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    • 专利摘要:
    4-hydroxyphenyl phosphonium salts with antiparasitic properties. The present invention provides a new series of compounds derived from 4-hydroxyphenyl phosphonium. Furthermore, the invention relates to a pharmaceutical composition comprising said compounds and to the use in the treatment or prevention of diseases caused by parasites, preferably trypanosomatid parasites. In particular, these compounds have demonstrated in vivo efficacy for the treatment of visceral leishmaniasis in a murine model. These diseases can occur in both humans and animals. (Machine-translation by Google Translate, not legally binding)
    公开号:ES2801674A1
    申请号:ES201930604
    申请日:2019-07-01
    公开日:2021-01-12
    发明作者:Christophe Dardonville；Díaz Eduardo José Cueto；Conde Francisco Gamarro；González José Ignacio Manzano；Martínez Ana Perea；Rodríguez José María Alunda；Durán Juan José Torrado；Molero Ana Isabel Olías
    申请人:Consejo Superior de Investigaciones Cientificas CSIC；Universidad Complutense de Madrid；
    IPC主号:
    专利说明:

    [0005] The present invention relates to 4-hydroxyphenylphosphonium derivatives and their potential in the treatment of diseases, both human and animal, caused by trypanosomatid parasites (leishmaniasis, trypanosomiasis). Therefore, the invention is framed in the pharmaceutical and veterinary sector.
    [0007] STATE OF THE TECHNIQUE
    [0009] Human leishmaniasis is a disease with worldwide distribution, the prevalence of which reaches 12 million cases. It appears mainly in tropical and subtropical areas, although it is also found in Mediterranean countries. Furthermore, there is a strong incidence of Leishmania infection in immunosuppressed patients, especially among AIDS patients (Alvar, J. et al. Adv. Parasitol. 2006 , 61, 223-74). Leishmania parasites produce different forms of the disease ranging from milder cutaneous leishmaniasis to visceral leishmaniasis, a much more severe form of the disease that is fatal if not treated properly. The Leishmania species that appears in the Mediterranean area and specifically in Spain is L. infantum, which produces cutaneous and visceral forms of the disease. Treatment options for treating leishmaniasis include the pentavalent antimonial derivatives (sodium stibogluconate and meglumine antimoniate), the antibiotic amphotericin B and its liposomal formulation AmBisome®, the antibiotic paromomycin, the alkylphosphocholine derivative miltefosine, and 8-aminoquinoline diamiditamaquine sinephosine pentamidine (Mishra, J. et al. Curr. Med. Chem. 2007 , 14, 1153-69). However, these drugs have several limitations such as their toxicity, difficult administration, high price and, most worryingly, the appearance of strains resistant to antimonials (Croft, SL et al. Clin Microbiol Rev 2006 , 19, 111- 26) and miltefosine (Srivastava, S. et al. Parasites & Vectors 2017 , 10, 49) In addition, the treatment of the most complex forms of the disease, as well as in cases of co-infection with HIV, is not very effective.
    [0010] The in vivo activity of tetraphenylphosphonium bromide on L. braziliensis (Hanson, WL et al. Technical reports ADA283312, 1994, 1-43) and the in vitro leishmanicidal activity of bisphosphonium salts derived from benzophenone (Luque-Ortega , JR et al. J. Med. Chem. 2010 , 53, 1788-98). In addition, phosphonium salts with activity on Trypanosoma brucei derived from benzophenone (Taladriz, A. et al. J. Med. Chem. 2012 , 55, 2606-2622), 4-hydroxybenzoate (Fueyo González, FJ et al. J. Med. Chem. 2017 , 60, 1509-1522; Ebiloma, GU et al. Eur. J. Med. Chem. 2018 , 150, 385-402), 4-hydroxybenzaldehyde and 4-hydroxybenzoic acid (Meco-Navas, A. et al. ACS Med. Chem. Lett. 2018 , 9, 923-928), triarylbenzylphosphonium halides (US Patent 4187300A). Also, triphenylphosphonium salts with activity on T. cruzi have been described (Long, TE et al. Bioorg. Med. Chem. Lett.
    [0011] 2012 , 22, 2976-2979; Cortes, LA et al. PLoS ONE 2015 , 10, e0136852). Furthermore, phosphonium salts with microbicidal activity have been described against the fungi Venturi inaequalis, Piricularia oryzae, Botrytis cinerea and Puccinia recondita, as well as against the protist Plasmopara vitícola (Patent DE3332716A).
    [0013] However, phosphonium salts derived from 1- (4-hydroxyphenyl) -oxoalkane with a long alkyl chain (> 14 carbons) between the benzoyl radical and the phosphonium group have not been described as leishmanicidal agents.
    [0015] An advantage of the compounds described in the present patent is that they are more stable against hepatic and plasma metabolism than the benzoate derivatives previously described. Furthermore, these compounds are more selective against Leishmania than the mentioned benzoates, and are effective on drug resistant strains.
