HK40107845A - Compound for preventing and/or treating neurodegenerative disease - Google Patents

Description

Compounds used for the prevention and/or treatment of neurodegenerative diseases

Technical Field

This invention relates to the field of medicine, and more particularly to compounds for the prevention and/or treatment of neurodegenerative diseases.

Background Technology

Alzheimer's disease is a common neurodegenerative disease characterized by memory decline, neuronal death, and other symptoms. Its main clinical markers are senile plaques and neurofibrillary tangles. The pathological changes in Alzheimer's disease are quite complex. Currently, countries around the world are researching effective treatments for Alzheimer's, but with limited success. There is currently no specific treatment or drug to reverse the progression of the disease. More than a decade ago, the US FDA approved five therapeutic compounds in two classes, including cholinesterase inhibitors and NMDA receptor antagonists, but these only temporarily improve symptoms and cannot halt the disease's progression. The main pathogenesis involves the deposition of β-amyloid protein in the brain tissue, abnormal cerebral vascular structure leading to activation of microglia and astrocytes in the brain, producing inflammatory mediators, increased free radical production in the brain tissue accompanied by weakened antioxidant capacity, and an imbalance between the acetylcholine and antiacetylcholine systems in the brain tissue. These factors result in damage to the structure and function of brain tissue, increased levels of inflammatory mediators in the hippocampus, and weakened antioxidant capacity in the hippocampus, ultimately inducing Alzheimer's disease. Pharmaceutical companies have developed various vaccines targeting the Abeta protein and phosphorylation inhibitors of several enzymes involved in its formation; however, these efforts have so far been ineffective. We believe that reducing the toxicity of this protein after the onset of disease is too late, and that therapeutic drugs should be developed by addressing the toxicity of the protein itself.

In recent years, the mechanism of action of RAC1 in the nervous system has gradually become a hot research area. Existing literature reports that dysregulation of physiological signals centered on RAC1 is associated with pathological changes in Alzheimer's disease and leads to age-dependent neurodegenerative diseases (Human Molecular Genetics, 2020, Vol. 29, No. 5). Furthermore, regulating RAC1 to increase LTP can enhance the brain's learning and memory abilities (PNAS, 2007, Vol. 104, No. 2). Therefore, RAC1 has become a highly promising target for the treatment and prevention of neurodegenerative diseases.

Summary of the Invention

The purpose of this invention is to provide a compound of Formula I, a method for its preparation, and its use in the prevention and/or treatment of neurodegenerative diseases.

In a first aspect, the present invention provides a compound, said compound being a compound of formula I or a stereoisomer thereof, a racemic mixture thereof, or a pharmaceutically acceptable salt thereof.

in,

Ring A is selected from the following group:

X ₁ is selected from the following group: O, S, NH, NR;

X ₂ is selected from the following group: O, NH, NR;

_R1 is selected from the following group:

R ₂ is selected from the following group: H, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C6 cycloalkyl;

_R3 and _R4 are independently selected from the group consisting of: H, D, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C6 cycloalkyl, or _R3 and _R4 together with the carbon to which they are attached to form a substituted or unsubstituted 3-7 membered cycloalkyl or a substituted or unsubstituted 3-7 membered heterocyclic group containing one or more heteroatoms selected from O, S or N;

Each of _R5 , _R6 , _R7 , _R8 , _R9 , and _R10 is independently selected from the group consisting of: H, D, halogen, trifluoromethyl, cyano, hydroxyl, amino, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C1-C6 alkyl- _NR16- , substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted C3-C6 cycloalkoxy, substituted or unsubstituted C3-C6 cycloalkyl- _NR16- ;

Each of _R11 , _R12 , _R13 , _R14 , and _R15 is independently selected from the group consisting of: H, D, halogen, trifluoromethyl, cyano, hydroxyl, amino, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C1-C6 alkyl- _NR16- , substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted C3-C6 cycloalkoxy, substituted or unsubstituted C3-C6 cycloalkyl- _NR16- ;

Each R ₁₇ and R ₁₈ is independently selected from the group consisting of: H, D, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy; or R ₁₇ and R ₁₈ together with the carbon atom to which they are attached form a substituted or unsubstituted 3-7 membered cycloalkyl group or a substituted or unsubstituted 3-7 membered heterocyclic group containing one or more heteroatoms selected from O, S or N;

Each R is independently a substituted or unsubstituted C1-C6 alkyl group;

Each of the substitutions independently refers to substitution by one or more substituents selected from the group consisting of: D, halogen, trifluoromethyl, cyano, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkyl-NR _16- , C3-C6 cycloalkyl, C3-C6 cycloalkoxy, C3-C6 cycloalkyl-NR _16- ;

Each R ₁₆ is independently selected from the group consisting of: H, C1-C6 alkyl groups;

The additional condition is: when _X1 is NH and _X2 is O, and _R3 , _R4 , _R17 , and _R18 are H, ring A is not...

In another preferred embodiment, the compound has the structure shown in Formula II:

Among them, rings A, _X1 , _X2 , _R1 , _R2 , _R3 , and _R4 are as defined above.

In another preferred embodiment, the compound has the structure shown in Formula II:

Among them, rings A, _X1 , _R1 , _R2 , _R3 , and _R4 are as defined above.

In another preferred embodiment, ring A is selected from the following group:

_R5 , _R6 , _R7 , _R8 , and _R9 are as defined above.

In another preferred embodiment, _X1 is selected from the group consisting of: O, S, NH, NR;

R is defined as above.

In another preferred embodiment, _X2 is selected from the group consisting of: O, NH, and NR;

R is defined as above.

In another preferred example, _R1 is selected from the following group:

_R11 , _R12 , _R13 , _R14 , _R15 , _R17 , and _R18 are defined as above.

In another preferred embodiment, _R2 is selected from the group consisting of: H, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C6 cycloalkyl.

Each of the substitutions independently refers to substitution by one or more substituents selected from the group consisting of: D, halogen, trifluoromethyl, cyano, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkyl-NR _16- , C3-C6 cycloalkyl, C3-C6 cycloalkoxy, C3-C6 cycloalkyl-NR _16- ;

Each R ₁₆ is independently selected from the group consisting of H, C1-C6 alkyl groups.

In another preferred embodiment, _R3 and _R4 are independently selected from the group consisting of H, D, halogens, substituted or unsubstituted C1-C6 alkyl groups, substituted or unsubstituted C1-C6 alkoxy groups, substituted or unsubstituted C3-C6 cycloalkyl groups, or _R3 and _R4 together with the carbon to which they are attached form substituted or unsubstituted 3-7 membered cycloalkyl groups or substituted or unsubstituted 3-7 membered heterocyclic groups containing one or more heteroatoms selected from O, S or N.

In another preferred embodiment, each of _R5 , _R6 , _R7 , _R8 , _R9 , and _R10 is independently selected from the group consisting of: H, D, halogen, trifluoromethyl, cyano, hydroxyl, amino, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C1-C6 alkyl- _NR16- , substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted C3-C6 cycloalkoxy, substituted or unsubstituted C3-C6 cycloalkyl- _NR16- ;

Each R ₁₆ is independently selected from the group consisting of H, C1-C6 alkyl groups.

In another preferred embodiment, each of _R5 , _R6 , _R7 , _R8 , _R9 , and _R10 is independently selected from the group consisting of: H, D, halogen, cyano, hydroxyl, amino, unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C1-C6 alkyl- _NR16- , substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted C3-C6 cycloalkoxy, substituted or unsubstituted C3-C6 cycloalkyl- _NR16- ;

Each R ₁₆ is independently selected from the group consisting of H, C1-C6 alkyl groups.

