• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    APAF-1 INHIBITING COMPOUNDS. The 2,5-piperazinadione derivatives of formula (I) are inhibitors of apoptotic peptidase activating factor 1 (Apaf-1), which are useful as active pharmaceutical principles for the prophylaxis and / or treatment of a pathological condition and / or physiological effect associated with an increase in apoptosis. (I)
    公开号:BR112012002134B1
    申请号:R112012002134-6
    申请日:2010-07-29
    公开日:2021-03-02
    发明作者:Àngel Messeguer Peypoch；Alejandra Moure Fernández；Daniel González Pinacho；Isabel Masip Masip；Enrique Perez Payá；Natividad Garcia Villar；Ester Monlleó Mas；Juanlo Catena Ruiz
    申请人:Spiral Therapeutics Inc；
    IPC主号:
    专利说明:

    The present invention relates to compounds for the prophylaxis and / or treatment of disorders caused by apoptosis cell death or for the prevention of degenerative processes caused by apoptosis cell death. State of the art
    Apoptosis, or programmed cell death, is a complex physiological phenomenon involved in maintaining cellular homeostasis. Apoptosis is regulated by several cellular control mechanisms due to its key role in maintaining health. Many pathologies are based on dysfunction of apoptosis. For this reason, an excess in cell death due to apoptosis can affect tissue functionality (for example, death of cardiomyocytes in cases of myocardial infarction), while an excessively inhibited apoptosis leads to uncontrolled cell survival (for example, neoplastic processes) . The cellular components that regulate apoptosis are in constant dynamic balance in a healthy cell. There are at least two well-characterized pathways for activating the apoptotic cascade of caspases. One of them, the extrinsic pathway, is activated by means of extracellular signaling and requires the participation of specific membrane receptors. The intrinsic pathway corresponds to cell stress, toxic agents, radiation, oxidizing agents, Ca2 + overload, DNA damage; it is activated in response to oncogenes and implies mitochondrial destabilization. In some pathophysiological conditions (for example, anoxia in organ cells to be transplanted, treatments with toxic substances), apoptosis increases and an excessive number of cells die, making the affected and impaired tissue function impossible. in some cases their survival.
    The molecular mechanisms of apoptosis induction lead to the activation of proteins with protease activity called caspases, also known as effectors of apoptosis. For these to be activated, the formation of a molecular complex called apoptosome is necessary. Apoptosome is formed by means of cytochrome c, pro-caspase-9 and apoptotic peptidase activation factor 1 (Apaf-1, Apoptotic Peptidase Activation Factor). It has been shown that inhibition of Apaf-1 inhibits the formation of an apoptosome complex and that it causes an inhibition of apoptosis (measured by activating caspase 3). In cellular assays in which apoptosis is induced by hypoxia (by reducing the concentration of oxygen in the air) or by chemical compounds, an increase in cell survival was observed when they were previously treated with apoptosis inhibitors.
    In a similar way, during the process of removal and transplantation of an organ, its cells are subjected to a condition of hypoxia that can lead to cell death thereby compromising the viability and functionality of the organ. For this reason, for example, only 70% of all corneas donated for transplant are suitable for implantation. This is due to the fact that cell death by apoptosis occurs during storage of the corneas. A similar situation occurs during kidney and heart transplants. There are solutions on the market for the transport of organs that exclusively provide buffered and sterile environments, as they do not contain any active molecules that are capable of preventing cell death by apoptosis.
    The study of the mechanisms involved in apoptosis allowed the identification of different potential pharmacological targets. For this reason, inhibitors have been designed that act at different levels of the apoptotic cascade, such as transcription factors, kinases, regulators of mitochondrial membrane permeabilization and inhibitors of the caspase family.
    Considering that the formation of the apoptosome is a key step in the apoptotic cascade and the consequent activation of caspases, the inhibition of Apaf-1 activation may have a greater impact on the inhibition of apoptosis than other studied pharmacological targets. There are indications in the scientific literature about the therapeutic implications of Apaf-1 inhibition. For this reason, the transduction of a negative dominant Apaf-1 by means of an adenovirus in a Parkinson's animal model was more effective than the transduction by adenovirus of a negative dominant of Caspase-1. Document W02007060524 describes the compounds derived from [1,4] diazepan-2,5-dione of the formula set out in
    followed as inhibitors of apoptosis.
    W02008009758 describes the compounds of the formula shown below as being inhibitors of UBC13-UEV interactions, which can be used in the preparation of pharmaceutical compositions for anti-tumor therapy or for the treatment and / or prophylaxis of diseases associated with metabolic routes, in which the UBC13 enzyme intervenes, with metabolic routes, in which the NF-kB transcription factor intervenes, or with routes in which PCNA or RAD6 intervenes. Although they can be considered as structurally similar to those of the present invention, they have a different use. R- (CRÍR2) q-CO-N (R3) -C (R4R5) -CO-NH2
    For this reason, it is desirable to provide new Apaf-1 inhibitory compounds. Description of the Invention
    The present invention provides new compounds derived from 2,5-piperazinadione of formula (I) which are endowed with Apaf-1 inhibitory activity.
    For this reason, a first aspect of the invention concerns the compounds of formula (I):
    and their pharmaceutically acceptable salts, where:
    RI and R2 are independently selected from -H, C1-5 alkyl, C2-5 alkenyl, - (CH2) 0-3 “cycloalkyl, - (CH2) 1-3-heterocycle, - (CH2) 0- 3-aryl, - (CH2) o-3 ~ heteroaryl, - (CH2) I-2-CH (aryl) 2, - (CH2) i-2- CH (aryl) (heteroaryl) e- (CH2) I- 2-CH (heteroaryl) 2,
    R3 is selected from -H, C1-5 alkyl, C2-5 alkenyl, - (CH2) 0-3-cycloalkyl, - (CH2) 1-3-heterocycle, - (CH2) 1-3-aryl , - (CH2) 1-3-heteroaryl, - (CH2) 1-3-CONR5R6, - (CH2) 1- 2-CH (aryl) 2, - (CH2) I-2-CH (aryl) (heteroaryl) e- (CH2) i-2-CH (heteroaryl) 2,
    R4 is selected from -H, C1-5 alkyl, - (CHR7) I-3-CO-NR5R6, - (CHR7) I-3-CO-OR5, - (CH2) i-3-NR5R6, - (CH2) 1-3- CO [NCHR7CO] mNH2 e- (CH2) i-3-C0 [NCHR7CO] mOR5, n is an integer selected from 1 and 2; m is an integer selected from 1, 2 and 3;
    R5 and R6 are independently selected from -H, C1-5 alkyl and- (CH2) 0-3-aryl,
    R7 is selected from -H, C1-6 alkyl, - (CH2) i-3-aryl and- (CH2) 1-3-heteroaryl, so that when m is greater than 1, the R7 substituents can be the same or different from each other, where the C1-6 alkyl, C2-5 alkenyl, cycloalkyl and heterocycle groups can be optionally substituted with one or more substituents selected independently from halogen, OR5, OCF3, SH, SR5, NR5R6 , NHCOR5; COOH, COOR5, OCOR5, aryl and heteroaryl, in which the aryl and heteroaryl groups can be optionally substituted with one or more substituents selected independently from halogen, CF3, OR5, OCF3, SH, SR5, NH2, NHCOR5; NO2, CN, COR5, COOR5, OCOR5, CONR5R6, - (CH2) 0_3NR5R6, SO2NH2, NHSO2CH3, C1-5 alkyl, aryl and heteroaryl, in which the heterocycle and heteroaryl groups can be optionally substituted on a secondary nitrogen atom with C1-5-alkyl, cycloalkyl or- (CH2) 0_3-aryl, with the proviso that when R2 is comprised of 2- (4-fluoro-phenyl) ethyl, R4 is comprised of -CH2-CO-NH2 and n is 1 , then: - if RI is comprised of 2- (4-fluoro-phenyl) ethyl, R3 is not 2- (4-methoxy-phenyl) ethyl, 2- (2-pyridyl) ethyl or 2- (2,4- dichlorophenyl) ethyl, and - if RI is comprised of 2- (2,4-dichlorophenyl) ethyl, R3 is not 2- (4-methoxy-phenyl) ethyl or 2- (2-pyridyl) ethyl.
    According to a particular embodiment of the invention, R1 is comprised of C1-5 alkyl or- (CH2) 0-3 aryl.
    According to another particular embodiment of the invention, R2 is comprised of C1-5 alkyl, - (CH2) 0_3-aryl, - (CH2) 0-3-heteroaryl or - (CH2) I-2-CH (aryl) 2 .
    According to another particular embodiment of the invention, R3 is comprised of -H, C1-5 alkyl, - (CH2) 1-3-heterocycle, - (CH2) 1-3-aryl or- (CH2) 1-3- heteroaryl.
    According to another particular embodiment of the invention, R4 is comprised of -H, - (CHR7) 1-3-CO-NR5R6, - (CHR7) 1-3-CO-OR5 or- (CH2) 1-3-CO [ NCHR7CO] mNH2.