    [0017] DESCRIPTION OF THE INVENTION
    [0019] The compounds described in the present invention may be useful for treating infections caused by trypanosomatid protozoa ( Leishmania and Trypanosoma).
    [0021] Therefore, a first aspect of the present invention refers to a compound of general formula (I), pharmaceutically acceptable salt, or solvate thereof.
    [0023] where:
    [0024] R1 is selected from OH and CH3;
    [0025] R2 and R3 are independently selected from C1-C6 alkyl, cycloalkyl, aryl, or heterocycle;
    [0026] X is a halogen.
    [0028] Hereafter you can refer to the compounds of formula (I) as compounds of the invention.
    [0030] The term "solvate" is intended to indicate a pharmaceutically acceptable solvate form of a specified compound that maintains the biological efficacy of said compound. Examples of solvates include compounds of the invention together with water, isopropanol, ethanol, methanol, DMSO (dimethylsulfoxide), ethyl acetate, acetic acid, or ethanolamine. The term "hydrate" is used when said solvent is water. The solvation methods are of general knowledge in the field.
    [0032] The term "pharmaceutically acceptable salt" refers to salts prepared from non-toxic pharmaceutically acceptable bases including inorganic bases and organic bases. Salts derived from inorganic bases include aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic, manganous, potassium, sodium, zinc salts and the like. Salts derived from pharmaceutically acceptable non-toxic organic bases include salts of primary, secondary and tertiary amines, substituted amines including natural substituted amines, cyclic amines, and basic ion exchange resins, such as arginine, betaine, caffeine, choline, N, N '-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethyl-morpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, morpholine, pipeline resphine polyamine, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, and Similar. When the compound of the present invention is basic, salts can be prepared from pharmaceutically acceptable non-toxic acids, including inorganic and organic acids. Such acids include acetic, benzenesulfonic, benzoic, camphorsulfonic, citric, ethanesulfonic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic, mandelic, methanesulfonic, mucic, nitric, pamophoric, pantothenic, acids. , sulfuric, tartaric, ptoluenesulfonic and the like.
    [0034] Preferred examples of pharmaceutically acceptable salts include ammonium, calcium, magnesium, potassium, and sodium salts, and those formed from maleic, fumaric, benzoic, ascorbic, pamoic, succinic, hydrochloric, sulfuric, bismethylene salicylic, methanesulfonic, ethanedisulfonic, propionic acids. , tartaric, salicylic, citric, gluconic, aspartic, stearic, palmitic, itaconic, glycolic, p-aminobenzoic, glutamic, benzenesulfonic, cyclohexylsulfamic, phosphoric and nitric.
    [0036] The term "alkyl" refers, in the present invention, to straight or branched saturated hydrocarbon chains having 1 to 6 carbon atoms, for example, methyl, ethyl, n-propyl, / -propyl, n-butyl , tere-butyl, sec-butyl, n-pentyl, n-hexyl, cyclohexyl Alkyl groups may be optionally substituted by one or more substituents such as alkynyl, alkenyl, halogene, hydroxyl, alkoxy, carboxyl, cyano, carbonyl, acyl , alkoxycarbonyl, amino, nitro or mercapto.
    [0038] By "halogen" is meant in the present invention a bromine, chlorine, iodine or fluorine atom.
    [0040] In a preferred embodiment, R1 is selected from OH and CH3, and more preferably R1 is OH.
    [0042] In another preferred embodiment, R2 and R3 are aryl, and more preferably phenyl.
    [0044] In another preferred embodiment, R2 and R3 are 1-naphthalenyl.
    [0046] In another preferred embodiment, R2 is 2-pyridyl and R3 is phenyl.
    [0047] In another preferred embodiment, the compounds of formula (I) are selected from the list comprising:
    [0049] - (15- (2,4-dihydroxyphenyl) -15-oxopentadecyl) triphenylphosphonium bromide ( 1 ) TAO99 - (16- (4-hydroxy-2-methylphenyl) -16-oxohexadecyl) triphenylphosphonium ( 2 ) bromide TAO100
    [0050] - (16- (4-hydroxy-2-methylphenyl) -16-oxohexadecyl) diphenyl (pyridin-2-yl) phosphonium ( 3 ) bromide TAO101
    [0051] - (16- (2,4-dihydroxyphenyl) -16-oxohexadecyl) tri (naphthalen-1-yl) phosphonium ( 4 ) bromide TAO118
    [0053] Another aspect of the present invention relates to a pharmaceutical composition comprising a compound of general formula (I)
    [0057] where:
    [0058] R1 is selected from OH and CH3;
    [0059] R2 and R3 are independently selected from C1-C6 alkyl, cycloalkyl, aryl, or heterocycle;
    [0060] X is a halogen.
    [0062] together with a pharmaceutically acceptable carrier.
