HK40003405A - Alkylpyrimidine derivatives for the treatment of viral infections and further diseases - Google Patents

Description

Alkyl pyrimidine derivatives for the treatment of viral infections and additional diseases

The present application is a divisional application of an invention patent having application date of 2013, 8/9, application number of 201380042284.4, entitled "alkyl pyrimidine derivatives for the treatment of viral infections and additional diseases".

The present invention relates to alkyl pyrimidine derivatives, processes for their preparation, pharmaceutical compositions, and their use in the treatment and/or therapy of diseases.

The present invention relates to the use of alkyl pyrimidine derivatives in the treatment of viral infections, immune or inflammatory disorders, and thus to the modulation or agonism of toll-like receptors (TLRs). Toll-like receptors are major transmembrane proteins characterized by an extracellular leucine-rich domain and a cytoplasmic extension containing a conserved region. The innate immune system can recognize pathogen-associated molecular patterns via these TLRs expressed on the cell surface of certain types of immune cells. Recognition of foreign pathogens activates cytokine production and upregulation of costimulatory molecules on phagocytes. This results in modulation of T cell behavior.

Most mammalian species have between ten and fifteen types of Toll-like receptors. Thirteen TLRs (simply named TLR1 to TLR13) have been identified in humans as well as in mice, and many of their equivalents have been found in other mammalian species. However, the equivalent forms of certain TLRs found in humans are not present in all mammals. For example, a gene encoding a protein similar to TLR10 in humans is present in mice, but appears to have been damaged at some point in the past by a retrovirus. On the other hand, mice express TLR11, TLR12, and TLR13, none of which are present in humans. Other mammals may express TLRs that have not been found in humans. Other non-mammalian species may have a TLR different from that of mammals, as evidenced by TLR14, which is found in takifugupuferfish. This can complicate the approach of using experimental animals as models of human innate immunity.

For a review of toll-like receptors see the journal articles below. Hoffmann, J.A, (Hoffmann, J.A.) Nature (Nature) 426, pp. 33-38, 2003; akira, S. (Archela, S.), Takeda, K. (Zhutian, K.), and Kaisho, T. (Beatly, T.), Annual Rev. immunology (Annual book), pp.21, 335-; ulevitch, R.J, (Eulveryki, R.J.), Nature Reviews: Immunology (review Nature: Immunology) 4, 512-.

Compounds showing activity on Toll-like receptors have been previously described, such as heterocyclic derivatives in WO 2000006577, adenine derivatives in WO 98/01448 and WO 99/28321, and pyrimidines in WO 2009/067081.

In the treatment of certain viral infections, regular injections of interferon (IFN α) may be administered, such as Hepatitis C Virus (HCV), (Fried et al, Peginte-alfa plus ribavirin for chronicmagnetic hepatitis C virus infection, PEGylated interferon- α in combination with ribavirin for chronic hepatitis C virus infection), N Engl J Med (New England journal of medicine) 2002; 347: 975-82) small molecule IFN, which is orally administered, provides the potential advantage of reduced immunogenicity and ease of administration.

Interferon α (eur. j. Cancer (journal of european Cancer) 46, 2849-57, and Cancer Res. (. Cancer research) 1992, 52, 1056) in combination with other drugs has also been given in the treatment of certain types of Cancer, and they are also vaccine adjuvants of interest due to the ability of TLR7/8 agonists to induce significant Th1 responses (hum. vaccines (human vaccine) 2010, 6, 322-.

However, there is a strong need for novel Toll-like receptor modulators with preferred selectivity and improved safety profile (profile) compared to the compounds of the prior art.

According to the present invention there is provided a compound of formula (I)

Or a pharmaceutically acceptable salt, solvate or polymorph thereof, wherein

R₁Is hydrogen, fluorine, hydroxy, amine, C_1-6Alkyl radical, C_1-6Alkylamino radical, C_1-6Alkoxy radical, C_3-7Cycloalkyl radical, C_4-7Heterocycle, aryl, bicyclic heterocycle, aralkyl, heteroaryl, heteroaralkyl, aryloxy, heteroaryloxy, carboxylic acid ester, carboxylic acid amide, each of which may be optionally substituted with one or more substituents independently selected from halogen, hydroxy, amino, C_1-6Alkyl, di- (C)_1-6) Alkylamino, (C)_1-4) Alkoxy radical- (C_1-4) Alkyl radical, C_1-6Alkylamino radical, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-6Cycloalkyl, carboxylic acid, carboxylic ester, carboxylic amide, heterocycle, aryl, alkenyl, alkynyl, aralkyl, heteroaryl, heteroaralkyl, or nitrile,

R₂is C each of which may be optionally substituted by one or more substituents_1-6Alkyl, these substituents being independently selected from the group consisting of halogen, hydroxy, amino, nitrile, carboxylic acid, carboxylic ester, carboxylic amide, C_1-3Alkyl radical, C_1-3Alkoxy or C_3-6Cycloalkyl, sulfone, sulfoxide, or nitrile,

with the proviso that N- (2-amino-5-phenethylpyrimidin-4-yl) -N-pentylamine is excluded.

