Mononuclear monovalent gold dihalide complexes, compositions thereof, and methods of making and using the same

Abstract

This invention discloses a mononuclear monovalent gold dihalogen complex, its composition, preparation method, and applications, belonging to the field of pharmaceutical technology. The complex has the general formula M[AuX2], where M is a pharmaceutically acceptable monovalent cation, and X is Cl, Br, or I. This complex has a very simple structure and excellent stability, and can penetrate various tissue-specific physiological barriers such as the blood-brain barrier. Pharmacodynamic experiments have confirmed that this compound can simultaneously achieve synergistic effects on multiple targets, including calcium homeostasis reset, abnormal protein depolymerization, chronic inflammation suppression, pathological calcification intervention, and restoration of cell signal transduction function, significantly prolonging the survival of SOD1 G93A ALS transgenic mice by 44.5%, with a safety window greater than 300. The compound of this invention has high safety and a wide therapeutic window, and can be used to prepare drugs for the prevention and / or treatment of various diseases such as neurodegenerative diseases, metabolic diseases, malignant tumors, chronic fatigue syndrome, age-related diseases, and immunodeficiency.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and in particular to mononuclear monovalent gold dihalogen complexes, their compositions, preparation methods, and applications. Background Technology

[0002] Chronic noncommunicable diseases (CCDs) have become the leading global health burden, encompassing a wide range of conditions including neurodegenerative diseases, metabolic diseases, malignant tumors, chronic inflammatory diseases, neurological dysfunctions, and diseases related to tissue fibrosis and pathological calcification. These diseases typically have a long course, complex causes, and are influenced by multiple factors, and currently, there is a lack of effective treatments in clinical practice that can address the root causes.

[0003] Current research confirms that the seemingly unrelated chronic diseases mentioned above actually share a common early pathological basis—cellular calcium homeostasis disorder and calcium overload. Calcium ions, as one of the most important second messengers in cells, participate in regulating almost all life activities, including neurotransmitter release, gene expression, cell proliferation and apoptosis, and muscle contraction. When cells are stimulated by persistent internal and external stresses (such as oxidative stress, inflammatory factors, and abnormal metabolites), the activity of calcium pumps on the cell membrane (such as plasma membrane calcium ATPase PMCA and sarcoplasmic reticulum calcium ATPase SERCA) decreases, and voltage-gated calcium channels and transient receptor potential channels open abnormally, leading to an abnormally high intracellular calcium ion concentration and a state of "calcium overload." Calcium overload then activates a series of calcium-dependent degrading enzymes (such as calpains and endonucleases), damages mitochondrial function, induces endoplasmic reticulum stress, and becomes a key initiating step in cell dysfunction, degeneration, and even death.

[0004] Meanwhile, abnormal cell signal transduction is another common pathological feature of the aforementioned diseases. During the progression of chronic diseases, three types of harmful substances gradually accumulate in the cellular microenvironment: ① abnormally folded protein aggregates (such as Aβ plaques and Tau protein tangles in Alzheimer's disease, α-synuclein aggregates in Parkinson's disease, and TDP-43 aggregates in ALS); ② fibrotic scars and inflammatory deposits produced by chronic inflammatory responses; and ③ microcrystals formed by abnormal calcium and phosphorus metabolism. These three types of pathological products work together to cover, block, or damage receptors, ion channels, gap junction proteins on the cell surface, and intracellular signal transduction complexes, leading to impaired transmission of neurotransmitters / neuropeptides, cytokines, and ion signals, thus disrupting intercellular communication and signal transduction networks.

[0005] Furthermore, the essence of aging is considered to be the progressive calcification, fibrosis, and abnormal protein accumulation in cells and tissues. With age, calcium metabolism disorders lead to pathological calcification of soft tissues (such as vascular calcification, heart valve calcification, and articular cartilage calcification), abnormal extracellular matrix remodeling leads to fibrosis, and protein homeostasis imbalance leads to the accumulation of various abnormally folded proteins. These age-related changes further exacerbate cell signaling disorders, creating a vicious cycle.

[0006] In the medical field, gold (especially gold ions) has a long history of medicinal use. Monovalent gold ions (Au) + ) due to its unique d 10 The electronic configuration allows for high-affinity and specific coordination binding with proteins containing active cysteine ​​residues in living organisms, thereby regulating the activity of various key proteins. Previous studies have shown that monovalent gold complexes possess anti-inflammatory, immunomodulatory, and thiol-containing protein-intervening functions, and have the potential to regulate calcium ion signaling pathways and penetrate tissue-specific physiological barriers (such as the blood-brain barrier). However, translating these theoretical advantages into stable, safe, and pharmaceutically viable drug molecules remains a pressing technical challenge in this field.

[0007] Currently, the existing technologies closest to the technical solution of this invention include the following categories: Category 1: Traditional organogold drugs—Auranofin. Auranofin is currently the only approved oral monovalent gold complex for clinical use. Its chemical structure is an organogold compound formed by the coordination of acetylated thioglucose and triethylphosphine gold, with the molecular formula C. 20 H 34 AuO9PS, with a molecular weight of 678.48, is primarily used to treat rheumatoid arthritis. However, aurinophene has several significant drawbacks: First, its complex structure, containing organophosphine and thioglycolic acid ligands, makes its metabolic pathways in vivo unclear, and the ligands themselves possess potential toxicity. Second, it exhibits poor selectivity for protein targets, resulting in significant side effects (commonly diarrhea, rash, stomatitis, liver and kidney damage, etc.). Third, due to its large molecular weight and lipid solubility, it struggles to effectively penetrate the blood-brain barrier, making it unsuitable for intervention in central nervous system diseases. Fourth, aurinophene cannot systematically reverse pathological calcification in soft tissues or repair overall abnormalities in cell signaling pathways; it can only alleviate the single pathological link of inflammation. The second category consists of trivalent gold compounds. Trivalent gold compounds such as gold trichloride (AuCl3) and sodium tetrachloroalloyate (Na[AuCl4]) are documented in chemical handbooks. However, trivalent golds possess strong oxidizing and cytotoxic properties, easily causing severe liver and kidney damage. Direct use in human treatment poses unacceptable safety risks and has never been developed as a therapeutic drug. Category 3: Nano gold (Au) 0Gold nanoparticles (typically 1-100 nm in diameter) are primarily used for chemical or physical gold plating, bioimaging, and drug delivery carriers due to their surface plasmon resonance effect and good biocompatibility. However, zero-valent gold nanoparticles lack specific targeting ability for intracellular calcium homeostasis-related proteins, cannot directly regulate intracellular calcium ion pathways, cannot intervene in pathological calcification processes, and cannot depolymerize abnormally folded protein aggregates. Their mechanism of action mainly relies on passive targeting (such as the EPR effect) or external physical stimulation (such as photothermal conversion), and does not belong to active, specific molecular pharmacological intervention. The fourth category: publicly disclosed simple monovalent gold halides. Inorganic chemistry handbooks record the basic chemical structures of simple monovalent gold halides such as potassium dichlorohydrin(I)ate (K[AuCl2]). However, these publicly available documents only describe them as general chemical reagents, without mentioning the development of any pharmaceutical salts (such as not salting them with pharmaceutically acceptable cations to produce pharmaceutical-grade raw materials), not providing industrial-scale preparation processes for stabilizing them and preventing disproportionation, not disclosing injectable, topical, or mucosal preparations containing such compounds, and not mentioning any pharmaceutical uses related to calcium homeostasis regulation, abnormal protein depolymerization, chronic inflammation suppression, or pathological calcification intervention. More importantly, known simple monovalent gold salts have serious instability problems: under normal storage conditions, free monovalent gold ions are highly susceptible to disproportionation (2Au). + →Au 3+ +Au 0 This process generates toxic trivalent gold and black elemental gold precipitates, leading to drug inactivation and the introduction of harmful impurities. This technical deficiency has long remained unresolved, making simple monovalent gold salts considered "untreatable." The fifth category: Existing clinical treatments. For the aforementioned chronic diseases, current clinical treatments are all symptomatic. For example, Alzheimer's disease mainly uses cholinesterase inhibitors or NMDA receptor antagonists, which can only partially improve cognitive symptoms and cannot stop neuronal degeneration; Parkinson's disease mainly uses levodopa supplementation, which only improves motor symptoms; rheumatoid arthritis uses anti-inflammatory drugs or immunosuppressants, which only control inflammatory activity; there are currently no approved specific drugs for ME / CFS. All these existing treatments can only intervene in a single pathological link (such as simply anti-inflammatory or simply supplementing neurotransmitters), and cannot simultaneously clear the three types of pathological products that cause abnormal cell signaling (abnormal protein aggregates, inflammatory scars and deposits, and calcium and phosphorus crystals), cannot fundamentally reset cellular calcium homeostasis, and cannot break the mutually reinforcing negative feedback loop between chronic diseases and aging.

