CN111956639A - Use of organic selenium compounds for treating diseases associated with neuroinflammation - Google Patents

Abstract

The invention provides the use of a compound of formula (I), R, in the manufacture of a medicament for the treatment or prevention of diseases and conditions associated with neuroinflammation_a‑R_b-c (w) -O-L-secn (i) wherein the variables are as defined in the specification.

Description

Use of organic selenium compounds for treating diseases associated with neuroinflammation

Technical Field

The present application relates to the field of pharmaceutical chemistry, in particular to the use of compounds of formula (I) for the preparation of a medicament for the treatment or prevention of diseases and conditions associated with neuroinflammation.

Background

Neuroinflammation can lead to nerve damage, produce a series of pathophysiological changes, and cause related neurological diseases and symptoms. Related diseases are mainly a group of nervous system diseases which mainly show disorders of behaviors and psychological activities.

Senile dementia is a typical neuroinflammation-related disease, which is a nervous system disease with various cognitive dysfunctions mainly caused by memory impairment, and can be divided into three types: primary dementia, vascular dementia, and other dementia. Primary dementias include Alzheimer's Disease (AD), Parkinson's Disease (PD), and pick's disease. The main pathological features of alzheimer's disease are β -like starch deposits, fiber tangles, loss of basal nuclear cholinergic neurons, extensive loss of hippocampal and cortical neurons, and synaptic changes. Recent studies have indicated that the appearance of activated microglia is another pathological feature of AD. Microglia are immune cells of the central nervous system, and play an important role in the central nervous system as resident immune and phagocytic cells. In the inflammatory process of the central nervous system, a variety of factors can induce the activation of microglia. Activated microglia can release a large number of neurotoxic factors including Nitric Oxide (NO), interleukin IL-1 β, Reactive Oxygen Species (ROS). Research shows that inhibiting neuroinflammation can become an important strategy for preventing and treating AD (CN 110638854A).

At present, neuroinflammation is generally considered to be an important cause of occurrence of various nervous system diseases. Machunyan et al indicate that neuroinflammation mechanism is the main cause of depression, and inflammation hypothesis considers that stress stimulation triggers inflammatory process and finally causes depression (Chinese medicine report, 2017,14(14): 33-35). Smith et al in 1991 suggested that inflammation could be involved in pathophysiological processes in depression (Barbara S Beltz, Michael F Tlustry, Jeanne L Benton, et al. omega-3 facial acids upper regulated adolest neurogenesis [ J ]. Neurosci Lett, 2007, 415 (2): 154-158.), which was also supported by a number of findings.

Parkinson's Disease (PD) is a degenerative disease of the central nervous system characterized mainly by progressive degeneration and loss of dopaminergic neurons in the substantia nigra pars compacta and formation of lewy bodies, and its clinical manifestations mainly include motor symptoms such as resting tremor, bradykinesia, rigidity of muscles and postural disorders, and non-motor symptoms such as depressed mood, anxiety, sleep disorders, cognitive disorders. A number of research data have shown that microglial and astrocyte-mediated inflammatory responses play an important role in the pathogenesis of PD. Microglia are important immune sensing and effector cells in the central nervous system, the substantia nigra region is one of brain regions with the most abundant distribution of microglia in the brain, and the substantia nigra dopaminergic neurons are extremely sensitive to microglia-mediated inflammatory responses. During the development and progression of neurodegenerative diseases, over-activated microglia can produce a range of inflammatory factors, such as IL-1 β, IFN- γ, NO, IL-6, TNF- α, and the like. On one hand, the inflammatory factors can cause damage and loss of neurons, and the damaged neurons can release NO, ROS and the like, act on microglia, promote the microglia to release the inflammatory factors, promote the further activation of the microglia, and form a vicious circle between the damage of the neurons and the activation of the microglia; on the other hand, these inflammatory factors can also activate astrocytes, causing them to release inflammatory factors to further damage neurons. Therefore, how to effectively inhibit the inflammatory response of nerve cells and reduce the excessive activation of glial cells becomes a potential target for treating neurodegenerative diseases (CN 110623974A).

With the recent intensive research on the pathogenesis of various nervous system disorders such as alzheimer's disease, parkinson's disease, multiple sclerosis, cerebral infarction, progressive muscular dystrophy, parkinson's disease and huntington's chorea, more and more studies have shown that chronic inflammatory responses in the brain mediated by microglial activation are one of the pathological features of nervous system disorders (CN 111116595A).

Although research has been conducted to elucidate a route for the treatment of diseases or conditions associated with neuroinflammation, the existing compounds including anti-inflammatory agents, particularly non-steroidal anti-inflammatory agents, still have problems of insufficient efficacy, limited safety, etc., and are far from meeting the increasing demand of human beings for the prevention and treatment of the related diseases. Therefore, the development of more effective and novel structural compounds has become an urgent need.

Disclosure of Invention

Through a large number of experimental researches, the inventor unexpectedly discovers a selenium-cyanogen-containing organic compound which has unexpected biological activity for preventing and treating diseases or symptoms related to neuroinflammation. The compounds are useful in the treatment and/or prevention of a variety of related diseases or conditions.

Based on the above findings, in a first aspect, the present invention provides a compound having the structure of the following formula (I):

R_a-R_b-C(W)-O-L-SeCN

(I)

or a pharmaceutically acceptable salt thereof, wherein

R_aRepresents aryl or heteroaryl, optionally substituted by one, two or three substituents selected from: phenyl, methylsulfonylphenyl C_1-4Alkyl, halobenzoyl, halophenylamino, benzoyl, C_1-4Alkylphenylamino, halogen, C_1-4Alkyl radical, C_1-4Alkoxy, halo C_1-4Alkoxy radical, C_1-4Alkyl acyl radical, C_1-2Alkylamino, halogeno C_1-4An alkyl group;

w is selected from O, S and Se;

R_bis a straight or branched alkylene or alkenylene chain of 1 to 4 carbon atoms, or is a direct covalent bond;

l is a straight or branched alkylene chain of 1 to 4 carbon atoms.

In the compounds of formula (I) according to the invention, preferably R_aSelected from unsubstituted or substituted phenyl, indenyl, naphthyl, phenylaminophenyl, benzoylphenyl, benzopyrryl, benzopyrrolidinyl and phenylpyrrolidooxacyclohexyl.

In the compounds of the formula (I) according to the invention, R is_aBy definition, the substituents are selected from phenyl, mesylphenyl C_1-4Alkyl, halophenyl-formyl, phenyl-formyl, C_1-4Alkylphenylamino, halogen, C_1-4Alkyl radical, C_1-4Alkoxy, halo C_1-4Alkyl, halo C_1-4Alkoxy, acyl, C_1-4Alkyl acyl, acyloxy, C_1-2Alkyl acyloxy, C_1-2An alkylamino group.

Preferably, R_aHeteroaryl in the definition is selected from the group consisting of benzopyrrole, benzopyrrolidine, benzocyclopentene, benzothiophene, benzofuranyl, benzimidazolyl, azabenzimidazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, imidazopyridinyl, isoxazolopyridinyl; particularly preferably, the heteroaryl group is selected from the group consisting of a benzopyrryl group, a benzopyrrolidinyl group and a phenylpyrrolidinooxeclocyclohexyl group.

In some preferred embodiments of the invention, R_aSelected from phenyl, phenylaminophenyl, benzoylphenyl, indenyl, benzopyrryl, benzopyrrolidinyl and benzopyrrolidino-oxacyclohexyl, and R_aOptionally substituted with one or two substituents selected from: halogen selected from fluorine, chlorine, bromine, iodine, phenyl, phenylamino-, benzoyl, methoxy, straight-chain or branched alkyl of 1 to 4 carbon atoms, C_1-4An alkoxy group.

