Spiranic compounds derived from oxindol-pirazolo [3,4-b] pyridinone and its therapeutic uses (Machin

专利摘要:
Espiranic compounds derived from oxindol-pyrazolo [3,4-b] pyridinone and its therapeutic uses. The present invention relates to oxindol-pyrazolo [3,4-b] pyridinone derivatives, which act as modulators of amp-activated protein kinase (ampk) and its use for the treatment and prevention of diseases or disorders regulated by ampk. Therefore, these compounds may be useful for the treatment of inflammatory, autoimmune, cardiovascular, neurological and cancer diseases. (Machine-translation by Google Translate, not legally binding)
公开号:ES2623082A1
申请号:ES201531786
申请日:2015-12-10
公开日:2017-07-10
发明作者:Ana Castro Morera；Pascual SANZ BIGORRA；Sergio QUESADA SÁNCHEZ；María Adelaida GARCÍA GIMENO；Francisco Javier Luque Garriga；Axel BIDON-CHANAL BADIA
申请人:Consejo Superior de Investigaciones Cientificas CSIC；Universitat Autonoma de Barcelona UAB；Universitat de Barcelona UB；Centro de Investigacion Biomedica en Red CIBER；
IPC主号:

专利说明:

The present invention relates to a series of compounds derived from oxindolpyrazolo [3,4-b] pyridinone and their use for the treatment of inflammatory, autoimmune, cardiovascular, neurological and cancer diseases. It also refers to pharmaceutical compositions containing said compounds. Therefore, the
The invention belongs to the field of pharmaceutical chemistry.
STATE OF THE TECHNIQUE
Obesity, type 2 diabetes, hypertension, cancer and cardiovascular diseases
15 involve serious changes in glucose or lipid metabolism and severely affect the health and quality of life of affected individuals. The growing prevalence of these diseases makes finding new pharmacological targets for treatment an urgent task.
20 In this context, serine / threonine protein kinase activated by AMP (AMPK) is a relevant biological target for this type of pathology, as it acts as a key sensor of cellular energy status, coordinating metabolic pathways and non-metabolic processes with the objective to balance the level of nutrient supply and energy demand. AMPK responds to changes in intracellular AMP levels,
25 stimulating routes associated with the generation of ATP and / or inhibiting processes that consume ATP. Consequently, the synthesis of glucose, lipids and proteins as well as cell growth are inhibited, while fatty acid oxidation and glucose reuptake are activated. [Hardie DG; Ross FA et al. 2012 Nature Reviews
13: 251-262]. Thus, compounds capable of regulating energy metabolism through
30 of the AMPK modulation constitute a promising strategy for the prevention and treatment of this type of diseases.
At the pharmacological level, the concept of AMPK as a potential target for treating metabolic syndrome has been supported by the discovery of two main classes of antidiabetic drugs, thiazolidinediones (rosiglitazone, troglitazone and pioglitazone) and biguanides (metformin and fenformin), which activate AMPK in cells in
cultivation and in vivo. Traditionally rosiglitazone is considered a PPARy agonist and exerts its antidiabetic effects through adipocyte differentiation. AMPK could participate in the antidiabetic effects of rosiglitazone (Kadowaki T. et al., The Journal of Clinicallvestigation 116: 1784-1792, 2006]. In turn, metformin could
5 activate AMPK in vitro and in vivo by inhibiting complex I {Zhou G. et al., The Journal of Clinical Investigation 108: 1167-1174, 2001], and the hypoglycemic effect could be completely blocked by inactivation of the LKB1 kinase , confirming the key role of AMPK in mediating the antidiabetic effect of metformin (Shaw RJ et al., Seienee (New York) NY 310: 1642-1646,2005].
10 Similarly, the action of AMPK on energy metabolism would be responsible for the beneficial effects that the activation of this kinase has on the heart, especially in the protection against damage caused by ischemia-reusion. It has also been seen that AMPK activation decreases hypertrophy
15 and the risk of heart failure, so the administration of AMPK activators would be recommended in these pathologies (Zaha VG and Young LH. 2012 Circus Res. 11 1: 800-814].
More recently, AMPK's participation in regulation has become evident
20 of metabolism, not only cellular but also bodily. It has been shown that the hormone leptin derived from adipocytes leads to a stimulation of AMPK and, therefore, an increase in the oxidation of fatty acids in skeletal muscles (Minokoshi Y. et al, 2002, Nature, 415, 339). Adiponectin, another hormone derived from adipocytes that leads to an improved carbohydrate and lipid metabolism,
25 stimulates AMPK in liver and skeletal muscles (Yamanauchi T. et al., 2002, Nature Medicine, 8, 1288]. Activation of AMPK in these circumstances does not appear to be due to an increase in AMP cell levels, but rather to phosphorylation of the catalytic subunit by one or more upstream kinases.
30 Based on the aforementioned metabolic consequences of AMPK activation, profound beneficial effects of AMPK activation in vivo can be expected. Thus, in the liver the gene expression of gluconeogenic enzymes would be diminished, which would reduce liver glucose production and improve global glucose homeostasis. On the other hand, direct inhibition and / or expression
Reduced key enzymes in lipid metabolism would reduce their synthesis and also increase fatty acid oxidation. This would lead to an improvement in glucose homeostasis and, due to a reduction in the accumulation of triglycerides in skeletal muscle, also an improved insulin action. Therefore, the combination of these effects in metabolic syndrome should significantly reduce the risk of acquiring cardiovascular diseases.
AMPK participates in the regulation of the PI3k / Akt! MTOR route. MTOR is a serinaltreonin kinase enzyme that regulates protein synthesis. AMPK negatively regulates the mTOR route at various levels. First, AMPK phosphorylates and activates the tuberous sclerosis 2 (TSC2) protein, which is an mTOR inhibitor. Secondly, AMPK phosphorylates and inactivates the raptor protein, a subunit of the complex in which mTOR is found, leading to inactivation of the complex. In this way, the activation of AMPK inhibits cell growth and protects cells from glucose-induced apoptosis (Burkewitz K, Zhang and and Mair
W.B. 2014 Cell. Metabolism 20: 10-25) The AMPK can also be a target
Therapeutic for many cancers that have constitutive action on the PI3K / Akt signaling pathway. The treatment of various cancer cell lines by AlGAR (5-amino-1-¡-3-D-ribofuranosyl-imidazol-4-carboxamide), an AMP analogue capable of stimulating AMPK, attenuates cell proliferation both in in vitro studies and in in vivo [Rattan R, Giri S, Singh AK, Singh 1. 2005 J. Biol. Ghem. 280: 39582
20 39593).
AMPK activation by AICAR reduces the expression of lipogenic enzymes
FAS and ACC, resulting in suppression of cell proliferation
cancer prostates Many cancer cells display a speed
25 markedly increased de novo fatty acid synthesis correlated with high levels of FAS. FAS inhibition suppresses the proliferation of cancer cells and induces cell death. Thus, the activation of AMPK and the inhibition of FAS activity is a target for pharmacological therapy of this type of cancer.
30 On the other hand, AlGAR is an activator of AMPK that exerts an anti-inflammatory effect by attenuating the production of cytokines and pro-inflammatory mediators. Likewise, AlGAR attenuates the progression of experimental autoimmune encephalomyelitis, limiting leukocyte infiltration through the blood brain barrier, so it has been suggested that AMPK activating agents act as anti-
35 inflammatory and can maintain a therapeutic potential in KrabbelTwitcher's disease [Giri S et al. 2008, J. Neurochem., 105: 1820-1833).
It has also been described that the activation of AMPK plays a neuroprotective role in different neurodegenerative diseases. Thus, its activation can exert a neuroprotective effect through the regulation of PGC-1 and UCP2 levels against brain damage induced during epilepsy states (Han et aL, 2011, Neurosci Lett
5 500: 133-138] or through its role in processes of excitoxicity and apoptosis inhippocampal neurons [Ullah el al., 2014, CNS Neurosci. Ther. 20: 327-338].
DESCRIPTION OF THE INVENTION
The authors of the present invention have found a family of compounds characterized by being potent modulators of AMPK. Therefore, the compounds of the invention may be useful in the treatment or prophylaxis of diseases that are related to the regulation of said enzyme.
In a first aspect, the present invention relates to a compound of general formula (1):
(one)
or their corresponding pharmaceutically acceptable isomers, salts or solvates, in which R1-Rs Y X are defined below.
R1 is selected from hydrogen, halogen, substituted or unsubstituted (C1-C10) alkyl,
ORa, SRa, COO (Ra), CF), N (Rah Y a substituted or unsubstituted aryl group (CS-C1S).
Preferably R1 is selected from chlorine, bromine and a substituted phenyl group.
R2 is selected from hydrogen, substituted or unsubstituted (C1-C10) alkyl and CORa. Preferably R2 is hydrogen.
R3 is selected from hydrogen, substituted or unsubstituted (C1-C1O) alkyl and aryl (Cs
elS) substituted or unsubstituted. Preferably R3 is selected from hydrogen, a phenyl group and 4-methoxyphenyl.
5 ~ and Rs are independently selected from hydrogen, substituted aryl (CS-C1S) orunsubstituted, halogen and nitro group (-N02). Preferably ~ is selected fromhydrogen and fluorine, and Rs is selected from hydrogen, fluorine, a nitro group and a groupphenyl.
10 X is a halogen, preferably fluorine. R ... is selected from hydrogen, substituted or unsubstituted (C 1 -C 0) alkyl and substituted or unsubstituted aryl (CselS).