In a first embodiment, the present invention relates to compounds of formula (I) wherein R is₁Is a heterocyclic ring and R₂Is C substituted, for example, by one hydroxy group_1-6An alkyl group.

In a second embodiment, the present invention provides a compound of formula (I), wherein R is₁Is hydrogen and wherein R₂Is C each of which may be optionally substituted by one or more substituents_1-6Alkyl, these substituents being independently selected from the group consisting of halogen, hydroxy, amino, nitrile, carboxylic acid, carboxylic ester, carboxylic amide, C_1-3Alkyl radical, C_1-3Alkoxy or C_3-6Cycloalkyl, sulfone, sulfoxide, or nitrile.

Compounds of formula (I) and pharmaceutically acceptable salts, solvates or polymorphs thereof have activity as pharmaceuticals, particularly as modulators of Toll-like receptors, especially TLR7 and/or TLR 8.

In a further aspect, the present invention provides a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt, solvate or polymorph thereof together with one or more pharmaceutically acceptable excipients, diluents or carriers.

Furthermore, the compound of formula (I) or a pharmaceutically acceptable salt, solvate or polymorph thereof according to the invention, or a pharmaceutical composition comprising said compound of formula (I) or a pharmaceutically acceptable salt, solvate or polymorph thereof, may be used as a medicament.

Another aspect of the invention is: the compound of formula (I) or a pharmaceutically acceptable salt, solvate or polymorph thereof, or the pharmaceutical composition comprising said compound of formula (I) or a pharmaceutically acceptable salt, solvate or polymorph thereof, may accordingly be used to treat any disorder involving the modulation of TLR7 and/or TLR 8.

The term "alkyl" refers to a straight or branched chain saturated aliphatic hydrocarbon containing the specified number of carbon atoms.

The term "halogen" refers to fluorine, chlorine, bromine or iodine.

The term "alkenyl" refers to an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon double bond.

The term "alkynyl" refers to an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon triple bond.

The term "cycloalkyl" refers to a carbocyclic ring containing the specified number of carbon atoms.

The term "alkoxy" refers to an alkyl group (carbon and hydrogen chain) bonded individually to an oxygen, such as, for example, a methoxy group or an ethoxy group.

The term "aryl" means an aromatic ring structure that may optionally include one or two heteroatoms selected from N, O and S (particularly selected from N and O). The aromatic ring structure may have 5, 6, or 7 ring atoms. In particular, the aromatic ring structure may have 5 or 6 ring atoms.

The term "aryloxy" refers to an aromatic ring structure. The aromatic group is solely bonded to oxygen, like for example phenol.

The term "heteroaryloxy" refers to an aromatic ring structure that may optionally include one or two heteroatoms selected from N, O and S. The aromatic group contains 5 to 7 ring atoms, one of which is individually bonded to oxygen, like for example hydroxypyridine.

The term "bicyclic heterocycle" refers to an aromatic ring structure as defined for the term "aryl" consisting of two fused aromatic rings. Each ring may optionally include a heteroatom selected from N, O and S (particularly selected from N and O).

The term "aralkyl" refers to an aromatic ring structure as defined for the term "aryl" optionally substituted with an alkyl group.

The term "heteroaralkyl" refers to an aromatic ring structure as defined for the term "heteroaryl" optionally substituted with an alkyl group.

"heterocycle" refers to a saturated or partially saturated molecule and includes, but is not limited to, tetrahydrofuran, dioxane, or other cyclic ethers. Heterocycles containing nitrogen include, for example, azetidine, morpholine, piperidine, piperazine, pyrrolidine, and the like. Other heterocycles include, for example, thiomorpholine, morpholine, and cyclic sulfones.

A "heteroaryl" group is a heterocyclic group that is aromatic in nature. These are monocyclic, bicyclic or polycyclic rings containing one or more heteroatoms selected from N, O or S. Heteroaryl groups can be, for example, imidazolyl, isoxazolyl, furanyl, oxazolyl, pyrrolyl, pyridonyl, pyridyl, pyridazinyl, pyrazinyl, thiophene, or quinoline.