    According to another particular embodiment of the invention, n is
    According to another particular embodiment of the invention, m is
    According to another particular embodiment of the invention, R5 is comprised of -H or I-C1-5 alkyl
    According to another particular embodiment of the invention, R6
    De is understood by -H. according to another particular embodiment of the invention, R7 is comprised of -H, C1-5 alkyl, - (CH2) 1-3 “aryl or- (CH2) 1-3-heteroaryl.
    A second aspect of the present invention relates to a compound of formula (I), or a pharmaceutically acceptable salt thereof for use as an active pharmaceutical ingredient, and particularly for use in the prophylaxis and / or treatment of a pathological condition and / or physiological associated with an increase in apoptosis, wherein the pathological and / or physiological condition associated with an increase in apoptosis is selected from organ or cell preservation, and particularly transplantation or conservation; prevention of cytotoxicity, and particularly cytotoxicity mediated by chemical substances, by physical agents such as radiation, acoustic trauma, burns or by biological agents such as hepatitis virus infection; pathologies due to hypoxia conditions, such as cardiac infarction or cerebral infarction; eye pathologies, such as lesions caused by eye surgery, age-related macular degeneration, diabetic retinopathy, retinitis pigmentosa or glaucoma; neurodegenerative diseases, such as Alzheimer's, Huntington's, Parkinson's or amyotrophic multiple sclerosis; diabetes, and particularly preservation of islets of Langerhans or cytotoxicity associated with diabetes such as, for example, nephrotoxicity; osteoarthritis; arthritis; inflammation or immunodeficiencies, such as AIDS-associated CD4 + T lymphocyte decrease.
    Another aspect of the present invention relates to the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof for the manufacture of a medicament intended for the prophylaxis and / or treatment of a pathological and / or physiological condition associated with an increase of apoptosis, and particularly one of the conditions mentioned above.
    Another aspect of the present invention relates to a method of prophylaxis and / or treatment of an individual or organ that suffers or is susceptible to suffering from a pathological and / or physiological condition associated with an increase in apoptosis, and particularly one of the conditions mentioned above. , which comprises administering to said individual or organ a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof together with sufficient amounts of pharmaceutically acceptable excipients.
    Preferred are the compounds of formula (I) and their pharmaceutically acceptable salts, and particularly the compounds of formula (I) described as examples or as intermediates.
    The compounds of the present invention can be used alone or in combination with one or more compounds that are useful for the prophylaxis and / or treatment of a pathological and / or physiological condition associated with an increase in apoptosis, such as preservation of organs or cells , and particularly transplantation or conservation; prevention of cytotoxicity, and particularly cytotoxicity mediated by chemical substances, by physical agents such as radiation, acoustic trauma, burns or by biological agents such as hepatitis virus infection; pathologies due to hypoxia conditions, such as cardiac infarction or cerebral infarction; eye pathologies, such as lesions caused by eye surgery, age-related macular degeneration, diabetic retinopathy, retinitis pigmentosa or glaucoma; neurodegenerative diseases, such as Alzheimer's, Huntington's, Parkinson's or amyotrophic multiple sclerosis; diabetes, and particularly preservation of islets of Langerhans or cytotoxicity associated with diabetes such as, for example, nephrotoxicity; osteoarthritis; arthritis; inflammation or immunodeficiencies, such as AIDS-associated CD4 + T lymphocyte decrease.
    The term "C1-5 alkyl", alone or in combination, means a straight or branched chain alkyl group that is endowed with 1 to 5 carbon atoms.
    The term "alkenyl-Cs-s", alone or in combination, means a straight or branched chain group that is endowed with 2 to 5 carbon atoms and that is endowed with one or more unsaturated bonds.
    The term "cycloalkyl", alone or in combination, refers to a stable monocyclic radical of 3 to 7 members, which is saturated or partially saturated, and which only consists of carbon and hydrogen atoms. Examples of cycloalkyls are as follows: cyclopropyl, cyclopentyl, cyclohexyl, 1-cyclohexyl, cycloeptyl.
    The term "heterocycle", alone or in combination, means a saturated or partially unsaturated heterocycle of 5 to 10 bonds, which contains one or more heteroatoms selected from nitrogen, oxygen and sulfur. For the purposes of this invention, the heterocycle can be a monocyclic or bicyclic ring system that can include condensed ring systems. Examples of heterocyclic groups are tetrahydrofuranyl (THE), dihydrofuranyl, dioxanyl, morphophilic, piperazinyl, piperidinyl, 1,3-dioxolanyl, imidazolidinyl, imidazolinyl, pyrrolidyl, pyrrolidinyl, tetrahydropyranyl, dihydropyranyl and ashydropyranyl.
    The term "aryl", alone or in combination, refers to a mono or polycyclic aromatic ring system containing carbon ring atoms. Preferred aryls are 5-10-membered monocyclic or bicyclic aromatic ring systems, such as ■ phenyl or naphthyl having optionally one or more substituents, preferably one to three independently selected from halogen, CF3, OH, 0R5, OCF3 , SH, SR5, NH2, NHCOR5; NO2, CN, COR5, COOR5, OCOR5, CONR5R6, - (CH2) o-3 NR5R6, SO2NH2, NHSO2CH3, C1-5 alkyl, aryl and heteroaryl.
    The term "heteroaryl", alone or in combination, refers to an aromatic or partially aromatic heterocycle that contains at least one ring heteroatom selected from O, S and N. Thus, heteroaryls include condensed heteroaryls to other classes of rings, such as aryls, cycloalkyls and heterocycles that are not aromatic. Examples of heteroaryl groups include: pyrrolyl, isoxazolyl, isothiazolyl, pyrazolyl, pyridyl, oxazolyl, thiazolyl, imidazolyl, triazolyl, furyl, thienyl, pyrimidyl, benzisoxazolyl, benzoxazolyl, indothobenzolyl, dihydrobenzolol, indidrobenzol quinazolinyl, naphthyridinyl, isobenzylfuranyl, benzimidazolyl, benzofuranyl, benzothienyl, quinolyl, indolyl, isoquinolyl, dibenzofuranyl, benzothiophenyl, tetrahydrobenzothiophenyl and the like.
    The term "optionally substituted with one or more substituents" means that a group can be unsubstituted or else substituted with one or more substituents, preferably with 1, 2, 3 or 4 substitutes, from the moment that said group be equipped with 1, 2, 3 or 4 positions that can be replaced. The term "pharmaceutically acceptable salts" means those salts that retain the efficiency and typical properties of free bases or free acids and that do not cause discomfort in a biological sense or in any other sense.
    According to the invention, the compounds of formula (I) and their pharmaceutically acceptable salts are useful for the prophylaxis and / or treatment of a pathological and / or physiological condition that is associated with an increase in apoptosis through its activity as inhibitors of Apaf-1.
    Unless otherwise defined, all technical and scientific terms used in this context have the same meaning as those commonly understood by the person skilled in the field of the invention. Methods and materials that are similar or equivalent to those described in this context can be used in the practice of the present invention. Throughout the description and claims the word "comprises" and its variants are not intended to exclude other technical characteristics, additives, components, stages or stereoisomers of the compounds involved. For those skilled in the art, other objects, advantages and features of the invention will be understood partly from the description and partly from the practice of the invention.
    The compounds of formula (I) can be prepared following different methods known to anyone skilled in the field of organic synthesis, and particularly through the general procedures illustrated in the following schemes. The starting materials for the preparation methods are either commercially available or can be prepared using methods in the literature. Unless otherwise indicated, the groups R1, R2, R3, R4, R5, R6 and R7 have the meaning described in general formula (I).
    The compounds of formula (I) can be obtained from the methods and schemes described below: Method A Scheme 1