    [0064] The pharmaceutically acceptable carriers that can be used in said compositions are substances known to those skilled in the art and commonly used in the preparation of therapeutic compositions.
    [0066] The compounds and compositions of this invention can be used with other drugs to provide combination therapy. The other drugs can be part of the same composition, or they can be provided as a separate composition for administration at the same time or at a different time.
    [0067] Therefore, in another preferred embodiment, the composition of the invention is characterized in that it can also comprise at least one other active principle, preferably an antiparasitic agent.
    [0069] Another aspect of the present invention refers to the use of a compound, of general formula (I), for the manufacture of a medicament.
    [0071] Yet another aspect of the present invention refers to the use of a compound of the invention, of general formula (I), for the preparation of a medicine for the prevention and / or treatment of diseases caused by parasites, preferably diseases caused by parasites. protozoa. These diseases occur in an animal, preferably a mammal, including humans.
    [0073] In a preferred embodiment, the diseases are diseases caused by a parasite of the genus Leishmania or Trypanosoma. And even more preferably the parasites are of the species L. aethiopica, L. amazonensis, L. arabica, L. archibaldi, L. aristedesi, L. braziliensis, L. chagasi ( syn. L. infantum), L. colombiensis, L. . deanei, L. donovani, L. enriettii, L. equatorensis, L. forattinii, L. garnhami, L. gerbili, L. guyanensis, L. herreri, L. hertigi, L. infantum, L. killicki, L. lainsoni , L. major, L. mexicana, L. naiffi, L. panamensis, L. peruviana, L. pifanoi, L. shawi, L. tarentolae, L. tropica, L. turanica, L. venezuelensis, Trypanosoma brucei, T. cruzi, T. congolense, T. equinum, T. equiperdum, T. evansi, T. vivax.
    [0075] In another preferred embodiment, the disease is selected from leishmaniasis, human African trypanosomiasis (sleeping sickness), animal trypanosomiasis, or American trypanosomiasis (Chagas disease).
    [0077] Another aspect of the present invention refers to a method of treatment and / or prevention of parasitic diseases or diseases caused by parasites that comprises the administration to an infected individual (in need thereof) of a therapeutically effective amount of at least one compound of the invention of formula (I), or of the pharmaceutical composition as described in the present invention. The individual can be an animal, preferably a mammal, including humans.
    [0079] In the sense used in this description, the expression "therapeutically effective amount" refers to the amount of the agent or compound capable of developing the therapeutic action determined by its pharmacological properties, calculated to produce the desired effect and, in general, will be determined, among other causes, due to the characteristics of the compounds, as well as the age, condition of the patient, the severity of the alteration or disorder, and the route and frequency of administration.
    [0081] Said therapeutic composition can be prepared in solid form or in suspension, in a pharmaceutically acceptable diluent. The therapeutic composition provided by this invention can be administered by any appropriate route of administration, for which said composition will be formulated in the appropriate pharmaceutical form for the chosen route of administration.
    [0083] Throughout the description and claims the word "comprise" and its variants are not intended to exclude other technical characteristics, additives, components or steps. For those skilled in the art, other objects, advantages and characteristics of the invention will emerge partly from the description and partly from the practice of the invention. The following examples and figures are provided by way of illustration, and are not intended to be limiting of the present invention.
    [0085] DESCRIPTION OF THE FIGURES
    [0087] Fig. 1. Synthetic route for the preparation of the derivatives of formula (I): (i) HBr 48%, 110 ° C; (ii) BF3-Et2O, 110 ° C; (iii) triphenylphosphine (for 1 and 2 ) or diphenyl-2-pyridylphosphine (for 3 ), CH3CN, 80 ° C, 5-18 days; (iv) tris (naphthalen-1-yl) phosphane (para 4 ), DMF, 110 ° C, 5 days.
    [0089] Fig. 2 . Plasma concentration obtained after oral administration of 20 mg / kg of compound 1 in 5-8% DMSO to BALB / c mice. The values correspond to the mixture of the plasmas obtained at the sampling times (n = 3).
    [0090] Examples
    [0092] The invention will be illustrated below by means of tests carried out by the inventors, which demonstrate the specificity and effectiveness of the compounds described in the present invention.
    [0094] The compounds of formula (I) that are synthesized below are outlined in Figure 1.
    [0096] Example 1. Synthesis of the compounds of the invention
    [0098] 16-bromohexadecanoic acid (5). In a Kimax screw cap tube, a solution of 16-hydroxyhexadecanoic acid (1.0 g, 3.67 mmol) in acetic acid (4 mL) and 48% aqueous HBr (4 mL) is stirred at 110 ° C for 41 h. After the reaction mixture has cooled, the precipitated supernatant is collected by filtration, washed with water and dried under vacuum, obtaining 1,671 g of crude product. Compound 5 is recrystallized from hexane, obtaining white crystals (1074 g, 87%). 1H NMR (400 MHz, CDCl3) 53.34 (t, J = 6.9 Hz, 2H), 2.28 (t, J = 7.5 Hz, 2H), 1.79 (p, J = 7.2 Hz, 2H), 1.57 (p, J = 7.5 Hz, 2H), 1.40-1.12 (m, 22H). 13C NMR (101 MHz, CDCh) 5 179.9, 34.03, 34.02, 32.9, 29.62, 29.60, 29.58, 29.54, 29.44, 29.42, 29.2, 29.1, 28.8, 28.2, 24.7.
