WO2020020101A1 - Benzisoselenazolidone amine compound, and preparation method therefor and use thereof - Google Patents

Abstract

A benzisoselenazolidone amine compound having a structure as represented by formula (I). The compound not only can inhibit the activity of PDE-4 or TNF-α, but also has the GPx activity and a cell ferroptosis inhibiting function, and thus can be used for treating diseases mediated by PDE-4, TNF-α, and/or impaired GPx function.

Description

Benzoisoselazolone amine compounds, preparation method and application thereof Technical field

The invention belongs to the technical field of medicine and relates to a class of benzoisoselazolidamine compounds having PDE-4 and / or TNF-α inhibitory activity. The invention also relates to methods of preparing these compounds, and these compounds as PDE-4 and / or in the treatment and / or prevention of diseases or conditions associated with the PDE-4 and / or TNF-α pathway in humans or other mammals Use of a TNF-α inhibitor.

Background technique

PDE-4 (Phosphodiesterase-4): Phosphodiesterases (PDEs) are a large family of two important messengers in the cell: cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP). ) Is hydrolyzed to inactive adenosine 5-phosphate (5-AMP) and guanosine 5-phosphate (5-GMP), of which PDE-4 is one of the important members of the phosphodiesterase family, and can selectively hydrolyze cAMP. PDE-4 is divided into four subtypes PDE-4A ～ D. Except for PDE-4C, the other subtypes are mainly distributed in airway smooth muscle cells and lymphocytes, macrophages, neutrophils, eosinophils, and eosinophils. Inflammatory cells and T cells such as basic granulocytes, monocytes, and epithelial cells. PDE-4 inhibitors can increase cAMP levels, by inhibiting multiple inflammatory mediators, inhibiting the up-regulation and expression of cell adhesion factors, inhibiting the activation of white blood cells, inducing apoptosis, and inducing the production of cytokines with inhibitory activity (such as white blood cells Interleukin-6) and induce the release of catecholamines and endogenous hormones to inhibit the activity of these immune cells and inflammatory cells, which can be used to treat diseases caused by inflammation, such as asthma, chronic obstructive pulmonary (COPD), Central nervous system diseases caused by damage to neurons caused by potential inflammation, such as rheumatoid arthritis, multiple sclerosis, Alzheimer's disease (AD) and Parkinson's disease (PD).

TNF-α (tumor necrosis factor): TNF-α is a type of cytokine with multiple biological effects discovered in the 1970s. It is mainly secreted by activated monocytes / macrophages / T cells and passes through the cell membrane. Specific receptor binding, such as by activating the three signal pathways of Caspase protease, JNK, and the transcription factor NF-κB, thus causing a number of different biological processes, and ultimately realizing its regulation of cell growth and apoptosis, tumor formation, immunity, and inflammation And biological functions such as stress response. Improper TNF-α production and continued activation of TNF-α signals will lead to systemic human pathological processes, including systemic inflammatory response syndrome, inflammatory bowel disease, rheumatoid arthritis, neurodegenerative diseases (multiple sclerosis) , Motor neuron disease, Alzheimer's disease, Parkinson), psoriasis, cerebral malaria, diabetes, tumors, osteoporosis, allograft rejection, multiple sclerosis, HBV, HCV and HIV, etc. (Brenner D.et.al. Regulation of tumor necrosis factor signaling: live or let die. Nat Rev Immunol. 2015, 15 (6), 362).

Apremilast (Otezla): an oral selective phosphodiesterase-4 (PDE-4) inhibitor, approved by the US FDA and European regulators for active psoriasis in late 2014 and early 2015, respectively Of arthritis (PSA) and moderate to severe plaque psoriasis (Plaque psoriasis). Apestide inhibits PDE-4 and increases the level of cyclic adenosine monophosphate (cAMP) in PDE-4 expressing cells, thereby activating protein kinase K (PKA) and phosphorylated cAMP response element binding protein (CREB) while inhibiting Nuclear factor-κB (NF-κB) drives gene transcription, which in turn reduces expression of pro-inflammatory mediators such as TNF-α, IFN-γ, IL-8, IL-2, and IL-23 and increases anti-inflammatory cytokines such as IL- 10 expressions. At present, in the treatment of psoriatic arthritis, a single administration of apostat is not inferior to that of secukinumab, adalimumab (Humel), etanercept (Enley), and infliximab. Antibiotics and other biologics are obviously better than other traditional medicines; but single-administration of albast is less effective than biologics in the treatment of psoriasis, and the advantages are not obvious compared with traditional medicines for psoriasis This is one of the main reasons why Apster is only approved for psoriasis treatment in Europe. In addition, the clinical side effects of taking aprost tablets still include diarrhea, nausea and headache. Therefore, there is an urgent need in the art for an improved structure of Apster derivatives to optimize their performance.

It is worth noting that selenium (in the form of selenocysteine) is glutathione peroxidase (GSH-Px), iodinated thyrosine, and mammalian thioredoxin reductase ( The important active center of TrxR) is also an essential trace element of the human body, which has a variety of beneficial effects on human health, such as enhancing the body's immunity, anti-oxidation and anti-tumor. Modern epidemiological studies have also shown that too low selenium content in plasma is a risk for cancer (liver cancer, stomach cancer, prostate cancer, lung cancer, colorectal cancer, etc.), cardiovascular and cerebrovascular diseases, psoriasis, osteoarthritis and AIDS. Factors (Marco Vinceti.et.al.Friend or Foe. The Current Current Epidemiologic Evidence on Selenium and Human Cancer Risk.J.Environ.Sci.Heal.2013, 31,305; Margaret P Rayman.The The Importance of Selenium The Tolerance Human Health. .2000,356,233). Based on the important role of selenium in autoimmune diseases, cardiovascular and cerebrovascular diseases, and tumors (Li Feng et al. Evidence-based medical research on the efficacy of selenium supplementation in the treatment of psoriasis vulgaris. Shanghai Medical Journal. 2017, 21, 6; Agnieszka B Serwin.et.al. Selenium status in psoriasis and relaxations to the duration of the situation and the situation of the situation. Nutrition. 2003, 19, 301; Marta Wacewicz.et.al.Concentration of the selenium, zinc / copper total antioxidant status and c-reactive protein in the patient of patients with psoriasis treated by narrow-band ultraviolet B phototherapy: A case-control study. JTrace Elem Med Bio. 2017, 44, 109.), combined with the basis of Apost's clinical application The present invention creatively designs and synthesizes a series of new benzoisoselazolidone amine compounds in order to further reduce the effectiveness of existing PDE-4 and / or TNF-α inhibitor drugs on the basis of improving the effectiveness of disease treatment. Toxic response, increase the comprehensive treatment index of the drug.

Summary of the Invention

The object of the present invention is to provide a new type of benzoisoselazolidamine structure type compound.

Another object of the present invention is to provide a method for preparing such compounds.

A further object of the present invention is a class of benzoisoselazolidamine structural compounds, which not only inhibit PDE-4 and / or TNF-α, but also inhibit iron death in normal cells and have prevention And / or use in a medicament for treating diseases related to PDE-4 and / or TNF-α overexpression.