    According to Method A, since it is deprotected from the fluorenomethyloxycarbonyl group, the amine II attached to the solid support is acylated with an acylating agent III, where X represents a leaving group, for example, a halogen and Y represents OH or halogen. When Y represents a halogen, for example, chloroacetyl chloride, the reaction can be carried out in the presence of a base, such as triethylamine. When Y represents -OH, for example, bromic acetic acid, the reaction can be carried out in the presence of a suitable coupling agent, for example, N, N'-diisopropylcarbodiimide. In both cases, the reaction can be carried out in an inert solvent that is capable of swelling the resin, such as N, AP-dimethylformamide or methylene chloride and at room temperature or under microwave irradiation to reduce the reaction time to a minimum. Then, amine IVa is coupled using a tertiary amine as a base. The reaction can be carried out at room temperature or under microwave irradiation.
    A carboxylic acid VI, where PG represents a protecting group, such as allyl, is reacted with amine V to obtain amide VII, using a coupling agent, such as, for example, the combination of 7Y, N'-diisopropylcarbodiimide and 1-hydroxybenzotriazole. An IVb amine is then added, via Michael's reaction using a base and a solvent, such as N, 2V-dimethylformamide or dimethyl sulfoxide to obtain intermediate VIII after excision of the resin using a mixture trifluoroacetic acid, dichloromethane and water. Intermediate VIII is cyclized (intermediate IX) and deprotected in a basic medium providing intermediate acid X.
    Intermediate X can alternatively be prepared in the solid phase according to Scheme 2, where amine V 'can be prepared from amine IVa either by means of a reducing amination reaction with a glyoxylate in THF-AcOH using a reducing agent such as NaBHaCN, or alternatively by means of alkylation with a bromoacetate or a bromoacetamide using a tertiary amine as the base. Subsequently, acid VI is coupled to obtain 5 amide VII '. An IVb amine is then added and through Michael's reaction and subsequent in situ cyclization produces intermediate ester IX, which through basic treatment provides compound X. Scheme 2
    Layout 3
    Obtaining a compound of formula I:
    A compound of formula Ia can be obtained from intermediate X by coupling with an amine attached to a solid support Va or Va ', obtained according to the methodology indicated above, in the presence of a coupling agent, such as, for example, the combination of HATU and HOBT. The compound of formula Ib can be obtained in a similar manner to the synthesis of compound Ia, except in the case of the solid base on which the solid starting support (Ilb) has a halogen group instead of an amino group, such as, for example, example, chlorotrithyl resin, obtaining an acid after excision from the resin. The Ic ester can be synthesized by means of esterification of the corresponding acid Ib by means of common esterification methods in organic synthesis, such as, for example, using methanol in an acidic medium such as sulfuric acid. In the case of Ib, the same can be achieved by saponification of the Ic ester. The compounds of formula Id can be obtained by reacting intermediate X with a primary amine IVc.
    An alternative strategy for obtaining the compounds of formula I can be carried out by acylating amine II with an amino acid of formula XI (Method B). Method B

    As is obvious to a person skilled in the field of the invention, it is possible to combine some of the steps of method A with some of the steps of method B to obtain a compound of formula I.
    Alternatively, it is possible to obtain the compounds of formula le and If as illustrated in the scheme described below. Layout 3

    Peptide XIII and pseudopeptide XIV that will be linked to acid X can be obtained by means of standard peptide synthesis reactions. For this reason, the amine resin (Z = NH2) OR chloride Ilb (Z = C1) can be reacted with a suitably protected amino acid (XII), using a suitable coupling agent.
    Optionally, the process can be repeated successively, previously deprotecting the amine, to obtain peptide XIII. Carboxylic acid X then reacts with XIII to obtain compound le. By combining the amino acid units (XIII) with a glycine unit (following method A or B), pseudopeptide XIV is obtained, which will lead to obtaining If in a manner similar to that previously described.
    The primary amines used IVa, IVb and IVc are found commercially available or can be obtained by known methods (March, Advanced Organic Chemistry, 1991, Ed. John Wiley & Sons) or by using, for example, the schemes described then. Layout 4