15 Where the substituents are selected from OR ', = 0, SR', SOR ', S02R', N02, NHR ', N (R'¡', = N-R ', NHCOR', N (COR'¡ ' , NHSO, R ', NR'C (= NR') NHR ', CN, halogen, C (O) R', COOR ', OC (O) R', CONHR ', CON (R') "alkyl (C , -C ,,) substituted or unsubstituted, substituted or unsubstituted (C2-C 10) alkenyl, substituted or unsubstituted alkynyl (e2-Cl0), substituted or unsubstituted aryl (CS-C1S) and heterocycle (CS-C1S ) replaced or
20 unsubstituted, where each R 'group is independently selected from H, OH, NO "NH" SH, CN, halogen, O, C (O) H, C (O) alkyl, COOH, alkyl (C, -C, ,) substituted or unsubstituted, substituted or unsubstituted (C2-Cl0) alkenyl, substituted or unsubstituted alkynyl (C2e1O), substituted or unsubstituted aryl (CS-C1S) and substituted or unsubstituted heterocycle (CselS).
The term "halogen", as understood in the present invention, includes fluorine, chlorine, bromine and iodine.
The term "alkyl" refers to a linear or branched hydrocarbon chain,
30, which has 1 to 10, preferably 1 to 6 and more preferably 1 to 4 carbon atoms. Optionally it may be substituted by at least one substituent described above.
The term "alkenyl" refers to a hydrocarbon chain containing at least one carbon-carbon double bond, linear or branched, having from 2 to 10, preferably from 2 to 6 and more preferably from 2 to 4 carbon atoms .
Optionally it may be substituted by at least one substituent described above.
The term "alkynyl" refers to a hydrocarbon chain that contains at least
5 a carbon-carbon triple bond, linear or branched, having 2 to 10,preferably from 2 to 6 and more preferably from 2 to 4 carbon atoms.Optionally it can be substituted by at least one described substituentpreviously.
10 The term "aryl" refers to an aromatic, monocyclic or polycyclic ring, having 6 to 18 carbon atoms. As examples, phenyl, naphthyl, anthranyl or phenanthryl are mentioned, with phenyl being preferred. Optionally it may be substituted by at least one substituent described above. When the aryl radical carries 2 or more substituents, the substituents may be the same or different.
The term "heteroaryl" refers to a 5-18 membered aromatic ring, optionally substituted by at least one substituent described above, having one or more heteroatoms selected from N, 0, and S as atoms in the ring. The heteroaryl can be a mono-, bi-or tricyclic ring system.
The term "substituted" unless otherwise specified, refers to the substitution by one or more substituents (for example 1, 2, 3 or 4) or any combination thereof from the list of substituents mentioned above. When two or more substituents are present, each substituent may be the same or
25 different.
The hydroxyl groups, thiols, amines, acids can be optionally protected. There are a large number of protecting groups of these functional groups, and they are well known to those skilled in the art. As a guide, see Protecting
30 groups, Kocienski, 2004, 3rd edition.
According to the present specification, any of the compounds defined above, that is, those compounds that respond to the general formula (1), can also be referred to herein as "compound or compounds of the
Invention ".
It should also be understood that the present invention encompasses all isomers of the compounds of formula (1), that is, all geometric, tautomeric and optical forms, and mixtures thereof (eg, racemic mixtures). When there are more chiral centers in the compounds of formula (1), the present invention includes within its
5 reach all possible diastereomers, including mixtures. The different isomeric forms can be separated or resolved from one another by conventional methods, or any given isomer can be obtained by conventional synthetic methods or by stereospecific, stereoselective or asymmetric synthesis.
The term "tautomer" or "tautomeric form", as used in the present invention, refers to structural isomers of different energies that are interconvertible via a low energy barrier. For example, protonic tautomers (also known as prototropic tautomers) that include interconversions by migration of a proton, such as keto-enol isomerizations
15 or imine-enamine. Valencia tautomers include interconversions by reorganization of some bond electrons.
The present invention also includes isotope-labeled compounds, which are identical to those cited in formula (1) except that one or more atoms have been replaced by an atom that has an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine, iodine and chlorine, such
as 2H, 3H, 11C, 14C, 18F, 1231 and 1251.
In this sense, within the scope of the present invention are the pharmaceutically acceptable salts of said compounds containing the aforementioned isotopes and / or other isotopes of other atoms. The isotope-labeled compounds of the present invention, for example those in which radioactive isotopes such as 3H or 14C are incorporated, are useful in drug and / or tissue distribution assays. Particularly preferred are tritium isotopes, i.e. 3H, and carbon-14, i.e. 14C, for their ease of preparation and detectability. The 11C and 18F isotopes are particularly useful in PET (positron emission tomography), and 1251 isotopes are particularly useful in SPECT 35 (single photon emission computed tomography), all useful in brain imaging. In addition, replacement with heavier isotopes such
such as deuterium, that is, 2H, can provide some therapeutic advantages that result from greater metabolic stability, for example, longer half-life in vivo ° lower dosage requirements, and therefore in some cases may be preferred. Isotopically labeled compounds of formula (1) can be
5 generally prepare by performing the procedures described in the following examples, substituting an isotopically unlabeled reagent for an easily available isotopically labeled reagent.
On the other hand, for pharmaceutical use, the salts mentioned above will be
10 physiologically and pharmaceutically acceptable salts. Pharmaceutically acceptable salts include those described by Serge, Bighley and Monkhouse, J. Pharm. Sci., 1977, 66, 1-19. Thus, the term "pharmaceutically acceptable salts" refers to salts prepared from non-toxic pharmaceutically acceptable bases including inorganic bases and organic bases. The salts derived from bases
Inorganic salts include aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic, manganous, potassium, sodium, zinc, and the like salts. 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 resins of
Basic ion exchange, 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, moñolina, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine,
Trimethylamine, tripropylamine, tromethamine, and the like. 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 acid, benzenesulfonic, benzoic, camphorsulfonic, citric, ethanesulfonic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric, isethionic,
30 lactic, maleic, malic, mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic and the like.
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, bismethylenesalicylic, methanesulfonic, ethanedisulfonic acids, propionic, tartaric, salicylic,
citric, gluconic, aspartic, stearic, palmitic, itaconic, glycolic, p-aminobenzoic,
glutamic, benzenesulfonic, cyclohexylsulfamic, phosphoric and nitric.
Also, the compounds of formula (1) may be in crystalline form as
5 free compounds or as solvates and both forms are within the scope of thepresent invention Solvation methods are generally known withinThe technique. Suitable solvates are pharmaceutically acceptable solvates. InA particular embodiment, solvates are hydrates.
The compounds of formula (1) or their salts or solvates are preferably in a pharmaceutically acceptable or substantially pure form. By "pharmaceutically acceptable form" is meant, among others, that they have a pharmaceutically acceptable level of purity excluding normal pharmaceutical additives such as diluents and carriers, and not including material considered toxic at levels
15 normal dosage. The purity levels for the active ingredient are preferably greater than 50%, more preferably, greater than 70%, more preferably, greater than 90%. In a preferred embodiment, they are greater than 95% of the compound of formula (1) or its salts, or solvates.
In a preferred embodiment, the compound of general formula (1) is selected from the list comprising:
3 '- (4-chloropheni 1) -5'-fluoro-2,6'-dioxo-1', 5 ', 6', 1'-tetrahydrospiro {indoline-3,4'-pyrazole {3,4b] pyrid i na] -5 '-ca rbon itrilo;
25 3 '- (4-chloropheni 1) -5,5'-difluoro-2,6'-dioxo-1', 5 ', 6', l'-tetrahydrospiro {indoli na-3,4'pyrazol {3, 4-b] pyridine] -5'-carbonitrile; 3 '- (4-chloropheni 1) -5-fluoro-2,6' -dioxo-5-nitro-1 ', 5', 6 ', 1'-tetrahydrospiro (indoline-3,4'pyrazol (3,4 -b] pyridine] -5'-carbonitrile; 3 '- (4-chlorophenyl 1) -5'-fluoro-2,6'-dioxo-5-phenyl-l', 5 ', 6', T-tetrahydrodrospiro [ Indolin-3.4 '
30 pyrazol [3,4-b] pyrid ina] -5'-carbonitrile; 5-chloro-3 '- (4-chlorophenyl) -5' -flu gold-2,6 '-dioxo-1', 5 ', 6', l'-tetrah idrospiro [i ndolin a-3,4 'pyrazole (3,4-b] pyrid ina] -5'-carbonitrile; 3 '- (4-chloropheni 1) -5', 6-difluor-2,6'-dioxo-1 ', 5', 6 ', 7 '-tetrahydrospiro (indoline-3,4'-pyrazol [3,4b] pyrid i na] -5'-ca rbonitrile;
3 '- (4-Bromophenyl) -5' -fluoro-2, 6'-dioxo-1 ', 5', 6 ', 1'-tetrahydrospiro [indoline-3,4'-pyrazole [3,4b] pyridine ] -5'-carbonitrile; and 3 '- (4-bromophenyl) -5', 6-difluoro-2,6'-dioxo-1 ', 5', 6 ', 7'-tetrahydrospiro [indoline-3,4'pyrazol [3, 4- b] pi rid ina] -5'-carbonitri lo.
Another aspect of the invention is a pharmaceutical composition comprising a
5 compound of formula (1), its pharmaceutically acceptable isomers, salts or solvates. In a preferred embodiment the pharmaceutical composition further comprises at least one pharmaceutically acceptable carrier.
The term "vehicle" refers to a diluent, adjuvant or excipient with which
10 administers the active substance. Such pharmaceutical vehicles may be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. Water or aqueous solutions of saline solution and aqueous solutions of dextrose and
15 glycerol, particularly for injectable solutions. Suitable pharmaceutical vehicles are described in "Remington's Pharmaceutical Sciences" by EW Martin, 1995. Preferably, the vehicles of the invention are approved by the regulatory agency of a state government or a federal government, or are listed in the United States Pharmacopoeia, in the European Pharmacopoeia or other Pharmacopoeia
20 recognized in general for use in animals, and more particularly in humans.