In another preferred embodiment, each of _R11 , _R12 , _R13 , _R14 , and _R15 is independently selected from the group consisting of: H, D, halogen, trifluoromethyl, cyano, hydroxyl, amino, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C1-C6 alkyl-NR _16- , substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted C3-C6 cycloalkoxy, substituted or unsubstituted C3-C6 cycloalkyl-NR _16- ;

Each R ₁₆ is independently selected from the group consisting of H, C1-C6 alkyl groups.

In another preferred embodiment, each _R17 and _R18 is independently selected from the group consisting of H, D, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, and C3-C6 cycloalkoxy; or _R17 and _R18 together with the carbon to which they are attached form a substituted or unsubstituted 3-7 membered cycloalkyl group or a substituted or unsubstituted 3-7 membered heterocyclic group containing one or more heteroatoms selected from O, S, or N.

In another preferred example, R ₁₇ is selected from the group consisting of H and D.

In another preferred embodiment, R ₁₈ is a C1-C6 alkyl group.

In another preferred embodiment, when ring A is H, _R3 , _R4 , _R17 and _R18 are not simultaneously H.

In another preferred embodiment, when ring A is ,

_R5 , _R6 , _R7 , _R8 , and _R9 are as defined above;

_R3 and _R4 are independently selected from the group consisting of H, C1-C6 alkyl groups;

R ₁₇ and R ₁₈ are independently selected from the group consisting of H, D, and C1-C6 alkyl groups;

Furthermore, _R3 , _R4 , _R17 , and _R18 are not all H at the same time.

In another preferred embodiment, when ring A is ,

_R5 , _R6 , _R7 , _R8 , and _R9 are as defined above;

_R3 and _R4 are H;

R ₁₇ and R ₁₈ are independently selected from the group consisting of H, D, and C1-C6 alkyl groups;

Furthermore, R ₁₇ and R ₁₈ are not both H.

In another preferred embodiment, when ring A is ,

_R5 , _R6 , _R7 , _R8 , and _R9 are as defined above;

_R3 is H;

_R4 is a C1-C6 alkyl group;

_R17 and _R18 are H.

In another preferred embodiment, ring A is selected from the following group:

_R5 , _R6 , _R7 , _R8 , _R9 , and _R10 are as defined above;

_R3 and _R4 are independently selected from the group consisting of: H, D, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C6 cycloalkyl, or _R3 and _R4 together with the carbon to which they are attached to form a substituted or unsubstituted 3-7 membered cycloalkyl or a substituted or unsubstituted 3-7 membered heterocyclic group containing one or more heteroatoms selected from O, S or N;

Each R ₁₇ and R ₁₈ is independently selected from the group consisting of: H, D, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy; or R ₁₇ and R ₁₈ together with the carbon atom to which they are attached form a substituted or unsubstituted 3-7 membered cycloalkyl group or a substituted or unsubstituted 3-7 membered heterocyclic group containing one or more heteroatoms selected from O, S or N;

Each of the substitutions independently refers to substitution by one or more substituents selected from the group consisting of: D, halogen, trifluoromethyl, cyano, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkyl-NR _16- , C3-C6 cycloalkyl, C3-C6 cycloalkoxy, C3-C6 cycloalkyl-NR _16- ;

Each R ₁₆ is independently selected from the group consisting of H, C1-C6 alkyl groups.

In another preferred embodiment, when ring A is ,

_R5 , _R6 , _R7 , and _R8 are as defined above;

_R3 and _R4 are independently selected from the group consisting of: H, D, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C6 cycloalkyl.

Each R ₁₇ and R ₁₈ is independently selected from the group consisting of: H, D, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, and C3-C6 cycloalkoxy.

Each of the substitutions independently refers to substitution by one or more substituents selected from the group consisting of: D, halogen, trifluoromethyl, cyano, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkyl-NR _16- , C3-C6 cycloalkyl, C3-C6 cycloalkoxy, C3-C6 cycloalkyl-NR _16- ;

Each R ₁₆ is independently selected from the group consisting of H, C1-C6 alkyl groups.

In another preferred embodiment, when ring A is ,

_R5 , _R6 , _R7 , and _R8 are as defined above;

_R3 and _R4 are independently selected from the following group: H, D, halogen, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl;

Each R ₁₇ and R ₁₈ is independently selected from the group consisting of: H, D, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, and C3-C6 cycloalkoxy.

In another preferred embodiment, ring A is

X ₁ is selected from the following groups: NH, NR;

_X2 is 0;

_R1 is

_R2 is H;

_R3 and _R4 are selected from the following group: H, C1-C6 alkyl;

_R5 , _R6 , _R7 , _R8 , and _R9 are each H;

Each of _R11 , _R12 , _R13 , _R14 , and _R15 is independently selected from the group consisting of H and halogens; each of _R17 and _R18 is independently selected from the group consisting of H, D, and C1-C6 alkyl groups.

Each R is an unsubstituted C1-C6 alkyl group.

In another preferred embodiment, ring A is X _{and 1} is NH;

_X2 is 0;

_R1 is _R2 is H;

_R3 and _R4 are selected from the following group: H, C1-C6 alkyl;

_R5 , _R6 , _R7 , _R8 , and _R9 are each H;

_R11 , _R12 , _R14 , and _R15 are each H;

R ₁₃ is a halogen, preferably F;

R ₁₇ is selected from the following groups: H, D;

R ₁₈ is selected from the following group: H, D, C1-C6 alkyl.

In another preferred embodiment, ring A is _X1 selected from the group consisting of: NH, NR;

_X2 is 0;

_R1 is _R2 is H;

_R3 and _R4 are H;

Each of _R5 , _R6 , _R7 , and _R8 is independently selected from the following group: H, halogens;

Each of _R11 , _R12 , _R13 , _R14 , and _R15 is independently selected from the following group: H, halogens;

Each _R17 and _R18 is independently selected from the group consisting of H and C1-C6 alkyl groups;

Each R is an unsubstituted C1-C6 alkyl group.

In another preferred embodiment, ring A is

X ₁ is NH;

_X2 is 0;

_R1 is

_R2 is H;

_R3 and _R4 are H;

_R5 is selected from the following group: H, halogens;

_R6 , _R7 , and _R8 are each H;

_R11 , _R12 , _R14 , and _R15 are each H;

R ₁₃ is a halogen, preferably F;

R ₁₇ is H;

R ₁₈ is selected from the following group: H, C1-C6 alkyl.

In another preferred embodiment, _R2 is selected from the group consisting of: H, halogen, C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkoxy, and C3-C6 cycloalkyl.

_R3 and _R4 are independently selected from the following group: H, D, halogen, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl.

In another preferred embodiment, ring A is

_R1 is

Each of _R5 , _R6 , _R7 , _R8 , and _R9 is independently selected from the group consisting of: H, halogen, trifluoromethyl, cyano, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, and C3-C6 cycloalkoxy.

Each of _R11 , _R12 , _R13 , _R14 , and _R15 is independently selected from the group consisting of: H, halogen, trifluoromethyl, cyano, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, and C3-C6 cycloalkoxy.

In another preferred embodiment, the compound is selected from the group consisting of:

In another preferred embodiment, the pharmaceutically acceptable salt is an inorganic acid salt or an organic acid salt;

The inorganic acid salts are selected from the following group: hydrochloride, hydrobromide, hydroiodide, sulfate, hydrogen sulfate, nitrate, phosphate, and acid phosphate;

The organic acid salts are selected from the group consisting of: formate, acetate, trifluoroacetate, propionate, pyruvate, glycolate, oxalate, malonate, fumarate, maleate, lactate, malate, citrate, tartrate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, salicylate, picrate, glutamate, ascorbate, camphorate, and camphorsulfonate.

A second aspect of the invention provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and one or more safe and effective amounts of the compounds described in the first aspect of the invention.