    An amine can be obtained by means of a Mitsunobu reaction starting from alcohol and potassium phthalimide in the presence of, for example, diethyl azodicarboxylate (DEAD) and triphenylphosphine in tetrahydrofuran as a solvent and later release with hydrazine hydrate. (Mitsunobu, J. Am. Chem. Soo. 1972, 94, 679-680)
    N-substituted glycines V and XI can be synthesized using some of the methods that are illustrated below, such as, for example, reducing amination of the corresponding glycine with a suitable aldehyde (Scheme 5) using reducing agents, such as NaBHo NaBHβCN or NaBH (AcO) 3 or by nucleophilic substitution of an ester with an R-NH2 amine (Scheme 6). Layout 5
    Layout 6
    Examples Abbreviations: AcOEt Ethyl acetate Brine Saturated NaCl solution DCM Dichloromethane DIC N, N '-di-isopropyl-carbodiimide DIPEA N, 7 7-diisopropyl-ethyl-amine DMF N, N-dimethi1-formamide DMSO Dimethyl- sulfoxide EDO 1- (3-dimethyl-amino-propyl) -3-ethyl-carbodiimide Eq. molar equivalent Et3N Triethyl-amine Fmoc 9-Fluorenyl-methoxy-carbonyl IPA Isopropyl alcohol HATU 2- (lH- hexafluror-phosphate) 7-azabenzotriazol-l-yl) -1,1,3,3-tetramethyl ~ uronium HOBT 1-Hydroxy-benzotriazole HPLC High-performance liquid chromatography HRMS High-resolution mass spectrometry MeOH Methanol PyBOP Hexotluoro-phosphate benzotriazole-l- iloxy-trispirrolidinophosphonium RP Reverse phase rt Ambient temperature tr Retention time UV Ultraviolet TFA Trifluoroacetic acid
    The following examples serve to better illustrate the invention, and should not be considered as limiting it.
    The nomenclature used in this document is based on the CFW_CHEMICAL_NAME function present in version 12 of Chemdraw for Excel. General data:
    The compounds were synthesized using an AM RAM polystyrene resin purchased from Rapp Polymere GmbH (Germany). In the reactions, polystyrene syringes were used with a polyethylene disk using an IKA Labortechnik digital HS501 agitator. In the reactions carried out by means of microwaves the model CEM Discover was used with 10 ml glass reactors.
    The products were analyzed by: • Method A: Using an RP-HPLC using a Hewlett Packard Series 1100 equipment (UV detector 1315A) using an X-Terra C18 reverse phase column (15 x 0.4 6 cm , 5 μm). The wavelength used for the detection of UV was 210 nm. Mixtures of CH3CN-H2O with 0.1% TEA under 1 ml / min. were used as a mobile phase. The analyzes were conducted with a gradient of 20% to 70% CH3CN (10 min), and from 70% to 100% (8 min). • Method B: The products were analyzed using an Agilent 1100 HPLC equipment, equipped with a variable wavelength UV detector and a model 1100 VL mass spectrometer. The wavelength used for UV detection was 210 nm, while the MS detector was operated in the positive electrospray ionization mode and performed a scan from 100 m / z to 1300. Regarding the chromatographic separation , the column used was a Kromasil 100 C18 (4.0 x 40 mm, 3.5 μm) adjusted to 50 ° C, and 5 μl was injected. For elution, one of the solvent gradients described below was followed: 5-100% B in 7 min, 5% B 7- 8.5 min. The flow rate of the mobile phase is 1.4 ml / min. Solvent A consists of 0.2% formic acid in water, while B is comprised of 0.2% formic acid in acetonitrile. • Method C: using HPLC-UV-MS equipment from Waters provided with a detector equipped with serial diodes and a mass spectrometer model EMD1000. The wavelength used for UV detection was 210 nm, while the MS detector was operated in the positive electrospray ionization mode and a scan of 100 to 1000 m / z was performed. Regarding the chromatographic separation, the column used was a Kromasil C18 (2.1 x 50 mm, 3.5 μm) adjusted to 50 ° C and 2 μl was injected. For the elution, the following gradient was followed: 5-100% B, 0-5 min; 100% B, 5-6.5 min .; 5% B, 6.5-8 min. The flow rate of the mobile phase is 0.5 ml / min.
    High resolution mass spectrometry was performed using UPLC-HRMS using a Waters Acquity UPLC equipment coupled to an orthogonal acceleration flight time mass spectrometer model LCT Premier XE by Waters. Chromatographic analysis was performed using a Waters Acquity C18 column (10 x 2.1 mm, 1.7 μm).
    Mixtures of CH3CN-H2O with 20 mM formic acid at 0.3 ml / min were used as the mobile phase. The analyzes were conducted with a variable gradient from 50% to 100% CH3CN in 6 min. Intermediate VI VI: (Z) -2-Butenedioic acid allylic ester
    To a solution of 2 g of maleic anhydride (20 mmol) in chloroform, 1.8 ml of allyl alcohol (26 mmol, 1.3 eq.) Were added. The reaction mixture was stirred at reflux for 5 h. The resulting solution was treated with IN HC1 and was extracted with chloroform. The organic extracts were washed with a saturated solution of sodium chloride, dried over anhydrous magnesium sulfate and filtered. The solvent was evaporated under reduced pressure, and the residue obtained was identified as intermediate VI as an oil (95% purity, 85% yield). Intermediates X Xl: 2- (4- (2,4-dichloro-phenethyl) -1- (3,3-diphenyl-propyl) -3,6-dioxo-piperazin-2-yl) acetic acid

    A mixture of 2 g of Fmoc-Rink Amide AM polystyrene resin (0.61 mmol / g resin, 1.22 mmol) and 12 ml of 20% piperidine in DMF was stirred in a microwave reactor for 2 min . under 35 ° C. The resin was filtered and washed with DMF (3 x 15 ml), isopropyl alcohol (3 x 15 ml) and DCM (3 x 15 ml). The resin was treated with a solution of bromoacetic acid (III, 840 mg, 5 eq.) And N, N'-diisopropylcarbodiimide (1.15 ml, 5 eq.) In DMF (12 ml). The reaction mixture was subjected to stirring for 2 min, under 60 ° C in a microwave reactor. The resin was filtered and washed with DMF (3 x 15 ml), isopropyl alcohol (3 x 15 ml) and DCM (3 x 15 ml). A solution of 2,4-di-chloro-phenethylamine (IVa, 1.035 ml, 5 eq.) And triethylamine (0.85 ml, 5 eq.) In 12 ml of DMF was added to the resin and the suspension was subjected to stirring. for 2 min, under 90 ° C, activated by microwave. The supernatant was removed and the reaction was repeated under the same conditions. The obtained resin V was filtered and washed with DMF (3 x 15 ml), isopropyl alcohol (3 x 15 ml) and DCM (3 x 15 ml). Then, the resin was treated with a solution of allyl ester of (Z) —2— butenedioic acid (VI, 957 mg, 5 eq.), HOBT (825 mg, 5 eq.) And DIC (770 μl, 5 eq. ) in DCM: DMF (2: 1, 123 ml). The reaction mixture was subjected to stirring at room temperature for 30 min. and was filtered. The resin was subjected to drying and washed with DMF (3 x 15 ml), isopropyl alcohol (3 x 15 ml) and DCM (3 x 15 ml). Then, a solution of 3,3-diphenylpropylamine (IVb, 1.29 g, 5 eq.) And triethylamine (0.85 ml, 5 eq.) In 12 ml of DMF was added to the resin and the suspension was subjected to stirring for 3 h at room temperature. The resin was filtered and the reaction was repeated for 16 h at the same temperature. The supernatant was removed and the resin was dried and washed with DMF (3 x 15 ml), isopropyl alcohol (3 x 15 ml) and DCM (3 x 15 ml). The excision from the solid phase was carried out by means of treatment with a 60: 40: 2 TFA / DCM / water mixture (20 ml) for 30 min. at room temperature. The reaction mixture was filtered and the solvents were evaporated under reduced pressure. Cyclization was then carried out by treating the residue obtained with 20 ml of dioxane for 1.5 h under reflux (monitoring the reaction by means of HPLC). A 1: 2 solution (9 ml) of 4N sodium hydroxide and allyl alcohol was then added and the mixture was stirred for 45 min. under reflux. The crude reaction product was acidified with 1N hydrochloric acid and the solvent was evaporated. The resulting solution was extracted with ethyl acetate (3 x 50 ml) and the organic extracts were washed with a saturated NaCl solution (2 x 100 ml), dried in anhydrous MgSO4 and evaporated under reduced pressure to obtain 450 mg of the desired product (X, 70% purity, 95% yield under 210 nm). HRMS (M + H) + calculated for C29H29C12N2O4, 539.1504, experimental, 539.1515.
    The compounds shown below were prepared following a method similar to that described in the previous example, but using different amines:

    X.13: 2- (4- (2,4-dichloro-phenethyl) -3,6-dioxo-l- (4- (trifluoromethyl) benzyl) piperazin-2-yl) acetic acid
    Step 1: Intermediate V '
    The amount of 7.55 ml of Et3N and 2.50 g (27 mmol) of bromoacetamide was added to a solution of 5 g (41 mmol) of 2- (2-pyridyl) ethylamine in 300 ml of dioxane. The resulting mixture is heated to reflux overnight. The solution is evaporated to dryness and purified on silica gel using a mixture of AcOEt: MeOH: NH3 (10: 1: 0.01) as an eluent, thus providing 2.39 g of intermediate V '. Method B: tr: 0.261, m / z: 180. Step 2: Intermediate VII '- The amount of 4.90 ml of Et3N, 3.24 g (24 mmol) of HOBT, 4.60 g (24 mmol) of EDC and the product from step 1 was added to a solution consisting of 2.31 g (16.0 mmol) of monoethyl ester of maleic acid in 100 ml of DMF. The suspension formed is kept under stirring at room temperature for 18 h. It is then treated with water and AcOEt is added, the organic phase is separated and the aqueous phase is extracted once more with AcOEt. The organic phases are grouped and washed successively with saturated NaHCO3 solution and brine. They are subsequently subjected to drying over anhydrous Na2SO4, the solvent is filtered and evaporated under reduced pressure.
    1.5 g of the compound identified as example VII'.13 are obtained. Method B: tr: 1.094, m / z: 306. Step 3: Intermediate IX
    The amount of 0.8 ml (5.89 mmol) of EtgN and 1.5 5 g of intermediate VII'.13 (4.91 mmol) was added to a solution of 2-thiophenylethylamine (0.63 ml, 5, 4 mmol) in 40 ml of dioxane and the resulting solution was stirred for 18 h under reflux. The solution was evaporated to dryness and purified by means of 10 column chromatography on silica gel, using a mixture of AcOEt: MeOH: NH3 (10: 1: 0.01) as an eluent, providing 510 mg of an oil identified as intermediate IX. 13 (2- (3,6-dioxo-4- (2- (pyridin-2-yl) ethyl.) - 1- (2- (thiophen-2-yl) ethyl) piperazin-2-yl) ethyl acetate ). 15 Method A: tr: 2.289, m / z: 416.
    The following intermediates were prepared in a manner similar to intermediate IX.13:


    Step 4: Intermediate X
    To a solution of 550 mg (1.32 mmol) of intermediate IX.13 in 15 ml of a MeOH: THF mixture (1: 3) was added 1.6 ml of an IN LiOH solution and the mixture was left under stirring at room temperature all night. Then it is diluted in AcOEt and washed with water, the aqueous phase is acidified with an IN solution of HCl to a pH = 7 and extracted with AcOEt. Finally, the organic phases are grouped, dried over anhydrous Na2SO4, filtered and the solvent is evaporated under reduced pressure. 330 mg of a colorless oil identified as intermediate X.13 are obtained. Method B: tr: 1,768, m / z: 388
    The following intermediates were prepared in a manner similar to intermediate X.13:



    Compounds of formula Ia a) Ia.1.2: N- (2-amino-2-oxoethyl) -N- (2,4-dichloro-phenethyl) - 2- (4- (2,4-dichloro- phenethyl) -1- (3,3-diphenyl-propyl) -3,6-dioxo-piperazin-2-yl) acetamide
    - On a suspension of Va resin (0.61 mmol / g resin, 0.17 mmol), with the appropriate amine and 'previously swollen with a 2: 1 DCM: DMF solution (3 ml), Xl acid (100 ml) was added mg, 1.1 eq.), HOBt (40 mg, 1.5 eq.), HATU (105 mg, 1.5 eq.) and DIPEA (95 μl, 3 eq.). The reaction mixture was subjected to stirring at room temperature for 16 h. The resin was subjected to drying and washed with DMF (3x3 ml), isopropyl alcohol (3x3 ml) and DCM (3x3 ml) and was subsequently treated with a 60: 40: 2 TFA / DCM / water (5 ml) mixture for 30 min at room temperature. The resin was filtered and the filtrate was evaporated under reduced pressure to obtain 73 mg of the desired compound (Ia.1.2, 51% yield, 91% purity). HRMS (M + H) + calculated for C39H39C14N4O4, 767.1725, experimental, 767.1741.




    o) Ia.2.1: W- (2-amino-2-oxoethyl) -N- (2,4-dichloro-phenethyl) - 2- (4- (2,4-dichloro-phenethyl) -1 - (4-fluoro-benzyl) -3,6-dioxo-piperazin-2-yl) acetamide
    X.2 acid (100 mg, 1 eq.) Was added to a solution of 2- (4-fluorobenzylamino) acetamide (IVc, 28 μl, 1 eq.), DIC (85 μl, 3 eq.) And triethylamine (80 μl, 3 eq.) in 2 ml of DCM and the reaction mixture was stirred at room temperature for 3 h. The crude reaction product was neutralized with NaOH and was extracted with DCM. The organic extracts were washed with saturated sodium chloride, dried under anhydrous MgSOSO and evaporated under reduced pressure to obtain 96 mg of the desired compound Ia.2.1.

    Compounds of formula lb a) Ib.1.2 Acid 2- (TV- (2,4-dichloro-phenethyl) -2- (4- (2,4-dichloro-phenethyl) -1- (3.3 -diphenylpropyl) -3,6-dioxo-piperazin-2-yl) acetamide) acetic

    A solution of bromoacetic acid (275 mg, 5 eq.) And DIPEA (345 μl, 5 eq.) In DMF (3 ml) was added to 200 mg of 2-chlorotrityl chloride resin (1.6 mmol / g Cl / g resin, 0.17 mmol) and the suspension was subjected to stirring at room temperature for 1 h. The resin was filtered and washed with DMF (3x3 ml), isopropyl alcohol (3x3 ml) and DCM (3x3 ml). The resin was then treated with methanol (3 ml) for 10 min. to remove unreacted Cl atoms. The supernatant was removed and the residue was washed with DCM (3x3 ml), isopropyl alcohol (3x3 ml) and DMF (3x3 ml). A solution of 2,4-di-chloro-phenethylamine (IVa, 340 μl, 5 eq.) And triethylamine (280 μl, 5 eq.) In 3 ml of DMF was then added to the resin and the suspension was subjected to stirring under room temperature for 3 h. After filtering and washing with DMF (3x3 ml), isopropyl alcohol (3x3 ml) and DCM (3x3 ml), acid X (100 mg, 1.1 eq.) Was added to the resin in the presence of HOBT (40 mg, 1.5 eq.), HATU (105 mg, 1.5 eq.) And DIPEA (95 μl, 3 eq.) In DCM: DMF 2: 1 (3 ml). The reaction mixture was stirred at room temperature for 16 h and was filtered. The resin was subjected to drying and washed with DMF (3x3 ml), isopropyl alcohol (3x3 ml) and DCM (3x3 ml). Finally, the resin was treated with a 5:95 TFA: DCM mixture (5 ml) for 30 min. at room temperature, obtaining a crude reaction product which was filtered. The filtrate solvent was under reduced pressure to obtain 60 mg of the desired compound (Ib.1.2, 42% yield, 91% purity). HRMS (M + H) + calculated for C39H38C14N3O5, 768,1576, experimental, 768,1573.


