CN111956639B

CN111956639B - Therapeutic use of organic selenium compound for neuroinflammation-related diseases

Info

Publication number: CN111956639B
Application number: CN202010915420.8A
Authority: CN
Inventors: 毕秀丽; 贺贤然; 李少磊
Original assignee: Shenzhen Fushan Biotechnology Co ltd; Liaoning University
Current assignee: Shenzhen Fushan Biotechnology Co ltd; Liaoning University
Priority date: 2020-09-03
Filing date: 2020-09-03
Publication date: 2023-06-09
Anticipated expiration: 2040-09-03
Also published as: CN111956639A

Abstract

The invention provides the use of a compound of formula (I) in the manufacture of a medicament for the treatment or prophylaxis of neuroinflammatory disorders and conditions, R _a ‑R _b -C (W) -O-L-SeCN (I) wherein the variables are as defined in the specification。

Description

Therapeutic use of organic selenium compound for neuroinflammation-related diseases

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 neuroinflammatory related diseases and symptoms.

Background

Neuroinflammation can lead to nerve injury, produce a range of pathophysiological changes, and lead to the occurrence of related neurological diseases and symptoms. The related diseases are mainly a group of nervous system diseases which are mainly marked by disturbance of behaviors and psychological activities.

Senile dementia is a typical neuroinflammation-related disease, which is a neurological disease of various cognitive dysfunctions mainly due to memory impairment, and can be divided into three types: primary dementia, vascular dementia, and other causes of dementia. Primary dementia includes alzheimer's disease (Alzheimers disease, AD), parkinson's Disease (PD) and pick's disease. The main pathological features of alzheimer's disease are amyloid β deposition, fiber tangles, basal nuclear cholinergic neuron loss, extensive loss of hippocampal and cortical neurons, and synaptic changes. Recent studies indicate 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 number of factors can induce activation of microglia. Activated microglia can release a number of neurotoxic factors including Nitric Oxide (NO), interleukin IL-1 beta, reactive Oxygen Species (ROS). Studies have shown that inhibition of neuroinflammation can be an important strategy for the prevention and treatment of AD (CN 110638854 a).

Currently, neuroinflammation is considered to be an important cause of occurrence of various neurological diseases. Ma Chunyan et al indicate that the neuroinflammatory mechanism is the main cause of depression, and that the inflammatory hypothesis suggests that stress stimulates the inflammatory process and ultimately leads to the onset of depression (chinese medical guide 2017,14 (14): 33-35). Smith et al, 1991, suggested that inflammation could be involved in the pathophysiological process of depression (Barbara S Beltz, michael F Tlucty, jeanne L Benton, et al omega-3fatty acids upregulate adult neurogenesis[J. Neurosci Lett,2007, 415 (2): 154-158.) and this theory was supported by a number of research results.

Parkinson's Disease (PD) is a central nervous system degenerative disease characterized mainly by progressive degeneration, 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, myotonia and posture disorders, and non-motor symptoms such as depressed mood, anxiety, sleep disorders, cognitive disorders. Numerous studies have shown that microglial and astrocyte-mediated inflammatory responses play an important role in the pathogenesis of PD. Microglial cells are important immunological receptors and effector cells in the central nervous system, the substantia nigra region is one of the most abundant brain regions of microglial cell distribution in the brain, and the substantia nigra dopaminergic neurons are extremely sensitive to microglial-mediated inflammatory responses. During the development and progression of neurodegenerative diseases, overactivated microglia can produce a range of inflammatory factors such as IL-1 beta, IFN-gamma, NO, IL-6, TNF-alpha, etc. These inflammatory factors can lead to injury and loss of neurons on the one hand, and the injured neurons release NO, ROS and the like, act on microglia, promote the release of inflammatory factors, promote the further activation of microglia, and thus form a vicious circle between neuronal injury and microglia activation; on the other hand, these inflammatory factors can also activate astrocytes, causing them to release inflammatory factors that further damage neurons. Thus, how to effectively inhibit the inflammatory response of nerve cells, alleviating the overactivation of glial cells is a potential target for the treatment of neurodegenerative diseases (CN 110623974 a).

With the recent intensive research into 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, huntington's chorea, etc., more and more research results indicate that microglial activation-mediated chronic inflammatory responses in the brain are one of the pathological features of nervous system disorders (CN 111116595 a).

Although research has been conducted to elucidate a pathway for the treatment of related neuroinflammatory disorders or conditions, the existing compounds, including anti-inflammatory agents, in particular non-steroidal anti-inflammatory agents, are generally subject to insufficient efficacy, limited safety, etc., and are far from meeting the ever-increasing demands of humans for the prevention and treatment of related disorders. Therefore, the development of better-acting, more novel structural types of pharmaceutical compounds has become an urgent need.

Disclosure of Invention

Through a great deal of experimental research, the inventor unexpectedly discovers that the organic compound containing the selenium cyanide has unexpected biological activity for preventing and treating related diseases or symptoms of 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 _a Represents aryl or heteroaryl optionally substituted with one, two or three substituents selected from: phenyl, methanesulfonylphenyl C _1-4 Alkyl, halobenzoyl, halophenylamino, benzoyl, C _1-4 Alkylphenylamino, halogen, C _1-4 Alkyl, C _1-4 Alkoxy, halo C _1-4 Alkoxy, C _1-4 Alkyl acyl, C _1-2 Alkylamino, halo C _1-4 An alkyl group;

w is selected from O, S and Se;

R _b a linear or branched alkylene or alkenylene chain of 1 to 4 carbon atoms, or a direct covalent bond;

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

In the compounds of formula (I) of the present invention, preferably R _a Selected from unsubstituted or substituted phenyl, indenyl, naphthyl, phenylaminophenyl, benzoylphenyl, benzopyrrolyl, benzeneAnd pyrrolidyl and benzopyrrolidocyclohexyl groups.

