a nitrogenous chromogenic compound, designated as formazan, that forms insoluble crystals that remain within the cell because the cell membranes are waterproof. If the formazan has been properly metabolized by the mitochondrial enzymes, it accumulates in the cell, it does not come out. The test molecules inhibited the production of formazan and, in particular, the compound 1 showed a CC50 equal to 324 μΜ, the compound 2 of 848 μΜ, and the compound 3 of 529 μΜ, respectively. The cytotoxicity assays were also performed on drugs already used in therapy such as Etravirian and Ripilvirine. In this case, Etravirine showed a CC50 of 55 μΜ while Ripilvirine of 34 μΜ. The cytotoxicity experiments were performed in duplicate and repeated twice. Assessment of the Antiviral Activity of Compounds 1 , 2 e 3
To assess the antiretroviral activity of the compounds, an in vitro experimental model of infection with HIV virus (human immunodeficiency virus 1 ) was used. HIV is the etiological agent of acquired immunodeficiency. The virus infects T lymphocytes, in particular CD4, and causes a cytopathic effect. To test the effect of molecules 1 , 2 and 3 on the in vitro infection with HIV virus, lymphomonocytes were isolated from peripheral blood of healthy individuals and exposed to infection with HIV. The HIV NL4-3 virus was prepared after transfection of a cell line of human epithelial origin, HEK293T, with a purified plasmid DNA containing the infectious clone of HIV NL4-3 (pNL4-3, AIDS reagent, Programm NIH, Bethesda, MD, Adachi et al., 1986). Subsequently, the HIV-1 virus was purified from 20-50 imL of the culture supernatant of transfected cells by clarification (3,000 revolutions per minute), and ultracentrifugation on sucrose continuous gradient (40,000 revolutions per minute). The purified virus fractions were obtained by further centrifugation on a discontinuous gradient of iodixanol. The fractions separated on gradient were recovered and the amount of virus was assayed by assessing the presence of HIV p24 viral nucleocapsid protein. The lymphomonocytes were pre- treated and not with molecules 1 , 2 and 3 at concentrations of 1 , 10, and 100 μΜ, respectively, for 2 hours at 37°C. Subsequently, microcultures were set up in which the cells were exposed to infection by a "cell free" method consisting in the presentation of 1 x105 lymphomonocytes to 250 ng of purified HIV NL4-3 virus. To promote virus adsorption, the cells and the virus were centrifuged for 1 hour at 400 rpm at room temperature. Then the adsorption phase was extended for another 3 hours in a humidified atmosphere at 37°C, in the presence of 5% carbon oxide. After this time, the excess virus, i.e. the one not adsorbed on the cells, was removed by washings and it was replaced by fresh culture medium containing the compounds 1 , 2 and 3 at the same concentrations used in the pre-treatment. After 72 hours culture, the infection was assessed by an ELISA method measuring the production of the p24 protein.
The experiments were performed in triplicate and repeated 3 times. As a positive control, the effect of the compounds 1 , 2, and 3 were compared to that of latest generation antiretrovirals recently used in the clinic: Etravirine and Ripilvirine. These two compounds are classified as NNRTIs, i.e. HIV non-nucleoside reverse transcriptase inhibitors, and were chosen because they are present in most drug cocktails currently used in anti-HIV therapy.
The results, expressed as ICso, i.e. concentration of compound that causes inhibition of infection in 50% of the cells, highlighted that the compound 1 was the most active in inhibiting the infection, with an ICso, calculated for the specific assay used, of around 80 μΜ, while the compounds 2 and 3 showed an ICso, always in the same assay, of 169 μΜ and 105 μΜ, respectively. In addition, the reference compounds Etravirine Ripilvirine showed an ICso of 9 and 10, respectively. The global activity of the compounds was expressed as "selectivity index" (SI), which defines as a whole the in vitro activity of a molecule as the ratio of the toxic concentration to the concentration at which the molecule produces an effect. The results show that the SI for the molecule 1 , with the assays used, is 4 while the SI for the compounds 2 and 3 is 5. Considering that reference drugs such as Etravirine and Ripilvirine have an SI, calculated by us using the same assays, of 6 and 3, respectively, it is believed that the test compounds have overall a favorable SI.