In the compounds of formula (I) according to the invention, W is preferably O or S; more preferably O.

In the compounds of the formula (I) according to the invention, L is preferably (CH)₂)₂Or (CH)₂)₃That is, L is a C2 or C3 chain, and the chain is not limited to a straight chain and may include a branched moiety.

In some particularly preferred embodiments of the invention, the compound of formula (I) is a specific compound selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.

In another aspect of the invention, there is also provided a pharmaceutical composition comprising a compound of the invention and optionally a carrier.

In yet another aspect of the invention, the above-described pharmaceutical composition may further comprise an additional active pharmaceutical ingredient, i.e., a second medicament suitable for treating the same or a related condition medically indicated to be treatable or preventable by administration of a compound of the present invention.

In yet another aspect of the invention, there is provided a process for preparing a compound of the invention, said process comprising reacting DCC (N, N' -dicyclohexylcarbodiimide) with compound R_a-R_b-C (W) -OH in an organic solvent, wherein the variables are as defined above. Preferably, said compound R_a-R_bThe molar ratio of-C (W) -OH to DCC is 1:1 to 1: 3. Preferably, the reaction is carried out under catalysis of 4-Dimethylaminopyridine (DMAP); preferably, DMAP and R_a-R_b-C (W) -OH in a molar ratio of 1:1 to 1: 2. The reaction is preferably carried out at a temperature below 0 ℃, for example at a temperature of-5 ℃.

Various embodiments of the present invention provide methods of using a compound, composition or combination of the present invention comprising administering to a patient in need of treatment of a malcondition thereof the compound, composition or combination at a dose, frequency and duration sufficient to provide a beneficial effect to the patient.

Drawings

FIG. 1 shows the modulatory effects of compounds 1-9 of the present invention on the production of ROS by LPS-activated BV2 cells.

FIG. 2 shows the modulatory effect of compounds 1-9 of the present invention on IL-1 β production by LPS-activated BV2 cells.

FIG. 3 shows the modulatory effects of compounds 1-9 of the present invention on IL-18 production by LPS-activated BV2 cells.

FIG. 4 is a graph showing the effect of organoselenium compounds of the present invention on the release of Nitrite from LPS-activated BV2 cells.

FIG. 5 is a graph showing the effect of organoselenium compounds of the present invention on the level of iNOS mRNA in LPS-activated BV2 cells.

FIG. 6a is a graph showing the effect of organoselenium compounds of the present invention on reference memory (number of cross-over times) in the Morris space exploration experiments of various groups of mice.

FIG. 6b is a graph showing the effect of organoselenium compounds of the present invention on reference memory (platform peripheral distance) in the Morris space exploration experiments of mice in each group.

FIG. 6c is a graph showing the effect of organoselenium compounds of the present invention on reference memory (target quadrant dwell time) in Morris space exploration experiments in various groups of mice.

Detailed Description

The term "treatment" is defined as the treatment or care of a patient for the purpose of combating a disease, condition, or disorder, which includes the administration of a compound of the present invention to prevent the onset of symptoms or complications, or to alleviate symptoms or complications, or to cure or eliminate the disease, condition, or disorder.

"treating" in the context of the present invention means alleviating the symptoms associated with a disorder or disease, or inhibiting the further development or worsening of these symptoms, or preventing the disease or disorder. Similarly, an "effective amount" or "therapeutically effective amount" of a compound of the invention as used herein refers to an amount of the compound that completely or partially alleviates the symptoms associated with the disorder or condition, or halts or slows further development or worsening of these symptoms, or prevents or provides prophylaxis for the disorder or condition.

When a group is defined as "covalent bond," it is meant that the groups immediately adjacent to the indicated group are attached to each other by a covalent bond.

Unless a particular stereochemistry or isomeric form is specifically indicated, the compounds of the present invention are intended to include all chiral, diastereomeric, racemic forms of the structure. The compounds used in the present invention may include any enriched or resolved degree of optical isomers of any or all asymmetric atoms as is apparent from the description. It is within the scope of the present invention that racemic and diastereomeric mixtures and individual optical isomers may be separated or synthesized so as to be substantially free of their enantiomers or diastereomers.

Generally, "substituted" means that an organic group, as defined herein, which contains one or more bonds to a hydrogen atom, is substituted with one or more bonds to a non-hydrogen atom, i.e., a substituent.

Substituents of the compounds of the invention (including R)_aSubstituents in the definition) include halogen, alkyl (preferably C)_1-4Alkyl, more preferably C_1-2Alkyl group), alkoxy group (preferably C)_1-4Alkoxy, more preferably C_1-2Alkoxy), halo C_1-4Alkoxy (preferably halo C)_1-2Alkoxy), amino, acyl, C_1-4Alkylacyl (preferably C)_1-2Alkyl acyl), acyloxy, C_1-4Alkanoyloxy (preferably C)_1-2Alkanoyloxy), carboxy, nitro, cyano, C_1-4Alkylamino (preferably C)_1-2Alkylamino), halo C_1-4Alkyl (preferably halogenated C)_1-2Alkyl group), C_1-4Alkylamino radical C_1-4Alkyl (preferably C)_1-2Alkylamino radical C_1-2Alkyl), hydroxy C_1-4Alkyl (preferably hydroxy C)_1-2Alkyl), hydroxy C_1-4Alkylamino (preferably hydroxy C)_1-2Alkylamino), C_3-6Cycloalkyl radical, C_2-4An alkenyl group.

In the definition of substituents of the present invention, the dash "-" represents a group or a bonding position where a substituent is attached to the rest of the structural formula. As will be appreciated by those skilled in the art, for the structures of formula (I), R_aHaving a bonding position, and R_bThere are two bonding sites at both ends.

Alkyl groups include straight and branched chain alkyl groups having from 1 to about 8 carbon atoms, or in certain embodiments from 1 to 4 and even more preferably from 1 to 2 carbon atoms. Examples of straight chain alkyl groups include those having 1 to 8, preferably 1 to 4, more preferably 1-2 carbon atoms, such as methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl. Examples of branched alkyl groups include, but are not limited to, isopropyl, isobutyl, sec-butyl, tert-butyl, neopentyl, isoamyl, and 2, 2-dimethylpropyl. Representative substituted alkyl groups may be substituted one or more times with any of the aforementioned groups, for example, amino, hydroxyl, cyano, carboxyl, nitro, thio, alkoxy, and halogen groups.

Cycloalkyl is a cyclic alkyl group such as, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. In preferred embodiments, the cycloalkyl group has 3 to 8 ring members, while in other embodiments the number of ring carbon atoms is 3 to 5, 6 or 7. Cycloalkyl groups further include polycyclic cycloalkyl groups such as, but not limited to, norbornyl, adamantyl, bornyl, camphene, isobornene, and carenyl, and fused rings such as, but not limited to, decahydronaphthyl and the like. Cycloalkyl also includes rings substituted with straight or branched chain alkyl as defined above. Representative substituted cycloalkyl groups may be mono-substituted or substituted more than once, such as, but not limited to, 2-, 2,3-, 2,4-, 2,5-, or 2, 6-disubstituted cyclohexyl or mono-, di-, or tri-substituted norbornyl or cycloheptyl, which may be substituted with, for example, amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halo groups. The term "cycloalkenyl", alone or in combination, denotes cyclic alkenyl groups.

Aryl is a cyclic aromatic hydrocarbon that does not contain heteroatoms. Thus, aryl groups include, but are not limited to, phenyl, azulenyl, heptenylyl (heptalenyl), biphenyl, dicyclopentadiene acenyl (indacenyl), fluorenyl, phenanthryl, triphenylenyl (triphenylenyl), pyrenyl, napthyl (napthylenyl), biphenylene, anthracenyl, and naphthyl. In certain embodiments, the aryl group contains 6 to 14 carbons in the ring portion of the group. The aryl group may be unsubstituted or substituted, as defined above. Representative substituted aryl groups may be monosubstituted or substituted more than once, such as but not limited to 2-, 3-, 4-, 5-, or 6-substituted phenyl or 2-8 substituted naphthyl, which may be substituted with carbon or non-carbon groups (such as those described above).

