Cuprous-responsive copper dithiocarbamate complex prodrug compound as well as preparation method and application thereof

Abstract

The invention belongs to the field of prodrug compounds, and particularly relates to a prodrug compound of a cuprous-responsive copper dithiocarbamate complex as well as a preparation method and application of the prodrug compound. According to the compound disclosed by the invention, after the organic ligand in the structure is coordinated with the cuprous, an oxidation bond breaking reaction catalyzed by metal ions can be carried out in the presence of oxygen, and the cuprous is converted into cuprous along with the conversion of the cuprous into the cuprous. Then, dithiocarbamic acid formed by breakage can be competitively combined with bivalent copper ions, so that the dithiocarbamic acid copper complex with high biological activity is converted. The design can play a role in related indications such as tumor prevention and treatment, metal ion metabolism abnormality and the like.

Description

Technical Field

[0001] The invention belongs to the field of pharmaceutical technology and relates to a monovalent copper-responsive copper dithiocarbamate complex prodrug compound and a preparation method and application thereof. Background Art

[0002] Traditional drug small molecules lack selectivity in killing cells, and the emergence of the concept of prodrugs allows drug molecules to maintain their inertness when circulating in the body, and transform into active drugs in a special microenvironment in the body to exert their effects. Studies have shown that in tumor diseases and metal metabolism-related diseases, abnormal metal ion concentrations are often accompanied. Accordingly, metal coordination compounds have great application potential in these disease areas. Since metal coordination compounds are usually composed of metal ions and organic ligands, exogenous metal ions are often artificially introduced during drug design. Metal ions and ligands are delivered simultaneously through metal complex prodrugs to achieve the spatiotemporal unity of the two and stable combination, thereby enhancing the efficacy of the drug. However, additional supplementation of exogenous metal ions may destroy the physiological activity of proteins, generate reactive oxygen species, cause oxidative stress and cell damage, and cause metabolic problems. Therefore, considering the inherent toxicity of transition metals, their inclusion in drug therapy must be carried out in minimal amounts.

[0003] Metal ion catalysis refers to the process in which metal ions act as catalysts in chemical reactions, forming complexes with reactants, changing the electronic structure and geometric configuration of the reactant molecules, and thus promoting the breaking and formation of chemical bonds. In most cases, metal complex prodrug molecules need to release ligands through enzyme catalysis or in response to tumor microenvironment characteristics, and then the ligands will seek metal ions to form active drugs. Metal-catalyzed prodrugs can chelate endogenous metal ions after entering the human body, produce metal-catalyzed cleavage reactions, break off ligand molecules, form metal complexes at specific locations, and exert drug efficacy. This strategy can avoid the introduction of exogenous metal ions while exerting the efficacy of metal complex drugs, thereby reducing the harm of metal ions to the human body. Therefore, metal-catalyzed drug activation strategies are of great significance. Summary of the invention

[0004] In order to overcome the shortcomings of the prior art, the present invention provides a prodrug compound of a copper dithiocarbamate complex responsive to monovalent copper. The compound is composed of a monovalent copper ligand and dithiocarbamate, and does not require the addition of additional metal ions, thereby avoiding the potential harm of metal ions to the human body; if and only if the compound is responsive to Cu + After coordination, in the presence of oxygen, metal ion-catalyzed oxidative bond breaking reaction can be performed to release the dithiocarbamic acid prototype. + Cu 2+ The free dithiocarbamate molecule formed is Cu2+ A strong chelating agent that can bind Cu 2+ Forms copper dithiocarbamate complexes with antitumor activity.

[0005] Purpose of the Invention

[0006] The main purpose of the present invention is to provide a monovalent copper-responsive copper dithiocarbamate complex prodrug compound and a preparation method and application thereof.

[0007] Technical Solution

[0008] A monovalent copper-responsive copper dithiocarbamate complex prodrug compound, characterized in that it is a compound represented by general formula I:

[0009] General formula I:

[0010] in, represents a dithiocarbamate group,

[0011] R1 and R2 can be C1-C 10 Alkyl, C2-C 10 Alkenyl, C2-C 10 Alkynyl, C1-C6 alkyl optionally substituted by one or more hydroxyl, amino, halogen, cyano, nitro, sulfonyl, aromatic ring, aromatic heterocycle and condensed ring aromatic group, or may be combined to form C3-C7 cycloalkyl, C3-C7 heterocycloalkyl, C6-C 10 Aryl, 5-10 membered heteroaryl, condensed ring aromatic group, C6-C 10 Heterocyclic group;

[0012] More preferably, they are the same or different C1-C6 alkyl groups independently of each other, or they may be combined to form a C3-C7 cycloalkyl group.

[0013] In some specific embodiments of the present invention, R1 and R2 are preferably two independent diethyl groups, which are called diethyldithiocarbamate groups.

[0014] It is a self-fragmenting structure; R3 can be N, O, S; R4 can be -H or -NO2;

[0015] In some specific embodiments of the present invention, R3 is preferably O, and R4 is preferably H, that is, the self-fragmenting structure is preferably the structure: .

[0016] Represents an organic ligand that recognizes metal ions, generally a multidentate ligand containing N, S, O coordination groups, X is C, N; R5, R6 can be two identical or different coordination groups containing N, S, O, or one is a multidentate coordination group and the other is hydrogen;

[0017] The R5 and R6 may include but are not limited to the following structures: , , , ;

[0018] In some specific embodiments of the present invention, R5 and R6 are preferably , that is, the organic ligand that recognizes the metal ion is preferably of the structure: .