A third aspect of the invention provides a use of the compound described in the first aspect of the invention for preparing a medicament for the prevention and/or treatment of neurodegenerative diseases.

In another preferred embodiment, the neurodegenerative disease is selected from the group consisting of: Alzheimer's disease, epilepsy, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, and spinocerebellar ataxia.

A fourth aspect of the invention provides the use of the compound described in the first aspect of the invention for preparing a medicament for the prevention and/or treatment of RAC1-related diseases.

In another preferred embodiment, the RAC1-related disease is a neurodegenerative disease.

It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here.

Detailed Implementation

Through long-term and in-depth research, the inventors unexpectedly prepared a compound with excellent pharmacokinetic properties that can effectively prevent and/or treat neurodegenerative diseases. Based on this, the inventors completed this invention.

the term

In this invention, unless otherwise specified, the terms used have the general meanings known to those skilled in the art.

In this invention, the term "halogen" refers to F, Cl, Br, or I.

In this invention, "C1-C6 alkyl" refers to a straight-chain or branched alkyl group comprising 1-6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, neopentyl, pterpentyl, or similar groups.

In this invention, the term "C2-C6 alkenyl" refers to a straight-chain or branched alkenyl group having 2-6 carbon atoms and containing a double bond, and includes, without limitation, vinyl, propenyl, butenyl, isobutenyl, pentenyl, and hexenyl groups.

In this invention, the term "C2-C6 ynyl" refers to a straight-chain or branched ynyl group having 2-6 carbon atoms and containing a triple bond, and includes, without limitation, ethynyl, propynyl, butynyl, isobutynyl, pentylyl, and hexynyl.

In this invention, the term "C3-C8 cycloalkyl" refers to a cyclic alkyl group having 3-8 carbon atoms on a ring, and non-limitingly includes cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, etc. The term "C3-C6 cycloalkyl" has a similar meaning.

In this invention, the term "C1-C6 alkoxy" refers to a straight-chain or branched alkoxy group having 1-6 carbon atoms, and includes, without limitation, methoxy, ethoxy, propoxy, isopropoxy, and butoxy. Preferably, it is a C1-C4 alkoxy group.

In this invention, the term "heterocyclic group" refers to a 4-8 membered heterocyclic group containing 1, 2, or 3 heteroatoms selected from N, O, and S, including (but not limited to) the following groups:

In this invention, the terms "aromatic ring" or "aryl" have the same meaning, and are preferably "C6-C10 aryl". The term "C6-C10 aryl" refers to an aromatic cyclic group with 6-10 carbon atoms that does not contain heteroatoms on the ring, such as phenyl, naphthyl, etc.

In this invention, the terms "aromatic heterocycle" or "heteroaryl" have the same meaning, referring to a heteroaromatic group containing one or more heteroatoms. For example, "C3-C10 heteroaryl" refers to an aromatic heterocycle containing 1 to 4 heteroatoms selected from oxygen, sulfur, and nitrogen, and 3 to 10 carbon atoms. Non-limiting examples include: furanyl, thiophene, pyridinyl, pyrazolyl, pyrroleyl, N-alkylpyrroleyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, etc. The heteroaryl ring may be fused to an aryl, heterocyclic, or cycloalkyl ring, wherein the ring connected to the parent structure is the heteroaryl ring. The heteroaryl group may be optionally substituted or unsubstituted.

In this invention, the term "halogenated" refers to being replaced by a halogen.

In this invention, the term "substitution" refers to the substitution of one or more hydrogen atoms on a specific group by a specific substituent. The specific substituent is the substituent described accordingly above, or the substituent appearing in the various embodiments. Unless otherwise specified, a substituted group may have a substituent selected from a specific group at any substituted site of that group, and the substituents may be the same or different at each position. Those skilled in the art will understand that the combinations of substituents contemplated in this invention are stable or chemically feasible combinations. Such substituents include, but are not limited to: halogens, hydroxyl groups, carboxyl groups (-COOH), C1-C6 alkyl groups, C2-C6 alkenyl groups, C2-C6 alkynyl groups, C3-C8 cycloalkyl groups, 3- to 12-membered heterocyclic groups, aryl groups, heteroaryl groups, C1-C8 aldehyde groups, C2-C10 acyl groups, C2-C10 ester groups, amino groups, C1-C6 alkoxy groups, C1-C10 sulfonyl groups, etc.

In this invention, the terms 1-6 refer to 1, 2, 3, 4, 5, or 6. Other similar terms each have a similar meaning independently. The term "multiple" refers to 2-6, such as 2, 3, 4, 5, or 6.

It should be understood that when a group exists simultaneously at multiple different positions in a compound, its definition at each position is independent and can be the same or different. That is, the term "selected from the following group:" and the term "each independently selected from the following group:" have the same meaning.

compound

This invention provides a compound, which is a compound of Formula I or a stereoisomer, racemate, or pharmaceutically acceptable salt thereof.

The groups are as defined above.

In another preferred embodiment, in the compound, any one of ring A, _X1 , _X2 , _R1 , _R2 , _R3 , and _R4 is independently the corresponding group in the specific compound of the present invention.

As used herein, the term "pharmaceutically acceptable salt" refers to a salt formed by the compounds of the present invention with an acid or base that is suitable for use as a medicine. Pharmaceutically acceptable salts include both inorganic and organic salts. A preferred class of salts are those formed by the compounds of the present invention with an acid. Suitable acids for forming salts include, but are not limited to: inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, and phosphoric acid; organic acids such as formic acid, acetic acid, trifluoroacetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, benzoic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, and naphthalenesulfonic acid; and amino acids such as proline, phenylalanine, aspartic acid, and glutamic acid.

Another preferred class of salts are salts formed by the compounds of the present invention with a base, such as alkali metal salts (e.g., sodium or potassium salts), alkaline earth metal salts (e.g., magnesium or calcium salts), ammonium salts (such as lower alkanol ammonium salts and other pharmaceutically acceptable amine salts), such as methylamine salts, ethylamine salts, propylamine salts, dimethylamine salts, trimethylamine salts, diethylamine salts, triethylamine salts, tert-butylamine salts, ethylenediamine salts, hydroxyethylamine salts, dihydroxyethylamine salts, trihydroxyethylamine salts, and amine salts formed from morpholine, piperazine, and lysine, respectively.

Preparation method

The following schemes and examples describe methods for preparing compounds of formula I. Starting materials and intermediates are purchased from commercial sources, prepared by known procedures, or otherwise described. In some cases, the order of steps in performing the reaction scheme may be altered to promote the reaction or avoid unwanted byproducts.

The preparation method of the compound of Formula I of the present invention is described in more detail below, but these specific methods do not constitute any limitation on the present invention. The compounds of the present invention can also be conveniently prepared by optionally combining various synthetic methods described in this specification or known in the art, such combinations can be easily performed by those skilled in the art.

Typically, in the preparation process, each reaction is carried out under inert gas protection, in a suitable solvent, at 0 to 150°C, and the reaction time is usually 2 to 24 hours.

The preferred preparation method is as follows:

Method 1:

Step 1: In a solvent (dichloromethane), under condensing agent (DCC, EDCI, etc.) and alkaline conditions (4-dimethylaminopyridine, diisopropylethylamine), SM1 reacts with Michaelis acid to generate compound M1.

Step 2: In an ultra-dry solvent (ethyl acetate, 1,4-dioxane, etc.), compound M1 is refluxed to generate compound M2.

Step 3: In a solvent (acetonitrile, toluene, N,N-dimethylformamide, etc.) under alkaline conditions (such as diisopropylethylamine, potassium carbonate, DBU, etc.), compound M2 reacts with SM2 to generate compound M3.

Step 4: In a solvent (ethyl acetate, dichloromethane, 1,4-dioxane, etc.) under acidic conditions (hydrochloric acid, trifluoroacetic acid, etc.), compound M3 reacts to produce T (i.e., compound I).