The present invention provides a new class of benzoisoselazolidone amine compounds, solvates, crystalline forms, stereoisomers, isotopic compounds, metabolites, or prodrugs. These compounds have a structure represented by Formula I:

In formula (I),

R ₁ , R ₂ , R ₃ and R ₄ are each independently selected from the group consisting of H, D, halogen, hydroxyl, nitro, cyano, carboxyl, seleno, mercapto, C ₁ to C ₈ alkylselenyl , C ₁ to C ₈ alkylselenyl C ₁ to C ₈ alkylamino, C ₂ to C ₈ alkenselenoyl, α-C ₁ to C ₈ alkylselenyl amino acid, α-C ₁ to C ₈ alkylselenyl formyl Amino acids, C ₀ to C ₈ alkylamine C ₁ to C ₈ alkylselenyl, C ₀ to C ₈ alkylcarbamoylselenyl, arylselenyl, C ₀ to C ₈ alkyloxy C ₁ to C ₈ alkylselenyl, C ₀ to C ₈ alkoxyformyl C ₁ to C ₈ alkylselenyl, C ₀ to C ₈ alkoxyformyl C ₁ to C ₈ alkoxy, halogenated C ₁ to C ₈ alkylselenyl, C ₁ to C ₈ alkylsulfonyl, C ₁ to C ₈ alkylsulfonamido, C ₀ to C ₈ alkylaminosulfonyl, C ₁ to C ₈ alkyl, halogenated C ₁ to C ₈ alkyl, halogenated C ₁ to C ₈ alkoxy, C ₀ to C ₈ alkethynyl, C ₁ to C ₈ alkoxy, C ₁ to C ₈ alkanoyloxy, C ₁ to C ₈ alkoxy C ₁ to C ₈ alkoxy, C ₁ to C ₈ alkoxy C ₁ to C ₈ alkyl, C ₁ to C ₈ alkyl amine, C ₀ to C ₈ alkyl amine C ₁ to C ₈ alkyl, aryl, aryl C ₁ to C ₈ alkyl amine C ₁ ~ C ₈ alkyl, amidino, guanidino, an arylsulfonyl group, an aryl sulfonyl group, an aryl carboxylic Group, C ₀ ~ C ₈ alkoxy selenium formyl, aryl C ₁ ~ C ₈ alkyl group, an aryl C ₁ ~ C ₈ alkyl amide, C ₁ ~ C ₈ alkoxy, formyl, C ₁ ~ C ₈ alkylamido , C ₀ ～ C ₈ alkylamino group, aromatic selenium C ₁ ～ C ₈ amide group, cyanoseleno C ₁ ～ C ₈ amide group, benzoselazole C ₁ ～ C ₈ alkyl amide group, benzoselazole C ₁ ～ C ₈ alkylsulfonamide group, C ₀ to C ₈ alkylcarbamoylselenyl group, C ₀ to C ₈ alkylaminoformamide group, C ₀ to C ₈ alkylaminoformyl group, C ₁ to C ₈ alkylaminoformyl group Acyloxy, arylaminoformamide, arylaminoformyl, arylaminoformyloxy, piperidinyl, piperazinyl, morpholinyl, pyrrolyl, pyrazolyl, imidazolyl, pyridyl, Pyrazinyl, quinolinyl, pyrimidinyl, pyrimidinylamino, thiazolyl, thienyl, furyl, pyrrolyl, or absent; wherein R ₁ , R ₂ , R _3, and R ₄ are phenyl groups or 1-4 groups selected from the group consisting of halogen, hydroxyl, nitro, cyano, amino, trifluoromethyl, carboxyl, halo C ₁ to C ₈ alkoxy, C ₁ to C ₈ alkoxy Substituted phenyl

R ₅ is: C ₁ to C ₈ alkylsulfonyl, C ₁ to C ₈ alkylcarbamoyl, cyano;

R ₆ and R ₇ are each independently selected from the group consisting of: halogenated C ₁ to C ₈ alkoxy, C ₁ to C ₈ alkoxy, halogenated C ₁ to C ₈ alkylselenyl, C ₁ to C ₈ alkylselenyl;

X is: C or Se; where W is C, at least one selenium-containing substituent exists in the substituents of R ₁ , R ₂ , R ₃ , R ₄ , R _6, and R ₇ ; when W is Se, R ₁ , R ₂ , R ₃ , R ₄ and R ₅ are any of the groups described above;

n = 1-4;

The dotted line is: chemical bond or absent.

Preferably, the present invention provides structural compounds, solvates, crystalline forms, stereoisomers, isotopic compounds, metabolites or prodrugs of the general formulae (I-a) and (I-b):

In formulae (I-a) and (I-b),

R ₁ , R ₂ , R ₃ and R ₄ are each independently selected from the group consisting of H, D, halogen, hydroxyl, nitro, cyano, carboxyl, seleno, mercapto, C ₁ to C ₈ alkylselenyl , C ₁ to C ₈ alkylselenyl C ₁ to C ₈ alkylamino, C ₂ to C ₈ alkenselenoyl, α-C ₁ to C ₈ alkylselenyl amino acid, α-C ₁ to C ₈ alkylselenyl formyl Amino acids, C ₀ to C ₈ alkylamine C ₁ to C ₈ alkylselenyl, C ₀ to C ₈ alkylcarbamoylselenyl, arylselenyl, C ₀ to C ₈ alkyloxy C ₁ to C ₈ alkylselenyl, C ₀ to C ₈ alkoxyformyl C ₁ to C ₈ alkylselenyl, C ₀ to C ₈ alkoxyformyl C ₁ to C ₈ alkoxy, halogenated C ₁ to C ₈ alkylselenyl, C ₁ to C ₈ alkylsulfonyl, C ₁ to C ₈ alkylsulfonamido, C ₀ to C ₈ alkylaminosulfonyl, C ₁ to C ₈ alkyl, halogenated C ₁ to C ₈ alkyl, halogenated C ₁ to C ₈ alkoxy, C ₀ to C ₈ alkethynyl, C ₁ to C ₈ alkoxy, C ₁ to C ₈ alkanoyloxy, C ₁ to C ₈ alkoxy C ₁ to C ₈ alkoxy, C ₁ to C ₈ alkoxy C ₁ to C ₈ alkyl, C ₁ to C ₈ alkylamino, C ₀ to C ₈ alkylamine C ₁ to C ₈ alkyl, aryl, aryl C ₁ to C ₈ alkylamine C ₁ ~ C ₈ alkyl, amidino, guanidino, an arylsulfonyl group, an aryl sulfonyl group, an aryl carboxylic Group, C ₀ ~ C ₈ alkoxy selenium formyl, aryl C ₁ ~ C ₈ alkyl group, an aryl C ₁ ~ C ₈ alkyl amide, C ₁ ~ C ₈ alkoxy, formyl, C ₁ ~ C ₈ alkylamido , C ₀ ～ C ₈ alkylamino group, aromatic selenium C ₁ ～ C ₈ amide group, cyanoseleno C ₁ ～ C ₈ amide group, benzoselazole C ₁ ～ C ₈ alkyl amide group, benzoselazole C ₁ ～ C ₈ alkylsulfonamido, C ₀ to C ₈ alkylcarbamoylselenyl, C ₀ to C ₈ alkylcarbamoylamide, C ₀ to C ₈ alkylaminoformyl, C ₁ to C ₈ alkylaminoformyl an acyloxy group, an aryl group carboxamido, carbamoyl aralkyl, aryloxy formyloxy group or absent; _{wherein, R 1, R 2, R} 3 and said R ₄ is phenyl or substituted aryl group 1-4 substituted by a group selected from halogen, hydroxy, nitro, cyano, amino, trifluoromethyl, carboxyl, halogenated C ₁ to C ₈ alkoxy, C ₁ to C ₈ alkoxy At least one selenium-containing substituent in the substituents R ₁ , R ₂ , R ₃ , R ₄ , R ₆ and R _{7 in} (Ib);

R ₅ is: C ₁ to C ₈ alkylsulfonyl, C ₁ to C ₈ alkylsulfonyl; carbamoyl, cyano;

R ₆ and R ₇ are each independently selected from the group consisting of: halogenated C ₁ to C ₈ alkoxy, C ₁ to C ₈ alkoxy, halogenated C ₁ to C ₈ alkylselenyl, C ₁ to C ₈ alkylselenyl;

The dotted line is: chemical bond or absent.

As used herein, the term "halogen" refers to fluorine, chlorine, bromine, and iodine.

As used herein, the term "halo" may be monohalo or polyhalo.

As used herein, the term "alkanesulfonyl" refers to a linear or branched or cyclic saturated hydrocarbon sulfonyl group having 3 to 8 carbon atoms.

As used herein, the term “C ₁ to C ₈ alkylselenyl” refers to a straight or branched chain or cyclic saturated hydrocarbon selenium group, the cyclic saturated hydrocarbon having 3 to 8 carbon atoms.

As used herein, the term "C ₀ ~ C ₈ alkoxy selenium carboxamido" refers to straight or branched chain or cyclic saturated alkane selenium carboxamido, a cyclic saturated hydrocarbon having 3 to 8 carbon atoms.

As used herein, the term "alkanesulfonamido" refers to a straight-chain or branched or cyclic saturated alkanesulfonamide group having 3 to 8 carbon atoms.

As used herein, the term "alkylaminosulfonyl" refers to an N-monosubstituted or disubstituted linear or branched or cyclic saturated alkaneaminosulfonyl group having 3 to 8 carbon atoms.