    [0100] 16-bromo-1- (2,4-dihydroxyphenyl) hexadecan-1-one (6a) In a Kimax tube (previously dried in the oven) add resorcinol (33 mg, 0.3 mmol) and 16-bromohexadecanoic acid 5 (104 mg, 0.3 mmol). Next, BF3- (Et2O) 2 (1.5 mL) is added under argon atmosphere. The tube is closed with the screw cap and the reaction mixture is stirred at 110 ° C for 2 h. The reaction mixture is then allowed to cool and is poured onto ice. The aqueous phase is extracted with EtOAc (25 mL). The organic phase is washed with brine (5 mL), dried (MgSO4), and evaporated to obtain a brown oil. The product is purified by chromatography on silica (10 g) eluting with hexane / EtOAc (100/0 ^ 90/10), obtaining 6a as a white amorphous solid (58 mg, 45%). HPLC (UV)> 90%. 1H NMR (300 MHz, Chloroform-d) 5 12.81 (s, 1H), 7.59 (d, J = 8.9 Hz, 1H), 6.40-6.20 (m, 2H), 6.11 (brs, 1H), 3.34 (t, J = 7.0 Hz, 2H), 2.82 (t, J = 7.5 Hz, 2H), 1.78 (p, J = 7.1 Hz, 2H), 1.65 (p, J = 7.3 Hz, 2H), 1.45-1.06 (m, 20H).
    [0101] 13C NMR (75 MHz, CDCI3) 5 205.6, 165.4, 162.7, 132.6, 114.0, 107.9, 103.7, 38.2, 34.3, 33.0, 29.76, 29.73, 29.67, 29.61,29.57, 29.52, 28.9, 28.3, 25.1.
    [0103] 16-bromo-1- (4-hydroxy-2-methylphenyl) hexadecan-1-one (6b). Compound 6b is obtained following the same procedure as for compound 6a starting from mcresol (30.6 mg, 0.28 mmol). The compound is purified by chromatography on silica with hexane / EtOAc (100/0 ^ 30/1) obtaining a white amorphous solid (15 mg, 13%). HPLC (UV)> 95%. 1H NMR (400 MHz, Chloroform-d) 5 12.36 (s, 1H), 7.57 (d, J = 8.3 Hz, 1H), 6.72 (d, J = 1.5 Hz, 1H), 6.63 (dd, J = 8.3, 1.5 Hz, 1H), 3.34 (t, J = 6.9 Hz, 2H), 2.89 - 2.84 (m, 2H), 2.28 (s, 3H), 2.25 - 2.18 (m, 2H), 1.83 - 1.74 (m, 2H ), 1.66 (p, J = 7.4 Hz, 2H), 1.59-1.50 (m, 2H), 1.39-1.17 (m, 18H). 13C NMR (101 MHz, CDCl3) 5 206.5, 162.8, 147.9, 130.0, 120.2, 118.7, 117.3, 38.4, 34.6, 34.2, 33.0, 29.74, 29.73, 29.68, 29.61, 29.59, 29.58, 29.49, 29.41, 29.29, 28.91 , 28.33, 25.14, 24.86.
    [0105] (15- (2,4-dihydroxyphenyl) -15-oxopentadecyl) triphenylphosphonium bromide (1) (TAO99).
    [0106] Add 6a (75 mg, 0.18 mmol) and Ph3P (51 mg, 0.19 mmol) in anhydrous acetonitrile (1.5 mL) to a Kimax screw-cap tube. The reaction mixture is stirred for 18 days at 80 ° C under argon atmosphere. The tube is then left in the refrigerator (4 ° C) for several days until a white precipitate appears. The supernatant is removed and a little acetonitrile is added to wash off the precipitate. The tube is centrifuged and the supernatant is removed. The same operation of washing the precipitate with Et2O is repeated. Product 1 is isolated as a white amorphous solid (85mg, 100%). HPLC> 95%, 1H NMR (400 MHz, Chloroform-d) 5 12.71 (s, 1H), 9.70 (s, 1H), 7.78 - 7.56 (m, 15H), 7.48 (d, J = 8.9 Hz, 1H) , 6.55 (dd, J = 8.9, 2.4 Hz, 1H), 6.33 (d, J = 2.4 Hz, 1H), 3.41 (td, J = 13.1, 12.6, 7.3 Hz, 2H), 2.74 (t, J = 7.5 Hz, 2H), 1.60 (q, J = 7.4 Hz, 2H), 1.56-1.42 (m, 4H), 1.30-1.05 (m, 20H). 13C NMR (101 MHz, Chloroform-d) 5 205.2, 165.3, 165.0, 135.3 (d, J = 3.0 Hz), 133.6 (d, J = 9.9 Hz), 132.2, 130.7 (d, J = 12.5 Hz), 118.2 (d, J = 86.0 Hz), 112.8, 108.7, 103.4, 37.9, 30.6 (d, J = 15.5 Hz), 29.5, 29.4, 29.3, 29.2, 29.14, 29.11, 29.08, 25.2 (d, J = 1.8 Hz) , 22.8 (d, J = 50.3 Hz), 22.7 (d, J = 4.5 Hz). HRMS (ESI +) m / z 609.3482 (C40H50PO3 requires: 609.3498).