In the compounds of formula (I) according to the invention, R is _a By definition, the substituents are selected from phenyl, methylsulfonylphenyl C _1-4 Alkyl, halophenyl formyl, phenyl formyl, C _1-4 Alkylphenylamino, halogen, C _1-4 Alkyl, C _1-4 Alkoxy, halo C _1-4 Alkyl, halogenated C _1-4 Alkoxy, acyl, C _1-4 Alkanoyl, acyloxy, C _1-2 Alkylacyloxy radicals, C _1-2 An alkylamino group.

Preferably, R _a Heteroaryl in the definition is selected from the group consisting of benzopyrrolyl, benzopyrrolidinyl, benzocyclopentenyl, benzothienyl, benzofuranyl, benzimidazolyl, azabenzimidazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, imidazopyridinyl, isoxazolopyridinyl; particularly preferably, the heteroaryl group is selected from the group consisting of benzopyrrolyl, benzopyrrolidinyl and benzopyrrolidoxine.

In some preferred embodiments of the invention, R _a Selected from phenyl, phenylaminophenyl, benzoylphenyl, indenyl, benzopyrrolyl, benzopyrrolidinyl and benzopyrrolidoxil, and R _a Optionally substituted with one or two substituents selected from the group consisting of: halogen selected from fluorine, chlorine, bromine, iodine, phenyl, phenylamino-, benzoyl, methoxy, straight-chain or branched alkyl of 1 to 4 carbon atoms, C _1-4 An 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 formula (I) according to the invention, L is preferably (CH) ₂ ) ₂ Or (CH) ₂ ) ₃ I.e. L is a C2 or C3 chain, and the chain is not limited to a straight chain, and may comprise branched moieties.

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, namely a second drug, which is suitable for the treatment of the same condition or related condition that is medically indicated to be treatable or preventable by administration of a compound of the invention.

In yet another aspect of the invention, a process for preparing a compound of the invention is provided, the process comprising reacting DCC (N, N' -dicyclohexylcarbodiimide) with compound R _a -R _b -a step of reacting C (W) -OH in an organic solvent, wherein the variables are as defined above. Preferably, said compound R _a -R _b The molar ratio of (C (W) -OH to DCC is from 1:1 to 1:3. Preferably, the reaction is carried out under catalysis of 4-Dimethylaminopyridine (DMAP); preferably, DMAP and R _a -R _b The molar ratio of (C (W) -OH is from 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 the compounds, compositions, or combinations of the present invention, comprising administering the compounds, compositions, or combinations to a patient in need of treatment for an adverse condition thereof at a dosage, frequency, and duration sufficient to provide a beneficial effect to the patient.

Drawings

FIG. 1 shows the modulation of LPS-activated BV2 cells by compounds 1-9 of the present invention.

FIG. 2 shows the modulation of LPS-activated BV2 cells by compounds 1-9 of the present invention to produce IL-1β.

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

FIG. 4 is a graph showing the effect of organic selenium compounds provided by the invention on release of Nitri by LPS-activated BV2 cells.

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

Fig. 6a is a graph showing the influence of the organic selenium compound provided by the invention on the reference memory (number of times of crossing) in Morris space exploration experiments of mice in each group.

Fig. 6b is a graph showing the influence of the organic selenium compound provided by the invention on the reference memory (platform peripheral distance) in the Morris space exploration experiment of each group of mice.

Fig. 6c is a graph showing the influence of the organic selenium compound provided by the invention on the reference memory (target quadrant stop time) in the Morris space exploration experiment of each group of mice.

Detailed Description

The term "treatment" is defined as the treatment and care of a patient for combating a disease, condition or disorder, which includes the administration of a compound of the 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 symptoms associated with a disorder or disease, or inhibiting further development or worsening of such 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 symptoms associated with the disorder or condition, or stops or slows further development or worsening of such symptoms, or prevents or provides prophylaxis of the disorder or condition.

When a group is defined as a "covalent bond," it is intended that groups directly adjoining the specified group are linked to each other by a covalent bond.

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

In general, "substituted" means that an organic group as defined herein, which includes 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 compounds of the invention (including R _a Substituents in the definition include halogen, alkyl (preferably C) _1-4 Alkyl, more preferably C _1-2 Alkyl), alkoxy (preferably C _1-4 Alkoxy, more preferably C _1-2 Alkoxy), halo C _1-4 Alkoxy (preferably halo C) _1-2 Alkoxy), amino, acyl, C _1-4 Alkanoyl (preferably C _1-2 Alkyl acyl), acyloxy, C _1-4 Alkylacyloxy groups (preferably C _1-2 Alkanoyloxy), carboxyl, nitro, cyano, C _1-4 Alkylamino (preferably C) _1-2 Alkylamino, halo C _1-4 Alkyl (preferably halogenated C) _1-2 Alkyl group, C _1-4 Alkylamino C _1-4 Alkyl (preferably C _1-2 Alkylamino C _1-2 Alkyl), hydroxy C _1-4 Alkyl (preferably hydroxy C _1-2 Alkyl group, hydroxy group C _1-4 Alkylamino (preferably hydroxy C) _1-2 Alkylamino group, C _3-6 Cycloalkyl, C _2-4 Alkenyl groups.