Method 2:

Step 1: In a solvent (dichloromethane, carbon tetrachloride, etc.), compound M3 reacts with N-bromosuccinimide (NBS) to generate compound M4.

Step 2: In an inert solvent (such as N,N-dimethylformamide, dioxane, dimethyl sulfoxide, etc.), under alkaline conditions (such as potassium carbonate, potassium phosphate, etc.), and in the presence of a catalyst and ligand (such as Pd(PPh ₃ ) ₄ ), compound M4 reacts with SM3 or SM3' to generate M5.

Step 3: In a solvent (ethyl acetate, dichloromethane, 1,4-dioxane, etc.) and under acidic conditions (hydrochloric acid, trifluoroacetic acid, etc.), compound M5 reacts to produce T (i.e., compound I).

Method 3:

Step 1: In a solvent (acetonitrile, toluene, N,N-dimethylformamide, etc.) and under alkaline conditions (such as diisopropylethylamine, potassium carbonate, DBU, etc.), compound SM4 reacts with SM2 to generate compound M6.

Step 2: In the solvent tetrahydrofuran, under acidic (benzoic acid) conditions, compound M6 reacts with (triphenylphosphine) ketene to generate T (i.e., compound I).

Method 4:

Step 1: In a solvent (dichloromethane), under condensing agent (DCC, EDCI, etc.) and alkaline conditions (4-dimethylaminopyridine, diisopropylethylamine), SM1 reacts with Michaelis acid to generate compound M1.

Step 2: In an ultra-dry solvent (ethyl acetate, 1,4-dioxane, etc.), compound M1 is refluxed to generate compound M2.

Step 3: In a solvent (toluene) under acidic (p-toluenesulfonic acid) conditions, compound M2 reacts with SM5 to produce compound M7.

Step 4: In a solvent (ethyl acetate, dichloromethane, 1,4-dioxane, etc.) and under acidic conditions (hydrochloric acid, trifluoroacetic acid, etc.), compound M7 reacts to produce T (i.e., compound I).

In the above formulas, _R1 , _R2 , _R3 , and _R4 are as described above.

Unless otherwise specified, all of the above starting materials can be purchased commercially or synthesized according to the reported literature.

Pharmaceutical Compositions and Administration

The pharmaceutical compositions of the present invention comprise, within a safe and effective range, the compound of the present invention or a pharmacologically acceptable salt thereof, and a pharmacologically acceptable excipient or carrier. "Safe and effective range" refers to an amount of the compound sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, more preferably, 10-1000 mg of the compound of the present invention per dose. Preferably, "one dose" is one capsule or tablet.

"Pharmaceutically acceptable carriers" refers to one or more compatible solid or liquid fillers or gelling substances that are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with and with the compounds of the present invention without significantly reducing the efficacy of the compounds. Examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (e.g.), wetting agents (such as sodium dodecyl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

The pharmaceutical composition is an injection, capsule, tablet, pill, powder, or granule.

There are no particular limitations on the administration of the compounds or pharmaceutical compositions of the present invention. Representative administration methods include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and local administration.

Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following components: (a) fillers or compatibilizers, such as starch, lactose, sucrose, glucose, mannitol, and silica; (b) binders, such as hydroxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and gum arabic; (c) humectants, such as glycerin; (d) disintegrants, such as agar, calcium carbonate, potato starch or cassava starch, alginate, certain complex silicates, and sodium carbonate; (e) slowing agents, such as paraffin; (f) absorption accelerators, such as quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) adsorbents, such as kaolin; and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium dodecyl sulfate, or mixtures thereof. Buffers may also be included in capsules, tablets, and pills.

Solid dosage forms such as tablets, sugar pills, capsules, pellets, and granules can be prepared using coatings and shells, such as casings and other materials known in the art. They may contain opacifying agents, and the release of the active compound or compound from such compositions can be delayed in a portion of the digestive tract. Examples of encapsulating components that can be used are polymeric substances and waxes. If necessary, the active compound may also be formed into microcapsules with one or more of the excipients described above.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active compound, liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, e.g., ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures of these substances.

In addition to these inert diluents, the composition may also contain auxiliaries such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and fragrances.

In addition to the active compound, the suspension may contain suspending agents such as ethoxylated isooctadecyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitol esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions, or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents, or excipients include water, ethanol, polyols, and suitable mixtures thereof.

Dosage forms of the compounds of the present invention for topical administration include ointments, powders, patches, sprays, and inhalers. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be necessary.

The compounds of this invention can be administered alone or in combination with other pharmaceutically acceptable compounds (such as drugs for neurodegenerative diseases).

The treatment method of the present invention can be used alone or in combination with other treatment methods or drugs.

When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to the mammal (such as a human) requiring treatment. The dosage administered is the pharmaceutically considered effective dose. For a person weighing 60 kg, the daily dose is typically 1–2000 mg, preferably 50–1000 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the patient's health condition, which are all within the scope of the skills of a skilled physician.

Compared with the prior art, the present invention has the following main advantages:

(1) The compounds of the present invention have better pharmacokinetic properties;

(2) The compounds of the present invention have better efficacy.

The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions as described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or as recommended by the manufacturer. Unless otherwise stated, percentages and parts are by weight.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as are familiar to those skilled in the art. Furthermore, any methods and materials similar to or equivalent to those described herein may be applied to the methods of this invention. The preferred embodiments and materials described herein are for illustrative purposes only.

Example 1

The compounds synthesized in this invention:

The experimental procedure is as follows:

The synthesis route is as follows:

1. Synthesis of Compound 2

In a 100 mL three-necked flask, compound SM1 (5 g, 1.0 eq), hippuric acid (2.98 g, 1.1 eq), DMAP (3.4 g, 1.5 eq), and solvent DCM (100 mL) were mixed thoroughly. The mixture was purged three times with nitrogen and stirred for 10 min under nitrogen protection in an ice bath. The internal temperature was controlled at 0-10 °C. 20 mL of a DCM solution containing DCC (4.2 g, 1.1 eq) was added dropwise, and the reaction was allowed to proceed at room temperature for 16 h. TLC was used to detect the complete reaction of the starting material. The reaction solution was filtered, and the filtrate was washed six times with 50 mL of 5% potassium hydrogen sulfate solution each time. The organic phase was dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain a white solid. 50 mL of petroleum ether was added, and the mixture was stirred thoroughly before filtration to obtain the target product as a brown solid (3.2 g, yield 43.4%). No further purification was required for the next reaction. LC-MS [M-1]: 391.1.

2. Synthesis of Compound 3

Compound 2 (3.2 g, 1.0 eq) was purged three times with nitrogen under nitrogen protection, and then 45 ml of ultra-dry 1,4-dioxane was added. The mixture was reacted at 100 °C for 2 h. TLC was used to determine the completeness of the reaction. The reaction solution was concentrated under reduced pressure to obtain 2.6 g of compound 3, which could be proceeded to the next step without further purification. LC-MS [M-1]: 289.1.

3. Synthesis of Compound 4

In a 100 mL three-necked flask, compound 3 (1.0 g, 1.0 eq), 4-fluorobenzyl bromide (520 μL, 1.2 eq), potassium carbonate (714 mg, 1.5 eq), and acetonitrile (15 mL) were mixed thoroughly and refluxed for 2 h. The reaction proceeds were monitored by TLC to determine the completeness of the reaction. The solid was removed by filtration, and the filtrate was purified by column chromatography to yield 120 mg of compound 4. LC-MS [M+1]: 399.1.