[0008] In summary, the existing technologies have the following common drawbacks: (1) Traditional gold drugs have complex structures and high ligand toxicity, resulting in unclear targets and large side effects. They cannot accurately act on calcium homeostasis and cell signal transduction pathways, and cannot penetrate tissue-specific physiological barriers such as the blood-brain barrier, making it difficult to reach deep lesions; (2) Trivalent gold compounds have excessive oxidizing properties, and nano-gold has no intracellular activity. Neither of them can achieve intracellular pathological repair, and there are serious safety hazards; (3) It is known that simple monovalent gold halides are only basic chemical substances. Due to the lack of technical defects such as instability, easy disproportionation and difficulty in drug formation, they cannot be used as drugs; (4) All existing solutions can only intervene in a single or a few pathological links, and cannot simultaneously achieve the three-in-one synergistic repair of abnormal protein depolymerization, chronic inflammation suppression, pathological calcification intervention and calcium homeostasis reset; (5) For core chronic diseases such as neurodegenerative diseases, metabolic diseases, malignant tumors, ME / CFS, etc., the existing technologies do not have any root-cause intervention methods, and can only delay the progression of symptoms, but cannot reverse the cause.

[0009] Therefore, there is an urgent need in this field to develop a class of novel drug molecules that are chemically stable, low in toxicity and safe, can be formulated, can penetrate tissue-specific physiological barriers, and can simultaneously intervene in calcium homeostasis disorders, abnormal protein aggregation, chronic inflammation and pathological calcification at multiple targets, and to provide feasible industrial preparation methods and clinical medication regimens for them. Summary of the Invention

[0010] The purpose of this invention is to provide a class of chemically stable, low-toxicity, safe, formulation-compatible mononuclear monovalent gold dihalogen complexes that can penetrate tissue-specific physiological barriers, their compositions, preparation methods, and applications, so as to at least partially solve the above-mentioned problems of the prior art.

[0011] To achieve the above objectives, the present invention provides a mononuclear monovalent gold dihalogen complex with the general formula M[AuX2]; Wherein, M is a pharmaceutically acceptable monovalent cation, including any one of sodium ion, potassium ion, lithium ion, ammonium ion, meglumine ion, arginine ion, lysine ion, histidine ion, tris(hydroxymethyl)aminomethane ion, and glucosamine ion; X includes any one of chloride ions, bromide ions, and iodide ions; Au has a +1 valence and exhibits a single-core structure.

[0012] Optionally, the complex has a linear coordination structure, wherein the Au atom is aligned linearly with the two X atoms at 180°. The complex is stable in solid state for more than 6 months under light-proof and sealed conditions at 25±2℃, with the purity of the main component decreasing by ≤1%.

[0013] Secondly, the present invention provides a method for preparing the above-mentioned mononuclear monovalent gold dihalogen complex, comprising the following steps: (1) Mix chloroauric acid with water for injection to prepare a trivalent gold solution; (2) The trivalent gold solution is mixed with a reducing agent for reduction, so that Au 3+ Restore to Au + A monovalent gold solution was obtained. (3) The monovalent gold solution is mixed with a stoichiometric amount of halide salt MX and stirred to form an M[AuX2] complex solution; (4) The M[AuX2] complex solution was concentrated under reduced pressure, cooled to crystallize, and filtered to obtain M[AuX2] crystals; (5) The M[AuX2] crystal is washed and dried to obtain M[AuX2] solid powder.

[0014] Optionally, in step (1), the concentration of the trivalent gold solution is 5~20 g / L.

[0015] Optionally, in step (2), the reduction step is carried out at 20~30℃, in the dark, and under mechanical stirring conditions, and the redox potential is controlled to be 0.8~1V (vs saturated calomel electrode SCE). The mechanical stirring speed is 200~400 r / min; The reducing agent includes any one of vitamin C solution, sodium ascorbate, and isoascorbic acid; The concentration of the monovalent gold solution is 3~15 g / L.

[0016] Optionally, in step (3), the stirring speed is 300~500 r / min, and the stirring time is 25~35 min; In step (4), the vacuum concentration is to concentrate the volume to 1 / 8 to 1 / 10 of the original volume at 30 to 40°C, and the cooling crystallization is to cool the volume to 0 to 4°C and let it stand for 10 to 15 hours to crystallize. In step (5), the washing is performed 1 to 3 times, the drying is vacuum drying until the moisture content is ≤0.5%, and the vacuum drying temperature is 25 to 30°C.

[0017] Thirdly, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of the above-mentioned mononuclear monovalent gold dihalogen complex, and a pharmaceutically acceptable carrier, diluent or excipient. The dosage forms of the pharmaceutical composition include injectable formulations, topical formulations, or mucosal formulations.

[0018] Optionally, the injectable formulation includes an intravenous injection solution, a subcutaneous injection solution, or an intrathecal injection solution; The topical preparations include sprays, gels, creams, disinfectants, or wound dressings; The topical preparation can be impregnated and loaded onto a medical carrier for use, the medical carrier including a medical cotton swab; The mucosal preparations include nasal sprays, mouthwashes, or throat sprays.

[0019] Fourthly, the present invention provides the use of the above-described mononuclear monovalent gold dihalogen complex, or the above-described pharmaceutical composition, in the preparation of a medicament for the prevention and / or treatment of the following diseases: Diseases with calcium homeostasis disorder, abnormal cell signaling, abnormal protein aggregation, chronic inflammation and / or pathological calcification as their pathological basis; The diseases mentioned include neurodegenerative diseases, metabolic diseases, malignant tumors, neurological dysfunction diseases, chronic inflammatory and fibrotic diseases, age-related degenerative changes, or diseases related to immunodeficiency and immune disorders.

[0020] In this invention, the application of the mononuclear monovalent gold dihalogen complex or the pharmaceutical composition in the prevention or improvement of malignant tumors includes inhibiting tumor-associated chronic inflammation, improving pathological calcification of the tumor microenvironment, enhancing radiotherapy and chemotherapy sensitivity, or reversing tumor drug resistance.

[0021] Optionally, the neurodegenerative diseases include Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), Huntington's disease, or vascular cognitive impairment; The metabolic diseases mentioned include type 2 diabetes, insulin resistance, or disorders of glucose and lipid metabolism. The neurological disorders mentioned include myalgic encephalomyelitis / chronic fatigue syndrome (ME / CFS), chronic intractable pain, sleep disorders, or cognitive decline; The chronic inflammatory and fibrotic diseases mentioned include rheumatoid arthritis, tissue fibrosis, or organ calcification; The age-related degenerative changes include organ dysfunction, skin aging, or decline in cell signaling function; The immunodeficiency and immune dysregulation-related diseases include acquired immunodeficiency, virus infection-associated immune failure, primary / secondary immune cell dysfunction, or systemic lupus erythematosus.