    Compounds of formula Ic Ic.1.2. 2- (TV- (2,4-di-chloro-phenethyl) -2- (4- (2,4-di-chloro-phenethyl) -1- (3,3-diphenylpropyl) -3, β- acetate dioxo- piperazin-2-yl) acetamide) methyl

    The mixture of acid Ib.1.2 (30 mg, 1 eq.), Methanol (7.5 ml) and H2SO4 (20 μl, 1 eq.) Was reacted for 15 h at room temperature. The crude reaction product was neutralized with NaOH and was extracted with DCM. The organic extracts were washed with saturated sodium chloride, dried over anhydrous MgSO4 and evaporated under reduced pressure to obtain 22 mg of the desired compound Ic.1.2 (72% yield, 86% purity). HRMS (M + H) + calculated for C ^HsgCl ^sOs, 782.1722, experimental, 5,782.1216.

































    Compounds of formula Id Id.1.2 N- (2,4-Di-chloro-phenethyl) -2- (4- (2,4-di-chloro-phenethyl) -1- (3,3-diphenylpropyl) -3, β-dioxo-piperazin-2-yl) acetamide

    Xl acid (100 mg, 1 eq.) Was added to a solution of 2,4-dichloro-phenethylamine (IVc, 28 μl, 1 eq.), DIC (85 μl, 3 eq.) And triethylamine (80 μl, 3 eq.) in 2 ml of DCM and the reaction mixture was stirred at 10 ° C for 3 h. The crude reaction product was neutralized with NaOH and was extracted with DCM. The organic extracts were washed with saturated sodium chloride, dried over anhydrous MgSO4 and evaporated under reduced pressure to obtain 96 mg of the desired compound Id.1.2 (yield 73%, purity 89%). HRMS (M + H) + calculated for C37H36C14N3O3, 710.1511, experimental, 710.1522.

