Heteroaryl is an aromatic ring compound containing 5 or more ring members, wherein one or more ring members are heteroatoms, such as, but not limited to N, O and S. Is named as C₂The heteroaryl group of the heteroaryl group may be a 5-membered ring having two carbon atoms and three heteroatoms, a 6-membered ring having two carbon atoms and four heteroatoms, or the like. Likewise, C₄The heteroaryl group may be a 5-membered ring having one heteroatom, a 6-membered ring having 2 heteroatoms, or the like. The sum of the number of carbon atoms plus the number of heteroatoms is equal to the total number of ring atoms. Heteroaryl groups include, but are not limited to, groups such as: pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridyl, thienyl, benzothienyl, benzofuranyl, indolyl, azaindolyl, indazolyl, benzimidazolyl, azabenzimidazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, imidazopyridinyl, isoxazolopyridyl, thianaphthyl, purinyl, xanthyl, adenine, guanine, quinolyl, isoquinolyl, tetrahydroquinolyl, quinoxalyl and quinazolinyl. Heteroaryl groups may be unsubstituted or may be substituted as described above. Representative substituted heteroaryl groups can be substituted one or more times with those groups described above.

Other examples of aryl and heteroaryl groups include, but are not limited to, phenyl, biphenyl, indenyl, naphthyl (1-naphthyl, 2-naphthyl), N-hydroxytetrazolyl, N-hydroxytriazolyl, N-hydroxyimidazolyl, anthracenyl (1-anthracenyl, 2-anthracenyl, 3-anthracenyl), thienyl (2-thienyl, 3-thienyl), furyl (2-furyl, 3-furyl), indolyl, oxadiazolyl, isoxazolyl, quinazolinyl, fluorenyl, xanthenyl, isoindolyl, benzhydryl, acridinyl, thiazolyl, pyrrolyl (2-pyrrolyl), pyrazolyl (3-pyrazolyl), imidazolyl (1-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl), triazolyl (1,2, 3-triazol-1-yl, 1,2, 3-triazol-2-yl, 1,2, 3-triazol-4-yl, 1,2, 4-triazol-3-yl), oxazolyl (2-oxazolyl, 4-oxazolyl, 5-oxazolyl), thiazolyl (2-thiazolyl, 4-thiazolyl, 5-thiazolyl), pyridyl (2-pyridyl, 3-pyridyl, 4-pyridyl), pyrimidinyl (2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, 6-pyrimidinyl), pyrazinyl, pyridazinyl (3-pyridazinyl, 4-pyridazinyl, 5-pyridazinyl), quinolinyl (2-quinolinyl, 3-quinolinyl, 4-quinolinyl, 5-pyridazinyl), 5-quinolyl group, 6-quinolyl group, 7-quinolyl group, 8-quinolyl group), isoquinolyl group (1-isoquinolyl group, 3-isoquinolyl group, 4-isoquinolyl group, 5-isoquinolyl group, 6-isoquinolyl group, 7-isoquinolyl group, 8-isoquinolyl group), benzo [ b ] furyl group (2-benzo [ b ] furyl group, 3-benzo [ b ] furyl group, 4-benzo [ b ] furyl group, 5-benzo [ b ] furyl group, 6-benzo [ b ] furyl group, 7-benzo [ b ] furyl group), 2, 3-dihydro-benzo [ b ] furyl group (2- (2, 3-dihydro-benzo [ b ] furyl group), 3- (2, 3-dihydro-benzo [ b ] furyl group), 4- (2, 3-dihydro-benzo [ b ] furyl), 5- (2, 3-dihydro-benzo [ b ] furyl), 6- (2, 3-dihydro-benzo [ b ] furyl), 7- (2, 3-dihydro-benzo [ b ] furyl), benzo [ b ] thienyl (2-benzo [ b ] thienyl, 3-benzo [ b ] thienyl, 4-benzo [ b ] thienyl, 5-benzo [ b ] thienyl, 6-benzo [ b ] thienyl, 7-benzo [ b ] thienyl), 2, 3-dihydro-benzo [ b ] thienyl, (2- (2, 3-dihydro-benzo [ b ] thienyl), 3- (2, 3-dihydro-benzo [ b ] thienyl), 4- (2, 3-dihydro-benzo [ b ] thienyl), 5- (2, 3-dihydro-benzo [ b ] thienyl), 6- (2, 3-dihydro-benzo [ b ] thienyl), 7- (2, 3-dihydro-benzo [ b ] thienyl), indolyl (1-indolyl, 2-indolyl, 3-indolyl, 4-indolyl, 5-indolyl, 6-indolyl, 7-indolyl), indazole (1-indazolyl, 3-indazolyl, 4-indazolyl, 5-indazolyl, 6-indazolyl, 7-indazolyl), benzimidazolyl (1-benzimidazolyl, 2-benzimidazolyl, 4-benzimidazolyl, 5-benzimidazolyl, etc.), 6-benzimidazolyl, 7-benzimidazolyl, 8-benzimidazolyl), benzoxazolyl (1-benzoxazolyl, 2-benzoxazolyl), benzothiazolyl (1-benzothiazolyl, 2-benzothiazolyl, 4-benzothiazolyl, 5-benzothiazolyl, 6-benzothiazolyl, 7-benzothiazolyl), carbazolyl (1-carbazolyl, 2-carbazolyl, 3-carbazolyl, 4-carbazolyl) and the like.

More particularly, aryl and heteroaryl groups may include phenyl, isoindolinyl, imidazolyl, oxazolyl, benzimidazolyl and benzoxazolyl groups; wherein any aryl or heteroaryl group may be unsubstituted, mono-substituted or, independently, poly-substituted, for example with a J group as defined herein.

The term "alkoxy" means that the oxygen atom is attached to an alkyl group as defined above, including cycloalkyl groups. Examples of linear alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, pentoxy, hexoxy, and the like. Examples of branched alkoxy groups include, but are not limited to, isopropoxy, sec-butoxy, tert-butoxy, isopentoxy, isohexoxy, and the like. Examples of cycloalkoxy groups include, but are not limited to, cyclopropoxy, cyclobutoxy, cyclopentoxy, cyclohexyloxy, and the like.

The term "halogen" as used herein includes fluorine, chlorine, bromine and iodine. "haloalkyl" includes monohaloalkyl and polyhaloalkyl (wherein all halogen atoms may be the same or different). Partially halogenated alkyl is "haloalkyl" within the meaning of the present disclosure. Examples of haloalkyl groups include trifluoromethyl, 1-dichloroethyl, 1, 2-dichloroethyl, 1, 3-dibromo-3, 3-difluoropropyl, and the like.

The term "acyl" as used herein refers to a group comprising a carbonyl moiety, wherein the group is bonded via the carbonyl carbon atom. The carbonyl carbon atom is also bonded to other carbon atoms, which may be part of an alkyl, aryl, aralkylcycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl, or the like. In the particular case where the carbonyl carbon atom is bonded to hydrogen, the group is "formyl", which is an acyl group (the term being as defined herein). Other examples include acetyl, benzoyl, phenylacetyl, pyridylacetyl, cinnamoyl, acryloyl, and the like. When a group containing a carbon atom bonded to a carbonyl carbon atom contains a halogen, the group is referred to as a "haloacyl". One example is trifluoroacetyl.