[0022] The mechanism of action of the compounds of this invention is as follows: The compounds of this invention can penetrate various tissue-specific physiological barriers (including the blood-brain barrier, blood-eye / retina / lens barrier, blood-testis barrier, blood-placental barrier, etc.) to reach the lesion directly. Through the high affinity and specific binding of monovalent gold ions to key functional proteins containing active cysteine ​​residues, they exert multi-target synergistic pharmacological effects. (1) Reset cellular calcium homeostasis: Specifically binds to the cysteine ​​residues of the active sites of plasma membrane calcium ATPase (PMCA) and sarcoplasmic reticulum calcium ATPase (SERCA) to repair the oxidatively inactivated calcium pump function; at the same time, it regulates the cysteine ​​oxidation modification state of voltage-gated calcium channels (VGCC) and transient receptor potential channels (TRPC3 / 6) to inhibit abnormal calcium influx, thereby reducing intracellular calcium overload from the perspective of "enhanced calcium excretion + reduced calcium influx" and restoring cellular calcium homeostasis.

[0023] (2) Unfolded proteins: a dual mechanism mainly involving direct binding to depolymerization, supplemented by indirect regulation by chaperone proteins: ü Direct mechanism: Monovalent gold ions preferentially bind to free cysteine ​​residues exposed on the surface of abnormally folded proteins (such as Cys173 / Cys175 of TDP-43, Cys291 / Cys322 of Tau protein, Cys132 of α-synuclein, and intermolecular disulfide bonds of Aβ polypeptide), breaking the covalent disulfide bonds that maintain the aggregate structure, while disrupting the hydrophobic interactions and hydrogen bond network between protein molecules, causing the dense, insoluble aggregates to depolymerize into soluble monomers; Indirect mechanism: By restoring the molecular chaperone activity of heat shock proteins Hsp70 and Hsp90 (repairing their oxidized and inactivated cysteine ​​residues), it assists the depolymerized proteins in proper folding or degradation via the ubiquitin-proteasome pathway, preventing re-aggregation.

[0024] (3) Inhibit chronic inflammation: It specifically binds to Cys179, the active site of IκB kinase β (IKKβ), to block the activation of the NF-κB signaling pathway; at the same time, it binds to Cys838 residues of the NLRP3 inflammasome to inhibit the assembly and activation of the inflammasome; in addition, it can also inhibit the oxidative modification of TREM2 receptor on the surface of microglia, reduce the excessive activation of microglia and the release of pro-inflammatory factors (IL-1β, TNF-α, IL-6), and block the chronic inflammatory cascade from upstream.

[0025] (4) Intervening in pathological calcification and improving abnormal calcium metabolism: On the one hand, it can embed itself inside the hydroxyapatite lattice, destroy the orderly growth and deposition of crystals, and inhibit the formation and progression of tissue microcalcification; on the other hand, it can bind to the cysteine ​​residues of the active center of matrix metalloproteinases MMP-2 / MMP-9, inhibit their excessive degradation of the extracellular matrix, reduce the formation sites of calcification core, and regulate osteogenic / osteoclast-related signaling pathways to improve systemic calcium metabolism disorders.

[0026] (5) Restore cell signal transduction function: Clear abnormal protein aggregates, inflammatory scars and calcium and phosphorus crystal deposits in cell signal transduction pathways; At the same time, it binds to the cysteine ​​residues of gap junction protein Cx43 to repair oxidatively damaged gap junction channels and restore the transmission of electrical and chemical signals between cells; In addition, it can regulate the oxidative state of cell adhesion molecules (NCAM, VCAM-1), improve cell adhesion and communication functions, and rebuild normal cell signaling networks.

[0027] Compared with the prior art, the present invention has at least the following advantages: (1) Extremely simple structure and high stability: The compound of the present invention can be stably stored for more than 6 months in both solid and aqueous solution states, which is significantly better than simple gold salts without coordination protection.

[0028] (2) High safety and wide therapeutic window: Acute toxicity tests show that LD50 is low. 50 The concentration was 32.7 mg / kg, with a safety window >65°, which is far higher than the clinical drug standard.

[0029] (3) Synergistic effect of multiple targets: In vitro and in vivo experiments have confirmed that the compound of the present invention can significantly reduce cellular calcium overload, depolymerize Aβ fibers, inhibit the release of inflammatory factors, restore neuronal communication, and prolong the survival of ALS transgenic mice by 44.5%.

[0030] (4) Penetrating the blood-brain barrier: Pharmacokinetic experiments have shown that the brain / plasma concentration ratio reaches 0.35, which can effectively act on central lesions.

[0031] (5) Green preparation and easy to industrialize: It uses neutral and mild reducing agents such as vitamin C to reduce, avoids strong reducing agents, eliminates the generation of nano gold and trivalent gold impurities, the process is safe and environmentally friendly, and the raw materials are readily available.

[0032] (6) Broad spectrum of medicinal salts: It can form salts with more than 10 pharmaceutically acceptable monovalent cations, and is suitable for different dosage forms with varying solubility, osmotic pressure and pharmacokinetic requirements. Attached Figure Description

[0033] Figure 1 This is a schematic diagram of the structure of compound M[AuX2] of the present invention (Note: The overall structure is an ionic complex, consisting of an external cation M). + Complexes with the inner sphere anion [AuX2] - Two parts; Inner sphere: Au atoms are located at the center, symmetrically connected to one X atom on each side, arranged in a strictly horizontal straight line, with bond angles marked at 180°; M + Place in [AuX2] - (The diagram above illustrates the interaction of ionic bonds).

[0034] Figure 2 This is a process flow diagram of the preparation method of the present invention.

[0035] Figure 3 One of the schematic diagrams of the mechanism of action of the compounds of this invention: Schematic diagram of resetting cellular calcium homeostasis (Note: The basic principle is to repair PMCA and SERCA calcium pumps → inhibit VGCC / TRPC calcium channels → reduce intracellular calcium overload); Figure 4 This is one of the schematic diagrams illustrating the mechanism of action of the compounds in this invention: a schematic diagram of abnormal protein depolymerization (Note: The basic principle is to directly bind to TDP-43, Tau, α-synuclein, and Aβ → break disulfide bonds and depolymerize protein aggregates; activate Hsp70 / Hsp90 molecular chaperones → promote correct folding and degradation; taking the depolymerization of abnormal TDP-43 folding as an example, the monovalent gold ions in the mononuclear monovalent gold dihalogen complex M[AuX2] of this invention can specifically target and bind to the free cysteine ​​sulfhydryl groups exposed on the surface of abnormally folded TDP-43 proteins, replacing and breaking the intermolecular disulfide bonds that maintain the structure of protein aggregates, destroying the intermolecular hydrophobic interactions and hydrogen bond network, causing the insoluble pathological protein aggregates to depolymerize into soluble protein monomers; at the same time, it activates Hsp70 and Hsp90 molecular chaperones to assist the proteins after depolymerization to achieve correct folding, or to complete the degradation of abnormal proteins through the ubiquitin-proteasome pathway, inhibiting the re-aggregation of abnormal proteins. The principle is the same for other proteins such as Tau, α-synuclein, and Aβ). Figure 5 One of the schematic diagrams of the mechanism of action of the compounds of the present invention: Schematic diagram of the inhibition of chronic inflammation (Note: The monovalent gold ion in the mononuclear monovalent gold dihalogen complex M[AuX2] of the present invention can block the IKKβ-NF-κB pathway, inhibit the NLRP3 inflammasome, reduce microglial cell overactivation → downregulate IL-1β, TNF-α, IL-6 inflammatory factors). Figure 6 One of the schematic diagrams of the mechanism of action of the compounds of this invention: Schematic diagram of intervention in pathological calcification (Note: The basic principle of 1 is to disrupt the growth of hydroxyapatite crystal lattice → inhibit soft tissue microcalcification; the basic principle of 2 is to regulate MMP-2 / MMP-9 and balance osteogenic / osteoclastic signals → improve systemic calcium metabolism disorder). Figure 7 This is one of the schematic diagrams illustrating the mechanism of action of the compounds in this invention: a schematic diagram of restoring cell signal transduction (Note: The basic principle is to clear the three major pathological deposits (abnormal proteins, inflammatory scars, and calcium phosphate crystals) → repair gap junction protein Cx43 → rebuild intercellular electrical / chemical signal communication). Figure 8 This is the time-concentration curve of sodium dichloroalloy(1) in mouse tissues, as shown in Experimental Example 5 of this invention. Figure 9 This is a distribution diagram of gold elements in various tissues 30 minutes after drug administration in Experimental Example 5 of this invention. Detailed Implementation

[0036] The present invention will be further illustrated below with reference to specific embodiments. The following descriptions are merely a few embodiments of the present invention and are not intended to limit the present invention in any way. Although the present invention is disclosed below with reference to preferred embodiments, it is not intended to limit the present invention. Any modifications or variations made by those skilled in the art without departing from the scope of the present invention, based on the disclosed technical content, are equivalent to equivalent implementations and fall within the scope of the technical solution.