In the definition of substituents in the present invention, a short "-" represents the bond position where a group or substituent is attached to the rest of the formula. As will be appreciated by those skilled in the art, for the structure of formula (I), R _a Having one bond-forming position, R _b There are two key-forming locations 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, even more preferably from 1 to 2 carbon atoms. Examples of the straight-chain alkyl group 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, isopentyl and 2, 2-dimethylpropyl. Representative substituted alkyl groups may be substituted one or more times with any of the above groups, such as amino, hydroxy, 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 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, isobornenyl, and carenyl, as well as fused rings such as, but not limited to, decahydronaphthyl, and the like. Cycloalkyl also includes rings substituted with straight or branched chain alkyl groups as defined above. Representative substituted cycloalkyl groups may be mono-substituted or more than one time substituted, 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 groups, which may be substituted with, for example, amino, hydroxy, cyano, carboxyl, nitro, thio, alkoxy, and halo groups. The term "cycloalkenyl" alone or in combination means cyclic alkenyl.

Aryl is a cyclic aromatic hydrocarbon that does not contain heteroatoms. Thus, aryl groups include, but are not limited to, phenyl, azulenyl, heptenyl (hepatanyl), biphenyl, dicyclopentadiene-phenyl (indacenyl), fluorenyl, phenanthryl, triphenylenyl, pyrenyl, naphtalenyl (napthalenyl), biphenylenyl, anthracenyl and naphthyl. In certain embodiments, aryl groups contain 6 to 14 carbons in the ring portion of the group. Aryl groups may be unsubstituted or substituted, as defined above. Representative substituted aryl groups may be monosubstituted or more than once substituted, 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 heteroatomsSuch as, but not limited to N, O and S. Designated C ₂ Heteroaryl groups of heteroaryl groups may be 5-membered rings having two carbon atoms and three heteroatoms, 6-membered rings having two carbon atoms and four heteroatoms, etc. Similarly, C ₄ Heteroaryl groups may be 5 membered rings with one heteroatom, 6 membered rings with 2 heteroatoms, etc. 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, imidazopyridyl, isoxazolopyridinyl, thianaphtyl, purinyl, xanthinyl, adenine, guanine, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, quinoxalinyl, and quinazolinyl. Heteroaryl groups may be unsubstituted or substituted with groups as described above. Representative substituted heteroaryl groups may be substituted one or more times with those 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, isoindanyl, 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-pyridyl, 5-pyrimidinyl, pyridazinyl (2, 5-pyridyl), pyrimidinyl (4-pyridyl), pyrimidinyl (4-pyridyl, 5-pyrimidinyl), pyrimidinyl (1, 2, 3-imidazolyl), pyrimidinyl (4-pyridyl) and other examples of aryl and heteroaryl groups including, such as aryl and heteroaryl groups, quinolinyl (2-quinolinyl, 3-quinolinyl, 4-quinolinyl, 5-quinolinyl, 6-quinolinyl, 7-quinolinyl, 8-quinolinyl), isoquinolinyl (1-isoquinolinyl, 3-isoquinolinyl, 4-isoquinolinyl, 5-isoquinolinyl, 6-isoquinolinyl, 7-isoquinolinyl, 8-isoquinolinyl), benzo [ b ] furanyl (2-benzo [ b ] furanyl, 3-benzo [ b ] furanyl, 4-benzo [ b ] furanyl, 5-benzo [ b ] furanyl, 6-benzo [ b ] furanyl, 7-benzo [ b ] furanyl), 2, 3-dihydro-benzo [ b ] furanyl (2- (2, 3-dihydro-benzo [ b ] furanyl), 3- (2, 3-dihydro-benzo [ b ] furanyl), 4- (2, 3-dihydro-benzo [ b ] furanyl), 5- (2, 3-dihydro-benzo [ b ] furanyl), 6-benzo [ b ] furanyl, 6-dihydro-benzo [ b ] furanyl, 7-benzo [ b ] furanyl), 2, 3-dihydro-benzo [ b ] furanyl, 3- (2, 3-dihydro-benzo [ b ] furanyl), 3- (2, 3-benzo [ b ] furanyl), 4- (2, 3-dihydro-benzo [ b ] furanyl), (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, 6-yl, 7-benzimidazolyl, 8-benzimidazolyl), benzoxazolyl (1-indolyl, 2-indolyl, 6-indolyl, 7-benzoxazolyl, 2-benzoxazolyl, 4-thiazolyl, etc. (1-benzimidazolyl, 2-indolyl, 4-thiazolyl, 4-benzoxazolyl, etc.).

More particularly, aryl and heteroaryl groups may include phenyl, isoindolylalkyl (isoindolylinyl), imidazolyl, oxazolyl, benzimidazolyl and benzoxazolyl; wherein any aryl or heteroaryl group may be unsubstituted, monosubstituted or independently polysubstituted, e.g. with J groups as defined herein.

The term "alkoxy" refers to an oxygen atom attached to an alkyl group as defined above, including cycloalkyl groups. Examples of straight chain 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, isopentyloxy, isohexyloxy, 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 groups are "haloalkyl" groups within the meaning herein. Examples of haloalkyl 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, aralkyl cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl, or the like. In the particular case where the carbonyl carbon atom is bonded to hydrogen, the group is a "formyl" group, which is an acyl group (the term is defined herein). Other examples include acetyl, benzoyl, phenylacetyl, pyridylacetyl, cinnamoyl, and acryl, and the like. When a group comprising a carbon atom bonded to a carbonyl carbon atom comprises a halogen, the group is referred to as a "haloacyl group". One example is trifluoroacetyl.

"salts" as are well known in the art include organic compounds such as carboxylic acids, sulfonic acids, or combinations of amines and counterions in ionic form. For example, an acid in anionic form may form a salt with: cations such as metal cations, e.g., sodium, potassium, etc.; ammonium salts such as NH ₄ ⁺ 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 an ion that has been approved for human use and is generally non-toxic, such as the hydrochloride salt or Sodium salt. "zwitterionic" is an internal salt, which can be formed, for example, in a molecule having at least two ionizable groups, one forming an anion and the other forming a cation, which are in equilibrium with each other. For example, an amino acid (e.g., glycine) may exist in a zwitterionic form. "zwitterionic" is a salt within the meaning herein.