    [0107] (16- (4-Hydroxy-2-methylphenyl) -16-oxohexadecyl) triphenylphosphonium (2) bromide (TAO100). In a Kimax tube, add 6b (6.9 mg, 0.016 mmol), Ph3P (4.2 mg, 0.016 mmol) and 0.6 mL of anhydrous DMF under argon atmosphere. The tube is closed with the screw cap and the reaction mixture is stirred at 80 ° C for 5 days. The solvent is evaporated and the product dissolved in MeOH is isolated by precipitation with Et2O / hexane at 4 ° C overnight. The supernatant is removed and the precipitate is washed successively with Et2O and hexane. Off-white amorphous solid (7.6 mg, 69%). HPLC> 95% .1H NMR (400 MHz, Chloroform-d) 512.36 (s, 1H), 7.84 - 7.74 (m, 6H), 7.72 (td, J = 7.3, 1.8 Hz, 3H), 7.63 (td, J = 7.7, 3.5 Hz, 6H), 7.57 (d, J = 8.2 Hz, 1H), 6.71 (d, J = 1.7 Hz, 1H), 6.63 (dd, J = 8.2, 1.7 Hz, 1H), 3.76 (dt , J = 13.2, 7.4 Hz, 2H), 2.87 (t, J = 7.5 Hz, 2H), 2.27 (s, 3H), 1.66 (p, J = 7.4 Hz, 2H), 1.60 - 1.50 (m, 2H) , 1.38-1.07 (m, 22H). 13C NMR (101 MHz, Chloroform-d) 5206.6, 162.8, 147.9, 135.1 (d, J = 3.0 Hz), 133.9 (d, J = 10.0 Hz), 130.6 (d, J = 12.5 Hz), 130.1, 120.3, 118.7 (d, J = 85.6 Hz), 118.6, 117.3, 38.4, 30.6 (d, J = 15.5 Hz), 29.71, 29.69, 29.67, 29.64, 29.57, 29.54, 29.45, 29.41, 29.3, 24.8, 23.0 (d, J = 49.3 Hz), 22.8 (d, J = 4.6 Hz), 22.1. HRMS (ESI +) m / z 607.3709 (C41H52PO2 requires: 607.3705).
    [0109] (16- (4-Hydroxy-2-methylphenyl) -16-oxohexadecyl) diphenyl (pyridin-2-yl) phosphonium (3) bromide TAO101. The same procedure is used as for the synthesis of compound 2 . A mixture of 6b (5.7 mg, 0.014 mmol) and Ph3P (3.7 mg, 0.014 mmol) in anhydrous acetonitrile (0.6 mL) is stirred at 80 ° C for 5 days under argon atmosphere. The solvent is evaporated and the product dissolved in MeOH is isolated by precipitation with Et2O at 4 ° C overnight. The supernatant is removed and the precipitate is washed successively with Et2O and hexane. Brown amorphous solid (4.6 mg, 48%). HPLC> 95%. 1H NMR (400 MHz, Chloroform-d) 512.36 (s, 1H), 8.81 (dt, J = 4.7, 1.5 Hz, 1H), 8.42 (ddt, J = 7.9, 5.9, 1.1 Hz, 1H), 8.09 (tdd , J = 7.9, 5.1, 1.8 Hz, 1H), 7.87 - 7.78 (m, 4H), 7.77 - 7.70 (m, 2H), 7.62 (tdd, J = 9.1, 4.5, 3.2 Hz, 5H), 7.57 (d , J = 8.0 Hz, 1H), 6.71 (dd, J = 1.6, 0.9 Hz, 1H), 6.63 (dd, J = 8.0, 1.6 Hz, 1H), 3.75 - 3.62 (m, 2H), 2.92 - 2.82 ( m, 2H), 1.72-1.46 (m, 4H), 1.37-1.07 (m, 22H). 13C NMR (101 MHz, Chloroform-d) 5206.6, 162.8, 151.9 (d, J = 19.1 Hz), 147.9, 145.4, 144.2, 138.8 (d, J = 10.3 Hz), 135.1 (d, J = 3.0 Hz), 134.1 (d, J = 9.6 Hz), 132.5 (d, J = 24.5 Hz), 130.5 (d, J = 12.6 Hz), 130.1, 128.1 (d, J = 3.5 Hz), 120.3, 118.6, 117.9 (d, J = 85.4 Hz), 117.3, 38.4, 30.6 (d, J = 15.2 Hz), 29.72, 29.70, 29.68, 29.63, 29.58, 29.56, 29.46, 29.32, 29.28, 24.8, 22.62, 22.59 (d, J = 13.6 Hz), 22.1, 22.0. HRMS (ESI +) m / z 608.3666 (C40H51PNO2 requires: 608.3657).