[0037] Unless otherwise specified, the raw materials used in the embodiments of the present invention are all purchased commercially and used directly without any special treatment.

[0038] Unless otherwise specified, the analytical methods in the embodiments all adopt conventional instrument or equipment settings and conventional analytical methods. Example 1

[0039] Preparation of raw materials for sodium dichloroalloy(I)ate (Na[AuCl2]) injection: 1. Take 10g of high-purity chloroauric acid (HAuCl4・3H2O) with a purity of 99.99% (4N grade) and dissolve it in 500mL of water for injection to obtain trivalent gold mother solution; 2. Under light protection and mechanical stirring (350 r / min), slowly add 0.1 mol / L vitamin C solution, controlling the potential at around 0.9 V (vs. saturated calomel electrode SCE), until the solution is completely colorless; 3. Add 2.5g of sodium chloride and stir for 30 minutes until completely dissolved; 4. Concentrate under reduced pressure at 35℃ to approximately 60mL, then slowly cool to 2℃ and allow to stand for 12 hours to crystallize. 5. After filtration, the sample was washed twice with cold water for injection and dried under vacuum at 28°C to constant weight, yielding 5.8 g of white sodium dichloroalloy(I) crystals with a purity of 99.7%. Example 2

[0040] Preparation of sodium dichloroalloy(I) injection solution: Mix 0.1 g (based on pure gold) of sodium dichloroalloy(I) prepared in Example 1 with 0.9 g of sodium chloride, add water for injection to 100 mL, stir to dissolve, adjust the pH to 7.0 with 0.1 mol / L sodium hydroxide, filter aseptically through 0.22 μm, fill into 10 mL ampoules, sterilize at 121 °C for 15 min to obtain sodium dichloroalloy(I) injection solution. Example 3

[0041] Preparation of sodium diiodoalloy(I)ate (Na[AuI2]) raw material: 1. Take 10g of high-purity chloroauric acid (HAuCl4・3H2O) with a purity of 99.99% (4N grade), dissolve it in 500mL of water for injection, and stir until completely dissolved to prepare trivalent gold mother solution; 2. Under conditions of light protection and mechanical stirring (350 r / min), slowly add 0.1 mol / L vitamin C solution, monitor the potential throughout the process, and control the potential to be stable at around 0.9 V (vs saturated calomel electrode SCE) until the solution is completely colorless, then stop adding the solution to complete the reduction of trivalent gold to monovalent gold. 3. Add 4.2g of sodium iodide to the reduced solution and stir continuously for 30 minutes until the solid is completely dissolved; 4. Concentrate the system under reduced pressure at 35°C to approximately 60 mL, then slowly cool it to 2°C at a rate of 1°C / min, and allow it to stand at low temperature for 12 hours to crystallize. 5. The crystals were collected by filtration, washed twice with water for injection pre-cooled to 2°C, and then vacuum dried at 28°C to constant weight to obtain pale yellow sodium diiodoalloy(I) crystals. The product weighed 6.9g and the purity was tested to be 99.6%. Example 4

[0042] Preparation of sodium diiodoalloy(I)ate (Na[AuI2]) external spray: 0.02 g (based on pure gold) of sodium diiodoalloy(I) prepared in Example 3 was mixed with 5 g of glycerol and 0.01 g of benzalkonium chloride. Purified water was added and stirred until completely dissolved, and the volume was adjusted to 100 mL. The pH of the system was adjusted to 6.8 using 0.1 mol / L sodium hydroxide solution. After sterile filtration through 0.22 μm, the solution was filled into 10 mL spray bottles and sterilized at 121 °C for 15 min to obtain sodium diiodoalloy(I) (Na[AuI2]) external spray, which can be used for wound repair and intervention of skin inflammation. Example 5

[0043] Preparation of gold iodide medical cotton swabs: Take the sodium diiodoalloy(I) topical spray prepared in Example 4, soak a medical degreased cotton swab in it, drain off the excess solution, seal it in a sterile aluminum foil bag, and sterilize it by irradiation to obtain iodine gold medical cotton swabs, which can be used for local intervention of oral ulcers, small skin wounds, and folliculitis. Example 6

[0044] Preparation of dichloroalloy(I)-glucuronide: 1. Take 10g of high-purity chloroauric acid (HAuCl4・3H2O) with a purity of 99.99% (4N grade) and dissolve it in 50mL of deionized water, stirring until completely dissolved.

[0045] 2. Under light-protected conditions and with mechanical stirring (350 r / min), slowly add 0.5 mol / L vitamin C solution dropwise, controlling the reaction temperature at 25℃, while monitoring the reaction system potential. Stop adding the solution when the potential stabilizes at 0.45V (vs. saturated calomel electrode SCE). Au 3+ Completely restored to Au + .

[0046] 3. Slowly add 0.42g of meglumine (N-methyl-D-glucosamine) to the reaction solution, protect from light, and stir mechanically (350r / min) for 30min. Adjust the pH to 6.5.

[0047] 4. Concentrate under reduced pressure until crystals precipitate, let stand at 4°C for 12 hours, filter, wash with a small amount of cold anhydrous ethanol, and dry under vacuum at 28°C to constant weight to obtain 10.3 g of white crystalline powder, with a yield of 92.1%.

[0048] 5. HPLC analysis showed that the product purity was 99.4%; after being stored in solid state at 25°C protected from light for 6 months, the purity remained at 99.0%, with no disproportionation or decomposition.

[0049] 6. This product can be used directly to prepare lyophilized powder for injection. After reconstitution, the clarity of the solution meets the requirements of the pharmacopoeia. Example 7

[0050] Preparation of arginine from dichloroalloy (I): 1. Take 10g of high-purity chloroauric acid (HAuCl4・3H2O) with a purity of 99.99% (4N grade) and dissolve it in 50mL of deionized water, stirring until completely dissolved.

[0051] 2. Under light-protected conditions and with mechanical stirring (350 r / min), slowly add 0.5 mol / L vitamin C solution dropwise, controlling the reaction temperature at 25℃, while monitoring the reaction system potential. Stop adding the solution when the potential stabilizes at 0.45V (vs. saturated calomel electrode SCE). Au 3+ Completely restored to Au + .

[0052] 3. Slowly add 0.38g of L-arginine to the reaction solution, protect from light, and react with mechanical stirring (350r / min) for 30min. Adjust the pH to 6.8.

[0053] 4. Concentrate under reduced pressure until crystals precipitate, let stand at 4°C for 12 hours, filter, wash with a small amount of cold anhydrous ethanol, and dry under vacuum at 28°C to constant weight to obtain 9.8 g of white crystalline powder, with a yield of 89.5%.

[0054] 5. HPLC analysis showed that the product purity was 99.3%; after being stored in solid state at 25°C protected from light for 6 months, the purity remained at 98.8%, with no disproportionation or decomposition.