The compounds and pharmaceutical compositions of this invention may be
employees alone or together with other drugs or active ingredients to provide a
combination therapy The other drugs or active ingredients may be part of the
The same pharmaceutical composition, or be provided as a separate pharmaceutical composition, for administration at the same time or at a different time. Examples of pharmaceutical compositions include any solid composition (tablets, pills, capsules, granules, etc.) or liquid (solutions, suspensions or emulsions) for oral, topical or parenteral administration.
In a preferred embodiment, the invention relates to the use of a compound of formula (1) as defined above, or a pharmaceutically acceptable tautomer, prodrug, salt or solvate, or of a pharmaceutical composition containing at least one compound of formula (1), as defined above, for the manufacture of a medicament, preferably for the prevention or treatment of diseases related to the activation of AMPK, and even more
preferably for the treatment or prevention of cardiovascular, inflammatory, autoimmune, neurological, metabolic syndrome and cancer diseases.
In another preferred embodiment, the present invention relates to the use of a compoundof formula (1) as defined above, or a tautomer, prodrug, salopharmaceutically acceptable solvate, or of a pharmaceutical composition thatcontains at least one compound of formula (1), as defined above,for the manufacture of a medicine for the treatment of type 1 and 2 diabetes,
10 obesity, inflammation, dyslipidemia, hypertension, hyperglycemia, hypertriglycerimidemia, insulin resistance, epilepsy and diseases of KrabbefTwitcher, Alzheimer, Parkinson and Huntington.
In another preferred embodiment, the present invention relates to the use of a compound
15 of formula (1) as defined above, or a pharmaceutically acceptable tautomer, prodrug, salo solvate, or of a pharmaceutical composition containing at least one compound of formula (1), as defined above, for the manufacture of a medication for the treatment of type 1 and type diabetes
2.
In another preferred embodiment, the present invention relates to the use of a compound of formula (1), as defined above, or a pharmaceutically acceptable tautomer, prodrug, salo solvate, or of a pharmaceutical composition containing at least one compound of formula (1), as defined above,
25 for the manufacture of a medicament for the treatment of epilepsy.
In another preferred embodiment, the present invention relates to the use of a compound of formula (1) as defined above, or a pharmaceutically acceptable tautomer, prodrug, salo solvate, or of a pharmaceutical composition which
30 contains at least one compound of formula (1), as defined above, for the manufacture of a medicament for the treatment of KrabbefTwitcher disease.
In another preferred embodiment, the present invention relates to the use of a compound 35 of formula (1) as defined above, or a pharmaceutically acceptable tautomer, prodrug, salo solvate, or a pharmaceutical composition which
Contains at least one compound of formula (1), as defined above, for the manufacture of a medicament for the treatment of Alzheimer's, Parkinson's and Huntington's diseases.
In a more preferred embodiment, the present invention relates to the use of a compound of formula (1) as defined above, or a pharmaceutically acceptable tautomer, prodrug, salt or solvate, or of a pharmaceutical composition containing at least one compound of formula (1), as defined above, for the manufacture of a medicament for the treatment of prostate and breast cancer.
Another aspect of the invention relates to a method of treating a disorder or disease related to the activation of AMPK, preferably selected from the metabolic syndrome and inflammatory, autoimmune, cardiovascular, neurological and cancer diseases, and more preferably type diabetes. 1 and 2, hyperglycemia, obesity, inflammation, epilepsy, prostate cancer, breast cancer and KrabbefTwitcher Alzheimer's, Parkinson's and Huntington's diseases, comprising administering to a subject the compound of the invention in a therapeutically effective amount, or a Pharmaceutical composition of the invention comprising the compound of the invention in a therapeutically effective amount.
The amount of compound of the invention, its pharmaceutically acceptable isomers, salts or solvates thereof, therapeutically effective to be administered (also referred to herein as therapeutically effective or effective amount), as well as its dosage to treat a pathological condition with Said compounds will depend on numerous factors, among which are the age, the patient's condition, the severity of the disease, the route and frequency of administration, the modulating compound to be used, etc.
Another aspect of the invention relates to a process for obtaining the compounds of general formula (1) comprising the following steps:
(a) condensation reaction between compound of formula (11) and an NH2 compound
NHR,
R,
in o
elevenwhere R1 and RJ are defined above;
(b) condensation reaction of the compound of formula (IV)
R,
or
IV
with the compound obtained in step (a) of formula (111) for 12-24 hours at 60
120 'C
R,
111where R1-Rs are defined above; Y
(c) reaction of the compound obtained in step (b) of formula (la) with a halogenating agent
N
~ O
.
where R1-Rs are defined above.
In a final aspect, the present invention relates to the compounds of general formula (la) or their corresponding pharmaceutically acceptable isomers, salts or solvates
~
N
~ o
.
wherein R1-Rs are defined above, with the proviso that when R1 is hydrogen ~ it is not hydrogen.
Throughout the description and the claims the word "comprises" 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 features of the invention will be derived partly from the description and partly from the practice of the
15 invention. The following examples and figures are provided by way of illustration, and are not intended to be limiting of the present invention.
DESCRIPTION OF THE FIGURES
FIG. 1. Effect of AMPK activation of compound 1. Example of the analysis of
Western Blot showing that compound 1 activates AMPK in a dose-dependent manner. This effect is confirmed by the increased dose-dependent phosphorylation of its ACC substrate, in a range of concentrations between 7.5-120 IJM. Hek293 cells are treated with 30 IJg of compound 1 for 1 hour. Lysed cells are analyzed by Western
10 810t using anti-phosphoThr172 AMPK alpha, anti-AMPKb1 (used as load control), anti-phosphoSer79ACC and anti-ACC (used as load control). Molecular weight markers are indicated to the right of the figure.
FIG. 2. Effect of AMPK activation of compounds 11 and 13. Example of the analysis
15 of Western 810t showing that compounds 11 and 13 activate AMPK in a dose-dependent manner. This effect is confirmed by the increased dose-dependent phosphorylation of its ACC substrate, in a concentration range between 5 -100 IJM. Hek293 cells are treated with 30 IJg of compounds 11 and 13 for 1 hour. Lysed cells are analyzed by
20 Western 810t using anti-phosphoThr172 AMPK alpha, anti-AMPKb1 (used as load control), anti-phosphoSer79ACC and anti-ACC (used as load control). Molecular weight markers are indicated to the right of the figure.
FIG. 3. Effect of AMPK activation of compound 12. Example of the analysis of
Western 810t showing that compound 12 activates AMPK in a dose-dependent manner. This effect is confirmed by the increased dose-dependent phosphorylation of its ACC substrate, in a concentration range between 5 -100 IJM. Hek293 cells are treated with 30 IJQ of compound 12 for 1 hour. Lysed cells are analyzed by Western
30 810t using anti-phosphoThr172 AMPK alpha, anti-AMPKb1 (used as load control), anti-phosphoSer79ACC and anti-ACC (used as load control). Molecular weight markers are indicated to the right of the figure.
FIG. 4. Effect of AMPK activation of compound 15. Example of Western Blot analysis showing that compound 15 activates AMPK in a dose-dependent manner. This effect is confirmed by the increased dose-dependent phosphorylation of your ACC substrate, in a range of concentrations.
5 between 5 -50 IJM. Hek293 cells are treated with 30 IJg ofcompound 15 for 1 hour. Lysed cells are analyzed by Western810t using anti-phosphoThr172 AMPK alpha, anti-AMPKb1 (used as control ofload), anti-phosphoSer79ACC and anti-ACC (used as load control). TheMolecular weight markers are indicated to the right of the figure.
10 EXAMPLES
The invention will now be illustrated by tests carried out by the inventors, which show the effectiveness of the product of the invention.
Example 1 General procedure of synthesis of the compounds of general formula m
The compounds of the general formula (1) were synthesized according to the synthetic route 20 which is collected in the following reaction scheme:
I o R'xx> = °
R. 6 N
,or, R''OyH
I. & (Al
in
'"
OR
'"
"
25 a) Hydrazine (1eq.), EtOH, 12h; b) Ethyl cyanoacetate (1 eq.), cat., water, 90 oC, 12h; e) Selectfluor (2 eq.), MeOH, 3h
General procedure for the synthesis of 5-amino-3-phenyl-1H-pyrazoles (111) To a solution of the corresponding benzoylacetonitrile (100 mg, 1 eq.) In 10 mL of 30 EtOH at reflux the corresponding hydrazine (1.5 eq.). The mixture is allowed to stir at reflux one night. Once the reaction is over, the solvent is removed at
reduced pressure and the dried residue is triturated in EhO, a precipitate appearing that is filtered, washed with cold Et20 and allowed to air dry.
3-amino-5- (4-chlorophenyl) -1 H-pyrazole (111-1)
5 Following the general procedure of synthesis of 5-amino-3-phenyl-1H-pyrazoles, 150 mg (66%) of a white solid are obtained. P.f. 169 'C.' H NMR (300 MHz, DMSOd,) ~ (ppm): 7.65 (d, J = 8.2 Hz, 2H, H2 '), 7.40 (d, J = 8.2 Hz , 2H, H3 '), 5.73 (s, 1 H, H4), 4.90 (s, 2H, NH,). HPLC: t R = 4.49 (99.9%). MS (ES, positive mode) miz 194.0 [M + H) '.
3-amino-5- (4-bromophenyl) -1H-pyrazole (111-2)
Following the general procedure of synthesis of 5-amino-3-phenyl-1H-pyrazoles, 74 mg (71%) of a white solid are obtained. P.f. 160-162 'c' H NMR (300 MHz, DMSO-d,) ~ (ppm): 11.72 (s, 1H, NH), 7.55 (m, 4H, H-Ar), 5.73 ( s, 1H, H4), 4.89 (s,
15 2H, NH,). HPLC: t R = 2.98 (99.9%). MS (ES, positive mode) miz 239.8 [M) '.