As used herein, the term "alkylaminoformyl" refers to an N-monosubstituted or disubstituted linear or branched or cyclic saturated alkaneaminoformyl group having 3 to 8 carbon atoms.

As used herein, the term "alkyl" refers to a straight or branched straight chain or branched or cyclic saturated hydrocarbon group, said cyclic saturated alkane having 3 to 8 carbon atoms.

As used herein, the term "alkoxy" refers to a linear or branched or cyclic saturated alkoxy group, said cyclic saturated alkane group having 3 to 8 carbon atoms.

As used herein, the term "alkethynyl" refers to a linear or branched or cyclic saturated hydrocarbon ethynyl, which is 3 to 8 carbon atoms.

As used herein, the term "alkanoyloxy" refers to a linear or branched or cyclic saturated hydrocarbon acyloxy group, said cyclic saturated alkane having 3 to 8 carbon atoms.

As used herein, the term "alkanoselenyl" refers to a linear or branched or cyclic saturated hydrocarbylselenyl group, which cyclic saturated alkane has 3 to 8 carbon atoms.

As used herein, the term "alkylamino" refers to an N-mono- or di-substituted linear or branched or cyclic saturated hydrocarbon amine group having 3 to 8 carbon atoms.

As used herein, the term "alkoxyformyl" refers to a linear or branched or cyclic saturated hydrocarbon oxyformyl which has 3 to 8 carbon atoms.

As used herein, the term "alkamido" refers to a linear or branched or cyclic saturated hydrocarbon amide, which is 3 to 8 carbon atoms.

As used herein, the term "alkylaminoformamide," is N-monosubstituted or disubstituted, and refers to a linear or branched or cyclic saturated hydrocarbon aminoformamide group, said cyclic saturated alkane having 3 to 8 carbons atom.

As used herein, the term "stereoisomers" refers to compounds that differ in chirality at one or more stereocenters. Stereoisomers include enantiomers and diastereomers.

As used herein, unless otherwise specified, the "piperazinyl, morpholinyl, pyrrolyl, pyrazolyl, pyrrolyl, imidazolyl, pyrimidylamino" substitution substitution sites are all on N.

As used herein, the "pyridyl, pyrimidinyl, thiazolyl, thienyl, furanyl, pyrazinyl, quinolinyl" unless otherwise specified, the substituted attachment sites are all on C.

In the case where the compounds described in the present invention have stereoisomers, the present invention includes all stereoisomers of the compounds.

The invention also includes deuterated compounds in which any one or more of the hydrogen atoms in the compound is replaced by its stable isotope deuterium.

As used herein, the term "metabolite" refers to an active substance produced after a change in the chemical structure of a drug molecule in vivo, which is generally a derivative of the aforementioned drug molecule, which may also be chemically modified.

As used herein and unless otherwise specified, the term "polymorph" refers to one or more crystal structures formed when the molecules are arranged differently in the lattice space during crystallization.

As used herein, the term "solvate" refers to a crystalline form of a compound of general formula (I), a crystalline form, a stereoisomer, an isotope compound, a metabolite, or a prodrug, which also includes one or more incorporated crystals Solvent molecules in the structure. The solvate may include a stoichiometric or non-stoichiometric amount of a solvent, and the solvent molecules in the solvent may exist in an ordered or non-ordered arrangement. A solvate containing a non-stoichiometric amount of a solvent molecule may result from the solvate losing at least one, but not all, of the solvent molecules. In a particular embodiment, a solvate is a hydrate, meaning that the crystalline form of the compound further includes water molecules, with water molecules as the solvent.

As used herein and unless otherwise specified, the term "prodrug" refers to a derivative of a compound containing a biologically reactive functional group such that the biologically reactive functional group can be cleaved from the compound or otherwise occur under biological conditions (in vitro or in vivo). React to provide the compound. Generally, a prodrug is inactive, or at least less active than the compound itself, so that its activity cannot be exerted until the compound is cleaved from a biologically reactive functional group. Bioreactive functional groups can be hydrolyzed or oxidized under biological conditions to provide the compounds. For example, a prodrug may include a biohydrolyzable group. Examples of biohydrolyzable groups include, but are not limited to, biohydrolyzable phosphates, biohydrolyzable esters, biohydrolyzable amides, biohydrolyzable carbonates, biohydrolyzable carbamates, and biohydrolyzable Ureide. For a review of prodrugs, see, for example, J. Rautio et al., Nature Drugs Discovery (2008) 7, 255-270 and Prodrugs: Challenges and Rewards (V. Stella et al. Ed., Springer, 2007).

The compounds of the general formula (I), solvates, crystalline forms, stereoisomers, isotopic compounds, metabolites or prodrugs of the invention may contain one or more asymmetric centers ("stereoisomers"). As used herein, the term "stereoisomers" refers to enantiomers, diastereomers, epimers, endo-exo isomers, atropis All stereoisomers including atropisomers, regioisomers, cis- and trans-isomers, etc. The "stereoisomers" herein also include "pure stereoisomers" and "enriched stereoisomers" or "racemates" of the aforementioned various stereoisomers. These stereoisomers can be separated, purified, and enriched by asymmetric synthesis methods or chiral separation methods (including but not limited to thin-layer chromatography, rotary chromatography, column chromatography, gas chromatography, high-pressure liquid chromatography, etc.), and can also be purified by It can be obtained by chiral resolution by bonding with other chiral compounds (chemical bonding, etc.) or salt formation (physical bonding, etc.).

Some of the preferred novel selenium-containing amine compounds of the present invention are shown below. These examples are only for further explanation of the present invention, and do not limit the scope of the present invention in any way.

Among them, it is well known that any stereocenter of any of the above-listed compounds may be in an absolute (R)-or (S) -configuration, or a racemic mixture of the two, if not explicitly stated. The invention relates to a racemic mixture of these compounds, a mixture enriched in any one enantiomer, and any isolated enantiomer. For the scope of the present invention, it should be understood that the racemic mixture refers to a 50%: 50% mixture of two R and S enantiomers, and the separated enantiomers should be understood as pure enantiomers (i.e. 100%) or a highly enriched mixture of certain enantiomers (purity ≥98%, ≥95%, ≥90%, ≥88%, ≥85%, ≥80%).

According to a second aspect of the present invention, a method for preparing the above-mentioned benzoselazole compound is provided, and the method includes the following methods.

The following abbreviations apply throughout the specification and examples:

The following abbreviations apply throughout the specification and examples:

Ac

AcOH

Base organic or inorganic base

DMF N, N-dimethylformamide

EA ethyl acetate

EtOH ethanol

EDC 1-Ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride

H ₂ O ₂ hydrogen peroxide

HOBt 1-hydroxybenzotriazole

LC-MS

NMR

Pd / CH ₂ palladium carbon hydrogen reduction system

TLC thin layer chromatography

V solution volume

The compound of formula I of the present invention can be prepared according to the following general method:

a) Synthetic route of divalent selenium compounds in the I-a structure series of the present invention

b) Synthetic route of tetravalent selenium compounds in the I-a structure series of the present invention

c) Synthetic route of I-b structural compounds of the present invention

The synthesis of the divalent selenium type compound in the structure series of the formula Ia can be based on the substituted 2-halobenzoic acid, and the intermediate can be obtained through the reaction of step (1) with (S) -2- (substituted acyl) -1-substituted phenylethylamine. Body a ', and then obtained by reacting step (2) with a selenium reagent;

The compounds of the formula I-a series can also be synthesized by directly using o-selenochlorobenzoyl chloride as the raw material and directly (S) -2- (methanesulfonyl) -1-substituted phenylethylamine to obtain the target compound;

In step (1), substituted 2-halobenzoic acid is used as a raw material, and DCC, EDCI, HOBT is used as a condensation agent under conventional condensation conditions, and triethylamine, pyridine, diisopropylethylamine, etc. are used as a base, and ( S) -2- (Methanesulfonyl) -1-substituted phenylethylamine is reacted in an organic solvent (rt ～ 120 ° C) to obtain intermediate a ′. The solvent used includes, but is not limited to, N, N-dimethylformamide , Dimethyl sulfoxide, acetonitrile, tetrahydrofuran, dichloromethane, chloroform, and ethyl acetate and other organic solvents (Reference: Heteroatom Chemistry. 2014, 35, 320);

In step (2), the intermediate a ′ is coupled to a ring reaction by selenization under the action of a selenization reagent such as selenium powder, LiSeSeLi, NaSeSeNa, KSeSeK, NaSeCN, etc., to obtain the target compound Ia. The solvents used include but not Limited to organic solvents such as N, N-dimethylformamide, dimethyl sulfoxide, acetonitrile, tetrahydrofuran, dichloromethane, chloroform, and ethyl acetate (References: Org. Lett. 2010, 12, 23; J. Org (Chem. 2017, 82, 3844; Tetrahedron. 2011, 67, 9565).