    Compounds of formula le.1 6-Amino-2- (2- (4- (2,4-dichloro-phenethyl) -1- (3,3-diphenylpropyl) -3, β-dioxo-piperazin-2 -yl) acetamide) hexanamide Rink amide-Fmoc resin (II, 500 mg, 0.305 mmol)
    it was deprotected with 5 ml of 20% piperidine in DMF with agitation in a microwave reactor for 2 min. at 60 ° C. The resin was filtered and washed with DMF (3 x 15 ml), isopropyl alcohol (3 x 15 ml) and DCM (3 x 15 ml). The amino acid Fmoc-L-Lys (Boc) -OH (XI, 286 mg, 2 eq.) Was then joined to the resin using HOBT (82 mg, 2 eq.) And DIC (96 μl, 2 eq.) In 5 ml of DMF. The mixture was subjected to stirring at room temperature for 1 h. The resin was filtered and washed with DMF (3 x 15 ml), isopropyl alcohol (3 x 15 ml) and DCM (3 x 15 ml). Once the Fmoc group was removed with 5 ml of 20% piperidine in DMF for 20 min., The resin was filtered and washed with DMF (3 x 15 ml), isopropyl alcohol (3 x 15 ml) and DCM (3 x 15 ml). Subsequently, the resin was treated with a solution of acid X (181 mg, 1.1 eq.), HATU (348 mg, 3 eq.), HOBT (123 mg, 3 eq.) And DIPEA (0.313 ml, 6 eq. ) in 5 ml of DMF. The reaction mixture was subjected to stirring at room temperature for 16 h. The resin was subjected to drying and washed with DMF (3x3 ml), isopropyl alcohol (3x3 ml) and DCM (3x3 ml) and was subsequently treated with an 80:20 mixture of TFA / DCM / water / triisopropyl silane: 2.5: 2.5 (5 ml) for 30 min. at room temperature. The resin was filtered and the filtrate was evaporated under reduced pressure. The obtained residue is purified by means of normal phase chromatography using a gradient of a mixture of dichloromethane-methanol-ammonia to obtain 15 mg of the desired compound (le.l, 8% yield, 100% purity). MS (M + H) + calculated for C35H4iCl2N5O4, 666.26, experimental, 666.40.
    Compounds of the formula If If.1.2 6-Amino-2- (2- (N- (2,4-dichloro-phenethyl) -2- (4- (2,4-dichloro-phenethyl) -1- (3,3-diphenylpropyl) -3,6-dioxo-piperazin- 2-11) acetamide) acetamide) hexanamide
    .
    The Rink amide-Fmoc resin (II, 800 mg, 0.42 mmol) was deprotected with 8 ml of 20% piperidine in DMF subjected to stirring in a microwave reactor for 2 min. under 60 ° C. The resin was filtered and washed with DMF (3 x 15 ml), isopropyl alcohol (3 x 15 ml) and DCM (3 x 15 ml). The amino acid Fmoc-L-Lys (Boc) -OH (XII, 497 mg, 2 eq.) Was attached to the resin using HOBT (143 mg, 2 eq.) And DIC (165 μl, 2 eq.) In 8 ml of DMF. The mixture was subjected to stirring at room temperature for 1 h. The resin was filtered and washed with DMF (3 x 15 ml), isopropyl alcohol (3 x 15 ml) and DCM (3 x 15 ml). After the Fmoc group was removed with 8 ml of 20% piperidine in DMF for 20 min., The resin was filtered and washed with DMF (3 x 15 ml), isopropyl alcohol (3 x 15 ml) and DCM (3 x 15 ml). The resin was treated with a solution of bromoacetic acid (III, 295 mg, 4 eq.) And DIC (0.33 ml, 4 eq.) In DMF: DCM 1: 2 (8 ml) and the mixture was subjected to stirring for 20 min. at room temperature. The resin was filtered and washed with DMF (3 x 15 ml), isopropyl alcohol (3 x 15 ml) and DCM (3 x 15 ml). A solution of 2,4-dichlorophenethylamine (IVd, 0.32 ml, 4 eq.) And triethylamine (0.295 ml, 4 eq.) In 12 ml of DMF was added to the resin and the suspension was stirred for 3 h at room temperature. The supernatant was removed and the reaction was repeated under the same conditions. The resin was filtered and washed with DMF (3 x 15 ml), isopropyl alcohol (3 x 15 ml) and DCM (3 x 15 ml) to obtain resin XIV, which was treated with an acid solution X (274 mg, 2 eq.), HATU (116 mg, 1.6 eq.), HOBT and DIPEA (0.295 ml, 3.2 eq.) in 8 ml of DMF. The reaction mixture was stirred at room temperature for 3 h. The resin was subjected to drying and washed with DMF (3x3 ml), isopropyl alcohol (3x3 ml) and DCM (3x3 ml) and was subsequently treated with an 80:20 mixture of TFA / DCM / water / triisopropyl silane: 2.5: 2.5 (5 ml) for 30 min. at room temperature. The resin was filtered and the filtrate was evaporated under reduced pressure. The residue obtained was purified by means of semi-preparation RP-HPLC using a gradient of an acetonitrile-water mixture to obtain 128 mg of the desired compound (If.1.2, 34% yield, with 99% purity ). HRMS (M + H) + calculated for C45H50CI4N6O5, 895.2675; experimental, 895.2648. Pharmacological Examples
    In vitro assay for inhibition of apoptosome formation Recombinant Apaf-1 produced in insect cells (rApaf-1) was incubated in the presence (under a concentration of 10 μM) or absence (as a control) of the compounds to be evaluated in the buffer. assay (20 mM Hepes-KOH pH 7.5, 10 mM KC1, 1.5 mM MgC12, 1 mM EDTA, 1 mM EGTA, 1 mM DTT, 0.1 mM PMSF) for 15 minutes at 30 ° C. The final concentration of rApaf-1 was 40 nM. Then, dATP / Mg (Sigma) and purified horse cytochrome c (Sigma) were added, reaching final concentrations of 100 μM and 0.1 μM, respectively. It was incubated for 60 minutes under 30 ° C and then recombinant procaspase-9 produced in E. coli (procaspase-9, 0.1 μM final concentration) was added and incubated for 10 minutes under 30 ° C before being incubated. add the fluorogenic substrate of caspase-9 Ac-LEDH-afc (50 μM final concentration). The total test volume was 200 μl. Caspase activity was monitored continuously by releasing afc at 37 ° C on a 5 Wallac 1420 Workstation (Àexc = 390 nm, Àem = 510 nm).
    Activity values for some compounds described in the examples are shown in the following table expressed as percent inhibition of Apaf-1.
    权利要求:
    Claims (5)
    [0001]
    1. Compound characterized from the group consisting of:
    [0002]
    Compound according to claim 1, characterized in that it is:
    [0003]
    Compound according to claim 1, characterized in that it is:
    [0004]
    4. Compound according to claim 1, characterized by being selected from the group consisting of:
    [0005]
    Compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, characterized in that it is for use as a pharmaceutical active ingredient.
    类似技术:
    公开号 | 公开日 | 专利标题
    BR112012002134B1|2021-03-02|apaf-1 inhibitory compounds
    WO1995003277A1|1995-02-02|Novel pyrrolidine derivative
    KR19980703726A|1998-12-05|indole derivatives as cGMP-PDE inhibitors
    JP2002520335A|2002-07-09|Dolastatin 15 derivative
    MX2014005638A|2014-11-25|Modulators of the g protein-coupled mas receptor and the treatment of disorders related thereto.
    JP2007527904A|2007-10-04|Pyrrolopyridine-2-carboxylic acid hydrazide compounds as glycogen phosphorylase inhibitors
    WO2007036131A1|2007-04-05|Carzole sulphamide derivatives and their preparation method
    TW200838502A|2008-10-01|Triazole derivative
    JP4927303B2|2012-05-09|Pharmaceutically active benzsulfonamide derivatives as inhibitors of protein Jun kinase
    ES2835851T3|2021-06-23|Indole amide compound as necrosis inhibitor
    MX2007015274A|2008-02-22|Indole derivatives having antitumor activity.
    WO1992018488A1|1992-10-29|Novel oxazinone derivative
    JP2007527903A|2007-10-04|Indole-2-carboxylic acid hydrazide compound
    Dahiya et al.2013|Synthetic studies on novel nitroquinazolinone analogs with antimicrobial potential
    CN108484640B|2020-09-15|Anti-tumor apoptosis protein inhibitor
    Friedrichsen et al.2001|Application of enzymatically stable dipeptides for enhancement of intestinal permeability. Synthesis and in vitro evaluation of dipeptide-coupled compounds
    BRPI0618837A2|2011-09-13|apoptosis inhibiting compounds
    NZ587824A|2012-05-25|Novel method for the production of sulphonylpyrroles as hdac inhibitors
    FR2941946A1|2010-08-13|SUBSTITUTED 3-BENZOFURANYL-INDOL-2-ONE-3-ACETAMIDOPIPERAZINE DERIVATIVES, THEIR PREPARATION AND THEIR THERAPEUTIC USE
    KR20100017964A|2010-02-16|1-benzylpyrazole derivatives, preparation thereof and therapeutic use thereof