General procedure for synthesis of 2,6'-dioxo-3'-phenyl-1 ', 5', 6 ', 7'
tetrahydrosp i ro {indoli na -3.4 I -pi razol [3, 4-b] Pyridi na} -5'-carbonitriIos (la)
The corresponding isatin (1 eq.) Is suspended in water (0.05 mLlmg) and the mixture is
20 add ethyl cyanoacetate (1 eq.) And the catalyst indicated in each case, stirring at 80 ° C for 1 h. After this time, the corresponding 3-amino5-phenyl-pyrazole (1 eq) is added, and stirred overnight at 90 ° C. Once the reaction is finished, the crude is purified according to the procedure indicated in each case.
25 3 '- (4-chlorophenyl) -2, 6'-dioxo-1', 5 ', 6', 7'-tetrahydrospiro [indoline-3, 4'-pyrazole [3,4-bjpyridinej5'-carbonitrile (la -one)
Following the general procedure of synthesis of 2,6'-dioxo-3'-phenyl-1 ', 5', 6 ', 7'tetrahydrospiro {indoline-3,4'-pyrazol [3,4-b] pyridine} - 5'-carbonitriles, was split from 340 mg of isatin, 581, 94 mg of 3-amino-5- (4-chlorophenyl) -1H-pyrazole and triethylamine (10 mol%) as catalyst. Once the reaction is finished, the crude is allowed to cool to t.a., a brown precipitate appearing that is filtered and washed with water and Et20, obtaining 666 mg (75%) of a brown solid. Mp.> 350 oC. 1H NMR (300 MHz, DMSO-d6) ~ (ppm): 12.92 (s, 1H, NH), 12.79 (s, 1H, NH), 11.44 (s, 1H, NH), 11, 19 (s, 1H, NH), 11.02 (s, 1H, NH), 10.95 (s, 1H, NH), 7.44-7.28 (m, 1H), 7.17-6, 76 (m, 8H, H-Ar), 35 6.73 -6.61 (t, J = 7.5 Hz, 1 H), 5.38 -5.25 (s, 1 H, H5 '), 4.96 -4.89 (s, 1 H, H5 '). HPLC
(H, O 15 ~ 95% MeCN, 0.2% HCOOH for 10 min): IR = 7.44 (99.9%). MS (ES, positive mode): miz 390.4 [M + H '].
3 '- (4-chlorophenyl) -5-fluoro-2, 6'-dioxo-1', 5 ', 6', 7'-tetrahydrospiro {indoline-3, 4'-pyrazole {3,45 b] pyridine] -5'-carbonylryl (la-2)
Following the general procedure of synthesis of 2,6'-dioxo-3'-phenyl-1 ', 5', 6 ', 7'tetrahydrospiro {indoline-3,4'-pyrazol [3,4-b] pyridine} - 5'-carbonitriles, starting from 200 mg of 5-fluoroisatin, 234.53 mg of 3-amino-5- (4-chlorophenyl) -1H-pyrazole and acetic acid (40% mol) as catalyst. Once the reaction is fine, the oil is allowed to cool to
10 t.a. The resulting precipitate is filtered, washed with water and allowed to dry, obtaining 290 mg (58%) of a brown solid. PI. > 350 oC. 'H NMR (300 MHz, DMSO-d6) Ó (ppm): 12.97 (s, 1 H), 12.84 (s, 1 H), 11.45 (s, 1 H), 11.25 ( s, 1 H), 11.06 (s, 1 H), 11.00 (s, 1H), 7.68-6.85 (m, 9H, H-Ar), 5.38 (s, 1H, H5 '), 4.97 (s, 1H, H5') HPLC: IR = 3.80 (95%). MS (ES, positive mode): miz 407.8 [M + H '].
15 3 '- (4-chlorophenyl) -2, 6'-dioxo-5-nitro-1', 5 ', 6', 7'-tetrahydrospiro {indoline-3,4 '-pyrazJ {3, 4b] pyridine] -5'-carbonylryl (la-3)
Following the general procedure of synthesis of 2,6'-dioxo-3'-phenyl-1 ', 5', 6 ', 7'tetrahydrospiro {indoline-3,4'-pyrazol [3,4-b] pyridine} - 5'-carbonitriles, split from 200 mg
20 of 5-nitroisatin, 197.53 mg of 3-amino-5- (4-chlorophenyl) -1H-pyrazole and triethylamine (40% mol) as catalyst. Once the reaction is finished, part of the solvent is removed under reduced pressure until leaving a volume approximately equal to half of the initial. When cooling to t.a., the appearance of a precipitate that is filtered and washed with water and DCM is observed, obtaining 140 mg (31%) of a red solid. Mp.> 350 oC.
25 'H NMR (300 MHz, DMSO-d6) Ó (ppm): 13.05 (s, 1H), 12.91 (s, 1H), 11.76 (s, 1H), 11, 67 (s, 1H), 11, 60 (s, 1 H), 11.33 (s, 1 H), 8.34 -6.81 (m, 9H), 5.62 (s, 1 H, H5 '), 5 , 11 (s, 1H, H5 '). HPLC: I ~ 3.80 (97%). MS (ES, positive mode): miz 435.4 [M + H '].
3 '- (4-chloropheniJ) -2, 6'-dioxo-5-phenyl-1 "5', 6 ', 7'-tetrahydrospiro {indo / ina-3, 4'-pyrazoJ {3, 430 b] pyridine ] -5'-carbonylryl (Ia-4)
Following the general procedure of synthesis of 2,6'-dioxo-3'-phenyl-1 ', 5', 6 ', 7'tetrahydrospiro {indoline-3,4'-pyrazol [3,4-b] pyridine} - 5 '· carbonitriles, starting from 35 mg of 5-phenylisatin, 30.36 mg of 3-amino-5- (4-chlorophenyl) -1 H-pyrazole and acetic acid (10 35 mol%) as catalyst. Once the reaction is finished, part of the
solvent under reduced pressure until leaving a volume approximately equal to half of the initial. It is allowed to cool to t.a and the resulting solid is filtered and washed with water and DCM, obtaining 21 mg (25%) of a white solid. Mp.> 350 oC. 1 H NMR (300 MHz, DMSO-d6) O (ppm): 12.96 (s, 1 H), 12.83 (s, 1 H), 11.45 (s, 1 H) 11.23 (s, 1 HOUR),
5 11.03 (s, 1H), 7.36-6.86 (m, 14H, H-Ar), 5.40 (s, 1H, H5 '), 4.96 (s, 1H, H5') . HPLC: t R = 4.36 (96%). MS (ES, positive mode): miz 465.6 (M + W].
5-chloro-3'-e4-chlorophenyl) -2, 6 '-dioxo-1 "5', 6 ', 7'-trahydrospiro [indoline-3, 4'-pyrazole [3, 4
b] pyridine] -5'-carbonitrile (la-5)
10 Following the general procedure of synthesis of 2,6'-dioxo-3'-phenyl-1 ', 5', 6 ', 7'tetrahydrospiro {indoline-3,4'-pyrazole (3,4-b] pyridine} -5'-carbonitriles, starting from 200 mg of 5-chloroisatin, 213.27 mg of 3-amino-5- (4-chlorophenyl) -1H-pyrazole and acetic acid (40% mol) as catalyst. the reaction, the solvent is removed at
The pressure is reduced and the residue is purified by column chromatography on silica gel (ethyl acetate: hexane), yielding 80 mg (17%) of a white solid. PI. > 350 oC. 'H NMR (300 MHz, DMSO-d6) O (ppm): 10.36 (s, 1H), 10.14 (s, 1H), 10.10 (s, 1H), 9.99 (s, 1H ), 7.59-6.98 (m, 14H, H-Ar), 5.00 (s, 1H, H5 '), 4.73 (s, 1H, H5'). HPLC: t R = 3.98 (98 'lo). MS (ES, positive mode): miz 424.4 (M ·].
3 '- (4-c1orophenyl) -6-fluor-2, 6'-dioxo-1', 5 ', 6', 7'-tetrahydrospiro [indoline-3, 4'-breath / [3, 4b] pyridine] -5'-carbonitrile (la-6)
Following the general procedure of synthesis of 2,6'-dioxo-3'-phenyl-1 ', 5', 6 ', 7'
Tetrahydrospiro {indoline-3,4'-pyrazole (3,4-b] pyridine} -5'-ca rbonitriles, starting from 90 mg of 6-fluoroisatin, 66.97 mg of 3-amino-5- (4 -chlorophenyl) -1H-pyrazole and piperidine (10% mol) as a catalyst Once the reaction is complete, the crude is purified by column chromatography on silica gel (ethyl acetate: hexane), obtaining 50 mg (36 ' lo) of a white solid. PI.> 350 oC. 'H NMR (300 MHz, DMSO-d6) O
30 (ppm): 12.95 (s, 1H), 12.83 (s, 1H), 11.47 (s, 1H), 11.23 (s, 1H), 11.20 (s, 1H), 11, 13 (s, 1 H), 7.45 -6.48 (m, 14H, H-Ar), 5.39 (s, 1 H, H5 '), 4.95 (s, 1 H, H5 '). HPLC: t R = 3.86 (96 'lo). MS (ES, positive mode): miz 408.3 (M + W].