Pharmaceutically acceptable salts of the compounds of formula (I) include acid addition salts as well as base salts thereof. Suitable acid addition salts are formed from acids which form non-toxic salts. Suitable base salts are formed from bases which form non-toxic salts.

The compounds of the present invention may also exist in unsolvated and solvated forms. The term "solvate" is used herein to describe a molecular complex comprising a compound of the invention and one or more pharmaceutically acceptable solvent molecules (e.g., ethanol).

The term "polymorph" refers to the ability of a compound of the invention to exist in more than one form or crystal structure.

The compounds of the invention may be administered as crystalline or amorphous products. The compounds may be obtained, for example, as solid plugs, powders or films by methods such as precipitation, crystallization, freeze drying, spray drying or evaporative drying. They may be administered alone or in combination with one or more other compounds of the invention or in combination with one or more other drugs. Typically, they will be administered as a formulation in combination with one or more pharmaceutically acceptable excipients. The term "excipient" is used herein to describe any ingredient other than one or more compounds of the present invention. The choice of excipient will depend largely on factors such as the particular mode of administration, the effect of the excipient on solubility and stability, and the nature of the dosage form.

The compounds of the present invention, or any subgroup thereof, may be formulated into different pharmaceutical forms for administration purposes. As suitable compositions, all compositions usually used for systemic administration can be cited. To prepare the pharmaceutical compositions of this invention, an effective amount of the particular compound, optionally in addition salt form, as the active ingredient is combined in intimate admixture with a pharmaceutically acceptable carrier, which carrier may take a wide variety of forms depending on the form of preparation desired for administration. Desirably, these pharmaceutical compositions are in unit dosage forms suitable for, e.g., oral, rectal, or transdermal administration. For example, in preparing the compositions in oral dosage form, any of the usual pharmaceutical media may be employed, such as, for example, water, glycols, oils, alcohols and the like in the case of oral liquid preparations (e.g., suspensions, syrups, elixirs, emulsions and solutions); or solid carriers such as starches, sugars, kaolin, diluents, lubricants, binders, disintegrating agents and the like in the case of powders, pills, capsules and tablets. Because of their ease of administration, tablets and capsules represent the most advantageous oral unit dosage form in which case solid pharmaceutical carriers are obviously employed. Also included are solid form preparations which can be converted to liquid form shortly before use. In compositions suitable for transdermal administration, the carrier may optionally include a penetration enhancer and/or suitable wetting agent, optionally in combination with small proportions of suitable additives of any nature which do not introduce significant deleterious effects on the skin. The additives may facilitate administration to the skin and/or may aid in the preparation of the desired composition. These compositions can be administered in different ways, e.g., as a transdermal patch, as a spot-on, as an ointment. The compounds of the invention may also be administered via inhalation or insufflation by means of the methods and formulations employed in the present field for administration by this means. Thus, in general, the compounds of the invention can be administered to the lungs in the form of a solution, suspension or dry powder.

It is particularly advantageous to formulate the above pharmaceutical compositions in unit dosage form for ease of administration and uniformity of dosage. Unit dosage form, as used herein, refers to physically discrete units suitable as unitary dosages, each unit containing a predetermined quantity of active ingredient calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. Examples of such unit dosage forms are tablets (including scored or coated tablets), capsules, pills, powder packets (powder packets), wafers, suppositories, injectable solutions or suspensions and the like, and segregated multiples thereof.

One of ordinary skill in the art of treatment of infectious diseases will be able to determine the effective amount from the test results presented below. In general, it is contemplated that the daily effective amount will be from 0.01mg/kg to 50mg/kg body weight, more preferably from 0.1mg/kg to 10mg/kg body weight. It may be appropriate to administer the required dose as two, three, four or more sub-doses at appropriate intervals throughout the day. The sub-doses may be formulated in unit dosage forms, for example each unit dosage form containing from 1mg to 1000mg, and in particular from 5mg to 200mg, of the active ingredient.

As is well known to those of ordinary skill in the art, the precise dose and frequency of administration will depend on the particular compound of formula (I) used, the particular condition being treated, the severity of the condition being treated, the age, weight and general physical condition of the particular patient, and other drugs that the individual may take. Furthermore, it is apparent that the effective amount may be decreased or increased depending on the response of the subject being treated and/or depending on the evaluation of the physician prescribing the compounds of the instant invention. The above mentioned ranges of effective amounts are therefore only indicative and are not intended to limit the scope or use of the invention in any way.