    [0111] (16- (2,4-dihydroxyphenyl) -16-oxohexadecyl) tri (naphthalen-1-yl) phosphonium bromide (4) (TAO118). In a Kimax tube, add 6a (53.6 mg, 0.13 mmol), tris (naphthalen-1-yl) phosphane (67.0 mg, 0.16 mmol) and 2.0 mL of anhydrous DMF under argon atmosphere. The tube is closed with the screw cap and the reaction mixture is stirred at 110 ° C for 5 days. The solvent is evaporated and the crude solid obtained is purified by chromatography on silica with hexane / EtOAc (0 ^ -20%), obtaining a mixture of 2 with tri (naphthalen-1-yl) phosphine oxide. The latter is removed by preparative plate chromatography (PTLC) eluting with EtOAc / MeOH (8/2). Pure compound 4 is an off-white solid (7mg, 6.4%). HPLC> 95%. 1H NMR (500 MHz, Methanol-d4) 58.43 (dd, J = 8.5, 1.5 Hz, 3H), 8.20 - 8.11 (m, 6H), 7.97 (d, J = 8.6 Hz, 3H), 7.78 - 7.61 (m , 7H), 7.54 - 7.47 (m, 3H), 6.33 (ddd, J = 8.8, 2.4, 0.6 Hz, 1H), 6.20 (d, J = 2.4 Hz, 1H), 4.00 (br, 2H), 2.88 ( t, J = 7.4 Hz, 2H), 1.68 (p, J = 7.4 Hz, 2H), 1.63-0.95 (m, 22H). HRMS (ESI +) m / z 759.3981 (C52H56O3P requires 759.3967).
    [0113] Example 2. Trials in vitro of the leishmanicidal activity of the compounds of the invention
    [0115] The compounds were evaluated in vitro against intracellular promastigote and amastigote forms of the parasite L. donovani (strain HU3) following the previously described MTT colorimetric method (Kennedy ML, et al. J Med Chem 2001 , 44, 4668-4676). Furthermore, its cytotoxicity on human THP-1 cells was evaluated and the selectivity index (IS) of each compound was calculated (Table 1).
    [0116] The compounds object of this patent show activities on intracellular promastigotes and amastigotes of L. donovani in the submicromolar range (0.07 - 0.28 pM), being respectively two and ten times more potent than the reference drugs amphotericin B and miltefosine in this essay. Furthermore, the compounds show selectivity towards Leishmania with selectivity indices (IS)> 20 against human THP-1 cells.
    [0117] Table 1.a
    [0119] Ref. Cmpto EC50 (pM)
    [0121] L. donovani IS “ L. donovani
    [0122] amastigotes THP-1 cells promastigotes
    [0123] intracellular
    [0125] TAO99 1 0.07 ± 0.02 0.09 ± 0.02 2.81 ± 0.35 31.2
    [0126] TAO100 2 0.09 ± 0.01 0.10 ± 0.01 2.45 ± 0.41 24.5
    [0127] TAO101 3 0.28 ± 0.05 0.10 ± 0.02 2.11 ± 0.12 21.1
    [0128] TAO118d 4 3.54 ± 0.36 1.25 ± 0.22 12.85 ± 2.30 10.3
    [0130] AmBe 0.13 ± 0.02 0.24 27.85 116.0
    [0131] MIL f 6.51 ± 0.35 0.91 ± 0.05 26.25 ± 2.58 28.8
    [0132] a Parasites and THP-1 cells were cultured in the presence of increasing concentrations of the compounds. Data represent the mean ± standard deviation of 3 independent experiments. b IS = selectivity index (EC50 THP-1 / EC50 parasite (amastigote form)). c not tested. d Sensitivity data obtained in L. infantum LEM3049. e Amphotericin B. f Miltefosine.
    [0134] Example 3. Activity of compound 1 against drug-resistant strains of L. infantum.
    [0136] No significant differences were found in sensitivity to compound 1 between the
    [0137] reference line drug-sensitive (LEM3049) and the strains LEM 3323 (resistant to
    [0138] Sb11), LEM 5159 (resistant to miltefosine) and LEM 2126 (resistant to miltefosine and
    [0139] paramomycin) (Table 2).