3 '- (4-bromofe ni /) - 2, 6' -dioxo-1 "5 ', 6', 7'-tetrahydrospiro [indoline-3, 4'-pyro zol [3,4-b] pyridine] 5 '-carbonitri / o (la-7)
Following the general procedure of synthesis of 2,6'-dioxo-3'-phenyl-1 ', 5', 6 ', 7'tetrahydrospiro {indoline-3,4'-pyrazol [3,4-b] pyridine} - 5'-carbonitriles, starting from 200 mg of isatin, 317 mg of 3-amino-5- (4-bromophenyl) -1H-pyrazole and piperidine (10% mol) as catalyst. Once the reaction is over, the crude is purified by silica gel column chromatography (ethyl acetate: hexane), obtaining 336 mg (57%) of a beige solid. PI. > 350 'C. 'H NMR (300 MHz, DMSO-d6) ~ (ppm): 12.95 (s, 1 H), 12.81 (s, 1 H), 11.20 (s, 1 H), 11.04 ( s, 1 H), 10.95 (s, 1 H), 10.44 (s, 1H), 7.55 -6.75 (m, 10H, H-Ar), 6.50 -6.46 ( t, J = 7.29 Hz, 4H), 5.34 (s, 1H, H5 '), 4.92 (s, 1H, H5'). HPLC (H, O 15 ~ 95% MeCN, 0.2% HCOOH for 10 min): tR =
10 7.44 (99.9%). MS (ES, positive mode): miz 390.0 [M + W).
3 '- (4-bromophenyl) -6-fluoro-2, 6'-dioxo-1', 5 ', 6', 7'-tetrahydrospiro [indoline-3,4 '-pyrazole [3, 4bjpiridinaj-S'- carbonitrile (la-8)
Following the general procedure of synthesis of 2,6'-dioxo-3'-phenyl-1 ', 5', 6 ', 7'
Tetrahydrospiro {indoline-3,4'-pyrazol [3,4-b] pyridine} -5'-ca rbonitriles, was started from 100 mg of 6-fluoroisatin, 144.19 mg of 3-amino-5- (4 -bromophenyl) -1 H-pyrazole and acetic acid {40% mol) as catalyst. Once the reaction is over, the crude is allowed to cool to
t.a. The resulting brown precipitate is filtered, washed with water and allowed to dry, obtaining 158 mg (58%) of a brown solid. Mp.> 350 oC. 1 H NMR (300 MHz,
20 DMSO-d6) ~ (ppm): 12.97 (s, 1H), 12.84 (s, 1H), 11.45 (s, 1H), 11.25 (s, 1H), 11.06 ( s, 1 H), 11.00 (s, 1 H), 7.68 -6.85 (m, 9H), 5.38 (s, 1 H, H5 '), 4.97 (s, 1 H , H5 '). HPLC: t R = 3.92 (98%). MS (ES, positive mode): miz 454.1 [M + 2H '].
General synthesis procedure for 5'-f1uoro-2,6'-dioxo-3'-phenyl-1 ', 5', 6 ', 7'
Tetrahydrospiro {indoli na-3,4'-pyrazol [3,4-b] pyridine} -5'-carbonitril os (1) The 2,6'-dioxo-3'-phenyl- derivative is dissolved in a flask 1 ', 5', 6 ', 7'tetrah idrospiro {indole and na-3,4, -pyrazol [3,4-b] pyrid ina} -5' -ca corresponding rbonitrile (1 eq.) And Selectfluor (2 eq.) in MeOH and stirred at reflux for 3 h. Once the reaction is finished, the solvent is removed under reduced pressure. The residue is filtered on gel.
30 of silica (DCM: MeOH 10: 1) to remove inorganic residues and the end product is separated into its diastereomeric pairs by semi-preparative HPLC made with a Sunfire C18 column of particle size 5 microns and dimensions 19x150 mm, using the gradient elution indicated in each case.
3 '- (4-chlorophenyl) -S' -fluoro-2, 6'-dioxo-1 ', 5', 6 ', 7'-tetrahydrospiro [indoline-3,4'-pyrazole [3,4b} pyridine} -5'-carbonitrile (Compounds 1 and 2)
Following the general procedure of synthesis of 5'-f1uoro-2,6'-dioxo-3'-fenill ', 5', 6 ', 7'-tetrahydrospiro {indoline-3, 4'-pyrazole [3,4-b ] pyridine} -5'-carbonitriles, starting from 200 mg of la-1 and 181.2 mg of Selectfluor, obtaining 60 mg (29%) of a white solid. The end product is separated into its diastereomeric pairs by HPLC
5 semi-preparative using an isocratic elution gradient H20-MeCN 31% -69%, 0.1% HCOOH for 60 min
Composite Structure MS (ES +) IHPLC
MS (ES +): MS (ES, positive mode): miz 408.2(M + W).HPLC (lsocratic H20-MeCN 31-69%, 0.1%HCOOH for 60 min): t R = 38.62 (99.9%).
MS (ES +): MS (ES, positive mode): miz 408.2 (M + W).
HPLC (Isocratic H20-MeCN 31-69%, 0.1% HCOOH for 60 min): t R = 41, 47 (99.9%).
1: They were 12.5 mg (6%). PI. > 350 'C.' H NMR (300 MHz, DMSO-d6) or (ppm):
10 7.23 (t, J = 7.8 Hz, 1H, H4), 7.12 (d, J = 8.1 Hz, 2H, H3 "), 6.93 (t, J = 7.70 Hz , 2H, H5, H6), 6.82 (d, J = 8.17 Hz, 2H, H2 "), 6.70 (t, J = 7.63 Hz, 1H, H7). HPLC (Isocratic H20-MeCN 31-69%, 0.1% HCOOH for 60 min): t R = 38.62 (99.9%). MS (ES +): MS (ES, positive mode): miz 408.2 (M + W). 2: They were 4 mg (2%). PI. > 350 'C.' H NMR (300 MHz, DMSO-d6) or (ppm): 7.26 (t, J = 7.5 Hz, 1H, H4), 7.19 (d, J = 8.4 Hz,
15 2H, H3 "), 6.98 (d, J = 7.9 Hz, 1H, H6), 6.87 (d, J = 8.4 Hz, 3H, H2", H5), 6.76 ( t, J = 7.6 Hz, 1H, H7). HPLC (Isocratic H20-MeCN 31-69%, 0.1% HCOOH for 60 min): t R = 41.47 (99.9%). MS (ES +): MS (ES, positive mode): miz 408.2 (M + W).
3 '- (4-chlorophenyl) -5, 5'-difluoro-2, 6'-dioxo-1 "5', 6 ', 7'-tetrahydrospiro [indoline-3, 4'-breath / [3,4-b ] pyridinaj-5'-carbonitri / o (Compounds 3 and 4)
20 Following the general procedure of synthesis of 5'-f1uoro-2,6'-dioxo-3'-fenill ', 5', 6 ', 7'-tetrahydrospiro {indoline-3,4'-pyrazole [3,4- b] pyridine} -5'-carbonitriles, starting from 50 mg of la-2 and 86.87 mg of Selectfluor, obtaining 40 mg (77%) of a brown syrup. The final product is separated into its diastereomeric partners by HPLC
semi-preparative using a gradient of isocratic elution H20-MeCN 31% -69%, 0.1% HCOOH for 40 min.
Compound 3 4 StructureMS (ES +) / HPLC MS (ES +): MS (ES, positive mode): mi z 426.4 [M + W]. HPLC (Isocratic H, O-MeCN 31-69 'lo, 0.1% HCOOH for 60 min): t R = 31, 39 (99.9' lo). MS (ES +): MS (ES, positive mode): miz 426.4 [M + W]. HPLC (Isocratic H, O-MeCN 31-69 'lo, 0.1% HCOOH for 60 min): t R = 36.34 (99.9' lo).
5 3: PI. > 350 oC. 'H NMR (300 MHz, DMSO-d6) Ó (ppm): 12.94 (s, 1H, NH), 11.51 (s, 1 H, NH), 7.20 (d, J = 8.3 Hz, 2H, H3 "), 7.12 (td, J = 9.1, 2.8 Hz, 1 H, H7), 6.96 (dd, J = 8.7, 4.4 Hz, 1 H , H4), 6.89 (d, J = 8.4 Hz, 2H, H2 "), 6.69 (dd, J = 8.2, 2.7 Hz, 1H, H6). HPlC (lsocratic H20-MeCN 31-69%, 0.1% HCOOH for 60 min): tR = 31, 39 (99.9 'lo). MS (ES +): MS (ES, positive mode): miz 426.4 [M + W]. 4: PI. > 350 oC. 'H NMR
10 (300 MHz, DMSO-d6) Ó (ppm): 13.12 (s, 1H, NH), 11.98 (s, 1H, NH), 11.44 (s, 1H, NH), 7.24 (d, J = 8.5 Hz, 2H, H3 "), 7.14 (td, J = 8.5, 2.3 Hz, 1H, H7), 7.00 (dd, J = 8.6, 4.4 Hz, 1H, H4), 6.91 (d, J = 8.4 Hz, 2H, H2 "), 6.72 (d, J = 8.1 Hz, 1H, H6). HPLC (Isocratic H, O-MeCN 31-69 'lo, 0.1% HCOOH for 60 min): t R = 36.34 (99.9' lo). MS (ES +): MS (ES, positive mode): miz 426.4 [M + H '].
15 3 '- (4-chlorophenyl) -5'-diffuoro-5-nitro-2, 6'-dioxo-1 "5', 6 ', 7'-tetrahydrospiro [indoline-3,4'pipe / {3, 4-bjpiridinaj-5'-carbonitri / o (Compounds 5 and 6)
Following the general procedure of synthesis of 5'-f1uoro-2,6'-dioxo-3'-fenill ', 5', 6 ', 7' -tetrahydrospiro {indoline-3,4'-pyrazole [3,4-b ] pyridine} -5 '-carbonitriles, split
20 of 50 mg of la-3 and 81.48 mg of Selecttluor, obtaining 40 mg (77%) of an orange syrup. The final product is separated into its diastereomeric pairs by semi-preparative HPlC using an isocratic elution gradient H20-MeCN 33% -70%, 0.1% HCOOH for 60 min.