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

"solvates" are similar compositions, except that a solvent other than water is used in place of water. For example, methanol or ethanol may form an "alkoxide," which may also be stoichiometric or non-stoichiometric.

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

In another aspect the invention provides a pharmaceutical composition comprising a compound of the invention and optionally a carrier or a pharmaceutical composition comprising in addition to a compound of the invention a further pharmaceutically active ingredient and optionally a carrier. The pharmaceutical compositions of the invention may be prepared by conventional techniques, for example as described in Remington: the Science and Practice of Pharmacy, 19 th edition, 1995, which is incorporated herein by reference. The composition may be presented in conventional form, for example in the form of a capsule, tablet, aerosol, solution, suspension or topical application.

Typical compositions comprise a compound of the invention and a carrier. For example, the active compound is typically admixed with a carrier, either diluted by the carrier, or enclosed within a carrier which may be in the form of an ampoule, capsule, sachet (sachets), paper or other container. When the active compound is admixed with a carrier, or when the carrier acts as a diluent, the carrier may be a solid, semi-solid, or liquid material that acts 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, saline solution, alcohols, polyethylene glycols, polyhydroxyethoxylated castor oil, peanut oil, olive oil, gelatin, lactose, terra alba, sucrose, dextrin, magnesium carbonate, sugar, cyclodextrin, amylose, magnesium stearate, talc, gelatin, agar, pectin, acacia, lower alkyl ethers of stearic acid or cellulose, silicic acid, fatty acids, fatty acid amines, fatty acid monoglycerides and diglycerides, pentaerythritol fatty acid esters, polyoxyethylene, hydroxymethyl cellulose and polyvinylpyrrolidone. Similarly, the carrier or diluent may comprise 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 react adversely with the active compounds. These additives may include wetting agents, emulsifying and suspending agents, salts which influence osmotic pressure, buffers and/or coloring substances, preservatives, sweeteners or flavoring agents. The composition may also be sterilized, if desired.

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

If a solid carrier is used for oral administration, the formulation may be tabletted, placed in hard gelatin capsules 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 include aqueous or oily suspensions which may be prepared using suitable dispersing or wetting agents and suspending agents. The injectable form may be in the solution phase or in the form of a suspension prepared with a solvent or diluent. Acceptable solvents or carriers include sterile water, ringer's solution, or isotonic saline solution. Alternatively, sterile oil may be employed as a solvent or suspending agent. Preferably, the oil or fatty acid is non-volatile, including natural or synthetic oils, fatty acids, monoglycerides, diglycerides or triglycerides.

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 injectable formulations, the formulations may optionally contain 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. The unit dosage form for injection may be in an ampoule or in a multi-dose container.

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 or slow release.

The compounds of the present invention are effective over a wide dosage range. For example, in the treatment of adults, a dosage 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. In selecting a patient treatment regimen, it may often be necessary to start with a higher dose and reduce the dose as the condition is controlled. The precise dosage will depend on 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, and the preferences and experience of the physician or veterinarian in charge.

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

Typically, dosage forms suitable for oral, nasal, pulmonary or transdermal administration include from about 125 μg to about 1250mg, preferably from about 250 μg 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, for example twice daily or three times daily. Alternatively, the dosage form may be administered less frequently than once daily, for example every other day or every week, if deemed appropriate by the prescribing physician.

The pharmaceutical composition may be in the form of a tablet, capsule, powder, granule, lozenge, liquid or jelly. Tablets and capsules for oral administration may be in a form suitable for unit dosage administration and may contain conventional excipients, examples of which are: binding agents such as syrup, gum arabic, gelatin, sorbitol, xanthan gum, polyvinylpyrrolidone (PVP); fillers such as lactose, sugar, corn flour, calcium phosphate, sorbitol or glycine; tablet lubricants such as magnesium stearate, silica, talc, polyethylene glycol or silica; disintegrants such as potato starch; acceptable lubricants such as sodium lauryl sulfate. The tablets may be coated according to methods known in conventional pharmaceutical practice. The oral liquid preparation can be formulated as aqueous or oily suspension, solution, emulsion, syrup or tincture, or can be formulated as a dry substance for reconstitution with water or other suitable carrier prior to use. These liquid preparations may contain conventional additives such as suspending agents (e.g., sorbitol, syrup, methylcellulose, glucose syrup, gelatin, hydrogenated edible oils and fats). Emulsifying agents (e.g. incubating phospholipids, sorbitol monooleate or gum arabic), non-aqueous carriers (including edible oils such as almond oil, rectified coconut oil, fats and oils such as glycerol, propylene glycol or ethanol), preserving agents (e.g. methyl or propyl parahydroxybenzoic acid or sorbic acid), and if desired, conventional flavoring agents or colorants.

The percentage of active substance in the pharmaceutical composition of the present invention is variable because the drug must be formulated in a proper ratio of dosage to achieve the desired therapeutic effect. In general, the pharmaceutical preparations according to the invention should be administered orally or by injection in amounts of 1 to 15 mg of active substance per 70kg of body weight per day. The following illustrative, but not limiting, examples are given.