"salts" as is well known in the art include organic compounds such as carboxylic acids, sulfonic acids or amines in ionic form in combination with a counter ion. For example, an acid in anionic form may form a salt with: cations such as metal cations, e.g., sodium, potassium, and the like; ammonium saltsSuch as NH₄ ⁺Or cations of various amines, including tetraalkylammonium salts such as tetramethylammonium, or other cations such as trimethylsulfonium, and the like. A "pharmaceutically acceptable" salt is a salt formed from ions that are approved for human use and are generally non-toxic, such as the hydrochloride or sodium salt. "zwitterions" are internal salts, which can be formed, for example, in molecules having at least two ionizable groups, one forming an anion and the other forming a cation, which are in equilibrium with one another. For example, an amino acid (e.g., glycine) can be present in zwitterionic form. "zwitterion" is a salt within the meaning herein.

A "hydrate" is a compound that exists in combination with a water molecule. The composition may contain a stoichiometric amount of water, such as a monohydrate or dihydrate, or may contain any amount of water.

A "solvate" is a similar composition except that water is replaced with a solvent different from water. For example, methanol or ethanol may form "alcoholates", which may also be stoichiometric or non-stoichiometric.

"tautomers" are two forms of matter that differ only in the position of a hydrogen atom in the molecular structure.

Another aspect of the present invention provides a pharmaceutical composition comprising a compound of the present invention and optionally a carrier or a pharmaceutical composition further comprising a further pharmaceutically active ingredient in addition to a compound of the present invention and optionally a carrier. The pharmaceutical compositions of the invention may be prepared by conventional techniques, for example as described in Remington: the method described in The Science and Practice of Pharmacy, 19 th edition, 1995, which is incorporated herein by reference. The compositions may be presented in conventional forms, such as capsules, tablets, aerosols, solutions, suspensions or topical application forms.

Typical compositions comprise a compound of the invention and a carrier. For example, the active compound is typically mixed with a carrier, or diluted by a carrier, or enclosed within a carrier which may be in the form of an ampoule, capsule, sachet (sachet), paper or other container. When the active compound is mixed with a carrier, or when the carrier serves as a diluent, the carrier can be a solid, semi-solid, or liquid material that serves as a carrier, excipient, or medium for the active compound. The active compound may be adsorbed on a particulate solid carrier (e.g. contained in a sachet). Some examples of suitable carriers are water, salt solutions, alcohols, polyethylene glycols, polyhydroxyethoxylated castor oil, peanut oil, olive oil, gelatin, lactose, terra alba, sucrose, dextrin, magnesium carbonate, sugars, cyclodextrins, amylose, magnesium stearate, talc, gelatin, agar, pectin, acacia, stearic acid or lower alkyl ethers of cellulose, silicic acid, fatty acids, fatty acid amines, fatty acid mono-and diglycerides, pentaerythritol fatty acid esters, polyoxyethylene, hydroxymethylcellulose and polyvinylpyrrolidone. Similarly, the carrier or diluent may include any sustained release material known in the art, such as glyceryl monostearate or glyceryl distearate alone or with a wax.

The formulations may be mixed with adjuvants which do not deleteriously react with the active compound. These additives may include wetting agents, emulsifying and suspending agents, salts for influencing osmotic pressure, buffering and/or coloring substances, preservatives, sweeteners or flavorings. The composition may also be sterilized, if desired.

The route of administration may be any route which is effective for transporting the active compounds of the invention to the appropriate or desired site of action, for example the oral, nasal, pulmonary, buccal, subcutaneous, intradermal, transdermal or parenteral routes, for example the rectal, depot (depot), subcutaneous, intravenous, intraurethral, intramuscular, intranasal, ophthalmic solutions or ointments route, the oral route being preferred.

If a solid carrier is used for oral administration, the formulation may be tableted, placed in a hard gelatin capsule as a powder or pellet, or it may be in the form of a lozenge or troche. If a liquid carrier is used, the formulation may be in the form of a syrup, emulsion, soft gelatin capsule, or sterile injectable liquid, such as an aqueous or non-aqueous liquid suspension or solution.

Injectable dosage forms typically comprise an aqueous or oily suspension, which may be formulated using suitable dispersing or wetting agents and suspending agents. Injectable forms may be in the form of a solution phase or a suspension prepared with a solvent or diluent. Acceptable solvents or carriers include sterile water, ringer's solution, or isotonic saline solution. Alternatively, sterile oils may be employed as a solvent or suspending agent. Preferably, the oil or fatty acid is non-volatile and comprises a natural or synthetic oil, a fatty acid, a monoglyceride, diglyceride, or triglyceride.

For injection, the formulation may also be a powder suitable for reconstitution with a suitable solution as described above. Examples of these include, but are not limited to, freeze-dried, spin-dried or spray-dried powders, amorphous powders, granules, precipitates or microparticles. For injections, the formulation may optionally include stabilizers, pH modifiers, surfactants, bioavailability modifiers, and combinations of these agents. The compounds may be formulated for parenteral administration by injection, for example by bolus injection or continuous infusion. Unit dosage forms for injection may be in ampoules or in multi-dose containers.

The formulations of the present invention may be designed to provide rapid, sustained or delayed release of the active ingredient after administration to a patient by methods well known in the art. Thus, the formulation may also be formulated for controlled release or slow release.

The compounds of the present invention are effective over a wide dosage range. For example, in the treatment of adults, a dose of about 0.05 to about 5000mg, preferably about 1 to about 2000mg, more preferably about 2 to about 2000mg per day may be used. Typical dosages are from about 10mg to about 1000mg per day. When selecting a patient treatment regimen, it may often be necessary to start with a higher dose and reduce the dose when the condition is controlled. The precise dosage will depend upon the activity of the compound, the mode of administration, the desired treatment, the form of administration, the subject to be treated and the weight of the subject to be treated, as well as the preferences and experience of the attending physician or veterinarian.

Typically, the compounds of the present invention are dispensed in unit dosage forms containing from about 0.05mg to about 1000mg of the active ingredient per unit dose and a pharmaceutically acceptable carrier.

In general, dosage forms suitable for oral, nasal, pulmonary or transdermal administration include from about 125 μ g to about 1250mg, preferably from about 250 μ g to about 500mg, more preferably from about 2.5mg to about 250mg of the compound in admixture with a pharmaceutically acceptable carrier or diluent.

The dosage form may be administered once daily, or more than once daily, e.g., twice daily or three times daily. Alternatively, the dosage form may be administered less frequently than once daily, for example every other day or weekly, if deemed appropriate by the prescribing physician.

The pharmaceutical compositions may be in the form of tablets, capsules, powders, granules, lozenges, liquids or gels. Tablets and capsules for oral administration may be in a form suitable for unit dose administration and may contain conventional excipients, such as: binders such as syrup, gum arabic, gelatin, sorbitol, tragacanth, polyvinylpyrrolidone (PVP); fillers such as lactose, sugars, corn flour, calcium phosphate, sorbitol or glycine; tablet lubricants such as magnesium stearate, silicon dioxide, talc, polyethylene glycol or silicon dioxide; disintegrants such as potato starch; acceptable lubricants such as sodium lauryl sulfate. The tablets may be coated according to known methods of conventional pharmaceutical practice. Oral liquid preparations may be in the form of aqueous or oily suspensions, solutions, emulsions, syrups or tinctures, or may be presented as a dry substance for reconstitution with water or other suitable vehicle before use. These liquid preparations may contain conventional additives such as suspending agents (e.g., sorbitol, syrup, methyl cellulose, glucose syrup, gelatin, hydrogenated edible fats and oils). Emulsifying agents (e.g. lecithin, sorbitol monooleate or acacia), non-aqueous vehicles (including edible oils such as almond oil, fractionated coconut oil, fats and oils such as glycerol, propylene glycol or ethanol), preservatives (e.g. methyl or propyl p-hydroxybenzoic acid or sorbic acid), and if desired conventional flavouring or colouring agents.