    [0141] Table 2. Leishmanicidal activity of compound 1 on intracellular promastigote and amastigote forms of clinical isolates of L. infantuma
    [0143] ECso (pM)
    [0145] Promastigotes Intracellular amastigotes L. infantum Cmpto 1 AmB Cmpto 1 AmB
    [0146] LEM3049 0.08 ± 0.03 0.06 ± 0.02 0.12 ± 0.06 0.17 ± 0.02
    [0147] LEM3323 0.10 ± 0.02 0.07 ± 0.03 0.15 ± 0.07 0.18 ± 0.02
    [0148] LEM5159 0.11 ± 0.02 0.05 ± 0.01 0.14 ± 0.02 0.16 ± 0.03
    [0149] LLM2126 0.12 ± 0.03 0.10 ± 0.02 0.14 ± 0.01 0.16 ± 0.01
    [0150] a Clinical isolates of L. infantum used: LEM 3049 as reference line; LEM 3323, resistant to Sbm; LEM 5159, resistant to miltefosine (MIL) and LEM 2126, resistant to MIL and paromomycin. Amphotericin B (AmB) was used as the reference drug. Data represent the mean ± standard deviation of 3 independent experiments.
    [0152] Example 4. In vivo efficacy in a murine model of visceral leishmaniasis The compound is poorly soluble in water, but is solubilized in 5-8% DMSO / water, 20% EtOH / water or can be formulated as a colloidal suspension in 5% Y -cyclodextrin in water for oral administration (Table 3).
    [0154] Table 3 . Thermodynamic solubility of compound 1 at 24 h
    [0159] Based on the pharmacokinetic, stability and absorption data, Compound 1 was administered orally in a BALB / c murine model of visceral leishmaniasis. Three groups of mice were established: (i) G1, uninfected mice, 30 pg of compound 1 per mouse for 4 days (once a day); (ii) G2, infected mice and -cyclodextrin in milliQ water, and (iii) G3, compound 1 infected mice following the same pattern described for G1.
    [0161] The parasite load in the target organs (liver and spleen) was determined 48 h after the end of treatment with compound 1 . In this way, it was observed a reduction in the load of 98.9% in spleen and 95.3% in liver when comparing the groups of mice infected and treated with compound 1 with those infected but not treated (Table 4).
    [0163] Table 4. In vivo activity of compound 1 in the murine model (BALB / c) of visceral leishmaniasis using Leishmania infantum.
    [0164] Passive load Study group
    [0166]
    [0168] G2 (Infected vehicle [5% Y -cyclodextrin] 4.92 ± 1.21a 607.25 ± 176.96 po for 4 days)
    [0169] G3 (Infected compound 1 0.18 ± 0.07 (95.3) c 6.47 ± 1.77 (98.9) 30 pg po for 4 days)
    [0171] aMean number of amastigotes per milligram of tissue. bp.o., orally. c The reduction percentage compared to G2 is reflected in the value in parentheses.
    [0173] The reduction observed in the parasite load in target organs after treatment with compound 1 at a low dose (1.5 mg / kg once daily for 4 days) in our mouse model is similar to that obtained with miltefosine at a much higher dose. (ie 40 mg / kg once a day for 5 days) in the case of said drug (Sebastián-Pérez, V. et al. Antimicrob. Agents Chemother. 2018 , 62, e00603-00618). Therefore, Compound 1 is more potent in vivo than the oral reference drug miltefosine.
    [0175] Example 5. Pharmacokinetics, bioavailability and hepatic metabolism
    [0176] The bioavailability of 1 after oral administration of 20 mg / kg of compound in 5-8% DMSO to BALB / c mice is very high (Cmax = 182 pM) and very rapid (Tmax = 1 h) (Table 5 and Figure 2).
    [0177] The hepatic metabolism of compound 1 is species dependent. The microsomal stability of 1 towards metabolism by cytochrome P450 (Phase I metabolism) and uridine glucuronosyl transferase (UGT) (Phase II metabolism) in the presence of NADPH and UDPGA was studied. Compound 1 was rapidly metabolized by mouse liver microsomes (CD-1) (intrinsic high clearance) with half-life <30 min, confirming the pharmacokinetic data observed in vivo. In contrast, 1 was metabolized slowly by human liver microsomes (low to moderate intrinsic clearance) with t1 / 2> 2 h, and very slowly by the human liver S9 fraction within 2 h of the reaction (Table 5). In comparison, a high intrinsic clearance of the control drug diclofenac was observed by human fractions under the same conditions (t1 / 2 <30 min).
    [0179] Table 5. Pharmacokinetics and hepatic metabolism
    [0181]
    [0184] Example 6. In vitro tests of the trypanocidal activity of the compounds of the invention.
    [0186] The compounds of the invention show activities in the nanomolar range against the sensitive strain (s427) and the drug-resistance strain B48 of the Trypanosoma brucei parasite (Table 6).