In step (3), when the substituted benzoyl chloride is ortho-SeCl, tertiary amines such as triethylamine, diisopropylethylamine, and pyridine under basic conditions, and (S) -2- (methanesulfonate) Acyl) -1-substituted phenylethylamine (reaction rt ~ 120 ° C) to obtain the target compound. The solvents used include, but are not limited to, N, N-dimethylformamide, dimethylsulfoxide, acetonitrile, tetrahydrofuran, and dichloromethane. , Chloroform and ethyl acetate (References: J. Med. Chem. 2013, 56, 9089);

Structure of formula Ia tetravalent selenium series type of compound is a divalent substituted benzisoselenazol as raw materials, [O ^-] obtained peroxidation reaction, the solvent includes, but not limited to tetrahydrofuran, methylene chloride, chloroform and ethyl acetate with Organic solvents such as esters, the reaction temperature is -20 ° C to 0 ° C. The peroxide reagent used includes but is not limited to H ₂ O ₂ , O ₃ , and m-chloroperoxybenzoic acid (References: J. Org. Chem. 2005, 70, 868; J. Org. Chem. 2005, 70, 5023);

The synthesis of compounds 40-45 in the compounds of the structural type of the formula Ib is based on the deacetyl aposite at the 4-position as the raw material, followed by reaction step (a) diselenization, step (b) reduction, step (c) to form an ether, and Step (d) is obtained by the deprotection reaction; the synthesis of compound 46-51 is obtained by using deacetyl aposite at the 4-position as the raw material and obtained through step (e);

The compound of the structure type of the formula Ib is based on the deacetyl aposite at the 4-position as a raw material and undergoes an acylation reaction with N-benzoselazole-amino acid or N-benzoselazole-taurine or N- Selenomorpholine-acetic acid was obtained (References: J. Org. Chem. 2004, 46, 53; J. Med. Chem. 2001, 1021).

Specifically, according to the method of the present invention, a series of different structural types of benzoisoselazolidone amine compounds are obtained through the methods shown in reaction formulas a), b) and c). The reaction process usually uses TLC and LC-MS to check the completion of the reaction. After the reaction is completed, it is generally extracted with solvents such as methyl tert-butyl ether, ethyl acetate or dichloromethane, and then washed with saturated sodium bicarbonate, water and saturated saline in order. After drying over anhydrous sodium sulfate or magnesium sulfate, the solvent was removed under reduced pressure at low temperature. Key intermediate and final products were confirmed by NMR and mass spectrometry.

According to a third aspect of the invention, the compound of formula I has the effect of inhibiting PDE-4 and / or TNF-α overexpression. Accordingly, they can be used as PDE-4 and / or TNF-α inhibitors for the treatment (including combination therapy) of PDE-4 and / or TNF-α overexpression causing related diseases, including asthma, arthritis, rheumatoid joints Inflammation, gouty arthritis, rheumatoid spondylitis, osteoarthritis and other arthritic conditions; sepsis, septic shock, endotoxin shock, Gram-negative sepsis, toxic shock syndrome, Acute Respiratory Distress Syndrome, Cerebral Malaria, Chronic Pulmonary Inflammatory Disease, Silicosis, Pulmonary Sarcoidosis, Bone Resorption Disease, Graft-versus-Host Response, Allograft Rejection, Fever and Myalgia Caused by Infection, Secondary to Infection Or cachexia of malignant tumors, cachexia secondary to human acquired immunodeficiency syndrome (AIDS), AIDS, HIV, HBV, ARC (AIDS-related complications), keloid formation, scar tissue formation, localized ileitis, ulcers Colitis, multiple sclerosis, Alzheimer's disease (AD) and Parkinson's disease (PD), type 1 diabetes, autoimmune diabetes, diabetes insipidus, systemic lupus erythematosus, bronchitis, chronic obstructive airway disease ,Cattle Dermatophytosis, Bechet's disease, allergic purpuric nephritis, chronic glomerulonephritis, inflammatory bowel disease, leukemia, allergic rhinitis, depression, multi-infarct dementia, or dermatitis.

Beneficial effect

The present invention designs and synthesizes a new class of benzoisoselazolidone-like compounds that have a significant inhibitory effect on PDE-4 and / or TNF-α. Compared to the existing PDE-4 inhibitor Apest, the benzoisoselazolidone inhibitor of the present invention can also improve mammals' diseases such as psoriasis caused by and / or exacerbated by lower selenium levels in the body, significantly improving the overall treatment effect.

detailed description

The present invention is further explained below with reference to specific embodiments, but the present invention is not limited. The experimental operation of the present invention is versatile and is not limited to the specific compounds mentioned in the following examples.

In the following preparation examples, ¹ H-NMR was measured with a Varian Mercury AMX300 type instrument. MS was determined using VG ZAB-HS or VG-7070 and Esquire 3000Plus-01005. All reaction solvents are re-distilled before use, and the anhydrous solvents used are obtained by drying in accordance with standard methods. Except for the instructions, all reactions were carried out under the protection of argon and followed by TLC. During post-treatment, they were dried by saturated saline and anhydrous sodium sulfate. The products were purified using column chromatography on silica gel (200-300 mesh) unless otherwise stated.

Example 1 Synthesis of Compound 1

synthetic route:

(1) Synthesis of Intermediate 1a

With stirring in an ice bath, a solution containing 2-bromo-3-nitrobenzoic acid (246 mg, 1 mmol), EDCI (384 mg, 2 mmol), DMAP (12 mg, 0.1 mmol) and N, N-diisopropylethyl (S) -2- (methylsulfonyl) -1- (3'-ethoxy-4'-methoxy) phenylethylamine (273 mg, 1 mmol), and then reacted at room temperature overnight. After the reaction was completed, ethyl acetate (2-20 ml) and 20 mL of water were added for extraction. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated to dryness under reduced pressure, and conventional silica gel column chromatography (V _{ethyl acetate} : V _{petroleum). Ether} = 1: 4 to 1: 1) to obtain Intermediate 1a (400 mg, yield 80%). HRMS-ESI: m / z calcd for C ₁₉ H ₂₁ BrN ₂ O ₇ S: 500.0253, found [M + H] ⁺ 501.0330; ¹ H NMR (500MHz, MeOD) δ7.89 (d, J = 7.4Hz, 1H ), 7.75 (d, J = 6.7Hz, 1H), 7.69-7.62 (m, 1H), 7.15-6.96 (m, 3H), 5.76-5.69 (m, 1H), 4.16-4.09 (m, 1H), 3.86 (s, 3H), 3.78–3.60 (m, 2H), 2.99 (s, 3H), 1.42 (t, J = 6.6Hz, 3H).

(2) Synthesis of intermediate 1b

To 20 ml of a tetrahydrofuran-methanol-water (10 ml / 8 ml / 2 ml) solution containing 1a (250 mg, 0.5 mmol) and ammonium chloride (214 mg, 4 mmol) was added zinc powder (162 mg, 2.5 mmol), and then the temperature was slowly raised to 50 ° C. The reaction was completed for 1 hour. After the reaction was completed, the mixture was filtered through celite, the filtrate was concentrated to dryness, and 20 ml of water was added. Then, ethyl acetate (20 mL × 2) was added for extraction. The organic phase was washed with saturated brine in sequence and dried over anhydrous sodium sulfate to obtain intermediate 1b. (240 mg, yield 100%). HRMS-ESI: m / z calcd for C ₁₉ H ₂₃ BrN ₂ O ₅ S: 470.0511, found [M + H] ⁺ 471.0588; ¹ H NMR (500MHz, CDCl ₃ ) δ7.14 (t, J = 7.7Hz, 1H), 7.01-6.83 (m, 5H), 5.70 (dd, J = 13.1, 6.4 Hz, 1H), 4.29 (brs, 2H), 4.13 (q, J = 6.9 Hz, 2H), 3.89 (s, 3H ), 3.78 (dd, J = 14.9, 6.4 Hz, 1H), 3.58 (dd, J = 14.9, 5.2 Hz, 1H), 2.69 (s, 3H), 1.49 (t, J = 6.9 Hz, 3H).