The percentage of active substance in the pharmaceutical composition of the present invention is variable because the pharmaceutical formulation must be formulated in a suitable proportion of the dosage to achieve the desired therapeutic effect. In general, the pharmaceutical preparations according to the invention should be administered orally or by injection in a dose of 1 to 15mg of active substance per 70kg of body weight per day. Some examples are given below by way of illustration and not by way of limitation.

The present invention provides, in various embodiments, methods of synthesizing the compounds of the invention herein. The general method comprises reacting R_a-R_bA compound represented by-C (W) -OH with 2-selenocyanolethanol or 3-selenocyanopropanol (NCSe (CH)₂)_nOH) (n is 2 or 3) in an organic solvent. The reaction can be carried out at low temperature, under catalysts (DCC and DMAP). In some preferred embodiments, the compound of formula R_a-R_bDissolving a (C) (W) -OH compound in tetrahydrofuran or dichloromethane, adding 4-dimethylaminopyridine, adding N, N' -Dicyclohexylcarbodiimide (DCC) under stirring, and reacting under stirring in ice bath to generate a target compound R_a-R_b-C(W)-O-L-SeCN。

Examples

The technical solution and applications of the present invention are further illustrated by the following specific examples:

examples of synthetic preparations

Example 1: preparation of Compound 1

The reaction formula is as follows:

a50 ml flask was charged with Compound II-a (100mg, 1.0eq) and 10ml of anhydrous tetrahydrofuran, and 2-selenoethanol (54mg, 1.2eq), DCC (62mg, 1.0eq) and DMAP (44mg, 1.2eq) were added in this order under ice-cooling, and the reaction was stirred for 12 hours and monitored by TLC for completion. The solvent was distilled off under reduced pressure, and the extract was separated by silica gel column chromatography using ethyl acetate (V): petroleum ether (V) ═ 1:3 as the mobile phase, gave a pale yellow solid, compound 1120 mg, in 86% yield.

Nuclear magnetic resonance¹H NMR(400MHz,CDCl₃):2.24(s,3H,-CH₃),2.88(s,3H,CH₃),3.16(t,2H,J＝8.00Hz,CH₂),3.64(s,2H,CH₂),3.85(t,2H,J＝8.00Hz,CH₂),6.55(t,1H,J＝8.00Hz,ArH),6.80(d,1H,J＝8.00Hz,ArH),7.11(d,1H,J＝4.00Hz,ArH),7.17(s,1H,CH),7.58(d,2H,J＝8.00Hz,ArH),7.73(d,2H,J＝8.00Hz,ArH).¹³C NMR(100MHz,CDCl₃):11.3,26.2,34.9,43.3,51.2,107.1(d,J＝24Hz),113.9(d,J＝22Hz),122.9,129.4,129.8,130.9,132.4,138.3,140.2,141.4,145.6,147.8(d,J＝9.5Hz),163.2,163.9,165.5,174.8.¹⁹F NMR(CDCl₃,376MHz):＝-112.1(s,F).

ESI-MS:471.4([M+1]⁺).

Example 2: preparation of Compound 2

The reaction formula is as follows:

a50 ml flask was charged with Compound II-b (100mg, 1.0eq) and 10ml of anhydrous tetrahydrofuran, and 2-selenoethanol (72mg, 1.2eq), DCC (82mg, 1.0eq) and DMAP (59mg, 1.2eq) were added in this order under ice-cooling, and the reaction was stirred for 12 hours and monitored by TLC for completion. The solvent was distilled off under reduced pressure, and the extract was separated by silica gel column chromatography using ethyl acetate (V): petroleum ether (V) ═ 1:3 as the mobile phase, gave a white solid, compound 2100 mg, 66% yield.

Nuclear magnetic resonance¹H NMR(400MHz,CDCl₃)：1.50(d,3H,J＝7.20Hz,CH₃),3.13(t,2H,J＝4.00Hz,CH₂),3.70(q,1H,J＝7.20Hz,CH),3.81(t,2H,J＝4.00Hz,CH₂),7.04-7.11(m,2H,ArH),7.31-7.42(m,4H,ArH),7.46-7.50(m,2H,ArH).¹³C NMR(100MHz,CDCl₃):18.3,24.2,48.4,53.8,114.7(d,J＝24Hz),124.2,129.1,129.3,129.5,130.4,131.8,136.4,144.5(d,J＝8Hz),159.8,162.5,165.3,178.6.¹⁹F NMR(CDCl₃,376MHz):＝-112.2(s,F).

ESI-MS:377.3([M+1]⁺).

EXAMPLE 3 preparation of Compound 3

The reaction formula is as follows:

a50 ml flask was charged with Compound II-c (100mg, 1.0eq) and 10ml of anhydrous tetrahydrofuran, and 2-selenoethanol (54mg, 1.2eq), DCC (62mg, 1.0eq) and DMAP (44mg, 1.2eq) were added in this order under ice-cooling, and the reaction was stirred for 12 hours and monitored by TLC for completion. The solvent was distilled off under reduced pressure, and the extract was separated by silica gel column chromatography using ethyl acetate (V): petroleum ether (V) ═ 1:3 as the mobile phase, gave a white solid, compound 3, 120mg, 84% yield.

Nuclear magnetic resonance¹H NMR(400MHz,CDCl₃):2.12(s,3H,CH₃),3.03(t,2H,J＝8.00Hz,CH₂),3.60(t,2H,J＝8.00Hz,CH₂),3.64(s,3H,CH₃),6.68(d,1H,J＝8.00Hz,ArH),6.86(d,1H,J＝8.00Hz,ArH),7.11(m,1H,ArH),7.60(d,2H,J＝8.00Hz,ArH),7.65(d,2H,J＝8.00Hz,ArH).¹³C NMR(100MHz,CDCl₃):14.2,26.3,33.5,56.7,101.1,103.7,112.9,114.6,116.4,129.4,130.4,131.3,131.8,133.4,134.2,139.4,157.3,166.7.

ESI-MS:476.8([M+1]⁺).

EXAMPLE 4 preparation of Compound 4

The reaction formula is as follows:

a50 ml flask was charged with Compound II-d (100mg, 1.0eq) and 10ml of anhydrous tetrahydrofuran, and 2-selenoethanol (72mg, 1.2eq), DCC (82mg, 1.0eq) and DMAP (59mg, 1.2eq) were added in this order under ice-cooling, and the reaction was stirred for 12 hours and monitored by TLC for completion. The solvent was distilled off under reduced pressure, and the extract was separated by silica gel column chromatography using ethyl acetate (V): petroleum ether (V) ═ 1:3 as the mobile phase, gave a pale yellow solid, compound 4, 100mg, 65% yield.

Nuclear magnetic resonance¹H NMR(400MHz,CDCl₃):1.48(d,3H,J＝8.00Hz,CH₃),2.12-2.16(m,2H,CH₂),3.03(m,2H,CH₂),3.68(q,1H,J＝8.00Hz,CH),7.40-7.46(m,3H,ArH),7.61-7.67(m,3H,ArH),7.76-7.82(m,3H,ArH).¹³C NMR(100MHz,CDCl₃):17.5,31.2,38.5,47.6,102.6,128.0,128.6,129.2,129.7,130.4,131.0,131.5,137.6,138.8,142.2,172.6,193.9.

ESI-MS:387.3([M+1]⁺).