[0055] 6. The osmotic pressure of this product's aqueous solution is close to that of human body fluids, making it suitable for preparing subcutaneous injection formulations. Example 8

[0056] Preparation of potassium dichloroalloy(I) acid (K[AuCl2]) raw material: 1. Take 10g of high-purity chloroauric acid (HAuCl4・3H2O) with a purity of 99.99% (4N grade), dissolve it in 500mL of water for injection, and stir until completely dissolved to obtain trivalent gold mother solution; 2. Under conditions of light protection and mechanical stirring (350 r / min), slowly add 0.1 mol / L vitamin C solution, monitoring the potential throughout the process and controlling the potential to be stable at around 0.9 V (vs. saturated calomel electrode SCE), until the solution is completely colorless, then stop adding the solution. 3. Add 3.1g of potassium chloride to the reaction solution and stir continuously for 30 minutes until the solid is completely dissolved; 4. Concentrate the system under reduced pressure at 35°C to approximately 60 mL, then slowly cool it to 2°C at a rate of 1°C / min, and allow it to stand at low temperature for 12 hours to crystallize. 5. The crystals were collected by filtration, washed twice with water for injection pre-cooled to 2°C, and then vacuum dried at 28°C to constant weight to obtain 6.0g of white potassium dichloroalloy(I) crystals, with a purity of 99.7%. Example 9

[0057] Preparation of lithium dichloroalloy(I) oxide (Li[AuCl2]) raw material: 1. Take 10g of high-purity chloroauric acid (HAuCl4・3H2O) with a purity of 99.99% (4N grade), dissolve it in 500mL of water for injection, and stir until completely dissolved to obtain trivalent gold mother solution; 2. Under light-protected and mechanically stirred (350 r / min) conditions, slowly add 0.1 mol / L vitamin C solution, monitor the potential throughout the process, and control the potential to be stable at around 0.9 V (vs saturated calomel electrode SCE) until the solution is completely colorless, then stop adding the solution to complete the reaction of reducing trivalent gold to monovalent gold. 3. Add 2.2g of lithium chloride to the reaction solution and stir continuously for 30min until the solid is completely dissolved to fully carry out the coordination salt formation reaction; 4. Concentrate the system under reduced pressure at 35°C to approximately 60 mL, then slowly cool it to 2°C at a rate of 1°C / min, and allow it to stand at low temperature for 12 hours to crystallize. 5. The crystals were collected by filtration, washed twice with water for injection pre-cooled to 2°C, and then vacuum dried at 28°C to constant weight to obtain 5.5g of white lithium dichloroalloy(I) crystals, with a purity of 99.6%. Example 10

[0058] Preparation of ammonium dichloroalloy(I) acid (NH4[AuCl2]) raw material: 1. Take 10g of high-purity chloroauric acid (HAuCl4・3H2O) with a purity of 99.99% (4N grade), dissolve it in 500mL of water for injection, and stir until completely dissolved to obtain trivalent gold mother solution; 2. Under light-protected and mechanically stirred (350 r / min) conditions, slowly add 0.1 mol / L vitamin C solution, monitor the potential throughout the process, and control the potential to be stable at around 0.9 V (vs saturated calomel electrode SCE) until the solution is completely colorless, then stop adding the solution to complete the reaction of reducing trivalent gold to monovalent gold. 3. Add 2.8g of high-purity ammonium chloride to the reaction solution and stir continuously for 30 minutes until the solid is completely dissolved, thus fully completing the monovalent gold coordination salt formation reaction; 4. Concentrate the system under reduced pressure at 35°C to approximately 60 mL, then slowly cool it to 2°C at a rate of 1°C / min, and allow it to stand at low temperature for 12 hours to crystallize. 5. The crystals were collected by filtration, washed twice with water for injection pre-cooled to 2°C, and then vacuum dried at 28°C to constant weight to obtain 5.7g of white ammonium dichloroalloy(I) crystals, with a purity of 99.6%. Example 11

[0059] Preparation of sodium dibromoalloy(I) acid (Na[AuBr2]) raw material: 1. Take 10g of high-purity chloroauric acid (HAuCl4・3H2O) with a purity of 99.99% (4N grade), dissolve it in 500mL of water for injection, and stir until completely dissolved to obtain trivalent gold mother solution; 2. Under light-protected and mechanically stirred (350 r / min) conditions, slowly add 0.1 mol / L vitamin C solution, monitor the potential throughout the process, and control the potential to be stable at around 0.9 V (vs saturated calomel electrode SCE) until the solution is completely colorless, then stop adding the solution to complete the reaction of reducing trivalent gold to monovalent gold. 3. Add 3.6g of sodium bromide to the reduced reaction solution and stir continuously for 30min until the solid is completely dissolved to fully carry out the coordination salt formation reaction; 4. Concentrate the system under reduced pressure at 35°C to approximately 60 mL, then slowly cool it to 2°C at a rate of 1°C / min, and allow it to stand at low temperature for 12 hours to crystallize. 5. Filter and collect the crystals, wash twice with water for injection pre-cooled to 2°C, and vacuum dry to constant weight at 28°C to obtain 5.6g of nearly white or slightly yellow sodium dibromoalloy(I) crystals, with a purity of 99.6%. Testing showed that the product, in its solid state, maintained a purity of 99.1% after 6 months of storage at 25°C protected from light, without disproportionation or decomposition; and in its aqueous solution state (0.1mg / mL, based on pure gold), maintained a purity of 98.6% after 6 months of storage at 25°C protected from light, without precipitation. Example 12

[0060] Mononuclear monovalent gold mixed dihalogen complex anion solution ([AuCl2)) - +[AuBr2] - Preparation of ) 1. Take 0.10g of high-purity chloroauric acid (HAuCl4・3H2O) with a purity of 99.99% (4N grade), add 10mL of sterile water for injection, stir to dissolve, and obtain a pale yellow trivalent gold mother liquor; 2. Add 0.05g of sodium bromide to the above solution and stir until completely dissolved to construct a chlorine-bromine mixed halogen system; 3. Under light-protected and stirring conditions, slowly add 0.02g of pharmaceutical-grade vitamin C powder in three portions, controlling the redox potential of the system at 0.9V (vs. saturated calomel electrode SCE). Stir until the solution changes from pale yellow to colorless and transparent. Au 3+ Completely restored to Au + ; 4. Continue adding sterile water for injection to bring the volume to 100 mL, stir well, and you will obtain the product containing [AuCl2]. - With [AuBr2] - Aqueous solutions of mixed mononuclear monovalent gold halide complexes; 5. After filtration through a 0.22μm sterile filter membrane for sterilization, the solution is sealed, protected from light, and refrigerated to obtain a stable active solution. The resulting solution has a pH of 7, is colorless and clear, without precipitation or disproportionation, and can be used directly in cell and animal experiments.

[0061] It should be noted that the mononuclear monovalent gold dihalogen complex described in this invention exists in aqueous solution form for in vivo administration, cell experiments, and practical applications. The substance that truly exerts its pharmacological activity is the mononuclear monovalent gold dihalogen complex anion ([AuX2)) formed after dissolution. - Therefore, protecting the stable aqueous solution of this type of active complex anion and its preparation method can more completely cover the actual pharmaceutical form and administration method of the present invention, making the scope of protection consistent with the actual pharmacological effects, experimental system and clinical application scenarios. At the same time, it also facilitates those skilled in the art to directly use the active solution to carry out pharmacological, pharmacokinetic, toxicological and formulation development research. Experimental Example 1

[0062] Active ingredient verification experiment: 1. Experimental Objective: To demonstrate that the pharmacological activity of the compound of this invention originates from monovalent gold ions (Au). + ), rather than halide ions (Cl) - (or other excipients).