Composite Structure MS (ES +) IHPLC MS (ES +): MS (ES, positive mode): mIz 453.3 [M + H '].
NO, HPlC (lsocratic H20-MeCN 30-70%, 0.1%
HCOOH for 60 min): t,. = 32.41 (99.9%). MS (ES +): MS (ES, positive mode): mIz 453.3 [M + H '].
NO,
HPlC (lsocratic H20-MeCN 30-70%, 0.1% HCOOH for 60 min): t,. = 36.74 (99.9%).
5: They were 4 mg (8%). PI. > 350 oC. 'H NMR (300 MHz, DMSO-d6) Ó (ppm): 13.22
(s, 1H, NH), 12.27 (s, 1H, NH), 8.24 (dd, J = 8.1, 1.5 Hz, 1H, H4), 7.54 (s, 1H, H1 ), 5 7.21 (d, J = 8.4 Hz, 2H, H3 "), 7.05 (m, 1 H, H7), 6.92 (d, J = 8.4 Hz, 2H, H2 "), 6.55 (m,
1H, H6). HPLC (lsocratic H20-MeCN 30-70%, 0.1% HCOOH for 60 min): t R = 32.41 (99.9%). MS (ES +): MS (ES, positive mode): miz 453.3 [M + H'J, 6: 2 mg (4%) were found. PI. > 350 oC. 'H NMR (300 MHz, DMSO-d6) Ó (ppm): 12.86 (s, 1H, NH), 11.65 (s, 1H, NH) 8.65 (dd, J = 8.1, 1 , 5 Hz, 1H, H4), 7.68 (s, 1H, H1), 7.32 (d, J = 8.4
10 Hz, 2H, H3 "), 7.28 (m, 1H, H7), 6.83 (d, J = 8.4 Hz, 2H, H2"), 6.77 (m, 1H, H6). HPLC (Isocratic H, O-MeCN 30-70%, 0.1% HCOOH for 60 min): t R = 36.74 (99.9%). MS (ES +): MS (ES, positive mode): mIz 453.3 [M + H ']. 3 '-e4-chlorophenyl) -5'-fluoro-2, 6' -dioxo-5-feniJ-1 ', 5', 6 ', 7'-tetrahydrospiro [indoline-3, 4'
pyrazol [3,4-b} pyridine} -5'-carbonitrile (Compounds 7 and 8)
15 Following the general procedure of synthesis of 5'-fluoro-2,6'-dioxo-3 '-fenill', 5 ', 6', 7'-tetrahydrospiro {indoline-3,4 '-pyrazole [3,4- b] pyridine} -5 '-carbonitriles, starting from 50 mg of la4 and 76.04 mg of Selectfluor, obtaining 50 mg (97%) of an orange oil. The final product is separated into its diastereomeric pairs by semi-preparative HPlC using an isocratic elution gradient H20-MeCN 40% -60%,
20 0.1% HCOOH for 60 min.
Composite Structure MS (ES +) fHPLC
MS (ES +): MS (ES, positive mode): miz 484.2 (M + W).
Ph
HPlC (lsocratic H20-MeCN 40-60%, 0.1% HCOOH for 60 min): t R = 22.73 (99.9%).
MS (ES +): MS (ES, positive mode): miz 484.2 (M + W).
Ph
HPlC (lsocratic H20-MeCN 40-60%, 0.1% HCOOH for 60 min): t R = 25.31 (99.9%).
7: They were 4 mg (8%). PI. > 350 'C.' H NMR (300 MHz, DMSO-d6) ~ (ppm): 13.21 (s, 1H, NH), 11.40 (s, 1H, NH), 7.47 (d, J = 8.2 Hz, 1H, H4), 7.41 -7.25 (m, 3H, H3 ", 5 H7), 7.08 (dd, J = 17.8, 8.8 Hz, 6H, Ph , H6), 6.88 (d, J = 8.2 Hz, 2H, H2 "). HPLC (lsocratic H20-MeCN 40-60%, 0.1% HCOOH for 60 min): t R = 22.73 (99.9%). MS (ES +): MS (ES, positive mode): miz 484.2 (M + Hl 8: 2 mg (4%) were found. PI.> 350 'C.' H NMR (300 MHz, DMSO-d6) ~ (ppm): 13.65 (s, 1 H, NH), 11, 35 (s, 1 H, NH), 7.88 (d, J = 8.2 Hz, 1H, H4), 7.61 - 7.40 (m, 3H, H3 ", H7), 7.12 (dd, J = 17.8, 8.8 Hz,
10 6H, Ph, H6), 6.69 (d, J = 8.2 Hz, 2H, H2 "). HPLC (Isocratic H, O-MeCN 40-60%, 0.1% HCOOH for 60 min): tR = 25.31 (99.9%) MS (ES +): MS (ES, positive mode): miz 484.2 (M + H '].
5-c / gold-3 '- (4-chlorophenyl} -5' -fluoro-2, 6'-dioxo-1 "5 ', 6', 7'-tet-rahidrospiro [indoline-3, 4'-breath / [ 3,4-b] pyridine] -5'-carbonitri / o (Compounds 9 and 10)
15 Following the general procedure of synthesis of 5'-fluoro-2,6'-dioxo-3'-fenill ', 5', 6 ', 7'-tetrahydrospiro {indoline-3,4' -pyrazole [3,4- b] pyridine} -5'-carbonitriles, starting from 60 mg of la-5 and 100.2 mg of Selectfluor, obtaining 60 mg (96%) of an orange oil. The final product is separated into its diastereomeric pairs by semi-preparative HPlC using an isocratic elution gradient H20-MeCN 36% -64%,
20 0.1% HCOOH for 60 min.
Compound
Structure
MS (ES +) IHPLC
MS (ES +): MS (ES, positive mode): mIz 443.4 [M + H '].
he
HPlC (Isocratic H20-MeCN 36-64%, 0.1% HCOOH for 60 min): t ,, = 21, 93 (99.9%).
MS (ES +): MS (ES, positive mode): miz 443.4 [M + H '].
he
HPlC (Isocratic H20-MeCN 36-64 EYE, 0.1%F
HCOOH for 60 min): t ,, = 25.42 (99.9%).
~ O
9: They were 4 mg (6%). PI. > 350 "C. 'H NMR (300 MHz, DMSO-d6) O (ppm): 13.24 (s, 1H, NH), 11.94 (s, 1H, NH), 11.38 (s, 1H , NH), 7.31 (dd, J = 8.4, 2.2 Hz, 1H, H6), 5.20 (d, J = 8.4 Hz, 2H, H3 "), 6.97 ( d, J = 8.4 Hz, 1H, H7), 6.88 (d, J = 8.5 Hz, 2H, H2 "), 6.79 (d, J = 2.2 Hz, 1H, H4) HPLC (Isocratic H, O-MeCN 36-64%, 0.1% HCOOH for 60 min): tR = 21.93 (99.9%) MS (ES +): MS (ES, positive mode): miz 443.4 [M + HT 10: 4 mg (6%) were found. PI.> 350 "C. 'H NMR (300 MHz, DMSOd6) O (ppm): 13.16 (s, 1 H, NH), 12.10 (s, 1 H, NH), 11.57 (s, 1 H, NH), 7.34 (dd, J = 8.4,
10 1.9 Hz, 1H, H6), 7.25 (d, J = 8.5 Hz, 1H, H7), 7.01 (d, J = 8.4 Hz, 2H, H3 "), 6, 90 (d, J = 8.4 Hz, 2H, H2 "), 6.81 (s, 1H, H4). HPLC (Isocratic H, O-MeCN 36-64%, 0.1% HCOOH for 60 min): t R = 25.42 (99.9%). MS (ES +): MS (ES, positive mode): miz 443.4 [M + H '].
3 '- (4-chlorophenyl) -5', 6-difluor-2, 6'-dioxo-1 ', 5', 6 ', 7'-tetrahydrospiro [indoline-3, 4'-pyrazole {3, 415 bjpiridinaj -5'-carbonitrile (Compounds 11 and 12)
Dissolve in a flask la-6 (50 mg, 0.123 mmol) and Selectfluor (86.87 mg, 0.246 mmol) in about 50 ml of MeOH and stir at reflux for 2 h. Once the reaction is finished, the solvent is removed under reduced pressure and the residue is purified by column chromatography on silica gel (ethyl acetate: hexane), obtaining
20 12 mg (23 EYE) of a white solid. The final product is separated into its diastereomeric pairs by semi-preparative HPlC using an isocratic elution gradient H20-MeCN 33% -67%, 0.1% HCOOH for 60 min.
Compound StructureMS (ES +) fHPLC
eleven MS (ES +): MS (ES, positive mode): miz 425.6 (M + W) HPLC (lsocratic H20-MeCN 33-67%, 0.1% HCOOH for 60 min): tR = 38.27 ( 99.9%).
12 MS (ES +): MS (ES, positive mode): miz 425.6 (M + W) HPLC (lsocratic H20-MeCN 33-67%, 0.1% HCOOH for 60 min): tR = 40.95 ( 99.9%).