(3) Synthesis of intermediate 1c

Add intermediate 1b (235mg, 0.5mmol) to a mixed solution of acetic anhydride (1mL) and pyridine (1ml), and then raise the temperature to 80 ° C for 2 hours. After the reaction is completed, add 1N hydrochloric acid solution to adjust the pH to 3, and then add Extraction with ethyl acetate (20mL × 2), the organic phase was sequentially washed with saturated brine, dried over anhydrous sodium sulfate, concentrated to dryness under reduced pressure and conventional silica gel column chromatography (Vethyl _acetate : V _{petroleum ether} = 1: 4 ～ 1: 1) to obtain Intermediate 1c (260 mg, yield 100%). HRMS-ESI: m / z calcd for C ₂₁ H ₂₅ BrN ₂ O ₆ S: 512.0617, found [M + H] ⁺ 513.0690; ¹ H NMR (500MHz, DMSO) δ9.53 (s, 1H), 9.11 (d , J = 8.6Hz, 1H), 7.65–7.41 (m, 2H), 7.23 (d, J = 7.5Hz, 1H), 7.18–6.95 (m, 3H), 5.60–5.47 (m, 1H), 4.03 ( dd, J = 13.7, 6.8 Hz, 2H), 3.75 (s, 3H), 3.68-3.50 (m, 2H), 2.96 (s, 3H), 2.09 (s, 3H), 1.33 (t, J = 6.9Hz , 3H).

(4) Synthesis of compound 1

To a DMF solution (2 mL) containing CuI (38 mg, 0.2 mmol), 1,10-o-diazaphenanthrene (39 mg, 0.2 mmol), a3 (256 mg, 0.5 mmol), selenium powder (0.8 g, 1 mmol), K ₂ CO ₃ (138 mg, 1 mmol), and then reacted at 110 ° C. overnight to completion under a nitrogen atmosphere. After the TLC reaction was completed, 20 ml of water was added, followed by extraction with ethyl acetate (20 mL × 2). The organic phase was sequentially washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The resulting filtrate was evaporated to dryness under reduced pressure and purified by silica gel column chromatography. (V _{ethyl acetate} : V _{petroleum ether} = 1: 4 to 1: 1) to obtain compound 1 (84 mg, yield 33%). HRMS-ESI: m / z calcd for C ₂₁ H ₂₄ N ₂ O ₆ SSe: 512.0520, found [M + H] ⁺ 513.0598; ¹ H NMR (500MHz, CDCl ₃ ) δ9.57 (s, 1H), 8.72– 7.44 (m, 3H), 7.12 (dd, J = 8.3, 2.1Hz, 1H), 7.04–6.94 (m, 2H), 5.99 (t, J = 6.8Hz, 1H), 4.18--4.04 (m, 3H) , 3.92 (s, 3H), 3.82 (dd, J = 14.6, 7.2 Hz, 1H), 2.97 (s, 3H), 2.25 (s, 3H) 1.49–1.42 (m, 3H).

Example 2 Synthesis of Compound 2

Under nitrogen and ice bath conditions, (254 mg, 1 mmol) of dichloroselenobenzoyl chloride (refer to J. Med. Chem. 2016, 59, 8125-8133) in dichloromethane solution was slowly added to A solution of 1- (3-ethoxy-4-methoxyphenyl) -2- (methylsulfonyl) ethylamine (273 mg, 1 mmol) and triethylamine (151 mg, 1.5 mmol) in dichloromethane (10 mL) ), And then continue to react to completion. After the TLC reaction was completed, 20 ml of water was added, and then dichloromethane (20 mL × 2) was added for extraction. The organic phase was sequentially washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The resulting filtrate was evaporated to dryness under reduced pressure and purified by silica gel column chromatography (V _acetone : V _{petroleum ether} = 1: 4 to 1: 1) to obtain compound 2 (432 mg, yield 95%). HRMS-ESI: m / z calcd for C ₁₉ H ₂₁ NO ₅ SSe: 455.0306, found [M + H] ⁺ 456.0378; ¹ H NMR (500MHz, CDCl ₃ ) δ8.04 (d, J = 7.9Hz, 1H) , 7.64--7.52 (m, 2H), 7.44 (t, J = 7.3 Hz, 1H), 7.06 (dd, J = 8.3, 1.9 Hz, 1H), 7.00 (d, J = 1.9 Hz, 1H), 6.91 ( d, J = 8.3 Hz, 1H), 5.98 (t, J = 6.8 Hz, 1H), 4.17-4.06 (m, 3H), 3.90 (s, 3H), 3.80 (dd, J = 14.8, 7.1 Hz, 1H ), 2.96 (s, 3H), 1.46 (t, J = 7.0 Hz, 3H). ¹³ C NMR (126 MHz, DMSO) δ 166.3, 149.6, 148.4, 140.1, 132.0, 131.5, 128.7, 127.9, 126.3, 126.2, 120.9, 113.2, 112.2, 64.4, 56.8, 55.9, 52.5, 41.8, 15.2.

Preparation of Example 3-6 The operation of Reference Example 1 was changed, and different substituted o-bromobenzoic acid was reacted with (S) -2- (methylsulfonyl) -1-substituted phenylethylamine; the preparation reference of Examples 7-14 The operation of Example 2 is as follows:

Example 15 Synthesis of Compound 15

(1) Synthesis of intermediate 15a

With stirring in an ice bath, acetyl chloride (316 mg, 4 mmol) was added to a tetrahydrofuran saturated sodium acetate mixed solution containing 2-amino-6-bromobenzoic acid (432 mg, 2 mmol) (volume ratio 1: 1, 5 ml). , Then warmed to room temperature and reacted for 0.5 hours; after the reaction was completed, 1N hydrochloric acid solution was added to adjust the pH to 3, and then ethyl acetate (20mL × 2) was added for extraction. The organic phase was sequentially washed with saturated brine, dried over anhydrous sodium sulfate, reduced The solution was concentrated to dryness to obtain intermediate 15a (520 mg, yield 100%). MS-ESI: m / z [ MH] - 256.0.

(2) Synthesis of intermediate 15b

With stirring in an ice bath, (S) -2- (methanesulfonyl)-was added to a DMF solution (2mL) containing 15a (258mg, 1mmol), EDCI (384mg, 2mmol), and DMAP (240mg, 2mmol). 1- (3'-cyclopentyloxy-4'-methoxy) phenylethylamine (313 mg, 1 mmol), and then reacted at room temperature overnight. After the reaction was completed, ethyl acetate (2-20 ml) and 20 mL of water were added for extraction. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated to dryness under reduced pressure, and conventional silica gel column chromatography (V _{ethyl acetate} : V _{petroleum). Ether} = 1: 4 to 1: 1) to obtain intermediate 15b (391 mg, yield 71%). MS-ESI: m / z [M + H] ⁺ 553.1.

(3) Synthesis of compound 15

To a solution (2mL) of acetonitrile containing CuI (38mg, 0.2mmol), 1,10-o-diazaphenanthrene (39mg, 0.2mmol), 15b (276mg, 0.5mmol), selenium powder (0.8g, 1mmol), K ₂ CO ₃ (138 mg, 1 mmol), and then reacted at 110 ° C. overnight to completion under a nitrogen atmosphere. After the TLC reaction was completed, 20 ml of water was added, followed by extraction with ethyl acetate (20 mL × 2). The organic phase was sequentially washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The resulting filtrate was evaporated to dryness under reduced pressure and purified by silica gel column chromatography. (V _{ethyl acetate} : V _{petroleum ether} = 1: 4 to 1: 1) to obtain compound 1 (76 mg, yield 27%). HRMS-ESI: m / z calcd for C ₂₄ H ₂₈ N ₂ O ₆ SSe: 552.0883, found [M + H] ⁺ 553.0962; ¹ H NMR (500MHz, CDCl ₃ ) δ9.53 (s, 1H), 8.71– 7.44 (m, 3H), 7.07-6.99 (m, 2H), 6.92 (d, J = 7.8Hz, 1H), 5.98 (t, J = 6.8Hz, 1H), 4.81-4.72 (m, 1H), 4.18 –4.05 (m, 1H), 3.86–3.80 (m, 4H), 2.97 (s, 3H), 2.69 (s, 3H), 1.99–1.56 (m, 8H).