[0063] 2. Experimental materials: Test sample: Sodium dichloroalloy(I)ate (Na[AuCl2], prepared in Example 1) Reference standard: Blank control: physiological saline for injection Comparative Example 1: 0.1 μmol / L sodium chloride (NaCl) solution Comparative Example 2: 0.1 μmol / L potassium chloride (KCl) solution Comparative Example 3: Mixed solution of 0.1 μmol / L sodium chloride and 0.1 μmol / L potassium chloride Cell line: SH-SY5Y cells Reagents and instruments: SH-SY5Y cells, Fluo-4 / AM calcium ion fluorescent probe, laser confocal microscope, ELISA reader, and other routine cell experiment reagents and instruments.

[0064] 3. Experimental Methods (1) A calcium overload model of SH-SY5Y cells was constructed and divided into a model group, a test sample group, comparative groups 1-3, and a blank control group.

[0065] (2) Each group was given the corresponding drug, with a final concentration of 0.1 μmol / L (based on solute). The blank control group was given an equal amount of physiological saline.

[0066] (3) After culturing for 24 hours, the intracellular calcium ion concentration and the positive rate of abnormal aggregation of TDP-43 were detected.

[0067] 4. The experimental results are shown in Table 1.

[0068] Table 1 Results of the Active Ingredient Validation Experiment Blank control group 100 2.3 Model group 327.5 68.7 <![CDATA[Test sample group (Na[AuCl₂])]]> 123.6 19.6 Comparative Example 1 (NaCl) 321.8 67.2 Comparative Example 2 (KCl) 325.1 68.1 Comparative Example 3 (NaCl + KCl) 323.4 67.9 Table 1 shows that chloride ions alone (NaCl, KCl, and mixtures thereof) had no effect on improving cellular calcium overload and TDP-43 aggregation, showing no significant difference from the model group. Only Na[AuCl2] solution containing monovalent gold ions significantly reduced intracellular calcium ion concentration and decreased abnormal TDP-43 aggregation. This fully demonstrates that all pharmacological activities of the compounds of this invention originate from monovalent gold ions (AuCl2). + Halogen ions only act as coordinating anions and have no pharmacological activity. Experiment Example 2

[0069] Stability studies of the compounds of this invention: 1. Experimental materials Test samples: Sodium dichloroalloy(I), potassium dichloroalloy(I), lithium dichloroalloy(I), ammonium dichloroalloy(I), sodium dibromoalloy(I), and sodium diiodoalloy(I) prepared in the embodiments of the present invention.

[0070] Reference standard: gold chloride (AuCl, a simple monovalent gold salt without coordination protection, known to be easily disproportionated and decomposed).

[0071] Solvent: Physiological saline for injection (pH 6.8±0.2).

[0072] 2. Experimental conditions Following the ICH Q1A(R2) guidelines for the stability of active pharmaceutical ingredients, the following settings are established: Long-term stability: 25℃±2℃, RH 60%±5%, protected from light and sealed; Solution stability: 25℃±2℃, protected from light, sealed under nitrogen protection; Accelerated stability: 40℃±2℃, RH 75%±5%, light-proof and sealed.

[0073] 3. Detection indicators Appearance, purity of main components (HPLC), trivalent gold (Au) 3+ ) content, elemental gold (Au) 0 ) Amount of precipitation, pH value.

[0074] 4. The long-term stability results (6 months) in the solid state are shown in Table 2.

[0075] Table 2. Statistical table of long-term stability results in solid state (6 months) <![CDATA[Na[AuCl2]]]> October White crystalline powder 99.7 0.02 0 January White crystalline powder 99.6 0.03 0 March White crystalline powder 99.5 0.05 0 June White crystalline powder 99.3 0.08 0 <![CDATA[K[AuCl2]]]> October White crystalline powder 99.6 0.02 0 January White crystalline powder 99.5 0.03 0 March White crystalline powder 99.4 0.06 0 June White crystalline powder 99.2 0.09 0 <![CDATA[Li[AuCl2]]]> October White crystalline powder 99.5 0.03 0 January White crystalline powder 99.4 0.04 0 March White crystalline powder 99.3 0.07 0 June White crystalline powder 99.1 0.10 0 <![CDATA[NH4[AuCl2]]]> October White crystalline powder 99.5 0.03 0 January White crystalline powder 99.4 0.04 0 March White crystalline powder 99.2 0.07 0 June White crystalline powder 99.0 0.11 0 <![CDATA[Na[AuBr2]]]> June White crystalline powder 99.1 0.08 0 <![CDATA[Na[AuI2]]]> October pale yellow crystalline powder 99.4 0.02 0 January pale yellow crystalline powder 99.3 0.04 0 March pale yellow crystalline powder 99.1 0.07 0 June pale yellow crystalline powder 98.9 0.10 0 Reference standard AuCl October Yellow crystalline powder 98.0 1.2 0.5 January Grayish-black powder (with a large amount of elemental gold precipitated) 62.3 21.5 14.2 March black powder 31.7 45.8 20.5 June Completely black and clumps up <10 >60 >25 As shown in Table 2, all the M[AuX2] solid compounds of the present invention showed a decrease in purity of the main component of <0.7% under long-term storage conditions of 6 months, with no obvious disproportionation or precipitation of elemental gold; while the reference standard AuCl underwent severe disproportionation and decomposition and turned black within 1 month, and almost completely failed after 6 months.

[0076] 5. The long-term stability results (6 months) of the aqueous solution are shown in Table 3. All compounds were prepared as 0.1 mg / mL (based on pure gold) physiological saline solutions and stored under nitrogen atmosphere and protected from light.

[0077] Table 3. Statistical table of long-term stability results of aqueous solution (6 months) <![CDATA[Na[AuCl2]]]> October colorless clear solution 99.7 6.8 0 January colorless clear solution 99.5 6.7 0 March colorless clear solution 99.2 6.6 0 June colorless clear solution 98.8 6.5 0 <![CDATA[K[AuCl2]]]> October colorless clear solution 99.6 6.8 0 January colorless clear solution 99.4 6.7 0 March colorless clear solution 99.1 6.6 0 June colorless clear solution 98.7 6.5 0 <![CDATA[Na[AuI2]]]> October pale yellow clear solution 99.4 6.8 0 January pale yellow clear solution 99.2 6.7 0 March pale yellow clear solution 98.9 6.6 0 June pale yellow clear solution 98.5 6.5 0 <![CDATA[Na[AuBr2]]]> June Aqueous solution 98.6 6.5 0 Reference standard AuCl October Yellow turbid solution 98.0 5.2 0.8 24h The substance is grayish-black and turbid, with black sediment. 45.2 4.1 32.7 7 days Completely blackened, with a large amount of sediment. <5 3.5 >80 As shown in Table 3, the aqueous solution of the compound of the present invention remains colorless / pale yellow and clear under light-proof and anaerobic conditions at pH 6.5~6.8. The purity of the main component decreases by <1.2% within 6 months, with no precipitation or disproportionation, and can be sterilely filtered through a 0.22μm filter membrane. The reference standard AuCl aqueous solution completely decomposes within 24 hours, producing a large amount of black elemental gold precipitate, and becomes completely ineffective after 7 days.

[0078] 6. Accelerated stability test results (40℃, 6 months): All M[AuCl2] compounds of this invention showed a decrease in main component purity of <1.5% and no significant disproportionation after 6 months under accelerated conditions; Sodium diiodoalloy(I) has a purity decrease of less than 2.0% after 6 months of accelerated processing. Its stability is slightly lower than that of chloride salts, but it still meets the requirements for pharmaceutical use. The reference standard AuCl completely decomposed within 3 days under accelerated conditions.

[0079] In summary, the compound of this invention is a coordination-saturated linear mononuclear complex, Au + Completely coordinated and protected by halide ions, d 10 The electronic configuration is stable and not prone to disproportionation; while the simple gold salt AuCl has no coordination protection, Au + When exposed, it is highly susceptible to disproportionation reactions: 2Au + →Au 3+ +Au 0 Therefore, its stability is extremely poor. All compounds of the general formula M[AuX2] of this invention, whether in solid or aqueous solution state, can be stably stored for more than 6 months under specified storage conditions, fully meeting the requirements for the production, storage and use of pharmaceutical preparations. Experimental Example 3

[0080] In vitro cell experiments of the compounds of this invention (using sodium dichloroalloy(I)ate (Na[AuCl2]) prepared in Example 1 as a representative example for verification): 1. Repair effect on calcium overload in SH-SY5Y cells Experimental objective: To verify the effect of the compound of this invention on reversing calcium overload in nerve cells.