    [0188] Table 6 .a
    [0189] Ref. Cmpto EC50 (pM)
    [0191] T. bruceib T. brucei B48c FRd
    [0194] Pente 0.0033 ± 0.0006 0.55 ± 0.03 168
    [0195] Dimin ^ 0.101 ± 0.005 1.33 ± 0.14 13.1 a L - o - s --- d - a - t - o - s --- r - e - p - r - e - s - e - n - ta - n --- l - a --- m - e - d - i - a --- ± --- d - e - s - v - ia - c - ió - n --- e - s - t - á - n - d - a - r - d - e --- 3 --- e - x - p - e - r - im --- e - n - t - o - s --- in - d - e - p - e - n - d - ie - n - t - e - s - - . - b Strain s427 of T. b. brucei. Strain B48 of T. b. brucei resistant to pentamidine and diminacene. FR = resistance factor (EC50 T. b. B48 / EC50 T. b. S427). e Pentamidine. f Diminacene.
    权利要求:
    Claims (15)
    [1]
    1. Compound of general formula (I), pharmaceutically acceptable salt, or solvate thereof

    [2]
    2. Compound according to claim 1, wherein R1 is OH.
    [3]
    3. Compound according to any of claims 1-2, wherein R2 and R3 are aryl, and more preferably phenyl.
    [4]
    4. Compound according to any of claims 1-2, wherein R2 and R3 are 1-naphthalenyl.
    [5]
    5. Compound according to any of claims 1-2, wherein R2 is 2-pyridyl and R3 is phenyl.
    [6]
    6. A compound according to claim 1 selected from the list comprising:
    - (15- (2,4-dihydroxyphenyl) -15-oxopentadecyl) triphenylphosphonium bromide ( 1 ) TAO99 - (16- (4-hydroxy-2-methylphenyl) -16-oxohexadecyl) triphenylphosphonium ( 2 ) bromide TAO100
    - (16- (4-hydroxy-2-methylphenyl) -16-oxohexadecyl) diphenyl (pyridin-2-yl) phosphonium ( 3 ) bromide TAO101
    - (16- (2,4-dihydroxyphenyl) -16-oxohexadecyl) tri (naphthalen-1-yl) phosphonium ( 4 ) bromide (TAO118)
    [7]
    7. Pharmaceutical composition comprising a compound of general formula (I)
    described according to any one of claims 1 to 6, together with a vehicle
    pharmaceutically acceptable.
    [8]
    8. Composition according to claim 7 that also comprises another active principle
    antiparasitic.
    [9]
    9. Compound of general formula (I) described according to any of the claims
    1 to 6 for use as a medicine.
    [10]
    10. Compound of general formula (I) for use as a medicine according to
    claim 9, wherein the drug is for the prevention and / or treatment of
    diseases caused by parasites.
    [11]
    11. Compound of general formula (I) for use as a medicine according to
    claim 10, where the disease is caused by protozoan parasites.
    [12]
    12. Compound of general formula (I) for use as a medicine according to
    claim 11, where the protozoan parasites are of the genus Leishmania or
    Trypanosoma.
    [13]
    13. Compound of general formula (I) for use as a medicine according to claim 12, where the parasites are of the species L. aethiopica, L. amazonensis, L. arabica, L. archibaldi, L. aristedesi, L. braziliensis, L. chagasi ( syn. L. infantum), L. colombiensis, L. deanei, L. donovani, L. enriettii, L. equatorensis, L. forattinii, L. garnhami, L. gerbili, L. guyanensis, L. herreri , L. hertigi, L. infantum, L. killicki, L. lainsoni, L. major, L. mexicana, L. naiffi, L. panamensis, L. peruviana, L. pifanoi, L. shawi, L. tarentolae, L tropica, L. turanica, L. venezuelensis, Trypanosoma brucei, T. cruzi, T. congolense, T. equinum, T. equiperdum, T. evansi, T. vivax.
    [14]
    14. Compound of general formula (I) for use as a medicine according to any of claims 9 to 13, wherein the diseases occur in animals, preferably in a mammal.
    [15]
    15. Compound of general formula (I) for use as a medicine according to any one of claims 9 to 14, wherein the disease is selected from leishmaniasis, human African trypanosomiasis (sleeping sickness), animal trypanosomiasis, American trypanosomiasis (Chagas disease) .
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    同族专利:
    公开号 | 公开日
    ES2801674B2|2021-09-29|
    WO2021001587A1|2021-01-07|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    US4187300A|1978-12-20|1980-02-05|The United States Of America As Represented By The Secretary Of The Army|Use of phosphonium salts in treatment of African trypanosomiasis|
    DE3332716A1|1983-09-10|1985-03-28|Hoechst Ag, 6230 Frankfurt|Novel quaternary phosphonium salts and their use as microbicidal agents|
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