EXAMPLE 5 preparation of Compound 5

The reaction formula is as follows:

a50 ml flask was charged with Compound II-e (100mg, 1.0eq) and 10ml of anhydrous tetrahydrofuran, and 2-selenoethanol (76mg, 1.2eq), DCC (86mg, 1.0eq) and DMAP (62mg, 1.2eq) were added in this order under ice-cooling, and the reaction was stirred for 12 hours and monitored by TLC for completion. The solvent was distilled off under reduced pressure, and the extract was separated by silica gel column chromatography using ethyl acetate (V): petroleum ether (V) ═ 1:3 as the mobile phase, gave a pale yellow solid, compound 5, 115mg, 73% yield.

Nuclear magnetic resonance¹H NMR(400MHz,DMSO):2.08(s,3H,CH₃),2.32(s,3H,CH₃),3.15(t,2H,J＝8.00Hz,CH₂),3.67(t,2H,J＝8.00Hz,CH₂),6.60–6.64(m,2H,ArH),6.96-6.99(m,1H,ArH),7.06-7.08(m,2H,ArH),7.15-7.17(m,1H,ArH),8.10(d,1H,J＝4.00Hz,ArH),9.67(s,1H,NH).¹³C NMR(100MHz,DMSO):13.8,21.2,23.8,48.4,114.8,117.7,119.2,124.4,126.2,126.8,131.1,131.4,134.8,137.3,138.6,147.2,172.5,174.6.

ESI-MS:374.1([M+1]⁺).

EXAMPLE 6 preparation of Compound 6

The reaction formula is as follows

A50 ml flask was charged with Compound II-f (100mg, 1.0eq) and 10ml of anhydrous tetrahydrofuran, and 2-selenoethanol (61mg, 1.2eq), DCC (70mg, 1.0eq) and DMAP (50mg, 1.2eq) were added in this order under ice-cooling, and the reaction was stirred for 12 hours and monitored by TLC for completion. The solvent was distilled off under reduced pressure, and the extract was separated by silica gel column chromatography using ethyl acetate (V): petroleum ether (V) ═ 1:3 as the mobile phase, gave a pale yellow solid, compound 6, 100mg, 69% yield.

Nuclear magnetic resonance¹H NMR(400MHz,CDCl₃):3.06-3.09(m,2H,CH₂),3.68-3.72(m,2H,CH₂),3.76(s,2H,CH₂),6.48–6.50(m,1H,ArH),6.25(d,1H,J＝8.00Hz,ArH),6.83-6.86(m,1H,ArH),7.03-7.06(m,1H,ArH),7.20-7.24(m,2H,ArH),7.53(d,2H,J＝8.00Hz,ArH),7.67(s,1H,NH).¹³C NMR(100MHz,CDCl₃):24.9,41.5,51.7,115.6,120.7,124.3,125.8,127.5,129.3,130.4,137.1,143.0,175.8.

ESI-MS:428.0([M+1]⁺).

EXAMPLE 7 preparation of Compound 7

The reaction formula is as follows

A50 ml flask was charged with Compound II-g (100mg, 1.0eq) and 10ml of anhydrous tetrahydrofuran, and 2-selenoethanol (52mg, 1.2eq), DCC (72mg, 1.0eq) and DMAP (51mg, 1.2eq) were added in this order under ice-cooling, and the reaction was stirred for 12 hours and monitored by TLC for completion. The solvent was distilled off under reduced pressure, and the extract was separated by silica gel column chromatography using ethyl acetate (V): petroleum ether (V) ═ 1:3 as the mobile phase, gave a pale yellow solid, compound 7, 120mg, 81% yield.

Nuclear magnetic resonance¹H NMR(400MHz,CDCl₃):0.88(t,3H,J＝8.00Hz,CH₃),1.30(t,3H,J＝8.00Hz,CH₃),1.92-2.14(m,2H,CH₂),2.75-2.83(m,4H,2×CH₂),3.12(q,2H,J＝8.00Hz,CH₂),3.16(q,2H,J＝8.00Hz,CH₂),3.65-3.71(m,2H,CH₂),3.92-4.00(m,2H,CH₂),6.94(d,1H,J＝8.00Hz,ArH),7.02(t,1H,J＝8.00Hz,ArH),7.36(d,1H,J＝8.00Hz,ArH).¹³C NMR(100MHz,CDCl₃):8.6,14.1,23.3,25.7,26.4,32.3,46.7,57.8,62.1,77.5,110.4,117.3,120.5,121.2,127.3,127.4,135.2,135.9,162.1,170.3.

ESI-MS:422.0([M+1]⁺).

EXAMPLE 8 preparation of Compound 8

The reaction formula is as follows

A50 ml flask was charged with Compound II-h (100mg, 1.0eq) and 10ml of anhydrous tetrahydrofuran, and 3-selenocyanopropanol (76mg, 1.2eq), DCC (86mg, 1.0eq) and DMAP (62mg, 1.2eq) were added in this order under ice-cooling and the reaction was stirred for 12 hours and monitored by TLC for completion. The solvent was distilled off under reduced pressure, and the extract was separated by silica gel column chromatography using ethyl acetate (V): petroleum ether (V) ═ 1:3 as the mobile phase, gave a pale yellow solid, compound 8, 120mg, 75% yield.

Nuclear magnetic resonance¹H NMR(400MHz,CDCl₃):2.11(s,3H,CH₃),2.10-2.22(m,2H,CH₂),2.28(s,3H,CH₃),3.15(t,2H,J＝8.00Hz,CH₂),3.48(q,2H,J＝8.00Hz,CH₂),6.48–6.50(m,1H,ArH),6.55(t,1H,J＝8.00Hz,ArH),6.82(d,1H,J＝8.00Hz,ArH),6.90(d,1H,J＝8.00Hz,ArH),7.00(t,1H,J＝8.00Hz,ArH),7.08(d,1H,J＝8.00Hz,ArH),7.12-7.18(m,1H,ArH),7.37(d,1H,J＝8.00Hz,ArH),¹³C NMR(100MHz,CDCl₃):13.9,21.4,26.4,32.5,37.8,105.7,114.2,115.4,116.3,122.3,124.8,126.3,127.2,131.2,132.3,137.9,138.3,146.5,171.4.

ESI-MS:388.0([M+1]⁺).

EXAMPLE 9 preparation of Compound 9

The reaction formula is as follows

A50 ml flask was charged with Compound II-i (100mg, 1.0eq) and 10ml of anhydrous tetrahydrofuran, and 3-selenocyanopropanol (57mg, 1.2eq), DCC (72mg, 1.0eq) and DMAP (51mg, 1.2eq) were added in this order under ice-cooling, and the reaction was stirred for 12 hours and monitored by TLC for completion. The solvent was distilled off under reduced pressure, and the extract was separated by silica gel column chromatography using ethyl acetate (V): petroleum ether (V) ═ 1:3 as the mobile phase, gave a pale yellow solid, compound 9, 105mg, 77% yield.

Nuclear magnetic resonance¹H NMR(400MHz,CDCl₃:0.98(t,3H,J＝8.00Hz,CH₃),1.35(t,3H,J＝8.00Hz,CH₃),1.80-1.92(m,2H,CH₂),2.08–2.13(m,2H,CH₂),2.38-2.41(m,1H,CH),2.77-2.82(m,5H,CH₂,CH₂,CH),2.30–2.38(m,1H,CH),3.09-3.16(m,1H,CH),3.50-3.57(m,1H,CH),4.00–4.12(m,2H,CH₂),6.72(brs,1H,NH),6.88-6.92(m,1H,ArH),7.00-7.05(m,1H,ArH),7.54(d,1H,J＝8.00Hz,ArH).¹³C NMR(100MHz,CDCl3):8.3,15.4,23.2,24.3,27.3,31.2,31.9,38.4,44.2,61.4,75.4,101.8,108.2,116.9,120.2,120.8,126.6,127.4,132.8,134.2,170.1.

ESI-MS:436.1([M+1]⁺).