The invention in various embodimentsMethods of synthesizing the compounds of the invention herein are provided. The general method includes reacting R _a -R _b The compound represented by-C (W) -OH is mixed with 2-selenocyanoethanol or 3-selenocyanopropanol (NCSe (CH) ₂ ) _n OH) (n is 2 or 3) in an organic solvent. The reaction may be carried out at low temperature over a catalyst (DCC and DMAP). In some preferred embodiments, a compound of formula R _a -R _b Dissolving a-C (W) -OH compound in tetrahydrofuran or dichloromethane, adding 4-dimethylaminopyridine, adding N, N' -Dicyclohexylcarbodiimide (DCC) under stirring, and stirring for reaction under ice bath condition to obtain a target compound R _a -R _b -C(W)-O-L-SeCN。

Examples

The technical scheme and application of the invention are further described below with reference to specific examples:

synthetic preparation example

Example 1: preparation of Compound 1

The reaction formula is as follows:

in a 50ml flask, compound II-a (100 mg,1.0 eq) and 10ml of anhydrous tetrahydrofuran were added, and 2-selenocyanoethanol (54 mg,1.2 eq), DCC (62 mg,1.0 eq) and DMAP (44 mg,1.2 eq) were added in this order under ice bath and stirred for 12 hours, and the reaction was complete by TLC. The solvent was removed by distillation under the reduced pressure, and the mixture was separated by column chromatography on silica gel using ethyl acetate (V): petroleum ether (V) =1:3 as mobile phase gave pale yellow solid, i.e. compound 1 120mg, 86% yield.

Nuclear magnetic resonance ¹ H NMR(400MHz,CDCl ₃ ):δ2.24(s,3H,-CH ₃ ),2.88(s,3H,CH ₃ ),3.16(t,2H,J＝8.00Hz,CH ₂ ),3.64(s,2H,CH ₂ ),3.85(t,2H,J＝8.00Hz,CH ₂ ),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). ¹³ C NMR(100MHz,CDCl ₃ ):δ11.3,26.2,34.9,43.3,51.2,107.1(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. ¹⁹ F NMR(CDCl ₃ ,376MHz):δ＝-112.1(s,F).

ESI-MS:471.4([M+1] ⁺ ).

Example 2: preparation of Compound 2

The reaction formula is as follows:

in a 50ml flask, compound II-b (100 mg,1.0 eq) and 10ml of anhydrous tetrahydrofuran were added, and 2-selenocyanoethanol (72 mg,1.2 eq), DCC (82 mg,1.0 eq) and DMAP (59 mg,1.2 eq) were added in this order under ice bath and stirred for 12 hours, and the reaction was complete by TLC. The solvent was removed by distillation under the reduced pressure, and the mixture was separated by column chromatography on silica gel using ethyl acetate (V): petroleum ether (V) =1:3 as mobile phase gave a white solid, compound 2 100mg, 66% yield.

Nuclear magnetic resonance ¹ H NMR(400MHz,CDCl ₃ )：δ1.50(d,3H,J＝7.20Hz,CH ₃ ),3.13(t,2H,J＝4.00Hz,CH ₂ ),3.70(q,1H,J＝7.20Hz,CH),3.81(t,2H,J＝4.00Hz,CH ₂ ),7.04-7.11(m,2H,ArH),7.31-7.42(m,4H,ArH),7.46-7.50(m,2H,ArH). ¹³ C NMR(100MHz,CDCl ₃ ):δ18.3,24.2,48.4,53.8,114.7(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. ¹⁹ F NMR(CDCl ₃ ,376MHz):δ＝-112.2(s,F).

ESI-MS:377.3([M+1] ⁺ ).

EXAMPLE 3 preparation of Compound 3

The reaction formula is as follows:

in a 50ml flask, compound II-c (100 mg,1.0 eq) and 10ml of anhydrous tetrahydrofuran were added, and 2-selenocyanoethanol (54 mg,1.2 eq), DCC (62 mg,1.0 eq) and DMAP (44 mg,1.2 eq) were added in this order under ice bath and stirred for 12 hours, and the reaction was complete by TLC. The solvent was removed by distillation under the reduced pressure, and the mixture was separated by column chromatography on silica gel using ethyl acetate (V): petroleum ether (V) =1:3 as mobile phase gave a white solid, compound 3, 120mg, 84% yield.

Nuclear magnetic resonance ¹ H NMR(400MHz,CDCl ₃ ):δ2.12(s,3H,CH ₃ ),3.03(t,2H,J＝8.00Hz,CH ₂ ),3.60(t,2H,J＝8.00Hz,CH ₂ ),3.64(s,3H,CH ₃ ),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). ¹³ C NMR(100MHz,CDCl ₃ ):δ14.2,26.3,33.5,56.7,101.1,103.7,112.9,114.6,116.4,129.4,130.4,131.3,131.8,133.4,134.2,139.4,157.3,166.7.

ESI-MS:476.8([M+1] ⁺ ).

EXAMPLE 4 preparation of Compound 4

The reaction formula is as follows:

in a 50ml flask, compound II-d (100 mg,1.0 eq) and 10ml of anhydrous tetrahydrofuran were added, and 2-selenocyanoethanol (72 mg,1.2 eq), DCC (82 mg,1.0 eq) and DMAP (59 mg,1.2 eq) were added in this order under ice bath and stirred for 12 hours, and the reaction was complete by TLC. The solvent was removed by distillation under the reduced pressure, and the mixture was separated by column chromatography on silica gel using ethyl acetate (V): petroleum ether (V) =1:3 as mobile phase gave pale yellow solid, compound 4, 100mg, 65% yield.

Nuclear magnetic resonance ¹ H NMR(400MHz,CDCl ₃ ):δ1.48(d,3H,J＝8.00Hz,CH ₃ ),2.12-2.16(m,2H,CH ₂ ),3.03(m,2H,CH ₂ ),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). ¹³ C NMR(100MHz,CDCl ₃ ):δ17.5,31.2,38.5,47.6,102.6,128.0,128.6,129.2,129.7,130.4,131.0,131.5,137.6,138.8,142.2,172.6,193.9.

ESI-MS:387.3([M+1] ⁺ ).