Preparation of the Compounds

Compounds of formula (I) were prepared according to scheme 1.

Preparation of example 1

Scheme 1:

synthesis of A-1

Will CH₃PPh₃Br (27.91g, 78.1mmol, 1.5 equiv.) was suspended in THF (70mL) and in N₂Stirred at-78 ℃ under an atmosphere. N-butyllithium (30mL, 75mmol, 1.44 equivalents, 2.5M in hexanes) was added dropwise over 20 minutes and stirred for an additional 0.5 hour, followed by the addition of 2-amino-4, 6-dichloro-5-formylpyrimidine [5604-46-6 as a suspension in THF (180mL)](10.0g, 52mmol, 1.0 equiv.). The cooling bath was removed and the mixture was stirred at room temperature for 2 hours. The reaction was cooled to-78 ℃ and NH was then slowly added₄Cl (saturated aqueous). The cooling bath was removed and the mixture was stirred for 1.5 hours. The organic layer was separated, washed with water and dried (Na)₂SO₄) The solid was removed by filtration, and the solvent of the filtrate was removed under reduced pressure. The crude product was purified by silica gel column chromatography using a petroleum ether/ethyl acetate gradient to provide a colorless oil, A-1(1.2 g).

¹HNMR (400MHz, chloroform-d) delta ppm 5.30(br.s.,2H),5.65(d,1H),5.82(d,1H),6.58(q,1H)

Preparation of B-1

A-1(1.0g, 5.26mmol), n-butylamine (0.39g, 5.26mmol) and Et₃N (0.53g, 5.26mmol, 1.0 equiv.) is refluxed in ethanol (10mL) for 12 hours. The solvent was removed under reduced pressure. The crude product was purified by column chromatography on silica gel using a petroleum ether/ethyl acetate gradient. The best fractions were combined and concentrated under reduced pressure to give B-1(300 mg).

LC-MS m/z＝227(M+H)

¹HNMR (400MHz, chloroform-d) δ ppm 0.95(t, J ═ 7.3Hz,3H),1.38(dq, J ═ 14.9,7.4Hz,2H),1.55(quin, J ═ 7.4Hz,2H),3.38(q, J ═ 7.3Hz,2H),4.75(br.s.,2H),5.39(br.s.,1H),5.5(m,2H),6.55(m,1H)

Preparation of example 1

To a solution of B-1(200mg, 0.88mmol, 1.0 equiv.) in methanol (5mL) was added 10% Pd/C (20mg) and reacted with H at 50 deg.C₂Gas (50Psi) mixing continued for 17 hours. Subjecting the crude product to preparative high performance liquid chromatography (C18 column, eluent: CH)₃CN/H₂O from 10/90 to 95/5, 0.05% HCl). The desired fractions were combined and concentrated under reduced pressure to afford 1(74 mg).

LC-MS m/z＝195(M+H)

¹HNMR (400MHz, chloroform-d) δ ppm 0.95(t, J ═ 7.3Hz,3H),1.20(t, J ═ 7.3Hz,3H),1.38(dq, J ═ 14.9,7.4Hz,2H),1.62(quin, J ═ 7.4Hz,2H),1.93(br.s.,1H),2.37(q, J ═ 7.3Hz,2H),3.40-3.63(m,2H),6.18(br.s.,1H),7.24(b, 2H), etcr.s.,1H),13.43(br.s.,1H)

Table 1. compounds of formula (I). All compounds were synthesized according to the procedure used to prepare example 1.

And (4) an analytical method. All compounds were characterized by LC-MS using the following method:

method 1 Agilent 1100LC-MS in positive ion mode was equipped with a YMC-PACK ODS-AQ, 50X2.0mm, 5 μm column maintained at 50 ℃. The following mobile phases and gradients were used to monitor at 220nm at 0.8mL/min over a total run time of 10 minutes:

method 2. Agilent 1100LC-MS in positive ion mode was equipped with a 5 μm column of YMC-PACK ODS-AQ, 50X2.0mm, held at 50 ℃. The following mobile phases and gradients were used to monitor at 220nm at 0.8mL/min over a total run time of 10 minutes:

biological activity of the compound of formula (I)

Description of the bioassay

Assessment of TLR7 and TLR8 activity

HEK293 cells transiently transfected with TLR7 or TLR8 expression vectors and NF κ B-1uc reporter constructs were used to assess the ability of these compounds to activate human TLR7(hTLR7) and/or TLR8(hTLR8) in a cellular reporter assay. In one instance, the TLR expression construct expresses the corresponding wild-type sequence or mutant sequence that includes a deletion in the second leucine rich repeat of the TLR. Such mutant TLR proteins have previously been shown to be more susceptible to agonist activation (US 7498409).