Example 16 Synthesis of Compound 16

Under nitrogen and ice bath conditions, a solution of (25 mg, 0.1 mmol) dichloroselenobenzoyl chloride in dichloromethane (1 mL) was slowly added to a solution containing (31 mg, 0.1 mmol) 1- (3-ethoxy-4 -Methoxyphenyl) -2- (methylsulfonyl) ethylamine (see US 2006/0084815 for the preparation method) and triethylamine (15 mg, 0.15 mmol) in a dichloromethane solution (2 mL), and then continue to react until complete. After the TLC detection reaction was completed, 5 ml of water was added, and then dichloromethane (5 mL × 2) was added for extraction. The organic phase was sequentially washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was evaporated to dryness under reduced pressure and purified by silica gel column chromatography (V _acetone : V _{petroleum ether} = 1: 4 to 1: 1) to obtain compound 16 (44 mg, yield 90%). HRMS-ESI: m / z calcd for C ₂₂ H ₂₅ NO ₅ SSe: 495.0619, found [M + H] ⁺ 496.0690; ¹ H NMR (500MHz, CDCl ₃ ) δ 8.06 (d, J = 7.9 Hz, 1H) , 7.64-7.52 (m, 2H), 7.44 (t, J = 7.3Hz, 1H), 7.08-6.99 (m, 2H), 6.91 (d, J = 7.8Hz, 1H), 5.97 (t, J = 6.8 Hz, 1H), 4.81--4.72 (m, 1H), 4.18--4.07 (m, 1H), 3.86--3.79 (m, 4H), 2.96 (s, 3H), 2.64 (s, 3H), 1.97--1.52 ( m, 8H).

Preparation of Examples 17-20 The operation of Reference Example 15 was used to change different substituted o-bromobenzoic acid and (S) -2- (methylsulfonyl) -1-substituted phenylethylamine or (S) -2- (amine methylamine). Acyl) -1-substituted phenylethylamine reaction; Examples 21-24 were prepared as described in Reference Example 16, and different substituted o-bromobenzoic acids were changed with 2- (methanesulfonyl) -1-substituted phenylethylamine or -(Carbamoyl) -1-substituted phenylethylamine, or 2- (cyano) -1-substituted phenylethylamine, the results of the examples obtained are as follows:

Example 25 Synthesis of Compound 25

With stirring in an ice bath, 30% hydrogen peroxide (0.12 mmol) was slowly added to a dichloromethane solution (2 mL) containing Compound 1 (45 mg, 0.1 mmol) and the reaction was continued for 2 h to completion. After the TLC detection reaction was completed, 5 ml of water was added, and then ethyl acetate (5 mL × 2) was added for extraction. The organic phase was sequentially washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The resulting filtrate was evaporated to dryness under reduced pressure and purified by silica gel column chromatography. (V _{ethyl acetate} : V _{petroleum ether} = 1: 1) to obtain compound 25 (36 mg, yield 77%). HRMS-ESI: m / z calcd for C ₁₉ H ₂₁ NO ₆ SSe: 471.0255, found [M + H] ⁺ 472.0730. ¹ H NMR (500 MHz, DMSO) δ 8.08 (d, J = 8.2 Hz, 1 H), 7.89 (d, J = 7.7 Hz, 1 H), 7.63 (t, J = 7.6 Hz, 1 H), 7.49 (t, J = 7.4Hz, 1H), 7.23 (s, 1H), 7.15–6.98 (m, 2H), 6.07 (t, J = 7.0Hz, 1H), 4.28–4.08 (m, 4H), 3.81 (s, 3H ), 2.93 (s, 3H), 1.37 (t, J = 6.9Hz, 3H).

The preparation of Examples 26 to 32 was performed with reference to Example 25. The obtained results of the examples are as follows:

Example 46 Synthesis of Compound 46

(1) Preparation method Step 1: Synthesis of N-benzoselazole-glycine

To a solution of glycine (751mg, 10mmol) and sodium carbonate (2.12g, 20mmol) in dichloromethane / water (50ml / 5ml) under vigorous stirring in an ice bath, 10mmol of 2-chloroselenyl-benzoyl chloride was added. (Obtained according to the preparation method of patent CN101016319A), after the dropwise addition is completed, and reacted at room temperature for 1 h. After the reaction was completed, 10 ml of water was added, the pH was adjusted to 4 with 1N HCl, and the organic phase was extracted with dichloromethane (50 ml x 2). The organic phase was sequentially washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The resulting filtrate was evaporated to dryness under reduced pressure and passed through silica gel. Purification by column chromatography (V _{ethyl acetate} : V _{petroleum ether} = 1: 1 to 2: 1), intermediate N-benzoselazole-glycine (1.85 g, 72%) was obtained, MS-ESI [M + H] ⁺ 258.0 (258.0). According to the method, alanine is used as a raw material to obtain N-benzoselazole-alanine, a key intermediate. According to the above method, taurine was used as a raw material to obtain other key intermediates N-benzoselazole-taurine (2.30 g, 75%), and MS-ESI [M + H] ⁺ 308.0 (308.0).

(2) Preparation method Step 2: Synthesis of compound 33

Under argon, HOBt (20 mg, 0.15 mmol) and EDC (29 mg, 0.15 mmol) were added to 1 ml of a DMF solution containing N-benzoselazole-glycine (26 mg, 0.1 mmol). After reacting for 1 hour, add 4-Deacetyl aposite (42 mg, 0.1 mmol), and the reaction was continued for 24 h at 80 ° C. After the reaction was completed, it was quenched by adding 2 ml of saturated ammonium chloride, and extracted with ethyl acetate (5 ml × 2). The organic phase was sequentially washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The resulting filtrate was evaporated to dryness under reduced pressure and subjected to silica gel chromatography. Purification (V _{ethyl acetate} : V _{petroleum ether} = 1: 2) gave compound 33 (37 mg, yield 56%). HRMS-ESI: m / z calcd for C ₂₉ H ₂₇ N ₃ O ₈ SSe: 657.0684, found [M + H] ⁺ 658.0762; ¹ H NMR (500MHz, CDCl ₃ ) δ 10.41 (s, 1H), 8.78 ( d, J = 8.0 Hz, 1H), 8.27 (d, J = 6.8 Hz, 1H), 8.13–8.09 (d, J = 7.6 Hz, 1H), 7.90–7.60 (m, 3H), 7.45–7.37 (m , 1H), 7.15--6.95 (m, 3H), 5.91--5.80 (m, 1H), 4.67 (s, 2H), 4.38--4.14 (m, 2H), 4.08--4.02 (q,

2H), 3.78 (s, 3H), 3.03 (s, 3H), 1.35 (t, J = 6.9Hz, 3H).

The preparation of Examples 34 to 40 was performed with reference to Example 33. The results of the examples are as follows:

Example 41 PDE-4 activity inhibition experiment

Methods According to Lance cAMP Assay: Weigh the test sample accurately, add DMSO to dissolve it, and mix thoroughly to make 100μM. The mother liquor was then diluted to 1 μM with DMSO, and then 4-fold diluted to 0.0038 nM. Add 2.5 μL substrate 20nM cAMP, 50nL compound DMSO solution to 384-well plate, and then add 2.5 μL PDE-4 enzyme buffer (1 × HBSS, 5mM Hepes pH 7.4, 3mM MgCl ₂ , 0.1% BSA) and incubate at room temperature for 90min. Add 5 μL Alexa

After incubating the 647-anti cAMP antibody for 30 minutes, adding 10 μL of detection reagent and incubating for 60 minutes, the LANCE signal was detected at 665 nm. The inhibition rate was calculated by the following formula, and the IC ₅₀ value was calculated from the inhibition rate using XLfit. Inhibition rate = [signal value (MAX)-signal value (sample)] x 100 / [signal value (MAX)-signal value (MIN)].

Note: MAX: blank control without enzyme; MIN: blank control without compound. See Table 1 for details.

Table 1. PDE-4 inhibitory activities

Note: A: <100nM; B: 100nM ～ 0.99μM; C:> 1μM.