11: 7.8 mg (15%) were found. PI, 270 oC. 'H NMR (400 MHz, DMSO-d6) or (ppm): 13.17 (s, 1H, NH), 11.92 (s, 1H, NH), 11.38 (s, 1H, NH), 7 , 21 (d, J = 8.5 Hz, 2H, H3 "), 6.93 (dd, J = 8.5, 5.3 Hz, 1 H, H4), 6.87 (d, J = 8 , 5 Hz, 2H, H2 "), 6.81 (dd, J = 8.9, 2.4
Hz, 1H, H7), 6.60-6.50 (m, 1H, H5). HPLC (Isocratic H20-MeCN 33-67%, 0.1%
5 HCOOH for 60 min): t ~ 38.27 (99.9%). MS (ES +): MS (ES, positive mode): miz 425.6 (M + W) .12: 4.0 mg (7%) were found. PI.> 350 oC. 'H NMR (400 MHz, DMSOd6) or (ppm): 7.25 (d, J = 8.4 Hz, 2H, H2 "), 7.22-7.14 (m, 1 H), 7.09-6.99 (m, 1 H ), 6.88 (d, J = 8.5 Hz, 2H, H3 "), 6.61-6.50 (m, 1 H). HPLC (Isocratico H, O-MeCN 3367%, 0.1% HCOOH for 60 min): t R = 40.95 (99.9%) MS (ES +): MS (ES, mode
10 positive): miz 425.6 (M + W].
3 '- (4-bromophenyl) -5'-fluoro-2, 6'-dioxo-1', 5 ', 6', 7'-tetrahydrospiro [indoline-3, 4'-pyrazo / [3,4b] pyridine ] -5'-carbonitrile (Compounds 13 and 14)
Dissolve in a la-7 flask (100 mg, 0.230 mmol) and Selecttluor (81, 58 mg, 0.238
mmol) in 50 mL of MeOH and stirred at reflux for 2 h. Once the
After the reaction, the solvent is removed under reduced pressure and the residue is purified by column chromatography on silica gel (ethyl acetate: hexane), obtaining 20 mg (19%) of a beige solid. The final product is separated into its diastereomeric pairs by semi-preparative HPLC using an isocratic elution gradient H20-MeCN 33-67%, 0.1% HCOOH for 50 min.
20 13: 4.5 mg (4 'lo) were found. PI. > 350 oC. 'H NMR (400 MHz, DMSO-d6) 5 (ppm): 7.22 (m, 3H, H-Ar), 6.93 (m, 2H), 6.75 (d, J = 8.5 Hz , 2H, H2 "), 6.70 (dt, J = 7.6, 1.1 Hz, 1 H, H5). HPLC (Isocratic H20-MeCN 33-67%, 0.1% HCOOH for 60 min) : tR = 31, 50 (99.9%) MS (ES +): MS (ES, positive mode): miz 452.0 [M ']. 14: They were
Compound StructureMS (ES +) / HPLC
13 Br ~ oMS (ES, positive mode): miz 452.0 [M ']. 14: 4.2 mg (4%) were found. HPLC (Isocratic H, O-MeCN 33-67 'lo, 0.1% HCOOH for 60 min): t R = 31.50 (99.9%).
14 BrMS (ES +): MS (ES, positive mode): miz 452.0 [M ']. HPLC (lsocratic H20-MeCN 33-67%, 0.1% HCOOH for 60 min): t R = 33.18 (99.9%).
5 4.2 mg (4 'lo). PI. > 350 oC. 'H NMR. (400 MHz, DMSO-d6) 5 (ppm): 7.32 (d, J = 8.3 Hz, 2H, H2 "), 7.27 (t, J = 7.4 Hz, 1H), 7, 21 (d, J = 7.8 Hz, 1H), 7.02 -6.91 (m, 1H), 6.79 (d, J = 8.4 Hz, 2H, H3 "), 6.49 - 6.35 (m, 1 H). HPLC (Isocratic H, O-MeCN 3367 'lo, 0.1% HCOOH for 60 min): t R = 33.18 (99.9%). MS (ES +): MS (ES, positive mode): miz 452.0 [M '].
10 3 '- (4-bromopheni /) - 5', 6-diffuoro-2, 6'-dioxo-I ', 5', 6 ', 7'-tetrahydrospiro [indophin-3,4'pirot / [3, 4-bjpiridinaj-5'-carbonitri / o (Compounds 15 and 16)
Dissolve in a la-8 spherical flask (70 mg, 0.155 mmol) and Selectfluor (82.25
mg, 0.232 mmol) in about 50 mL of MeOH and stirred at reflux for about 4 h. A
once the reaction is over, the solvent is removed under reduced pressure and the residue is
15 purified by column chromatography on silica gel (ethyl acetate: hexane), obtaining 30 mg (41%) of a white solid. The final product is separated into its diastereomeric pairs by semi-preparative HPLC using an isocratic elution gradient H20-MeCN 35% -40%, 0.1% HCOOH for 60 min.
Compound StructureMS (ES +) IHPLC
fifteen FMS (ES, positive mode): miz 471, 1 [M + W] HPlC (Isocratic H20-MeCN 35-40%, 0.1% HCOOH for 60 min): t, r30.7 (99.9%).
16 FMS (ES +): MS (ES, positive mode): mlz471, 1 [M + W]. HPlC (Isocratic H20-MeCN 35-40%, 0.1% HCOOH for 60 min): t, r 33.5 (99.9%).
15: They were 9.5 mg (13%). PI. > 350 oC. H NMR (400 MHz, DMSO-d6) and (ppm): 13.14 (s, 1H, NH), 11.85 (s, 1H, NH), 11.40 (s, 1H, NH), 7, 33 (d, J = 8.4 Hz, 2H, H3 "), 6.91 (dd, J = 8.5, 5.3 Hz, 1H, H4), 6.85 -6.74 (m, 3H , H2 ", H7), 6.53 (ddd, J = 9.8, 8.4, 2.5 Hz, 1 H, H5). HPlC (Isocratic H20-MeCN 35-40%, 0.1% HCOOH for 60
5 min): t R = 30.7 (99.9%). MS (ES +): MS (ES, positive mode): miz 471.1 [M + W]. 16: 1 mg (1%) was found. PI. > 350 oC. 'H NMR (400 MHz, DMSO-d6) and 7.49-7.07 (m, 4H), 7.06-6.66 (m, 3H). HPLC (Isocratic H, O-MeCN 35-40%, 0.1% HCOOH for 60 min): t, r 33.5 (99.9%). MS (ES +): MS (ES, positive mode): miz 471, 1 [M + H '].
Example 2 Measures of AMPK activation in cells in culture by the compounds of the general formula (IJ
cell line and treatments. the cell line used for treatments with
15 different reagents tested were HEK293T (human kidney embryonic cells). the cells were grown in DMEM medium (Dulbecco's Modified Eagle's Medium) with 25 mM glucose supplemented with 10% inactivated fetal bovine serum, 2 mM glutamine 100 units / ml penicillin and 100 IJg / ml streptomycin in a humid atmosphere at 3r C with 5% CO2. the cells were seeded on plates of
20 60 mm (p.60) to obtain a 70-80% confluence and washed in Krebs Ringer buffer (KRB: 12.5 mM NaCl, CaCl, 15 mM, KH, 2PO, 0.5 mM, 3 mM KCI , NaHCO, 2.5 mM, 0.5 mM MgS04, 10 mM HEPES pH 7.4, 95: 5 O2 / C02) at 3r C. Then, they were treated for 1 h at 3r C in the culture stove, adding to each of them the appropriate amounts of the compounds of general formula (1) 1, 11, 12, 13, and 15,
(5 mM stock in DMSO) dissolved in KRB / 25 mM glucose to reach
final concentrations indicated in the Figures. As activation control was used
5 mM fenformin.
5 Example3Obtaining HEK293T cell extracts
After the corresponding treatments, the supernatant was removed and quickly frozen in liquid N2. The plates were processed one by one, and kept on ice, first adding the cold lysis buffer. The composition of the buffer was as follows: 10 mM Tris pH 7.4, 15 mM EDTA pH 8.0, 50 mM NaF, Na, P, O, 15 mM, 0.6 M sucrose, 15 mM 2-Mercaptoethanol, a mixture of protease inhibitors without EDTA (Roche) and 1 mM PMSF. The cells were collected in lysis buffer with the help of a scraper and were lysed by syringes of 24
15 Gx5 / 8 "4 times each sample. A small amount was reserved to measure the amount of protein by Bradford and the rest was added with a loading buffer for electrophoresis and boiled for 5 min, kept at 2020 ° C until use. Protein concentration was determined by the Bradford method, with the Bradford Protein Assay Reagent Bio-Rad (BioRad).
20 Example 4 Protein Analysis by Western Blot
Protein extracts were analyzed by SDS-PAGE in gels of 8 or 10% of
25 acrylamide and 1.5 mm thick. 30 IJg of protein were loaded and transferred to a PVDF (Millipore) membrane for 1.5 h at 100 V. Blocking was done with 5% skim milk in TBS-T for 1 h at room temperature. Immunodetection was performed by incubating the primary antibody overnight at 4 ° C. After three washes at room temperature with TBS-T of 10 min each,
30 incubated with their corresponding secondary antibody conjugated to HRP. After three 10 min washes at room temperature with TBS-T, the membranes were developed with ECL plus (Pierce) and processed with a FUJI LAS 3000 (Fujifilm). The antibodies used were: Anti-pAMPKaThr172, anti-AMPK ~ 1 / ~ 2, anti-ACC and anti-pACCser79 from Cell Signaling Technology (Danvers, MA, USA)
35 diluted 1/1000. The secondary goat anti-rabbit HRP antibody from Santa Cruz Technology at a dilution 1/5000 or 1/10000. Detection of bands with the Anti-pAMPKaThr172 and anti-pACCser79 antibodies (AMPK substrate) with respect to their respective loading controls (anti-AMPK ~ 1 / p2 and anti-ACC) was taken as indicative of AMPK activation.
5 Example 6Results Description dose-response effect of the compounds of formulageneral you)
The activation status of the AMPK complex correlates with the levels of the form
10 phosphorylated of the catalytic subunit AMPKa pT172. As seen in the first panel of Figures 1-4, both the treatment of HEK293T cells with increasing doses of compounds 1, 11, 12, 13 and 15 increased the endogenous levels of AMPKa pT172, being indicative of the increased status of activation of the AMPK complex. The activation achieved in some cases was similar to that achieved
15 with fenformin, a compound with recognized AMPK activating capacity.