Biological Activity assay

Example 10 Regulation of ROS production by LPS-activated BV2 cells by organic selenium Compounds

The method comprises the following steps: serum-free medium is used for diluting DCFH-DA (ratio 1:1000), the final concentration is 10 muM, 20min before the drug treatment is finished, ROSUP is added into a positive control group (ratio 1:1000), the original medium in a 6-well plate is removed, 1ml of DCFH-DA diluent is added, a cell culture box is incubated for 20min, the cells are washed for 3 times by the serum-free cell culture solution, the cells are digested by trypsin, the cells are centrifuged at 4 ℃ and 1200rpm for 5min, PBS1X blows off the cells, the cells are washed again to prepare cell suspension, and 100 muL of each hole of a 96 black well plate is added. And detecting the values under the excitation wavelength of 502nm and the emission wavelength of 530nm of the fluorescence microplate reader.

Intracellular ROS levels, mainly H2O2 levels, were measured using the DCFH-DA method, and the results are shown in FIG. 1. As can be seen from the results of fig. 1, the ROS level in the Lipopolysaccharide (LPS) group was significantly increased compared to the Control group (Control group), which was close to the positive Control sulindac group (ROSUP). After adding the organic selenium compounds 1 to 9 (compounds 1 to 9 correspond to 1 to 39, 1 to 40, 1 to 43, 1 to 44, 1A to 38, 1A to 39, 1A to 40, 1A to 46 and 1A to 49 in the figure respectively), the ROS level can be found to be remarkably reduced after being treated for 48 hours at the concentrations of 3 mu M and 30 mu M, and the compound has better effect than the positive medicine group sulindac.

EXAMPLE 11 Regulation of IL-1 β production by LPS-activated BV2 cells by organic selenium compounds

The method comprises the following steps: balancing the sample and the kit to room temperature, and designing plate hole arrangement and a standard product in advance; a secondary hole; a blank well. And (3) diluting the standard substance, adding 40 mu l of sample diluent into the sample hole to be detected, and then adding 10 mu l of sample. 50 μ l of deionized water was added to the blank wells. Add 50. mu.l of standard to the standard well. Mix well, cover with sealing plate membrane, incubate 30min at 37 ℃. Washing liquid is prepared, 20ml of 20x concentrated washing liquid is added with deionized water to reach the constant volume of 40 ml. After incubation, 300. mu.l of wash solution was added to each well. After washing, the liquid is discarded, and the solution is dried. 50 μ l of enzyme-labeled reagent was added to each well, and no blank wells were added. Incubate at 37 ℃ for 30min and wash the plate again as above. 50. mu.l of the developing solution A was added to each well, and 50. mu.l of the developing solution B was added thereto. After gently shaking and developing for 10min at 37 ℃ in the dark, 50. mu.l of stop solution was added and the color immediately changed from blue to yellow. And placing the ELISA plate on a reading of an ELISA reader, and determining the concentration.

The content of IL-1 beta released by the activated BV2 cells was determined by ELISA. IL-1 β is a pro-inflammatory factor in the inflammatory response, and when the inflammatory signaling pathway is activated, a large amount of neuroinflammatory factors are released, causing cell damage, and the results are shown in FIG. 2. As can be seen from the results of FIG. 2, the IL-1. beta. level was significantly increased in the LPS group as compared with the Control group (Control group). After adding the organic selenium compounds 1 to 9 (compounds 1 to 9 correspond to 1 to 39, 1 to 40, 1 to 43, 1 to 44, 1A to 38, 1A to 39, 1A to 40, 1A to 46 and 1A to 49 in the figure respectively), the IL-1 beta level can be found to be obviously reduced after being treated for a period of time at the concentrations of 3 mu M and 30 mu M, and the compound has better effect than the maslinic acid as a positive medicine.

EXAMPLE 12 Regulation of IL-18 production by LPS-activated BV2 cells by organoselenium compounds

The method comprises the following steps: balancing the sample and the kit to room temperature, and designing plate hole arrangement and a standard product in advance; a secondary hole; a blank well. And (3) diluting the standard substance, adding 40 mu l of sample diluent into the sample hole to be detected, and then adding 10 mu l of sample. 50 μ l of deionized water was added to the blank wells. Add 50. mu.l of standard to the standard well. Mix well, cover with sealing plate membrane, incubate 30min at 37 ℃. Washing liquid is prepared, 20ml of 20x concentrated washing liquid is added with deionized water to reach the constant volume of 40 ml. After incubation, 300. mu.l of wash solution was added to each well. After washing, the liquid is discarded, and the solution is dried. 50 μ l of enzyme-labeled reagent was added to each well, and no blank wells were added. Incubate at 37 ℃ for 30min and wash the plate again as above. 50. mu.l of the developing solution A was added to each well, and 50. mu.l of the developing solution B was added thereto. After gently shaking and developing for 10min at 37 ℃ in the dark, 50. mu.l of stop solution was added and the color immediately changed from blue to yellow. And placing the ELISA plate on a reading of an ELISA reader, and determining the concentration.

The content of IL-18 released by the activated BV2 cells was determined by ELISA. IL-18 is also a proinflammatory factor in the inflammatory response, and the results are shown in FIG. 3. As can be seen from the results of FIG. 3, the IL-18 level was significantly increased in the LPS group as compared with the Control group (Control group). After adding the organic selenium compounds 1 to 9 (compounds 1 to 9 correspond to 1 to 39, 1 to 40, 1 to 43, 1 to 44, 1A to 38, 1A to 39, 1A to 40, 1A to 46 and 1A to 49 in the figure respectively), the IL-18 level can be found to be remarkably reduced after being treated for a period of time at the concentrations of 3 mu M and 30 mu M, and the compound has better effect than the positive medicine group sulindac.

Example 13 Effect of organic selenium Compounds on the Release of Nitrite from LPS-activated BV2 cells

The method comprises the following steps: and standing the Griess I and Griess II reagents for 30-40min at normal temperature. Cell supernatants and standards were added at 50. mu.l/well in 96-well plates, respectively. (the standard concentration is 0, 1,2, 5, 10, 20, 40, 60, 100. mu.M), 50. mu.l of Griess I is added into each well, mixed evenly, and then 50. mu.l of Griess II is added into each well, and mixed evenly. The reaction was carried out for 5-10min in the absence of light, and the OD value of each well was measured at 540 nm. And (5) making a standard curve and calculating the corresponding concentration of the Nitrite.

The content of nitrite in the cell culture medium was measured by the Griess method, and the results are shown in FIG. 4. As can be seen from the results of FIG. 4, after adding the organoselenium compounds 1 to 9 of the present invention (compounds 1 to 9 correspond to 1-39, 1-40, 1-43, 1-44, 1A-38, 1A-39, 1A-40, 1A-46 and 1A-49 in the figure, respectively), it was found that the amount of Nitrite released was reduced at different concentration gradients (3. mu.M, 10. mu.M, 30. mu.M and 100. mu.M). Among them, the compound 1, the compound 2, the compound 5 and the compound 7 have better action effects.

Example 14 Effect of organic selenium Compounds on the level of iNOS mRNA in LPS-activated BV2 cells

The method comprises the following steps: and extracting total RNA, adding 1mL of Trizol into the collected cell sediment, and mixing uniformly. Adding 0.2ml chloroform into the centrifuge tube, shaking vigorously for 15s, standing for 3min, centrifuging at 4 deg.C and 12,000rpm for 15min, and transferring the upper aqueous phase into a new RNase-free centrifuge tube. Adding isopropanol with the same volume, and mixing uniformly. After standing for 10min, the mixture was centrifuged at 12,000rpm at 4 ℃ for 15 min. The supernatant was removed, 1mL of 75% ethanol was added to wash the precipitate, the precipitate was centrifuged at 12,000rpm at 4 ℃ for 3min, and the supernatant was discarded. And placing the ventilating part of the centrifugal pipe for 5-10 min. After air drying, adding 30-100 mu L DEPC water, fully dissolving RNA, determining the absorption values at 260nm and 280nm by an ultramicro nucleic acid protein detector, and determining the RNA quality with OD260/280 between 1.8-2.0. According to the specification of a cDNA synthesis kit, genome DNA reaction is removed, the obtained RNA is subjected to reverse transcription, primer design is made in advance, and RT-qPCR, pre-denaturation, annealing, extension and melting curve are carried out for 40 cycles.