Example 42 TNF-α activity inhibition experiment

Peripheral blood from healthy volunteers was collected and collected using EDTA anticoagulation tubes. The blood was diluted 5 times with 1640 medium (Gibco, catalog number 11875-093, USA) and added to a 96-well cell culture plate (Costar, catalog number 3599, USA), and then 10 μl of the general formula (I ) The compound was treated with DMSO (Sigma, catalog number D2650, USA) solution. The final concentration of the compound was 100 nM, and the final concentration of DMSO was 0.2%. After incubating in a 5% CO ₂ incubator at 37 ° C. for 60 min, 10 μl of LPS (Sigma, Catalog No. L-2880, USA) was added to the reaction system to a final concentration of 10 ng / ml, and then at 37 ° C., 5% CO ₂ After 6 hours of incubation under conditions. The supernatant was collected and the TNF-α content was determined by an ELISA method (BD Biosciences, catalog number 555212, USA). The plate reader was used to detect the intensity of the absorbed light, the OD ₄₅₀ nm value was taken, the OD ₆₅₀ nm value was used as a reference, and the solution control group containing 0.2% DMSO medium was used as 0% inhibition. Record the raw data and standard curve. XL-fit software was used to draw a four-parameter drug inhibition curve and calculate the inhibition rate of each compound, as shown in Table 2.

Table 2. TNF-α inhibitory activity

Compound	TNF‐α inhibition rate (%)	Compound	TNF‐α inhibition rate (%)	Compound	TNF‐α inhibition rate (%)
1	<50	16	<50	29	＞ 50
4	≥50	17	<50	32	<50
9	<50	18	<50	34	≥50
10	≥50	20	≥50	35	<50

12	<50	twenty three	<50	37	≥50
14	<50	26	<50	Apster	<50

Example 43: Glutathione Peroxidase (GPx) Activity Experiment

Experimental method: The GPX activity of the compound of the general formula (I) of the present invention is studied by spectrophotometry. Mix glutathione (2.0mM), EDTA (1mM), glutathione disulfide reductase (1.7units mL ^-1 ), and nicotinamide adenine dinucleotide phosphate oxidase (NADPH; 0.4mM) In 0.1M potassium phosphate buffer at pH = 7.5. At room temperature (25 ° C), the sample to be tested (50 μm) was added to the above mixture, and then H ₂ O ₂ , tBuOOH or Cum-OOH (1.6 mM) were added to start the reaction. The initial reduction rate (v ₀ ) is calculated by measuring the oxidation rate of NADPH. The molar extinction coefficient (6.22mM ^-1 cm ^-1 ) is used to represent NADPH. The absorbance of the reaction system at 340nm is continuously detected. Each initial rate is measured at least. 3 times. Among them, the calibration determination of peroxidase activity subtracts the background response between peroxidase and glutathione. The experimental results are shown in Table 3.

Table 3. Antioxidant effects of compounds of formula I on pseudo-Gpx4

The experimental results show that benzoisoselazolidamine quasi-glutathione peroxidase (GPx) activity is significantly better than that of the positive control ebselen.

Example 44 Protective effect of benzoisoselazolidamine compounds on Erastin-induced iron death in HT22

Drugs: Tested benzoisoselazolidamine compounds, Erastin, DMSO dissolved.

Reagents: CCK-8 kit and DEME medium were purchased from Sigma; mouse HT22 hippocampal cells (Shanghai Jiaotong University).

CCK-8 experiment: HT22 cells were cultured in a 37 ° C incubator containing 5% CO ₂ and grown in DMEM medium containing 10% serum. Then culture HT22 cells in a 96-well plate, pre-treat the drug for 2 hours (5 μM), and add 0.5 μmol / L Erastin for 8 hours; then add 10 μL of CCK-8 solution to each well, and incubate in the incubator for 3 hours. Read the absorbance at 450 nm on a microplate reader. The cell survival rate was calculated according to the following formula: cell survival rate% = (treatment group-blank control group) / (control group-blank control group) * 100%. The experiment was repeated three times.

Experimental results: As shown in Figure 2, Erastin, an iron death enhancer, can cause apoptosis in HT22 cells. Benzoisoselazolidamine compounds can significantly reduce the damage of HT22 cells caused by Eratin and improve the cell survival rate.

Claims (10)

1. A benzoisoselazolidone amine compound, a solvate, a crystalline form, a stereoisomer, an isotope compound, a metabolite, or a prodrug of the structure represented by the general formula (I):

   

   In formula (I),

   R ₁ , R ₂ , R ₃ and R ₄ are each independently selected from the group consisting of H, D, halogen, hydroxyl, nitro, cyano, carboxyl, seleno, mercapto, C ₁ to C ₈ alkylselenyl , C ₁ to C ₈ alkylselenyl C ₁ to C ₈ alkylamino, C ₂ to C ₈ alkenselenoyl, α-C ₁ to C ₈ alkylselenyl amino acid, α-C ₁ to C ₈ alkylselenyl formyl Amino acids, C ₀ to C ₈ alkylamine C ₁ to C ₈ alkylselenyl, C ₀ to C ₈ alkylcarbamoylselenyl, arylselenyl, C ₀ to C ₈ alkyloxy C ₁ to C ₈ alkylselenyl, C ₀ to C ₈ alkoxyformyl C ₁ to C ₈ alkylselenyl, C ₀ to C ₈ alkoxyformyl C ₁ to C ₈ alkoxy, halogenated C ₁ to C ₈ alkylselenyl, C ₁ to C ₈ alkylsulfonyl, C ₁ to C ₈ alkylsulfonamido, C ₀ to C ₈ alkylaminosulfonyl, C ₁ to C ₈ alkyl, halogenated C ₁ to C ₈ alkyl, halogenated C ₁ to C ₈ alkoxy, C ₀ to C ₈ alkethynyl, C ₁ to C ₈ alkoxy, C ₁ to C ₈ alkanoyloxy, C ₁ to C ₈ alkoxy C ₁ to C ₈ alkoxy, C ₁ to C ₈ alkoxy C ₁ to C ₈ alkyl, C ₁ to C ₈ alkyl amine, C ₀ to C ₈ alkyl amine C ₁ to C ₈ alkyl, aryl, aryl C ₁ to C ₈ alkyl amine C ₁ ~ C ₈ alkyl, amidino, guanidino, an arylsulfonyl group, an aryl sulfonyl group, an aryl carboxylic Group, C ₀ ~ C ₈ alkoxy selenium formyl, aryl C ₁ ~ C ₈ alkyl group, an aryl C ₁ ~ C ₈ alkyl amide, C ₁ ~ C ₈ alkoxy, formyl, C ₁ ~ C ₈ alkylamido , C ₀ ～ C ₈ alkylamino group, aromatic selenium C ₁ ～ C ₈ amide group, cyanoseleno C ₁ ～ C ₈ amide group, benzoselazole C ₁ ～ C ₈ alkyl amide group, benzoselazole C ₁ ～ C ₈ alkylsulfonamide group, C ₀ to C ₈ alkylcarbamoylselenyl group, C ₀ to C ₈ alkylaminoformamide group, C ₀ to C ₈ alkylaminoformyl group, C ₁ to C ₈ alkylaminoformyl group Acyloxy, arylaminoformamide, arylaminoformyl, arylaminoformyloxy, piperidinyl, piperazinyl, morpholinyl, pyrrolyl, pyrazolyl, imidazolyl, pyridyl, Pyrazinyl, quinolinyl, pyrimidinyl, pyrimidinylamino, thiazolyl, thienyl, furyl, pyrrolyl, or absent; wherein R ₁ , R ₂ , R ₃ and R ₄ are phenyl groups 1-4 groups selected from the group consisting of halogen, hydroxyl, nitro, cyano, amino, trifluoromethyl, carboxyl, halo C ₁ to C ₈ alkoxy, C ₁ to C ₈ alkoxy Substituted phenyl

   R ₅ is: C ₁ to C ₈ alkylsulfonyl, C ₁ to C ₈ alkylcarbamoyl, carboxyl, cyano;

   R ₆ and R ₇ are each independently selected from the group consisting of: halogenated C ₁ to C ₈ alkoxy, C ₁ to C ₈ alkoxy, halogenated C ₁ to C ₈ alkylselenyl, C ₁ to C ₈ alkylselenyl;