EXAMPLE 5 preparation of Compound 5

The reaction formula is as follows:

in a 50ml flask, compound II-e (100 mg,1.0 eq) and 10ml of anhydrous tetrahydrofuran were added, and 2-selenocyanoethanol (76 mg,1.2 eq), DCC (86 mg,1.0 eq) and DMAP (62 mg,1.2 eq) were added in this order under ice bath and stirred for 12 hours, and the reaction was complete by TLC. The solvent was removed by distillation under the reduced pressure, and the mixture was separated by column chromatography on silica gel using ethyl acetate (V): petroleum ether (V) =1:3 as mobile phase gave pale yellow solid, compound 5, 115mg, 73% yield.

Nuclear magnetic resonance ¹ H NMR(400MHz,DMSO):δ2.08(s,3H,CH ₃ ),2.32(s,3H,CH ₃ ),3.15(t,2H,J＝8.00Hz,CH ₂ ),3.67(t,2H,J＝8.00Hz,CH ₂ ),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). ¹³ C NMR(100MHz,DMSO):δ13.8,21.2,23.8,48.4,114.8,117.7,119.2,124.4,126.2,126.8,131.1,131.4,134.8,137.3,138.6,147.2,172.5,174.6.

ESI-MS:374.1([M+1] ⁺ ).

EXAMPLE 6 preparation of Compound 6

The reaction is as follows

In a 50ml flask, compound II-f (100 mg,1.0 eq) and 10ml of anhydrous tetrahydrofuran were added, and 2-selenocyanoethanol (61 mg,1.2 eq), DCC (70 mg,1.0 eq) and DMAP (50 mg,1.2 eq) were added in this order under ice bath and stirred for 12 hours, and the reaction was completed by TLC. The solvent was removed by distillation under the reduced pressure, and the mixture was separated by column chromatography on silica gel using ethyl acetate (V): petroleum ether (V) =1:3 as mobile phase gave pale yellow solid, compound 6, 100mg, 69% yield.

Nuclear magnetic resonance ¹ H NMR(400MHz,CDCl ₃ ):δ3.06-3.09(m,2H,CH ₂ ),3.68-3.72(m,2H,CH ₂ ),3.76(s,2H,CH ₂ ),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). ¹³ C NMR(100MHz,CDCl ₃ ):δ24.9,41.5,51.7,115.6,120.7,124.3,125.8,127.5,129.3,130.4,137.1,143.0,175.8.

ESI-MS:428.0([M+1] ⁺ ).

EXAMPLE 7 preparation of Compound 7

The reaction is as follows

In a 50ml flask, compound II-g (100 mg,1.0 eq) and 10ml of anhydrous tetrahydrofuran were added, and 2-selenocyanoethanol (52 mg,1.2 eq), DCC (72 mg,1.0 eq) and DMAP (51 mg,1.2 eq) were added in this order under ice bath and stirred for 12 hours, and the reaction was completed by TLC. The solvent was removed by distillation under the reduced pressure, and the mixture was separated by column chromatography on silica gel using ethyl acetate (V): petroleum ether (V) =1:3 as mobile phase gave pale yellow solid, compound 7, 120mg, 81% yield.

Nuclear magnetic resonance ¹ H NMR(400MHz,CDCl ₃ ):δ0.88(t,3H,J＝8.00Hz,CH ₃ ),1.30(t,3H,J＝8.00Hz,CH ₃ ),1.92-2.14(m,2H,CH ₂ ),2.75-2.83(m,4H,2×CH ₂ ),3.12(q,2H,J＝8.00Hz,CH ₂ ),3.16(q,2H,J＝8.00Hz,CH ₂ ),3.65-3.71(m,2H,CH ₂ ),3.92-4.00(m,2H,CH ₂ ),6.94(d,1H,J＝8.00Hz,ArH),7.02(t,1H,J＝8.00Hz,ArH),7.36(d,1H,J＝8.00Hz,ArH). ¹³ C NMR(100MHz,CDCl ₃ ):δ8.6,14.1,23.3,25.7,26.4,32.3,46.7,57.8,62.1,77.5,110.4,117.3,120.5,121.2,127.3,127.4,135.2,135.9,162.1,170.3.

ESI-MS:422.0([M+1] ⁺ ).

EXAMPLE 8 preparation of Compound 8

The reaction is as follows

In a 50ml flask, compound II-h (100 mg,1.0 eq) and 10ml of anhydrous tetrahydrofuran were added, and 3-selenocyanuropanol (76 mg,1.2 eq), DCC (86 mg,1.0 eq) and DMAP (62 mg,1.2 eq) were sequentially added under ice bath and stirred for 12 hours to react completely by TLC. The solvent was removed by distillation under the reduced pressure, and the mixture was separated by column chromatography on silica gel using ethyl acetate (V): petroleum ether (V) =1:3 as mobile phase gave pale yellow solid, compound 8, 120mg, 75% yield.

Nuclear magnetic resonance ¹ H NMR(400MHz,CDCl ₃ ):δ2.11(s,3H,CH ₃ ),2.10-2.22(m,2H,CH ₂ ),2.28(s,3H,CH ₃ ),3.15(t,2H,J＝8.00Hz,CH ₂ ),3.48(q,2H,J＝8.00Hz,CH ₂ ),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), ¹³ C NMR(100MHz,CDCl ₃ ):δ13.9,21.4,26.4,32.5,37.8,105.7,114.2,115.4,116.3,122.3,124.8,126.3,127.2,131.2,132.3,137.9,138.3,146.5,171.4.

ESI-MS:388.0([M+1] ⁺ ).