Claims

1 . A compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of HIV infection
wherein
Ri represents H, (C1-C10) linear or branched alkyl, an allyl group, -Si(Ci-C6 alkyl)3, aryl, heteroaryl, -CH2COOR5, -ChteCONReRz, -C(S)NR6R7, -CORe, -SO2R6, -SORe; R2 represents H, (C1-C10) linear or branched alkyl, aryl, heteroaryl, COOR5;
R3, R4 represent, each one independently, H, (C1-C6) linear or branched alkyl, -(CH2)n-;
R5 represents H, (C1-C6) linear or branched alkyl;
R6, R7 represent, each one independently, H, (C1-C6) linear or branched alkyl, aryl, heteroaryl.
2. Compound of formula (I) or a pharmaceutically acceptable salt thereof for use according to claim 1 , wherein R3, R4 are each independently H.
3. Compound of formula (I) or a pharmaceutically acceptable salt thereof for use according to claim 1 or 2, wherein the aryl is phenyl, the heteroaryl is selected from furyl, pyrrolyl, thiophenyl, and pyridyl.
4. Compound of formula (I) or a pharmaceutically acceptable salt thereof for use according to any one of the claims 1 -3, wherein Ri is a (C1-C6) linear or branched alkyl, an allyl or a -CH2COOR5 group wherein R5 represents a (C1-C6) linear or branched alkyl.
5. Compound of formula (I) or a pharmaceutically acceptable salt thereof for use according to any one of the claims 1 -4, wherein R2 is phenyl, optionally substituted.
6. Compound of formula (I) or a pharmaceutically acceptable salt thereof for use according to any one of the claims 1 -5 selected from
7. Pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier for use according to any one of the claims 1 -6.
8. Pharmaceutical composition for use according to claim 7 further comprising an HIV agent preferably chosen from integrase inhibitors, reverse transcriptase inhibitors (NRTI, NtRTIs, NNRTIs), protease inhibitors (PI), fusion inhibitors, inhibitors of the co-receptor involved in entry of the virus, and/or a chemotherapy agent, and mixtures thereof.
9. Process for the production of a compound of formula (I) or a pharmaceutically acceptable salt thereof comprising reacting an isocyanate a) of formula
R1 N=C=0 a)
wherein Ri represents a substituent as defined in claim 1 , with a propargylamine b) of formula
wherein R2, R3, and R4 represent substituents as defined in claim 1 , in the presence of carbon oxide, a catalyst, preferably Pd based, Kl, and an organic solvent, preferably DME.
10. Process according to claim 9, wherein R3, R4 are H.
1 1 . Process according to claim 9 or 10, wherein Ri is a (Οι-Οβ) linear or branched alkyl, an allyl or a -CH2COOR5 group wherein R5 represents a (C1-C6) linear or branched alkyl.
12. Process according to any one of the claims 9-1 1 , wherein R2 is phenyl, optionally substituted.
13. Process for the production of a compound of formula (I) or a pharmaceutically acceptable salt thereof com rising providing a substituted urea of formula
wherein Ri, R2, R3, and R4 are as defined in claim 1 or 2,
adding carbon oxide in the presence of a catalyst, preferably Pd based, Kl, and an organic solvent, preferably DME.
14. Process according to claim 9 or 13, wherein said catalyst is Pdl2, said organic solvent is dimethoxyethane, and wherein the reaction temperature is comprised in the range from 30 to 150°C, preferably from 70 to 100°C.
EP17718413.2A 2016-03-04 2017-03-03 Use of 2-oxo-2h-pyrrol-1(5h)-carboxamide derivatives as anti-hiv agents and process for the production thereof Withdrawn EP3423053A1 (en)

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PCT/IB2017/051261 WO2017149511A1 (en) 2016-03-04 2017-03-03 Use of 2-oxo-2h-pyrrol-1(5h)-carboxamide derivatives as anti-hiv agents and process for the production thereof

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