[0081] Experimental materials: SH-SY5Y cells, Na[AuCl2], Fluo-4 / AM calcium ion fluorescent probe, calcium chloride.

[0082] Experimental methods: (1) Seed SH-SY5Y cells into 96-well plates and cultured until confluence reached 80%; (2) Calcium chloride at a final concentration of 10 mmol / L was added to the model group and the drug administration group to induce calcium overload, while the blank control group was given an equal amount of culture medium; (3) Na[AuCl2] was added to the drug administration groups at final concentrations of 0.01 μmol / L, 0.1 μmol / L, and 1 μmol / L, respectively, and the groups were cultured for 24 h. (4) Add Fluo-4 / AM probe and incubate for 30 min. Detect intracellular calcium ion fluorescence intensity using laser confocal microscopy.

[0083] The experimental results (n=6, x±s) are shown in Table 4.

[0084] Table 4. Calcium ion fluorescence intensity of different groups and comparison data with the model group. Blank control group 100.0±5.2 - Model group 342.7±18.5 - 0.01 μmol / L dosing group 268.3±12.7 *<0.05 0.1 μmol / L dosing group 125.6±8.3 ***<0.001 1 μmol / L dosing group 112.4±6.9 ***<0.001 Statistical analysis: One-way ANOVA was used. The treatment group and the model group showed statistically significant differences, which were dose-dependent (Table 4).

[0085] 2. Regarding Aβ 1-42 Depolymerization of fibers Experimental objective: To verify the depolymerization ability of the compound of this invention on β-amyloid fibrils.

[0086] Experimental materials: recombinant human Aβ 1-42 Protein, Na[AuCl2], thioflavin T (ThT) fluorescent dye.

[0087] Experimental methods: (1) In vitro induction of Aβ 1-42 Proteins form mature filaments (incubate at 37°C for 7 days). (2) Add Na[AuCl2] to a final concentration of 0.1 μmol / L and continue incubation for 24 h; (3) Add ThT dye and use a fluorescence spectrophotometer to detect the fluorescence intensity at 440nm excitation / 485nm emission wavelength.

[0088] The experimental results (n=6, x±s) are shown in Table 5.

[0089] Table 5 shows the Aβ 1-42 Statistical table of fiber depolymerization experimental data Blank control group 100.0±4.8 - Aβ fiber group 876.3±32.5 - Drug administration group 215.7±15.2 75.4±2.1 Statistical analysis: Compared with the Aβ fiber group, the drug-treated group showed *** P <0.001, indicating a significant depolymerization effect (Table 5).

[0090] 3. Inhibitory effect on LPS-induced release of inflammatory factors from BV2 microglia. Experimental objective: To verify the anti-inflammatory activity of the compounds of this invention.

[0091] Experimental materials: BV2 microglia, Na[AuCl2], lipopolysaccharide (LPS), ELISA kit.

[0092] Experimental methods: (1) BV2 cells were seeded in 24-well plates and cultured until confluence reached 70%; (2) The model group and the drug administration group were stimulated with LPS at a final concentration of 1 μg / mL, while the blank control group was given an equal amount of culture medium; (3) The drug-treated group was given Na[AuCl2] at a final concentration of 0.1 μmol / L and cultured for 24 h; (4) Collect the supernatant and detect the levels of IL-1β, TNF-α and IL-6 by ELISA.

[0093] The experimental results (n=6, x±s, pg / mL) are shown in Table 6.

[0094] Table 6. Cytokine Detection Data Blank control group 12.5±2.1 28.7±3.5 35.2±4.1 Model group 187.3±15.6 325.4±21.8 412.7±28.3 Drug administration group 42.6±5.3 78.5±8.2 96.3±10.5 Statistical analysis: Compared with the model group, *** P <0.001, the release of all three inflammatory factors was significantly inhibited (Table 6).

[0095] 4. Restorative effect on signal transduction function between damaged neurons Experimental objective: To verify the effect of the compound of this invention on the repair of neuronal gap connection communication. Experimental materials: primary rat cortical neurons, Na[AuCl2], scratch-labeled dye tracing (SLDT) kit Experimental methods: (1) Primary cortical neurons were cultured for 14 days and then induced with calcium chloride at a final concentration of 10 mmol / L for 24 hours. (2) The drug-treated group was given Na[AuCl2] at a final concentration of 0.1 μmol / L and cultured for another 24 h. (3) The SLDT method was used to detect the gap connection communication function between neurons and to calculate the dye diffusion distance.

[0096] The experimental results (n=6, x±s) are shown in Table 7.

[0097] Table 7. Data on dye diffusion distance for different groups Blank control group 187.5±12.3 Damage group 62.3±7.5 Drug administration group 156.7±10.2 Statistical analysis: Compared with the injury group, *** P <0.001 indicates a significant recovery in signal transmission between neurons (Table 7). Experiment Example 4

[0098] Animal in vivo efficacy experiments of the compounds of this invention (SOD1 G93A ALS mouse model, using sodium dichloroalloy(I)ate (Na[AuCl2]) prepared in Example 1 as a representative for verification): 1. Experimental objective: To verify the in vivo therapeutic effect of the compound of this invention on ALS.

[0099] 2. Experimental animals: SPF-grade SOD1 G93A transgenic mice (B6SJL-Tg(SOD1*G93A)1Gur / J), half male and half female, 8 weeks old.

[0100] 3. Experimental groups (n=15): Wild-type control group: Non-transgenic littermates were given an equal volume of physiological saline; Model group: SOD1 G93A mice, given an equal volume of physiological saline; Low-dose administration group: SOD1 G93A mice were intraperitoneally injected with 0.1 mg / kg Na[AuCl2] once daily; High-dose administration group: SOD1 G93A mice were injected intraperitoneally with 0.5 mg / kg Na[AuCl2] once daily; 4. Dosage time: Start administering the drug when the mice show clinical symptoms (hind limb weakness) and continue until the mice die.

[0101] 5. Observation indicators: Weight changes: Weigh yourself once a week; Motor function assessment: The BBB scoring method was used, and assessments were conducted twice a week. Survival period: Record the time from the onset of the disease to death of the mice.

[0102] The experimental results are shown in Table 8.

[0103] Table 8 Survival Time Statistics Model group 128±6 — Low-dose administration group 162±8 26.6 High-dose administration group 185±10 44.5 Statistical analysis: Compared with the model group, *** P <0.001 indicates a significant increase in survival (Table 8).

[0104] Motor function: The BBB score of mice in the drug-treated group decreased significantly slower than that in the model group. Four weeks after drug administration, the BBB score of mice in the high-dose group remained above 15 (compared to only 6 in the model group), and their motor function was significantly improved.

[0105] Weight changes: The model group mice experienced rapid weight loss after the onset of the disease, while the weight loss rate of the drug-treated group mice was significantly slowed down. The high-dose group mice were still able to maintain more than 85% of their normal weight 8 weeks after drug administration. Experimental Example 5

[0106] Pharmacokinetic and tissue distribution studies of the compounds of this invention (using sodium dichloroalloy(I)ate (Na[AuCl2]) prepared in Example 1 as a representative example for verification): Experimental objective: To verify that the compounds of this invention can penetrate the blood-brain barrier and be effectively enriched in brain tissue.

[0107] Experimental animals: SPF grade C57BL / 6 mice, male, 20~25g.

[0108] Experimental methods: 1. Mice were injected with Na[AuCl2] via the tail vein at a dose of 1 mg / kg; 2. Mice were sacrificed at 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 8 h, and 24 h after drug administration (n=5 / time point); 3. Collect plasma, brain tissue, heart, liver, kidney and other tissues, and determine the gold content using atomic absorption spectrophotometry.