EXAMPLE 9 preparation of Compound 9

The reaction is as follows

In a 50ml flask, compound II-i (100 mg,1.0 eq) and 10ml of anhydrous tetrahydrofuran were added, and 3-selenocyanuropanol (57 mg,1.2 eq), DCC (72 mg,1.0 eq) and DMAP (51 mg,1.2 eq) were added in this order under ice bath and stirred for 12 hours, and the reaction was completed by TLC. The solvent was removed by distillation under the reduced pressure, and the mixture was separated by column chromatography on silica gel using ethyl acetate (V): petroleum ether (V) =1:3 as mobile phase gave pale yellow solid, compound 9, 105mg, 77% yield.

Nuclear magnetic resonance ¹ H NMR(400MHz,CDCl ₃ :δ0.98(t,3H,J＝8.00Hz,CH ₃ ),1.35(t,3H,J＝8.00Hz,CH ₃ ),1.80-1.92(m,2H,CH ₂ ),2.08–2.13(m,2H,CH ₂ ),2.38-2.41(m,1H,CH),2.77-2.82(m,5H,CH ₂ ,CH ₂ ,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,CH ₂ ),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). ¹³ C NMR(100MHz,CDCl3):δ8.3,15.4,23.2,24.3,27.3,31.2,31.9,38.4,44.2,61.4,75.4,101.8,108.2,116.9,120.2,120.8,126.6,127.4,132.8,134.2,170.1.

ESI-MS:436.1([M+1] ⁺ ).

Biological Activity assay

EXAMPLE 10 Regulation of LPS-activated BV2 cells to produce ROS by organoselenium compounds

The method comprises the following steps: the DCFH-DA (proportion 1:1000) is diluted by serum-free culture medium, the final concentration is 10 mu M, the positive control group (proportion 1:1000) is added with the ROSUP 20min before the end of the drug treatment, the original culture medium in the 6-hole plate is removed, 1ml of DCFH-DA diluent is added, the cell culture box is incubated for 20min, the cells are washed 3 times by serum-free cell culture liquid, the cells are digested by trypsin, centrifuged for 5min at 1200rpm at 4 ℃, the PBS1X blows off the cells, the cells are re-washed to prepare cell suspension, and 100 mu l of each hole of the 96 black hole plate is added. The fluorescence microplate reader detects values at an excitation wavelength of 502nm and an emission wavelength of 530 nm.

Intracellular ROS content, mainly H2O2 levels, were detected 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 of the Lipopolysaccharide (LPS) group was significantly increased compared to the Control group (Control group), which is close to that of the positive Control sulindac group (ropp). After adding the organic selenium 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, respectively in the figures), treatment at concentrations of 3. Mu.M and 30. Mu.M for 48 hours, it was found that ROS levels were significantly down-regulated and had better effects than the positive drug group sulindac.

EXAMPLE 11 organic selenium Compounds modulation of LPS activated BV2 cells to IL-1 beta

The method comprises the following steps: balancing the sample and the kit to room temperature, and designing plate hole arrangement and standard substances in advance; a secondary hole; blank holes. The standard was diluted and 40. Mu.l of sample dilution was added to the wells to be tested, followed by 10. Mu.l of sample. 50 μl deionized water was added to the blank wells. 50 μl of standard was added to the wells. Mixing, covering with sealing plate membrane, and incubating at 37deg.C for 30min. A washing solution was prepared, and 20ml of 20x concentrated washing solution was added to deionized water to a volume of 40ml. After incubation 250-300. Mu.l of wash solution was added to each well. And (5) discarding liquid after washing, and spin-drying. 50 μl of enzyme-labeled reagent was added per well, and blank wells were not added. Incubate at 37℃for 30min and wash the plate again with the above procedure. Mu.l of the color-developing solution A and 50. Mu.l of the color-developing solution B were added to each well. After gentle shaking and development at 37℃for 10min in the dark, 50. Mu.l of stop solution were added, and the color was changed from blue to yellow immediately. The ELISA plate was placed on the ELISA reader for concentration measurement.

The content of IL-1β released by activated BV2 cells was examined by ELISA. IL-1β 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 in fig. 2, the IL-1β level was significantly increased in the LPS group compared to the Control group (Control group). After adding the organic selenium 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, respectively in the figures), treatment at concentrations of 3. Mu.M and 30. Mu.M for a period of time, it was found that IL-1β levels were significantly down-regulated and had better effects than the positive drug group sulindac.

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

The amount of IL-18 released by activated BV2 cells was measured by ELISA. IL-18 is also a pro-inflammatory factor in the inflammatory response, and the results are shown in FIG. 3. As can be seen from the results in FIG. 3, the IL-18 level was significantly increased in the LPS group compared to the Control group (Control group). After adding the organic selenium 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, respectively in the figures), treatment at concentrations of 3. Mu.M and 30. Mu.M for a period of time, it was found that the IL-18 levels were significantly down-regulated and had better effects than the positive drug group sulindac.

Example 13 Effect of organoselenium Compounds on LPS-activated BV2 cell Release of Nitri

The method comprises the following steps: griess I and Griess II reagent are kept stand at normal temperature for 30-40min. Cell supernatants and standards were added separately in 96-well plates at 50 μl/well. (standard concentrations of 0,1,2,5, 10, 20, 40, 60, 100. Mu.M) 50. Mu.l Griess I was added to each well, mixed well, and then 50. Mu.l Griess II was added to each well, mixed well. The reaction was kept away from light for 5-10min and OD per well was measured at 540 nm. Standard curves were made and the corresponding nitite concentrations were calculated.

Nitrite content in the cell culture medium was measured by Griess method, and the results are shown in FIG. 4. As can be seen from the results of FIG. 4, after the addition of the organic selenium 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, respectively, in the figures), it was found that the amount of Nitri released was reduced at different concentration gradients (3. Mu.M, 10. Mu.M, 30. Mu.M, 100. Mu.M). Among them, compound 1, compound 2, compound 5, and compound 7 have better action and effect.