4. Synthesis of compound T-01

Compound 4 (120 mg, 1.0 eq) was added to a 100 mL round-bottom flask and dissolved in 2.5 mL of DCM by stirring. Trifluoroacetic acid (120 μL, 5.0 eq) was added dropwise, and the mixture was reacted at room temperature for 4 h. The reaction mixture was monitored by TLC to determine the completeness of the reaction. The reaction solution was adjusted to pH 7-8 with saturated Na₂CO₃, extracted with dichloromethane, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The solution was then purified using a preparative chromatography plate to yield 60 mg of compound T-01. The HPLC purity was 99.7%. LC-MS[M+1]: 299.1. 1H NMR (400MHz, Chloroform-d) δ8.52 (dt, J＝4.9, 1.5Hz, 1H), 7.61 (td, J＝7.7, 1.9Hz, 1H), 7.42-7.35 (m, 2H), 7.21-7.07 (m, 4H ),6.23(s,1H),5.13(d,J＝1.7Hz,1H),5.06-4.91(m,2H),4.56(dd,J＝10.5 ,3.1Hz,1H),3.38(dd,J＝14.9,3.1Hz,1H),2.83(dd,J＝14.8,10.4Hz,1H).

Following the method used for compound T-01, the following compounds were synthesized:

Example 2

The compounds synthesized in this invention:

The experimental procedure is as follows:

The synthesis route is as follows:

1. Synthesis of Compound 2

In a 100 mL three-necked flask, compound SM1 (1 g, 1.0 eq), 4-fluorobenzyl bromide (1.14 g, 1.0 eq), DBU (1.2 g, 1.3 eq), and solvent acetonitrile (15 mL) were added and mixed thoroughly. After purging with nitrogen three times, the mixture was allowed to react at room temperature under nitrogen protection. The reaction was monitored by TLC to ensure the starting material was completely reacted. The solvent was removed by vacuum distillation, and the mixture was extracted with EA and water in a two-phase process. The EA layer was washed with saturated NaCl solution, dried with anhydrous sodium sulfate, and the EA was removed by rotary evaporation to obtain 1.2 g of product. ¹ H NMR (400MHz, DMSO) δ7.34 (dd, J＝8.6, 5.7Hz, 2H), 7.28-7.13 (m, 7H), 5.07 (s, 2H), 4.2 9(dt,J＝7.7,5.7Hz,1H), 2.96(dd,J＝13.7,5.3Hz,1H), 2.85(dd,J＝13.7,7.8Hz,1H).

2. Synthesis of compound T-29

Compound 2 (300 mg, 1.0 eq), (triphenylphosphine) ketene (378 mg, 1.1 eq), benzoic acid (12.5 mg, 0.1 eq), and THF (9 ml) were added to a 100 ml three-necked flask and mixed thoroughly. The mixture was purged three times with nitrogen, and then reacted at room temperature under nitrogen protection. TLC was used to determine the completeness of the reaction. The mixture was then subjected to two-phase extraction with water and EA. The EA layer was washed with saturated NaCl solution, dried over anhydrous sodium sulfate, and the EA was removed by rotary evaporation to obtain 700 mg of product. After purification, 87 mg of product was obtained with an HPLC purity of 97.7%.

¹ H NMR (400MHz, DMSO) δ7.69-7.55(m,2H),7.45-7.26(m,5H),7.21(dd,J＝12.6,6.2Hz,2H),5.44(s,1H),5.31(dd ,J＝5.7,4.4Hz,1H),5.17(dd,J＝33.8,11.7Hz,2H),3.24(dd,J＝14.5,4.1Hz,1H),2.93(dd,J＝14.5,6.4Hz,1H).

Example 3

The compounds synthesized in this invention:

The experimental procedure is as follows:

I. Synthesis of intermediate SM2

The synthesis route is as follows:

1. Synthesis of Compound 2

Compound 1 (1 g, 1.0 eq) was added to a 100 mL three-necked flask, purged three times with nitrogen, and then dissolved in ultra-dry THF (30 mL) under nitrogen protection. The mixture was stirred in an ice bath, maintaining an internal temperature of 0-10 °C. Lithium aluminum deuteride (476 mg, 2.0 eq) was added in portions, and the reaction was allowed to proceed overnight at room temperature. TLC analysis confirmed the reaction was complete. The reaction was quenched by adding saturated ammonium chloride solution in an ice bath. The mixture was filtered through diatomaceous earth, dried over anhydrous sodium sulfate, and evaporated to dryness to obtain 780 mg of a colorless liquid. ^¹H NMR (400 MHz, Chloroform-d) δ 7.38–7.30 (m, 2H), 7.09–7.00 (m, 2H).

2. Synthesis of compound SM2

Compound 2 (780 mg, 1.0 eq) and dichloromethane (25 mL) were added to a 100 mL three-necked flask. After purging with nitrogen three times, the mixture was stirred under nitrogen protection in an ice bath. Phosphorus tribromide (638 μL, 1.1 eq) was added dropwise, and the reaction was carried out at room temperature for 3 h. The reaction was monitored by TLC to determine the completeness of the starting material. The mixture was quenched with water, extracted with dichloromethane, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and purified by column chromatography to give 800 mg of compound SM2. ^¹H NMR (400 MHz, Chloroform-d) δ 7.43–7.32 (m, 2H), 7.09–6.97 (m, 2H).

II. Synthesis of Compound T-45

The synthesis route is as follows:

Referring to Example 1, compound T-45 was obtained. ^¹H NMR (400MHz, Chloroform-d) δ 7.43–7.35 (m, 2H), 7.34–7.26 (m, 3H), 7.22–7.16 (m, 2H), 7.15–7.07 (m, 2H), 5.45–5.32 (m, 1H), 5.07 (d, J = 1.6 Hz, 1H), 4.26 (dd, J = 9.6, 3.6 Hz, 1H), 3.23 (dd, J = 13.6, 3.6 Hz, 1H), 2.63 (dd, J = 13.6, 9.6 Hz, 1H).

Following the method used for compound T-45, the following compounds were synthesized:

Example 4

The compounds synthesized in this invention:

The experimental procedure is as follows:

I. Synthesis of intermediate SM1

The synthesis route is as follows:

1. Synthesis of Compound 3

50 ml of THF solvent was placed in a 250 ml three-necked flask, purged three times with nitrogen, and then kept under nitrogen protection. The flask was heated to -10 °C in an ice-salt bath. A mixture of _TiCl₄ (4.64 ml, 1.5 eq) and DCM (1 ml) was slowly added dropwise. After the addition was complete, the mixture was stirred in an ice-salt bath for 20 min. Compound 1 (3.3 ml, 1.0 eq) was added, and the mixture was stirred in an ice-salt bath for 10 min. Compound 2 (5 g, 1.1 eq) was added, _and the mixture was stirred in an ice-salt bath for 30 min. Pyridine (4.5 ml, 2.0 eq) was added, and the mixture was allowed to react overnight at room temperature. The reaction was confirmed to be complete by TLC. The reaction was quenched with saturated _NH₄Cl₄ solution in an ice bath. The mixture was extracted five times with EA, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain 5.5 g of compound 3. LC-MS [M+1]: 265.1.

2. Synthesis of Compound 4

Compound 3 (5.5 g, 1.0 eq) and MeOH (55 ml) were added to a 250 ml round-bottom flask. After purging with nitrogen three times and under nitrogen protection, sodium methoxide (227 mg, 0.2 eq) was added, and the mixture was reacted overnight at 70 °C. The reaction was confirmed to be complete by TLC. Heating was stopped, and the mixture was cooled to room temperature. The solvent was then evaporated, and the product was purified by column chromatography to obtain 4.6 g of compound 4. LC-MS [M+1]: 297.1.