   X is: C or Se; where W is C, at least one selenium-containing substituent exists in the substituents of R ₁ , R ₂ , R ₃ , R ₄ , R _6, and R ₇ ; when W is Se, R ₁ , R ₂ , R ₃ , R ₄ and R ₅ are any of the groups described above;

   n = 1-4;

   The dotted line is: chemical bond or absent.
2. The compound of general formula (I), solvate, crystalline form, stereoisomer, isotope compound, metabolite, or prodrug according to claim 1, characterized in that the compound is as general formula (Ia) and (Ib ) Shown:

   

   In formulae (I-a) and (I-b),

   R ₁ , R ₂ , R ₃ and R ₄ are each independently selected from the group consisting of H, D, halogen, hydroxyl, nitro, cyano, carboxyl, seleno, mercapto, C ₁ to C ₈ alkylselenyl , C ₁ to C ₈ alkylselenyl C ₁ to C ₈ alkylamino, C ₂ to C ₈ alkenselenoyl, α-C ₁ to C ₈ alkylselenyl amino acid, α-C ₁ to C ₈ alkylselenyl formyl Amino acids, C ₀ to C ₈ alkylamine C ₁ to C ₈ alkylselenyl, C ₀ to C ₈ alkylcarbamoylselenyl, arylselenyl, C ₀ to C ₈ alkyloxy C ₁ to C ₈ alkylselenyl, C ₀ to C ₈ alkoxyformyl C ₁ to C ₈ alkylselenyl, C ₀ to C ₈ alkoxyformyl C ₁ to C ₈ alkoxy, halogenated C ₁ to C ₈ alkylselenyl, C ₁ to C ₈ alkylsulfonyl, C ₁ to C ₈ alkylsulfonamido, C ₀ to C ₈ alkylaminosulfonyl, C ₁ to C ₈ alkyl, halogenated C ₁ to C ₈ alkyl, halogenated C ₁ to C ₈ alkoxy, C ₀ to C ₈ alkethynyl, C ₁ to C ₈ alkoxy, C ₁ to C ₈ alkanoyloxy, C ₁ to C ₈ alkoxy C ₁ to C ₈ alkoxy, C ₁ to C ₈ alkoxy C ₁ to C ₈ alkyl, C ₁ to C ₈ alkyl amine, C ₀ to C ₈ alkyl amine C ₁ to C ₈ alkyl, aryl, aryl C ₁ to C ₈ alkyl amine C ₁ ~ C ₈ alkyl, amidino, guanidino, an arylsulfonyl group, an aryl sulfonyl group, an aryl carboxylic Group, C ₀ ~ C ₈ alkoxy selenium formyl, aryl C ₁ ~ C ₈ alkyl group, an aryl C ₁ ~ C ₈ alkyl amide, C ₁ ~ C ₈ alkoxy, formyl, C ₁ ~ C ₈ alkylamido , C ₀ ～ C ₈ alkylamino group, aromatic selenium C ₁ ～ C ₈ amide group, cyanoseleno C ₁ ～ C ₈ amide group, benzoselazole C ₁ ～ C ₈ alkyl amide group, benzoselazole C ₁ ～ C ₈ alkylsulfonamide group, C ₀ to C ₈ alkylcarbamoylselenyl group, C ₀ to C ₈ alkylaminoformamide group, C ₀ to C ₈ alkylaminoformyl group, C ₁ to C ₈ alkylaminoformyl group an acyloxy group, an aryl group carboxamido, carbamoyl aralkyl, aryloxy formyloxy group or absent; _{wherein, R 1, R 2, R} 3 and said R ₄ is phenyl or substituted aryl group 1-4 substituted by a group selected from halogen, hydroxy, nitro, cyano, amino, trifluoromethyl, carboxyl, halogenated C ₁ to C ₈ alkoxy, C ₁ to C ₈ alkoxy At least one selenium-containing substituent in the substituents R ₁ , R ₂ , R ₃ , R ₄ , R ₆ and R _{7 in} (Ib);

   R ₅ is: C ₁ to C ₈ alkylsulfonyl; carbamoyl, cyano;

   R ₆ and R ₇ are each independently selected from the group consisting of: halogenated C ₁ to C ₈ alkoxy, C ₁ to C ₈ alkoxy, halogenated C ₁ to C ₈ alkylselenyl, C ₁ to C ₈ alkylselenyl;

   The dotted line is: chemical bond or absent.
3. The benzoisoselazolone amine compound, solvate, crystalline form, stereoisomer, isotope compound, metabolite, or prodrug according to claim 1, wherein the compound is selected from the group consisting of:

   

   

   
4. A method for preparing a benzoisoselazolidone amine compound according to any one of claims 1-3, obtained by the following method:

   a) Synthetic route of divalent selenium compounds in the I-a structure series of the present invention

   

   b) Synthetic route of tetravalent selenium compounds in the I-a structure series of the present invention

   

   c) Synthetic route of I-b structural compounds of the present invention

   

   The synthesis of divalent selenium compounds in the structure series of formula Ia can be based on the substitution of 2-halobenzoic acid. First, step (1) is condensed with (S) -2- (substituted acyl) -1-substituted phenylethylamine. The intermediate a 'is obtained through the reaction, and then the target compound is obtained through the reaction with step S (2) to form a ring with the selenium reagent. The divalent selenium type compound in the structure series of the formula Ia can also be synthesized directly with (S)- 2- (substituted acyl) -1-substituted phenylethylamine is reacted to obtain the target compound;

   Structure of formula Ia tetravalent selenium series type of compound to divalent substituted benzisoselenazol as raw materials, [O ^-] obtained peroxidation;

   The synthesis of the compound of formula I-b structure type is based on the 4-position deacetyl aposite as a raw material, and is obtained by condensation of step (e) with a selenoyl reagent.
5. The benzoisoselazolidone amine compound, solvate, crystalline form, stereoisomer, isotope compound, metabolite or prodrug according to any one of claims 1-3 in the preparation or prevention of PDE-4 and And / or use in medicine for diseases caused by TNF-α overexpression.
6. The benzoisoselazolidone amine compound, solvate, crystalline form, stereoisomer, isotope compound, metabolite or prodrug according to any one of claims 1-3 in the preparation or prevention of glutathione Application of medicine for diseases related to iron death caused by deficiency or deficiency of peroxidase.
7. A method for treating patients with related diseases caused by overexpression of PDE-4 and / or TNF-α, comprising administering to the patient a therapeutically effective amount of a benzoisoselazolidamine compound, a solvate according to any one of claims 1-3 , Crystalline form, stereoisomer, isotope compound, metabolite, or prodrug.
8. A method for treating patients with cell iron death-related diseases caused by insufficient or missing glutathione peroxidase, comprising administering to the patient a therapeutically effective amount of a benzoisoselazolidone compound according to any one of claims 1-3 , Solvates, crystalline forms, stereoisomers, isotopic compounds, metabolites, or prodrugs.
9. Benzoisoselazolidamine compounds, solvates, crystalline forms, stereoisomers, isotope compounds according to any one of claims 1-3 as inhibitors of PDE-4 and / or TNF-α and / or iron death , Metabolites, or prodrugs.
10. Use, method or benzoisoselazolidamine compound according to claim 5, 6, 7, 8 or 9 wherein said diseases include asthma, arthritis, rheumatoid arthritis, gouty arthritis, rheumatoid Spondylitis, osteoarthritis and other arthritic conditions; sepsis, septic shock, endotoxin shock, gram-negative sepsis, toxic shock syndrome, acute respiratory distress syndrome, chronic cerebral malaria Pulmonary inflammatory disease, silicosis, pulmonary sarcoidosis, bone resorption disease, graft-versus-host response, allograft rejection, fever and myalgia caused by infection, cachexia secondary to infection or malignancy, secondary to human Cachexia of acquired immunodeficiency syndrome (AIDS), AIDS, HIV, HBV, ARC (AIDS-related complex syndrome), keloid formation, scar tissue formation, localized ileitis, ulcerative colitis, multiple sclerosis, Alz Heim disease (AD) and Parkinson's disease (PD), type I diabetes, autoimmune diabetes, diabetes insipidus, systemic lupus erythematosus, bronchitis, chronic obstructive airway disease, psoriasis, Bechet's disease, and similar Allergic purpura nephritis, chronic glomerulonephritis, inflammatory bowel disease, leukemia, allergic rhinitis, depression, multi-infarct dementia or dermatitis.

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