EXAMPLE 14 Effect of organoselenium on LPS-activated BV2 cell iNOS mRNA levels

The method comprises the following steps: total RNA was extracted and the collected cell pellet was added to 1mL Trizol and mixed well. 0.2ml chloroform was added to the centrifuge tube, vigorously shaken for 15s, allowed to stand for 3min, and centrifuged at 12,000rpm at 4℃for 15min, and the upper aqueous phase was transferred to a new RNase-free centrifuge tube. Adding equal volume of isopropanol, and mixing. After 10min of standing, centrifugation was carried out at 12,000rpm at 4℃for 15min. The supernatant was removed, 1mL of 75% ethanol was added to wash the precipitate, and the precipitate was centrifuged at 12,000rpm at 4℃for 3min, and the supernatant was discarded. Placing the centrifuge tube at a ventilation position for 5-10 min. After airing, adding 30-100 mu L DEPC water to fully dissolve RNA, and determining the absorption values at 260nm and 280nm by an ultra-trace nucleic acid protein detector to determine the quality of the RNA, wherein OD260/280 is between 1.8 and 2.0. According to the specification of the cDNA synthesis kit, removing genome DNA reaction, carrying out reverse transcription on the obtained RNA, designing a primer in advance, and carrying out RT-qPCR, pre-denaturation, annealing, extension, melting curve and 40 cycles.

Inducible Nitric Oxide Synthase (iNOS) is a catalytic enzyme for the production of active nitrogen in the body. When the body is damaged, the expression level of iNOS is up-regulated, releasing a large amount of NO. The effect of different concentrations of 9 organoselenium compounds on the iNOS transcription level of LPS-activated BV2 cells was thus examined experimentally using the quantitative PCR method, the results of which are shown in figure 5. As can be seen from the results of FIG. 5, after cells were treated with the organic selenium 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, respectively, in the figures), it was found that the expression level of iNOS in BV2 cells induced by LPS was significantly reduced.

Example 15 Effect of organic selenium Compounds on improvement of mice model of cognitive behavioral disorders

The method comprises the following steps: single administration of LPS 2 μg ventricle causes inflammatory injury, which can cause cognitive dysfunction in mice. The ventricular injection method is as follows: after the 0.3% pentobarbital of the mice is injected and anesthetized in the abdominal cavity, the ventriculus administration is carried out by a mouse positioning instrument, the bregma is taken as an origin, the injection is carried out by uniformly and rightwards opening by 1.0mm and the depth is 2.0mm by 0.5mm, the injection quantity and the injection speed are respectively 2 mu l and 1 mu l/min, and the injection is stopped for 3min after the injection. LPS (Sigma, lot GJ 43739) was dissolved in artificial cerebrospinal fluid (artificial cerebrospinalfluid, ACSF).

70C 57 mice were divided into a blank group, LPS 2. Mu.g group, LPS 2. Mu.g+Compound 1 (20 mg/kg body weight) group, LPS 2. Mu.g+Compound 6 (20 mg/kg body weight) group, LPS 2. Mu.g+Compound 4 (20 mg/kg body weight) group, LPS 2. Mu.g+Compound 5 (20 mg/kg body weight) group and LPS 2. Mu.g+Compound 7 (20 mg/kg body weight) group, 7 groups in total, 10 each. After 5 days of Morris water maze space availability training, on day 6, single administration of 2 μg of LPS to the ventricle of each brain except for the blank group; while each group was given compound 1, compound 6, compound 4, compound 5 and compound 7 in combination, respectively, except for the model group. 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 performs water maze space exploration experiment and working memory detection.

Influence on Morris water maze working memory: as shown in Table 1, compared with the blank control group, the stage incubation period of the mice on day 1 after LPS administration of 2. Mu.g was significantly prolonged (P < 0.05), and the stage incubation period of the mice in each of the administration groups of Compound 1, compound 6, compound 4, compound 5 and Compound 7 was significantly shortened, and the difference was statistically significant. Day 3 LPS2 μg group, mice had significantly reduced stage latency compared to day 1; the other groups also have a reduced trend, and the difference has a 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 working memory capacity reduction of 1d of mice after 2 mu g LPS ventricular administration, and the effect can be sustained to 3d of mice after administration.

Table 1 stage latency of mice of each group in Morris water maze working memory experiments

^a :P＜0.05,vs LPS

Fig. 6 a-6 b show the effect on reference memory in a Morris water maze space exploration experiment. As can be seen from the results of fig. 6, compared with the control group, the number of times of the mice pass through the platform, the peripheral distance of the platform and the percentage of the stay distance in the target quadrant are all obviously reduced (P < 0.05), and each administration group of the compound 1, the compound 6, the compound 4, the compound 5 and the compound 7 can increase the number of times of the mice pass through the platform, the peripheral distance of the platform and the percentage of the stay distance in the target quadrant to different extents, which suggests that the compound 1, the compound 6, the compound 4, the compound 5 and the compound 7 can improve the reduction of the reference memory capacity of the mice 24h after the administration of 2 μg LPS ventricle.

Claims

1. A compound of formula (I)

R _a -R _b -C(W)-O-L-SeCN

(I)

Or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment or prevention of neuroinflammation-related disorders and symptoms,

wherein the compound of formula (I) is selected from the following compounds:

2. the use of claim 1, wherein the neuroinflammation-related disorders and symptoms are selected from the group consisting of alzheimer's disease, parkinson's disease, depression, anxiety, obsessive-compulsive disorder, and schizophrenia.

3. The use of claim 1 or 2, wherein the neuroinflammation-related disorder and symptom is alzheimer's disease.