Inducible Nitric Oxide Synthase (iNOS) is a catalytic enzyme for the production of body active nitrogen. When the body is injured, the expression level of iNOS is up-regulated to release a large amount of NO. Therefore, the effect of different concentrations of 9 organic selenium compounds on the transcription level of iNOS in BV2 cells activated by LPS was experimentally detected by using a quantitative PCR method, and the result is shown in FIG. 5. As can be seen from the results in FIG. 5, after the cells were treated with the organic selenium compounds 1 to 9 (compounds 1 to 9 correspond to 1 to 39, 1 to 40, 1 to 43, 1 to 44, 1A to 38, 1A to 39, 1A to 40, 1A to 46 and 1A to 49 in the figure, respectively), the LPS-induced expression level of iNOS in BV2 cells was significantly reduced.

Example 15 improvement of model of cognitive behavioral disorders in mice by organic selenium Compounds

The method comprises the following steps: a single administration of LPS2 μ g into the ventricles causes inflammatory damage that can cause cognitive dysfunction in mice. The ventricular injection method is as follows: after the mouse is anesthetized by 0.3 percent pentobarbital intraperitoneal injection, the ventricles of the brain are administrated by a mouse locator, the bregma is taken as an origin, 0.5mm is backwards, the injection is uniformly and laterally opened to the right by 1.0mm, the depth is 2.0mm, the injection amount and the injection speed are respectively 2 mu l and 1 mu l/min, and the needle is stopped for 3min after the injection. LPS (Sigma, lot No. GJ43739) was dissolved in artificial cerebrospinal fluid (ACSF).

70C 57 mice were divided into 7 groups of 10 mice each, namely, a blank control group, an LPS 2. mu.g + compound 1(20mg/kg body weight) group, an LPS 2. mu.g + compound 6(20mg/kg body weight) group, an LPS 2. mu.g + compound 4(20mg/kg body weight) group, an LPS 2. mu.g + compound 5(20mg/kg body weight) group and an LPS 2. mu.g + compound 7(20mg/kg body weight) group. After 5 days of Morris water maze space acquisition training, on day 6, 2 μ g of LPS was administered once to the ventricles of the brain except for the blank group; while in addition to the model group, compound 1, compound 6, compound 4, compound 5 and compound 7 were administered in combination for each group, respectively. Compound 1, compound 6, compound 4, compound 5 and compound 7 were dissolved in sterile PBS solution and administered to mice by intraperitoneal injection at a dose of 20mg/kg body weight for 3 consecutive days. After 3 days of administration, each group of mice was subjected to water maze space exploration experiment and working memory detection.

Effect on Morris water maze working memory: as shown in Table 1, the mice fed with LPS 2. mu.g had a significantly longer stage incubation period (P < 0.05) at day 1 compared to the control blank, and the mice fed with each of compound 1, compound 6, compound 4, compound 5 and compound 7 had a significantly shorter stage incubation period, with statistical differences. Compared with the first day, the incubation period of the mice in the LPS2 mug group is obviously reduced on the 3 rd day; the other groups have the same trend of reduction, and the difference has statistical significance, which suggests that the compound 1, the compound 6, the compound 4, the compound 5 and the compound 7 can obviously improve the reduction of the working memory capacity of the mice after 1d of administration caused by the 2 mu gLPS ventricular administration, and the effect can be continued to 3d after the administration.

TABLE 1 staging incubation period in Morris Water maze working memory experiment for each group of mice

^a:P＜0.05,vs LPS

FIGS. 6 a-6 b show the effect of reference memory on the Morris water maze space exploration experiment. As can be seen from the results in fig. 6, the mouse crossing times, the peripheral path of the platform, and the percentage of the target quadrant staying path were all significantly decreased (P < 0.05) in the 2 μ g LPS group compared to the control group, and each of the administration groups of compound 1, compound 6, compound 4, compound 5, and compound 7 was able to increase the mouse crossing times, the peripheral path of the platform, and the percentage of the target quadrant staying path to different extents, suggesting that compound 1, compound 6, compound 4, compound 5, and compound 7 were able to improve the decrease in the reference memory capacity of the mouse 24h after the administration of 2 μ g LPS ventricles.

Claims (8)

1. A compound of formula (I)

R_a-R_b-C(W)-O-L-SeCN

(I)

Or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment or prevention of diseases and conditions associated with neuroinflammation,

wherein the content of the first and second substances,

R_arepresents aryl or heteroaryl, optionally substituted by one, two or three substituents selected from: phenyl, methylsulfonylphenyl C_1-4Alkyl, halobenzoyl, halophenylamino, benzoyl, C_1-4Alkylphenylamino, halogen, C_1-4Alkyl radical, C_1-4Alkoxy, halo C_1-4Alkoxy radical, C_1-4Alkyl acyl radical, C_1-2Alkylamino, halogeno C_1-4An alkyl group;

w is selected from O, S and Se;

R_bis a straight or branched alkylene or alkenylene chain of 1 to 4 carbon atoms, or is a direct covalent bond;

l is a straight or branched alkylene chain of 1 to 4 carbon atoms.

2. The use of claim 1, wherein R_aSelected from the group consisting of unsubstituted or substituted phenyl, indenyl, naphthyl, phenylaminophenyl, benzoylphenyl, benzopyrryl, benzopyrrolidinyl and phenylpyrrolidooxacyclohexyl,

wherein the substituent is selected from phenyl and methylsulfonylphenyl C_1-4Alkyl, halogenated phenylFormyl, phenylformyl, C_1-4Alkylphenylamino, halogen, C_1-4Alkyl radical, C_1-4Alkoxy, halo C_1-4Alkyl, halo C_1-4Alkoxy, acyl, C_1-4Alkyl acyl, acyloxy, C_1-2Alkyl acyloxy, C_1-2An alkylamino group; and is

Wherein the heteroaryl group is selected from the group consisting of a benzopyrroliyl group, a benzopyrrolidinyl group, and a phenylpyrrolidinooxeclocyclohexyl group.

3. Use according to claim 1 or 2, wherein W is O.

4. Use according to any one of claims 1 to 3, wherein R_aSelected from: phenyl, phenylaminophenyl, benzoylphenyl, indenyl, benzopyrryl, benzopyrrolidinyl and benzopyrrolidino-oxacyclohexyl, and R_aOptionally substituted with one or two substituents selected from: halogen selected from fluorine, chlorine, bromine, iodine, phenyl, phenylamino, benzoyl, methoxy, straight or branched alkyl of 1 to 4 carbon atoms, C_1-4An alkoxy group.

5. The use according to any one of claims 1 to 4, wherein L is (CH)₂)₂Or (CH)₂)₃。

6. The use of claim 1, wherein the compound of formula (I) is a compound selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.

7. The use according to any one of claims 1 to 6, wherein the neuroinflammation-related diseases and symptoms are selected from the group consisting of Alzheimer's disease, Parkinson's disease, depression, anxiety, obsessive compulsive disorder and schizophrenia.

8. The use according to any one of claims 1 to 6, wherein the neuroinflammation-related disease or condition is Alzheimer's disease.

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