[0109] Experimental results: (1) Blood-brain barrier penetration: like Figure 8 As shown, the gold concentration in brain tissue reached its peak (125 ng / g) 30 minutes after administration, and the brain / plasma concentration ratio was 0.35, proving that the compound of the present invention can effectively penetrate the blood-brain barrier and reach therapeutic concentrations in brain tissue.

[0110] (2) Organizational distribution: like Figure 9 As shown, the drug is mainly distributed in the liver, kidneys, and brain tissue. Most of the drug is excreted through the kidneys after 24 hours, with no significant tissue accumulation. Experimental Example 6

[0111] Acute toxicity studies of the compounds of this invention (using sodium dichloroalloy(I)ate (Na[AuCl2]) prepared in Example 1 as a representative example): Experimental objective: To evaluate the acute toxicity and safety window of the compounds of this invention.

[0112] Experimental animals: SPF-grade ICR mice, half male and half female, 18-22g.

[0113] Experimental methods: 1. Mice were randomly divided into 5 groups, with 10 mice in each group. Each group of animals was injected intravenously with different doses of sodium dichloroisocyanurate solution, calculated based on pure gold, at doses of 5 mg / kg, 10 mg / kg, 20 mg / kg, 40 mg / kg, and 80 mg / kg respectively. The test drug solution was prepared using sterile water for injection, and the administration volume for each group was uniformly 10 mL / kg. 2. Observe continuously for 14 days and record the number of mice that died, changes in body weight, activity status, coat color, food intake and other poisoning-related symptoms.

[0114] Experimental results: (1) LD 50 Measurement: Median lethal dose (LD50) of Na[AuCl2] administered via tail vein injection in mice 50 The concentration was 32.7 mg / kg (95% confidence limit: 28.5~37.6 mg / kg).

[0115] (2) Safety window calculation: The effective therapeutic dose of the compound of this invention is 0.1~0.5 mg / kg, and the safety window (LD50) is... 50 The minimum effective dose (MPD) is greater than 300, which is far higher than the safety window standard (>10) of commonly used clinical drugs, proving that the drug of this invention has high safety and low toxicity, indicating that the clinical use risk of this product is extremely low and has a broad therapeutic window.

[0116] (3) Symptoms of poisoning: Mice in the low-dose group (≤10mg / kg) showed no signs of poisoning and gained weight normally; mice in the high-dose group (≥40mg / kg) showed a transient decrease in activity, which returned to normal after 24 hours, with no delayed toxicity.

[0117] In summary, the mononuclear monovalent gold dihalogen complex and its pharmaceutical composition provided by this invention have a simple and controllable preparation process, making them suitable for large-scale industrial production. The compound exhibits good stability, can be formulated into various dosage forms, and has shown significant therapeutic effects and extremely high safety in multiple disease models, demonstrating promising clinical application prospects and industrial applicability.

[0118] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them. Those skilled in the art should understand that modifications can be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A mononuclear monovalent gold dihalogen complex, characterized in that, Its general formula is M[AuX2]; Wherein, M is a pharmaceutically acceptable monovalent cation, including any one of sodium ion, potassium ion, lithium ion, ammonium ion, meglumine ion, arginine ion, lysine ion, histidine ion, tris(hydroxymethyl)aminomethane ion, and glucosamine ion; X includes any one of chloride ions, bromide ions, and iodide ions; Au has a +1 valence and exhibits a single-core structure.

2. The mononuclear monovalent gold dihalogen complex according to claim 1, characterized in that, The complex has a linear coordination structure, wherein the Au atom is aligned linearly with two X atoms at 180°. The complex is stable in solid state for more than 6 months under light-proof and sealed conditions at 25±2℃, with the purity of the main component decreasing by ≤1%.

3. The method for preparing the mononuclear monovalent gold dihalogen complex according to claim 1 or 2, characterized in that, Includes the following steps: (1) Mix chloroauric acid with water for injection to prepare a trivalent gold solution; (2) The trivalent gold solution is mixed with a reducing agent for reduction, so that Au 3+ Restore to Au + A monovalent gold solution was obtained. (3) The monovalent gold solution is mixed with a stoichiometric amount of halide salt MX and stirred to form an M[AuX2] complex solution; (4) The M[AuX2] complex solution was concentrated under reduced pressure, cooled to crystallize, and filtered to obtain M[AuX2] crystals; (5) The M[AuX2] crystal is washed and dried to obtain M[AuX2] solid powder.

4. The method for preparing mononuclear monovalent gold dihalogen complex according to claim 3, characterized in that, In step (1), the concentration of the trivalent gold solution is 5~20 g / L.

5. The method for preparing mononuclear monovalent gold dihalogen complexes according to claim 3, characterized in that, In step (2), the reduction step is carried out at 20~30℃, in the dark, and under mechanical stirring conditions, and the redox potential is controlled at 0.8~1V (vs saturated calomel electrode SCE). The mechanical stirring speed is 200~400 r / min; The reducing agent includes any one of vitamin C solution, sodium ascorbate, and isoascorbic acid; The concentration of the monovalent gold solution is 3~15 g / L.

6. The method for preparing mononuclear monovalent gold dihalogen complex according to claim 3, characterized in that, In step (3), the stirring speed is 300~500 r / min, and the stirring time is 25~35 min; In step (4), the vacuum concentration is to concentrate the volume to 1 / 8 to 1 / 10 of the original volume at 30 to 40°C, and the cooling crystallization is to cool the volume to 0 to 4°C and let it stand for 10 to 15 hours to crystallize. In step (5), the washing is performed 1 to 3 times, the drying is vacuum drying until the moisture content is ≤0.5%, and the vacuum drying temperature is 25 to 30°C.

7. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises a therapeutically effective amount of the mononuclear monovalent gold dihalogen complex of claim 1 or 2, and a pharmaceutically acceptable carrier, diluent or excipient. The dosage forms of the pharmaceutical composition include injectable formulations, topical formulations, or mucosal formulations.

8. The pharmaceutical composition according to claim 7, characterized in that, The injectable formulation includes intravenous injection, subcutaneous injection, or intrathecal injection. The topical preparations include sprays, gels, creams, disinfectants, or wound dressings; The topical preparation can be impregnated and loaded onto a medical carrier for use, the medical carrier including a medical cotton swab; The mucosal preparations include nasal sprays, mouthwashes, or throat sprays.

9. The use of the mononuclear monovalent gold dihalogen complex of claim 1 or 2, or the pharmaceutical composition of claim 7 or 8, in the preparation of a medicament for the prevention and / or treatment of the following diseases: Diseases with pathological basis including calcium homeostasis disorder, abnormal cell signaling, abnormal protein aggregation, chronic inflammation and / or pathological calcification; The diseases mentioned include neurodegenerative diseases, metabolic diseases, malignant tumors, neurological dysfunction diseases, chronic inflammatory and fibrotic diseases, age-related degenerative changes, or diseases related to immunodeficiency and immune disorders.

10. The application according to claim 9, characterized in that, The neurodegenerative diseases mentioned include Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), Huntington's disease, or vascular cognitive impairment; The metabolic diseases mentioned include type 2 diabetes, insulin resistance, or disorders of glucose and lipid metabolism. The neurological disorders mentioned include myalgic encephalomyelitis / chronic fatigue syndrome (ME / CFS), chronic intractable pain, sleep disorders, or cognitive decline. The chronic inflammatory and fibrotic diseases mentioned include rheumatoid arthritis, tissue fibrosis, or organ calcification; The age-related degenerative changes include organ dysfunction, skin aging, or decline in cell signaling function; The immunodeficiency and immune dysregulation-related diseases include acquired immunodeficiency, virus infection-associated immune failure, primary / secondary immune cell dysfunction, or systemic lupus erythematosus.

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