    WO2011057477A1|2011-05-19|Tetrapeptide analogs, preparation method and use thereof
    WO2016107541A1|2016-07-07|Pyrrole amide compound, preparation method therefor, and use thereof
    JP2012521395A|2012-09-13|Anticancer compounds, their preparation and their therapeutic use
    KR20210025001A|2021-03-08|Crystalline form of thiophene derivatives
    LE2014|N-GLYOXYLAMIDES AS VERSATILE PRECURSORS FOR PEPTIDE MIMICS AND HETEROCYCLES
    同族专利:
    公开号 | 公开日
    NZ598125A|2014-05-30|
    CN102574818A|2012-07-11|
    JP5694320B2|2015-04-01|
    BR112012002134A2|2016-05-31|
    EP2460798B1|2019-02-27|
    US9040701B2|2015-05-26|
    IN2012DN01378A|2015-06-05|
    AU2010277505A1|2012-03-08|
    MX2012001339A|2012-03-07|
    AU2010277505B2|2016-01-14|
    EA201270215A1|2012-11-30|
    US20120122868A1|2012-05-17|
    CN102574818B|2015-02-25|
    ES2727711T3|2019-10-18|
    BR112012002134B8|2021-05-25|
    CA2769408A1|2011-02-03|
    KR20120052354A|2012-05-23|
    WO2011012746A3|2011-07-14|
    WO2011012746A2|2011-02-03|
    JP2013500314A|2013-01-07|
    TR201907804T4|2019-06-21|
    EA021838B1|2015-09-30|
    CA2769408C|2017-09-05|
    EP2460798A4|2013-02-27|
    KR101786761B1|2017-10-18|
    EP2460798A2|2012-06-06|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    US3560503A|1968-09-18|1971-02-02|Council Scient Ind Res|Di-lower alkyl-substituted octahydropyrazinopyrimidinones|
    GB1284582A|1969-11-10|1972-08-09|Council Scient Ind Res|Substituted piperazinopyrimidinones|
    JPH04234375A|1990-12-27|1992-08-24|Ajinomoto Co Inc|New 2,5-dioxopiperazine compound and its production|
    WO1992019595A1|1991-05-07|1992-11-12|Merck & Co., Inc.|Fibrinogen receptor antagonists|
    JPH05117246A|1991-10-23|1993-05-14|Ajinomoto Co Inc|New 2,5-dioxopiperazine compound, its production and production of alpha-l-aspartyl-l-phenylalanine methyl ester derivative|
    WO1996000391A1|1994-06-23|1996-01-04|Affymax Technologies N.V.|Methods for the synthesis of diketopiperazines|
    AT277913T|2001-01-19|2004-10-15|Pharmacopeia Inc|BISARYL DERIVATIVES WITH FSH RECEPTOR MODULATING ACTIVITY|
    AT520981T|2002-10-02|2011-09-15|Dmi Biosciences Inc|DIAGNOSIS AND MONITORING OF DISEASES|
    WO2005052177A1|2003-11-28|2005-06-09|Kyowa Hakko Kogyo Co., Ltd.|Process for producing dipeptide|
    ES2296484B1|2005-11-23|2009-04-01|Fundacion De La Comunidad Valenciana Centro De Investigacion Principe Felipe |PHARMACEUTICAL COMPOSITION TO INHIBIT APOPTOSIS.|
    ES2293834B1|2006-07-20|2009-02-16|Consejo Superior Investig. Cientificas|COMPOSED WITH INHIBITING ACTIVITY OF UBC13-UEV INTERACTIONS, PHARMACEUTICAL COMPOSITIONS THAT INCLUDE IT AND ITS THERAPEUTIC APPLICATIONS.|AU2006259415B2|2005-06-15|2012-08-30|Massachusetts Institute Of Technology|Amine-containing lipids and uses thereof|
    WO2010053572A2|2008-11-07|2010-05-14|Massachusetts Institute Of Technology|Aminoalcohol lipidoids and uses thereof|
    ES2734973T3|2009-12-01|2019-12-13|Translate Bio Inc|Administration of mRNA for the increase of proteins and enzymes in human genetic diseases|
    WO2012027675A2|2010-08-26|2012-03-01|Massachusetts Institute Of Technology|Poly, their preparation, and uses thereof|
    WO2012135025A2|2011-03-28|2012-10-04|Massachusetts Institute Of Technology|Conjugated lipomers and uses thereof|
    RU2013154295A|2011-06-08|2015-07-20|Шир Хьюман Дженетик Терапис, Инк.|COMPOSITIONS OF LIPID NANOPARTICLES AND METHODS FOR DELIVERY OF mRNA|
    KR102272498B1|2011-10-27|2021-07-06|메사추세츠 인스티튜트 오브 테크놀로지|Amino acid derivatives functionalized on the n-terminal capable of forming drug incapsulating microspheres|
    WO2013078413A1|2011-11-22|2013-05-30|The United States Of America, As Represented By The Secretary, Department Of Health And Human Services|Modulators of lipid storage|
    WO2013185067A1|2012-06-08|2013-12-12|Shire Human Genetic Therapies, Inc.|Nuclease resistant polynucleotides and uses thereof|
    ES2443538B1|2012-07-19|2014-11-26|Consejo Superior De Investigaciones Cientificas |APAF1 inhibitor beta-lactam compounds|
    EP2882706A1|2012-08-13|2015-06-17|Massachusetts Institute of Technology|Amine-containing lipidoids and uses thereof|
    PT2968586T|2013-03-14|2018-11-13|Ethris Gmbh|Cftr mrna compositions and related methods and uses|
    EA201591229A1|2013-03-14|2016-01-29|Шир Хьюман Дженетик Терапис, Инк.|METHODS OF CLEANING MATRIX RNA|
    WO2014179562A1|2013-05-01|2014-11-06|Massachusetts Institute Of Technology|1,3,5-triazinane-2,4,6-trione derivatives and uses thereof|
    KR102096796B1|2013-10-22|2020-05-27|샤이어 휴먼 지네틱 테라피즈 인크.|Lipid formulations for delivery of messenger rna|
    EA201992208A1|2013-10-22|2020-07-31|Транслейт Био, Инк.|TREATMENT OF PHENYLKETONURIA USING mRNA|
    EP3501605A1|2013-10-22|2019-06-26|Translate Bio, Inc.|Mrna therapy for argininosuccinate synthetase deficiency|
    JP6571679B2|2014-04-25|2019-09-04|トランスレイト バイオ， インコーポレイテッド|Method for purifying messenger RNA|
    EP3587409A1|2014-05-30|2020-01-01|Translate Bio, Inc.|Biodegradable lipids for delivery of nucleic acids|
    EP3160959A4|2014-06-24|2018-02-28|Translate Bio, Inc.|Stereochemically enriched compositions for delivery of nucleic acids|
    EP3164379A1|2014-07-02|2017-05-10|Massachusetts Institute of Technology|Polyamine-fatty acid derived lipidoids and uses thereof|
    EP3310764A1|2015-06-19|2018-04-25|Massachusetts Institute of Technology|Alkenyl substituted 2,5-piperazinediones and their use in compositions for delivering an agent to a subject or cell|
    AU2018224326A1|2017-02-27|2019-09-19|Translate Bio, Inc.|Novel codon-optimized CFTR mRNA|
    CA3063531A1|2017-05-16|2018-11-22|Translate Bio, Inc.|Treatment of cystic fibrosis by delivery of codon-optimized mrna encoding cftr|
    AU2019256527A1|2018-04-19|2020-11-12|Spiral Therapeutics, Inc.|Inner ear drug delivery devices and methods of use|
    KR20210060480A|2018-08-24|2021-05-26|트랜슬레이트 바이오 인코포레이티드|Method for purifying messenger RNA|
    US10561736B1|2019-01-09|2020-02-18|Spiral Therapeutics, Inc.|Apoptosis inhibitor formulations for prevention of hearing loss|
    CA3131378A1|2019-02-25|2020-09-03|Spiral Therapeutics Inc.|Intratympanic injector devices and needles for delivery of drugs and methods of use|
    法律状态:
    2018-01-23| B07D| Technical examination (opinion) related to article 229 of industrial property law [chapter 7.4 patent gazette]|
    2018-04-10| B06F| Objections, documents and/or translations needed after an examination request according [chapter 6.6 patent gazette]|
    2018-06-05| B25A| Requested transfer of rights approved|Owner name: LOOP THERAPEUTICS, INC. (US) |
    2019-05-28| B07E| Notice of approval relating to section 229 industrial property law [chapter 7.5 patent gazette]|Free format text: NOTIFICACAO DE ANUENCIA RELACIONADA COM O ART 229 DA LPI |
    2019-07-02| B06T| Formal requirements before examination [chapter 6.20 patent gazette]|
    2019-10-01| B25A| Requested transfer of rights approved|Owner name: SPIRAL THERAPEUTICS INC (US) |
    2020-09-08| B06A| Notification to applicant to reply to the report for non-patentability or inadequacy of the application [chapter 6.1 patent gazette]|
    2020-12-22| B09A| Decision: intention to grant [chapter 9.1 patent gazette]|
    2021-03-02| B16A| Patent or certificate of addition of invention granted|Free format text: PRAZO DE VALIDADE: 10 (DEZ) ANOS CONTADOS A PARTIR DE 02/03/2021, OBSERVADAS AS CONDICOES LEGAIS. |
    2021-05-25| B16C| Correction of notification of the grant|Free format text: PRAZO DE VALIDADE: 20 (VINTE) ANOS CONTADOS A PARTIR DE 29/07/2010 OBSERVADAS AS CONDICOES LEGAIS. PATENTE CONCEDIDA CONFORME ADI 5.529/DF |
    优先权:
    申请号 | 申请日 | 专利标题
    ES200901757|2009-07-30|
    ESP200901757|2009-07-30|
    PCT/ES2010/000349|WO2011012746A2|2009-07-30|2010-07-29|Apaf-1 inhibitor compounds|
    [返回顶部]