3. Synthesis of Compound 5

Compound 4 (4.6 g, 1.0 eq), MeOH (46 ml), and Pd/C (460 mg, 0.1 eq) were added to a 100 ml reactor. After purging with hydrogen three times, the mixture was reacted under hydrogen protection at 70 °C for 5.5 h. Heating was stopped, and the mixture was cooled to room temperature. TLC was used to confirm the complete reaction of the starting materials. After adding diatomaceous earth and filtering, the filtrate was evaporated to dryness to give 5.19 g of compound 5. LC-MS [M+1]: 299.1.

4. Synthesis of Compound 6

Compound 5 (4.19 g, 1.0 eq), HCl (3 M) (104 mL, 25.0 eq), and glacial acetic acid (42 mL, 50.0 eq) were added to a 250 mL round-bottom flask. After purging with nitrogen three times, the mixture was reacted overnight at 125 °C under nitrogen protection. TLC analysis showed that the reaction proceeded to completion. Heating was stopped, and the mixture was cooled to room temperature. Water was added, and the mixture was filtered. The filtrate was evaporated to dryness to give 4.69 g of compound 6. LC-MS [M+1]: 181.1.

5. Synthesis of compound SM1

Compound 6 (4.69 g, 1.0 eq), THF (47 ml), and _H₂O (47 ml) were added to a 250 ml round-bottom flask. After purging with nitrogen three times, the mixture was kept under nitrogen protection and stirred in an ice bath for 10 min. NaOH (4.2 g, 4.0 eq) was added, and stirring was continued in an ice bath for 5 min. Boc anhydride (6.68 ml, 1.1 eq) was added, and the mixture was allowed to react overnight at room temperature. The reaction was confirmed by TLC to be complete. After evaporating the solvent THF, an appropriate amount of DCM was added, and the pH was adjusted to 4–5 with 1 N HCl. The mixture was stirred thoroughly for 10 min, and then separated. The DCM phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain 2 g of compound SM1. LC-MS [M+1]: 281.1.

II. Synthesis of compounds T-73 and T-74

The synthesis route is as follows:

Referring to Example 1, compound 5 was obtained, and after preparation and resolution, compounds T-73 and T-74 were obtained. LC-MS [M+1]: 313.1.

Following the methods used for compounds T-73 and T-74, the following compounds were synthesized:

Example 5

The compounds synthesized in this invention:

The experimental procedure is as follows:

The synthesis route is as follows:

1. Synthesis of Compound 2

In a 250 mL three-necked flask, compound SM1 (10 g, 1.0 eq), Michaelis acid (5.97 g, 1.1 eq), DMAP (6.9 g, 1.5 eq), and solvent DCM (100 mL) were mixed thoroughly. The mixture was purged three times with nitrogen and stirred for 10 min under nitrogen protection in an ice bath. The internal temperature was controlled at 0-10 °C. 50 mL of a DCM solution containing DCC (8.55 g, 1.1 eq) was added dropwise, and the reaction was allowed to proceed at room temperature for 16 h. TLC was used to detect the complete reaction of the starting materials. The reaction solution was filtered, and the filtrate was washed six times with 50 mL of 5% potassium hydrogen sulfate solution each time. The organic phase was dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain a white solid. 50 mL of petroleum ether was added, and the mixture was stirred thoroughly before filtration to obtain 14.3 g of the target product as a pale yellow solid. No further purification was required for the next reaction. LC-MS [M-1]: 390.1.

2. Synthesis of Compound 3

Compound 2 (14.3 g, 1.0 eq) was added to a 100 mL three-necked flask, purged three times with nitrogen, and then, under nitrogen protection, 172 mL of ultra-dry 1,4-dioxane was added. The mixture was reacted at 100 °C for 2 h. TLC was used to determine the completeness of the reaction. The reaction solution was concentrated under reduced pressure to obtain 11.2 g of compound 3, which could be proceeded to the next step without further purification. LC-MS [M-1]: 288.1.

3. Synthesis of Compound 4

Compound 3 (2.0 g, 1.0 eq), toluene (20 mL), p-fluorobenzylamine (790 μL, 1.0 eq), and p-toluenesulfonic acid (catalytic amount) were added to a 100 mL three-necked flask. After thorough mixing, the mixture was purged three times with nitrogen and then reacted at 100 °C for 5 h under nitrogen protection. The reaction was monitored by TLC to determine the completeness of the reaction. The mixture was extracted with water and ethyl acetate, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain 965 mg of compound 4. LC-MS[M+1]: 397.1. 1H NMR (400MHz, Chloroform-d) δ7.24 (td, J＝6.7, 6.3, 3. 3Hz,3H),7.19-7.15(m,2H),7.10(dd,J＝8.5,5.4Hz,2H),7.04-6.97(m,2H),4 .67(dd,J＝8.7,3.3Hz,1H),4.63(s,1H),4.45(s,1H),4.06(dd,J＝5.3,1.7Hz, 2H), 3.52 (dd, J＝13.7, 3.3Hz, 1H), 2.93 (dd, J＝13.7, 8.7Hz, 1H), 1.59 (s, 9H).

4. Synthesis of compound T-81

Compound 4 (100 mg, 1.0 eq) was added to a 100 mL three-necked flask and dissolved with 2.0 mL of DCM by stirring. Trifluoroacetic acid (193 μL, 10.0 eq) was added dropwise under ice bath conditions, and the reaction was carried out at room temperature for 3 h. The reaction proceeded as determined by TLC. The mixture was extracted with saturated sodium bicarbonate solution and ethyl acetate. The organic phase was washed with saturated sodium chloride and dried over anhydrous sodium sulfate. After purification, 30 mg of compound T-81 was obtained. The HPLC purity was 97.1%.

LC-MS[M+1]: 297.1. 1H NMR (400MHz, Chloroform-d) δ7.30 (dd, J＝7.8, 6.1Hz, 2H), 7.25 (s, 1H), 7.23-7.17 (m, 4H), 7.02 (t, J＝8.6Hz, 2H), 5.28 (s, 1H), 4.67 (d, J = 1.5Hz, 1H), 4.61 (t, J = 5.4Hz, 1H), 4.23 (dd, J = 8.6, 5.6Hz, 1H), 4.15 (d, J = 5.3Hz, 2H), 3.05-2.77 (m, 2H).

Following the method used for compound T-81, the following compounds were synthesized:

Example 6

The compounds synthesized in this invention:

The experimental procedure is as follows:

The synthesis route is as follows:

1. Synthesis of Compound 4

Referring to Example 5, compound 4 was obtained.

2. Synthesis of Compound 5

Compound 4 (178 mg, 1.0 eq) was added to a 100 mL three-necked flask, purged three times with nitrogen, and then dissolved in ultra-dry DMF under nitrogen protection. The mixture was stirred for 10 min in an ice bath, followed by the addition of 60% sodium hydride (36 mg, 2.0 eq). After stirring for another 15 min, iodomethane (31 μL, 1.1 eq) was added, and the mixture was reacted at room temperature for 3 h. The reaction was monitored by TLC to determine the completeness of the reaction. The mixture was extracted with water and ethyl acetate, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain 70 mg of compound 5. LC-MS [M+1]: 411.2.

3. Synthesis of compound T-82

Compound 5 (70 mg, 1.0 eq) was added to a 100 mL three-necked flask and dissolved in 2.0 mL of DCM by stirring. Trifluoroacetic acid (131 μL, 10.0 eq) was added dropwise under ice bath conditions, and the reaction was carried out at room temperature for 3 h. The reaction proceeded as determined by TLC. The mixture was extracted with saturated sodium bicarbonate solution and ethyl acetate. The organic phase was washed with saturated sodium chloride and dried over anhydrous sodium sulfate. After purification, 30 mg of compound T-82 was obtained. The HPLC purity was 95.4%. ¹ H NMR(400MHz,Chloroform-d)δ7.31(q,J＝8.6,8.0Hz,3H),7.20(dd,J＝8.6,4.5Hz,4H),7.07(t,J＝8.6Hz,2H),5.28(s,1H),4.79-4.72(m,1H),4.4 8(d,J＝15.7Hz,1H),4.42(dd,J＝9.8,3.0Hz,1H),4.33(d,J＝15.7Hz,1H),3.27(dd,J＝13.9,3.0Hz,1H),2.89(s,3H),2.62(dd,J＝13.8,9.8Hz,1H).