The second panel shows the analysis of the endogenous level of AMPKJ31, which was included as a load control to demonstrate that the increase in AMPKa pT172 levels were not due to an increase in the total levels of the enzyme complex.
20 The third panel shows the phosphorylation status of the enzyme acetyl CoA carboxylase (ACC), which is a substrate of the AMPK protein kinase, so an increase in AMPK activity increases the phosphorylation state of this substrate ( ACC pS79). As noted, the phosphorylation status of ACC increases
25 in parallel to the levels of AMPKa pT172.
The fourth panel indicates the total ACC levels as a load control, to validate that the change in ACC levels pS79 are due to the phosphorylation of the protein and not to an increase in its total levels.
All these data indicate that the compounds of general formula (1) 1, 11, 12, 13 and 15 are capable of inducing in vivo activation of the AMPK complex, at the indicated doses.

权利要求:
Claims (26)
[1]
1. Compound of formula (1)
R,
N
x
or
(one)
5 or its pharmaceutically acceptable isomers, salts or solvates, wherein: R 1 is selected from hydrogen, halogen, substituted or unsubstituted alkyl (Cl-Cm), ORa, SRa, COO (Ra), CF3, N (Rah. Y a substituted or unsubstituted aryl (CS-C1S) group; R2 is selected from hydrogen, substituted or unsubstituted (Cl-Cl0) alkyl, and
10 COR ;; R3 is selected from hydrogen, substituted or unsubstituted (Cl-Cl0) alkyl, and substituted or unsubstituted aryl (CS-C1S); ~ and Rs are independently selected from hydrogen, substituted or unsubstituted aryl (CS-C1S), halogen, and nitro group;
15 X is a halogen; and Ra is selected from hydrogen, substituted or unsubstituted alkyl (Cl-Cl0), and substituted or unsubstituted aryl (CS-C1S).
[2]
2. A compound according to claim 1, wherein X is fluorine.
[3]
3.-Compound according to any of the preceding claims, wherein R1 is chlorine, bromine, or a substituted or unsubstituted aryl group (CS-C1S).
[4]
4. A compound according to claim 3, wherein R1 is chlorine, bromine, or a substituted phenyl group.
[5]
5.-Compound according to any of the preceding claims, wherein R2 is hydrogen.
[6]
6. A compound according to any of the claims hydrogen, a phenyl group, or 4-methoxyphenyl.
[7]
7.-Compound according to any of the claims selected from hydrogen and fluorine.
[8]
8.-Compound according to any of the claims
P201 531786
above, where R3 is
above, where R4 is
above, where Rs is
10 selects from hydrogen, fluorine, a nitro group, and a phenyl group.
[9]
9. A compound according to claim 1, wherein the compound is selected from the list consisting of: 3 '- (4-chlorophenyl) -5'-flu oro-2, 6' -d ioxo-1 ', 5', 6 ', 7'-tetrah id rospiro [indolina-3,4'-pyrazole [3,4
15 b] pyridine] -5'-carbonitrile; 3 '- (4-chlorophenyl) -5.5' -diflu gold-2,6'-dioxo-1 ', 5', 6 ', 1'-tetrahydrospiro [indoline-3,4'pyrazole [3, 4- b) pyridine) -5'-carbon itrile; 3 '- (4-chlorof in il) -5-flu oro-2,6'-dioxo-5-n itro-1', 5 ', 6', l'-tetrah id rospi ro [indol i na-3 , 4'pyrazol (3,4-b] pyridi na] -5'-carbon itrile;
20 3 '- (4-chlorophenyl) -5' -fluoro-2,6 '-dioxo-5-phenyl-1', 5 ', 6', T-tetrahydrospiro (indoline-3,4, pyrazole (3, 4 -b] pyridi na] -5 '-carbon itri lo; 5-chloro-3' - (4-chlorophenyl) -5 '-f1uoro-2,6'-dioxo-1', 5 ', 6', 1 '-tetrahydrospiro [indoline-3,4'pyrazol (3,4-b] pyridine] -5'-carbonitrile; 3' - (4-chlorophenyl) -5 ', 6-diflu or -2,6'-dioxo- 1 ', 5', 6 ', T-tetrahydrospiro [indoline-3,4'
25 pyrazol (3, 4-b] pyridi na] -5'-carbon itrile; 3 '- (4-bromophenyl) -5' -fluoro-2,6'-dioxo-1 ', 5', 6 ', 1'-tetrahydrospiro [indoline-3,4'pyrazol (3,4-b] pyridine] -5'-carbonitrile; and 3 '- (4-bromophenyl) -5', 6-difl uoro-2,6'- dioxo-1 ', 5', 6 ', T-tetrahydrospiro [indoline-3,4'pyrazol (3,4-b] pyridi na] -5'-ca rbonitrile.
[10]
10. A pharmaceutical composition comprising a compound of formula (1), or its pharmaceutically acceptable isomers, salts or solvates as defined in any one of claims 1 to 9, and at least one pharmaceutically acceptable carrier.
[11]
11. Use of a compound of formula (1), or its pharmaceutically acceptable isomers, salts or solvates as defined in claims 1 to 9, for the preparation of a medicament.
[12]
12. Use of a compound of formula (1), or its pharmaceutically acceptable isomers, salts or solvates as defined in claims 1 to 9, for the preparation of a medicament for the prevention or treatment of diseases related to AMPK activation.
[13]
13. Use according to claim 12, wherein the diseases are selected from cardiovascular, inflammatory, autoimmune, neurological, metabolic syndrome and cancer.
[14]
14. Use according to claim 13, wherein the diseases are selected from type 1 and type 2 diabetes, obesity, inflammation, dyslipidemia, hypertension, hyperglycemia, hypertriglycerimidemia, insulin resistance, epilepsy and the diseases of KrabbelTwitcher, Alzheimer's, Parkinson and Huntington.
[15]
15. Use according to claim 14, wherein the disease is type 1 diabetes, epilepsy, KrabbelTwitcher disease, Alzheimer, Parkinson or Huntington.
[16]
16. Use according to claim 13, for the preparation of a medicament for the prevention or treatment of prostate and breast cancer.
[17]
17. Process for obtaining the compound of general formula (1) as defined in any of claims 1 to 9 comprising the following steps:
(a) condensation reaction between compound of formula (11) and an NH2NHRJ compound
"
or
"
P201 531786
wherein R1 and R3 are defined in claim 1;
(b) condensation reaction of the compound of formula (IV) R,
5 IV with the compound obtained in step (a) of formula (111) R,
111 for 12-24 hours at 60-120 ° C; where R1. R2. R3. ~ And Rs are defined in claim 1; Y
10 (c) reaction of the compound obtained in step (b) of formula (la)
l. with a halogenating agent; where R1. R2. R3 ~ And Rs are defined in claim 1.
[18]
18. Process according to claim 17, wherein the halogenating agent of step (c) is selectfluor.
[19]
19.-Compound of formula (the)
R,
<N N
or
R, /
.
or SUS pharmaceutically acceptable isomers, salts or solvates, wherein: R1 is selected from hydrogen, halogen, substituted or unsubstituted (C1-C10) alkyl, ORa, SRa, COO (Ra), CF3, N (Rah, and a aryl group substituted or not
5 substituted R2 is selected from hydrogen, substituted or unsubstituted (C1-ClO) alkyl, and CORa; RJ is selected from hydrogen, substituted or unsubstituted (C1-C10) alkyl, and substituted or unsubstituted aryl;
R4 and Rs are independently selected from hydrogen, aryl, halogen, and nitro group; Ra is selected from hydrogen, substituted or unsubstituted (C1-C10) alkyl, and substituted or unsubstituted aryl; with the proviso that when R1 is hydrogen, ~ it is not hydrogen.
[20]
20.-Compound according to claim 19, wherein X is fluorine.
[21]
21.-Compound according to claims 19 and 20, wherein R1 is chlorine, bromine, or a substituted or unsubstituted aryl group.
[22]
22.-Compound according to claim 21, wherein R1 is chlorine, bromine, or a substituted phenyl group.
[23]
23.-Compound according to claims 19 to 22, wherein R is hydrogen. 25
[24]
24.-Compound according to claims 19 to 23, wherein R3 is hydrogen, a phenyl group, or 4-methoxyphenyl.
[25]
25.-Compound according to claims 19 to 24, wherein R4 is selected from hydrogen and fluorine.
[26]
26. A compound according to claims 19 to 25, wherein Rs is selected from hydrogen, fluorine, a nitro group, and a phenyl group.

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ES2623082B1|2018-06-20|

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WO2017098073A1|2017-06-15|

引用文献:

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公开号 | 申请日 | 公开日 | 申请人 | 专利标题

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WO2011069298A1|2009-12-11|2011-06-16|F. Hoffmann-La Roche Ag|Novel cyclopropane indolinone derivatives|

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EP3010905A1|2013-06-20|2016-04-27|Boehringer Ingelheim International GmbH|Spiro-substituted oxindole derivatives having ampk activity|

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ES201531786A|ES2623082B1|2015-12-10|2015-12-10|SPIRANICAL COMPOUNDS DERIVED FROM OXINDOL-PIRAZOLO [3,4-b] PIRIDINONA AND ITS THERAPEUTIC USES|ES201531786A| ES2623082B1|2015-12-10|2015-12-10|SPIRANICAL COMPOUNDS DERIVED FROM OXINDOL-PIRAZOLO [3,4-b] PIRIDINONA AND ITS THERAPEUTIC USES|

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PCT/ES2016/070868| WO2017098073A1|2015-12-10|2016-12-09|Spiranic derivative compounds of oxindole-pirazolo[3,4-b]pyridinone and therapeutic uses thereof|