Briefly, HEK293 cells were grown in medium (DMEM supplemented with 10% FCS and 2mM glutamine). For transfection of cells in 10cm dishes, cells were dispersed with trypsin-EDTA, transfected with CMV-TLR7 or a mixture of TLR8 plasmid (750ng), NF-. kappa.B-luc plasmid (375ng) and transfection reagents, and humidified 5% CO at 37 deg.C₂Incubate under atmosphere overnight. Transfected cells were then dispersed with trypsin-EDTA, washed in PBS and resuspended to 1.67x 10 in culture medium⁵Density of individual cells/mL. Thirty microliters of cells were then dispensed into each well in a 384-well plate, where 10 μ Ι _ of compound (in 4% DMSO) had been present. At 37 deg.C, 5% CO₂After 48 hours of incubation, luciferase activity was determined by adding 15 μ l of Steady Lite Plus substrate (PerkinElmer) to each well and read out on a ViewLux ultraHTS microplate imager (PerkinElmer). Dose response curves were generated from measurements performed in quadruplicates. The minimum effective concentration (LEC) value for each compound, defined as the trigger-out-of-assay target, was determinedQuasi-deviation is at least twice the concentration of the effect.

In parallel, a similar dilution series of compound (10. mu.L of compound in 4% DMSO) and 30. mu.L per well of cells transfected individually with the NF-. kappa.B-luc reporter construct (1.67X 10)⁵Individual cells/mL). At 37 deg.C, 5% CO₂Luciferase activity was determined 6 hours after the next incubation by adding 15 μ l of Steady Lite Plus substrate (perkin elmer) to each well and reading on a ViewLux ultraHTS microplate imager (perkin elmer). The counter screen number is reported as LEC.

In the HEK293 TOX assay described above, all compounds showed CC50>24 μ M.

Claims (5)

1. A compound of formula (I)

(I)

Or a pharmaceutically acceptable salt thereof, wherein

R₁Is hydrogen, fluorine, hydroxy, amino, C_1-6Alkyl radical, C_1-6Alkylamino radical, C_1-6Alkoxy radical, C_3-7Cycloalkyl radical, C_4-7Heterocyclyl, aryl, bicyclic heterocyclyl, aralkyl, heteroarylOptionally substituted with one or more substituents independently selected from halogen, hydroxy, amino, C_1-6Alkyl, di- (C)_1-6) Alkyl-amino, (C)₁-₄) Alkoxy radical- (C_1-4) Alkyl radical, C_1-6Alkylamino radical, C_1-6Alkoxy radical, C_3-6A cyclo-alkyl group, a carboxyl group, a carboxylate group, a carboxylic acid amide group, a heterocyclic group, an aryl group, an alkenyl group, an alkynyl group, an aralkyl group, a heteroaryl group, a heteroaralkyl group, or a nitrile group,

R₂is C each of which may be optionally substituted by one or more substituents_1-6Alkyl, these substituents are independently selected from halogen, hydroxy, amino, carboxy, carboxylate, carboxamide, C_1-3Alkyl radical, C_1-3Alkoxy or C_3-6Cycloalkyl, sulfone, sulfoxide, or nitrile groups,

with the proviso that N- (2-amino-5-phenethylpyrimidin-4-yl) -N-pentylamine is excluded.

2. The compound of claim 1, wherein R₁Is a heterocyclic radical and R₂Is C substituted by hydroxy_1-6An alkyl group.

3. The compound of claim 1, wherein R₁Is hydrogen and wherein R₂Is C each of which may be optionally substituted by one or more substituents₁-₆Alkyl, these substituents are independently selected from halogen, hydroxy, amino, carboxy, carboxylate, carboxamide, C_1-3Alkyl radical, C_1-3Alkoxy or C_3-6A cycloalkyl group, a sulfone group, a sulfoxide group, or a nitrile group.

4. A pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, as claimed in any one of claims 1-3, together with one or more pharmaceutically acceptable excipients, diluents or carriers.

5. Use of a compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1-3, or a pharmaceutical composition according to claim 4, in the manufacture of a medicament for the treatment of a disorder in which TLR7 modulation and/or TLR8 modulation is implicated.