Following the method used for compound T-82, the following compounds were synthesized:

Experimental Example 1: Pharmacokinetic Study

1. Drug preparation

Accurately weigh approximately 10 mg of the sample to be tested, add 10% of the converted total volume of DMSO to dissolve it, and then slowly add 90% of the total volume of 0.5% MC solvent while stirring. Sonicate and vortex to mix thoroughly to obtain a solution of the preparation that is considered to be homogeneous, with a concentration of 1 mg/mL. Prepare fresh immediately before use.

Pipe 0.2 mL of the sample into a 1.5 mL centrifuge tube and store at -80 °C for analysis of the concentration of the drug solution.

2. Animal preparation

Animals were housed in rat cages and fasted for at least 10 hours starting the day before the experiment, but water was allowed. On the day of the experiment, each animal was weighed and marked on its tail. Blank blood samples were collected before drug administration. Blood was collected via tail vein.

3. Administration

Route of administration: oral gavage (p.o.)

Dosage concentration: 1 mg/ml

Dosage: 10 mg/kg

Dosage volume: 10 mL/kg

Procedure: Hold the rat upright with your left hand wearing a bite-proof glove, insert the gavage needle into the throat through the mouth, and insert the needle when you feel no obvious resistance. Then inject the drug into the stomach.

4. Sample collection

Whole blood (0.1-0.2 ml) was collected from test animals at 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 12 h, and 24 h after drug administration. The blood was placed in EDTA-Na ₂ anticoagulant tubes, inverted 3-4 times to mix, and centrifuged at 2000g for 5 min at 4℃ to separate the supernatant plasma. The plasma was then promptly transferred to -80℃ for storage until analysis. Blood was collected via tail vein.

5. Sample Analysis and Data Processing

5.1 Sample Analysis

A quantitative detection method for analytes was established using Shimadzu liquid chromatography and Triple Quad™ ⁶⁵⁰⁰⁺ AB mass spectrometry. The concentration of the parent drug in plasma was analyzed. Analytical results were controlled for variation using quality control samples, with an accuracy between 80% and 120%.

5.2 Data Processing

The main pharmacokinetic parameters were calculated using a non-compartmental model in Winnonlin Phoenix software. These parameters included the area under the curve (AUC(0-t) and AUC(0-∞)), elimination half-life (T _{_1/2} ), maximum plasma concentration (C _{_max} ), and time to reach maximum plasma concentration (T _{_max}).

The pharmacokinetic data of the test samples obtained through the above tests are shown in Table 1.

Table 1

Therefore, compounds T-47, T-51, and T-95 possess excellent pharmacokinetic properties.

Compound 50561, as described in patent WO2019029273A1, was synthesized and has the following structural formula:

The pharmacokinetic data of the test samples obtained through the above tests are shown in Table 2.

Table 2

Compound numbering Tmax(h) Cmax(ng/ml) AUClast(h*ng/ml) Compound 50561 1.33 838 2499

As can be seen from Tables 1 and 2, the compounds of the present invention have better pharmacokinetic properties.

Experimental Example 2: Brain Pharmacokinetic Study

1. Drug preparation

Administration solvent: 5% DMSO + 10% Solutol HS15 + 85% physiological saline

Formulation preparation process: Weigh an appropriate amount of powder, add an appropriate volume of DMSO, vortex, then add an appropriate volume of Solutol solution, vortex, and add an appropriate amount of physiological saline. Vortex and sonicate to form a homogeneous and clear formulation solution.

Before administration, collect two samples of the preparation and the remaining preparation after administration, and store them in an environment of 2-8℃ until they are sent out.

2. Laboratory animals

The laboratory animals are housed in the animal facility of Suzhou Xihua New Drug Development Co., Ltd. (License No.: SYXK(Su)2021-0019). The animal facility is equipped with an air conditioning system, has good ventilation, and maintains an indoor temperature between 20 and 26°C and a humidity between 40% and 70%. Artificial lighting is used in the animal facility, with 12 hours of light and 12 hours of darkness (except when working lights are needed for experimental operations or cleaning). The laboratory animals have free access to food and water.

Rats were purchased and fed normally for at least 3 days. Those rats that passed a veterinary examination and showed satisfactory physical condition were included in this experiment. Each rat was marked with a tail number. Animals in the oral administration group were fasted overnight the day before administration and resumed feeding 4 hours later, with free access to water. The source and number of animals used in this experiment are shown in Table 3.

All procedures performed on animals during the experiment must comply with the relevant Standard Operating Procedures (SOPs) for laboratory animal handling of Suzhou XiHua New Drug Development Co., Ltd., and be approved by the Laboratory Animal Welfare and Ethics Review Committee (IACUC) of Suzhou XiHua New Drug Development Co., Ltd.

Table 3. Sources and Number of Experimental Animals

Note: 3 animals per group.

3. Administration

Route of administration: oral gavage (p.o.)

Dosage concentration: 1 mg/ml

Dosage: 10 mg/kg

Dosage volume: 10 mL/kg

Dosage frequency: single dose

Before administration, check the state of the preparation and ensure its homogeneity by vortexing, stirring or shaking. Calculate the theoretical volume of administration for each SD rat in each group according to the following formula.

4. Sample collection and processing

Samples were taken from the experimental rats at 0.5h, 1h, 2h, 4h, 6h, 8h, and 24h after administration.

Plasma: At each time point, 0.15 mL of whole blood was collected from the jugular vein and placed in test tubes containing the anticoagulant EDTA- _K2 (3 μL, 15% EDTA- _K2 solution). The tubes were placed on moist ice and centrifuged within 1 hour (2,000 g, 2–8 °C, 10 min). Plasma was then collected. The plasma was stored in pre-chilled centrifuge tubes, flash-frozen on dry ice, and subsequently stored at -60 °C or lower in an ultra-low temperature freezer until LC-MS/MS analysis was performed.

Cerebrospinal fluid (CSF): Rats were euthanized using the carbon dioxide method. ~50 μL of CSF was collected via puncture. Using a 1 mL syringe, with the bevel facing upwards and the needle tip nearly horizontal, the CSF was inserted into the subarachnoid space. The needle was fixed in place, and the CSF was slowly aspirated. Samples were stored on dry ice for 30 minutes after collection, and then transferred to an environment of -90 to -60°C.

Brain tissue: Cardiac perfusion was performed first, followed by brain tissue harvesting. Under deep terminal anesthesia, the heart was perfused with approximately 8 ml of normal saline to flush out any remaining blood from the brain tissue. After harvesting, the brain tissue was gently washed once with frozen normal saline, dried, weighed, and then transferred to an environment of -90 to -60°C for cryopreservation.

5. Sample Analysis and Data Processing

5.1 Sample Analysis

A quantitative detection method for the analyte was established using Shimadzu liquid chromatography and Triple Quad™ ⁶⁵⁰⁰⁺ AB mass spectrometry. The concentration of the parent drug in the sample was analyzed. The analytical results were controlled for variation using quality control samples, with an accuracy between 80% and 120%.

5.2 Data Processing

Pharmacokinetic parameters were calculated using Winnonlin Phoenix 8.1.0.3530, including Tmax, Cmax, AUC(0-t), AUC(0-∞), T1/2, and MRT(0-∞). The pharmacokinetic data of the test samples obtained through these tests are shown in Table 4.

Table 4

Brain tissue/plasma total drug ratio (B/P) = Total drug concentration in brain tissue / Total drug concentration in plasma

Therefore, it can be concluded that compound T-47 has good blood-brain barrier permeability.

Experimental Example 3: Electrophysiological LTP Recording

1. Laboratory animals

strain C57BL/6J gender male rat age 6-8wks old

2. Experimental Grouping

Group number name N 1 Control 3 2 T-01 3

3. Experimental Methods

3.1 Brain slice preparation:

C57BL/6J mice, aged 6-8 weeks, were anesthetized and quickly decapitated. The scalp was cut open to remove the skull and dura mater. The whole brain was quickly removed and placed in artificial cerebrospinal fluid (ACSF) saturated with 95% _O2 and 5% _CO2 at 0-4°C for slight cooling. The cerebellum and one-third of the forebrain were removed, and the cerebrum was bisected along the midline. The hippocampus was separated from the ventromedial side along the cortical margin, and the brain tissue block containing the hippocampus was fixed to a carrier dish with glue. Brain slices with a thickness of 400 micrometers were cut coronally using a vibratory microtome. The brain slices were placed in an incubation tank immersed in a nylon mesh below the liquid surface, continuously filled with mixed gas, and incubated in a constant temperature water bath at 34°C for 0.5 h. Then, they were incubated at room temperature (26±1)°C until use, and experiments were started after 2-3 h.

3.2 Ex vivo brain slice potential recording:

Under direct visualization with a surgical microscope, the bipolar tungsten wire stimulation electrode with its tip exposed is placed on the Schaffer side path in the CA3 region. The stimulation electrode is connected to the stimulator via an isolator, and the recording electrode is connected to the digital-to-analog converter via a microelectrode amplifier and data acquisition software.

3.3 Electrophysiological Recording:

Before recording field excitatory postsynaptic potentials (fEPSPs), the brain slice is transferred to the recording tank, and a mixed gas (95% _O2 , 5% _CO2 ) is continuously circulated through the cerebrospinal fluid in the water bath. The stimulating electrode is inserted into the CA3 region of the hippocampus, and the recording electrode into the CA1 region. As the target site approaches, the insertion depth of both electrodes is slowly and precisely adjusted, with a 1 ms pulse width applied every 10–20 s to adjust the stimulation intensity until the optimal fEPSP is achieved. The electrode positions are then fixed, and the stimulation intensity at the optimal fEPSP location, which elicits 30–40% of the maximum response, is used as the baseline fEPSP intensity for recording. Recording continues for 20–30 minutes to establish a stable baseline. Afterward, perfusion drug administration (initial exploratory dose of 30 μM) is performed for 20–30 minutes (depending on drug efficacy). If the drug can enhance or reduce fEPSP, wash with normal ACSF for 40-60 minutes to observe whether the enhancement effect can be maintained for a long time; if it cannot significantly change fEPSP, stop the experiment. Data is acquired, amplified, displayed, and stored on a computer using an electrophysiological signal acquisition and processing system, and the experimental data and images are processed and output using Clampfit software.

4. Data Statistics and Analysis:

All experimental data are expressed as mean ± standard error (Mean ± SEM). All statistical analyses used were one-way ANOVA. Significance was indicated by: *p≤0.05, **p≤0.01, ***p≤0.001. p>0.05 indicates no significance.

The electrophysiological LTP recording data of the test samples obtained through the above tests are shown in Table 5.

Table 5

As shown in Table 5, compound T-01 significantly enhanced electrical signal transduction in brain slices (p≤0.05), indicating statistical significance. Compared with the control compound, it showed higher activation of LTP. Using the same detection method, compounds T-36, T-47, T-51, T-54, T-73, T-74, T-75, T-76, T-79, T-80, T-99, and T-100 of this invention also exhibited good activation effects on LTP.

All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims (10)

1. A compound, characterized in that the compound is a compound of formula I or a stereoisomer, racemate, or pharmaceutically acceptable salt thereof. in, Ring A is selected from the following group: X ₁ is selected from the following group: O, S, NH, NR; X ₂ is selected from the following group: O, NH, NR; _R1 is selected from the following group: R ₂ is selected from the following group: H, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C6 cycloalkyl; _R3 and _R4 are independently selected from the group consisting of: H, D, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C6 cycloalkyl, or _R3 and _R4 together with the carbon to which they are attached to form a substituted or unsubstituted 3-7 membered cycloalkyl or a substituted or unsubstituted 3-7 membered heterocyclic group containing one or more heteroatoms selected from O, S or N; Each of _R5 , _R6 , _R7 , _R8 , _R9 , and _R10 is independently selected from the group consisting of: H, D, halogen, trifluoromethyl, cyano, hydroxyl, amino, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C1-C6 alkyl- _NR16- , substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted C3-C6 cycloalkoxy, substituted or unsubstituted C3-C6 cycloalkyl- _NR16- ; Each of _R11 , _R12 , _R13 , _R14 , and _R15 is independently selected from the group consisting of: H, D, halogen, trifluoromethyl, cyano, hydroxyl, amino, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C1-C6 alkyl- _NR16- , substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted C3-C6 cycloalkoxy, substituted or unsubstituted C3-C6 cycloalkyl- _NR16- ; Each R ₁₇ and R ₁₈ is independently selected from the group consisting of: H, D, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy; or R ₁₇ and R ₁₈ together with the carbon atom to which they are attached form a substituted or unsubstituted 3-7 membered cycloalkyl group or a substituted or unsubstituted 3-7 membered heterocyclic group containing one or more heteroatoms selected from O, S or N; Each R is independently a substituted or unsubstituted C1-C6 alkyl group; Each of the substitutions independently refers to substitution by one or more substituents selected from the group consisting of: D, halogen, trifluoromethyl, cyano, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkyl-NR _16- , C3-C6 cycloalkyl, C3-C6 cycloalkoxy, C3-C6 cycloalkyl-NR _16- ; Each R ₁₆ is independently selected from the group consisting of: H, C1-C6 alkyl groups; The additional condition is: when _X1 is NH and _X2 is O, and _R3 , _R4 , _R17 , and _R18 are H, ring A is not... 2. The compound according to claim 1, characterized in that the compound has the structure shown in Formula II: Among them, rings A, _X1 , _R1 , _R2 , _R3 , and _R4 are as defined in claim 1. 3. The compound of claim 1, wherein ring A is selected from the group consisting of: _R5 , _R6 , _R7 , _R8 , and _R9 are as defined in claim 1. 4. The compound according to claim 1, wherein _R2 is selected from the group consisting of: H, halogens, C1-C6 alkyl, C1-C6 alkoxy, and C3-C6 cycloalkyl; _R3 and _R4 are independently selected from the following group: H, D, halogen, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl. 5. The compound of claim 1, wherein ring A is... _R1 is Each of _R5 , _R6 , _R7 , _R8 , and _R9 is independently selected from the group consisting of: H, halogen, trifluoromethyl, cyano, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, and C3-C6 cycloalkoxy. Each of _R11 , _R12 , _R13 , _R14 , and _R15 is independently selected from the group consisting of: H, halogen, trifluoromethyl, cyano, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, and C3-C6 cycloalkoxy. 6. The compound of claim 1, wherein the compound is selected from the group consisting of: 7. A pharmaceutical composition, characterized in that it comprises a pharmaceutically acceptable carrier and one or more safe and effective amounts of the compound of claim 1. 8. Use of the compound of claim 1, characterized in that it is used to prepare a medicament for the prevention and/or treatment of neurodegenerative diseases. 9. The use as described in claim 8, wherein the neurodegenerative disease is selected from the group consisting of: Alzheimer's disease, epilepsy, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, and spinocerebellar ataxia. 10. Use of the compound of claim 1, characterized in that it is used to prepare a medicament for the prevention and/or treatment of RAC1-related diseases.