[0019] Cu + With organic ligand After coordination occurs, in the presence of O2, the molecular structure of Formula I undergoes oxidative cleavage, initiating a 1,6-elimination reaction of the self-fragmented structure, releasing free dithiocarbamate groups, accompanied by Cu + Cu 2+ The released dithiocarbamate competes for the Cu 2+ A metal coordination reaction occurs to form a copper dithiocarbamate complex with high cytotoxicity, the general structural formula of which is: .

[0020] Compared with the prior art, the present invention has the following beneficial results:

[0021] The monovalent copper-responsive copper dithiocarbamate complex prodrug compound provided by the present invention is composed of a monovalent copper ligand and dithiocarbamate, and does not require the addition of additional metal ions, thereby avoiding the potential harm of metal ions to the human body; if and only if the compound is responsive to Cu + After coordination, in the presence of oxygen, metal ion-catalyzed oxidative bond breaking reaction can be performed to release the dithiocarbamic acid prototype. + Cu 2+ The free dithiocarbamate molecule formed is Cu 2+ It is a strong chelating agent that can form metal complexes with copper ions with antitumor activity.

[0022] Compared with traditional copper dithiocarbamate complex prodrug compounds, the prodrug compound provided by the present invention does not carry external copper ions, can recognize and chelate endogenous monovalent copper, and is accompanied by oxidative cleavage to achieve recovery of drug activity, and has good clinical practicality.

[0023] The prodrug compound disclosed in the present invention can utilize endogenous monovalent copper to achieve site-specific activation, thereby avoiding the potential toxicity caused by the introduction of exogenous metal ions, and has good application prospects in related indications such as tumor prevention and treatment and abnormal metal ion metabolism. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] The specific embodiments of the present invention are further described in detail below with reference to the accompanying drawings, wherein:

[0025] Figure 1 Schematic diagram of the structure of TZD in Example 1.

[0026] Figure 2 is the mass spectrum of TZD in Example 1.

[0027] Figure 3 It is the nuclear magnetic hydrogen spectrum of TZD in Example 1.

[0028] Figure 4 It is the NMR carbon spectrum of TZD in Example 1.

[0029] Figure 5 The sample was analyzed by high performance liquid chromatography for the generation of Cu(DTC)2 after TZD was co-incubated with monovalent copper (CuCl) in Example 2. It shows that when TZD is co-incubated with divalent copper (CuCl2), no oxidative bond breaking occurs, and no Cu(DTC)2 is generated; after TZD is co-incubated with monovalent copper (CuCl), oxidative bond breaking occurs, accompanied by the generation of Cu(DTC)2, and the longer the incubation time, the more significant the bond breaking.

[0030] Figure 6 The TZD response to Cu in Example 1 + Schematic diagram of the possible mechanism of the formation of Cu(DTC)2. DETAILED DESCRIPTION

[0031] The preferred embodiments are explained in conjunction with the accompanying drawings. It should be understood that the preferred embodiments described herein are only used to illustrate and explain the present invention, and are not used to limit the present invention. For example, the concentration and volume of the solution used can be adjusted as needed.

[0032] Example 1: Preparation of the prodrug compound TZD of the monovalent copper-responsive diethyldithiocarbamate copper complex

[0033] (1) The preparation process of TZD compound is as follows:

[0034]

[0035] 1) Preparation of intermediate compound 1: 2,6-pyridine dimethanol (5 g, 35.93 mmol) was dissolved in 48% HBr (50 mL) and heated to reflux at 105 °C for 15 h. After the reaction was completed, the pH of the reaction system was adjusted to 12-14 with 4 M NaOH solution, then extracted with ethyl acetate, washed with saturated brine and pure water, and the organic phase was collected, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 4:1).

[0036] 2) Preparation of intermediate compound 2: Intermediate compound 1 (1.79 g, 6.76 mmol, 1 eq), p-hydroxybenzyl alcohol (0.92 g, 7.41 mmol, 1.1 eq) and anhydrous potassium carbonate (0.93 g, 6.73 mmol, 1 eq) were dissolved in 30 mL of acetone and heated under reflux at 70 °C for 10 h. After the reaction was completed, the mixture was filtered, and the filtrate was concentrated and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1.5:1) to obtain the product.

[0037] 3) Preparation of intermediate compound 3: Intermediate compound 2 (1.08 g, 3.5 mmol, 1 eq), 2,2'-dipicolylamine (0.628 g, 3.15 mmol, 0.9 eq) and anhydrous potassium carbonate (0.483 g, 3.49 mmol, 1 eq) were dissolved in 20 mL of anhydrous acetonitrile, and stirred at room temperature overnight under nitrogen protection. After the reaction was completed, the mixture was filtered, and the filtrate was concentrated and purified by silica gel column chromatography (dichloromethane / methanol = 13:1) to obtain the product.

[0038] 4) Preparation of intermediate compound 4: Intermediate compound 3 (380 mg, 0.89 mmol, 1 eq) was dissolved in 10 mL of thionyl chloride, 3 drops of N,N-dimethylformamide were added as a catalyst, and the mixture was heated under reflux at 65 °C overnight under nitrogen protection. After the reaction was completed, the mixture was dried and used directly in the next step.

[0039] 5) Preparation of TZD: Intermediate compound 4 (250 mg, 0.56 mmol, 1 eq) and sodium diethyldithiocarbamate (192 mg, 1.12 mmol, 2 eq) were dissolved in acetonitrile, and the sodium diethyldithiocarbamate solution was slowly dripped into the solution of intermediate compound 4 and stirred at room temperature for 4 h. After the reaction was completed, the white insoluble matter was removed by filtration, the filtrate was collected and concentrated, redissolved in dichloromethane, extracted with water, and the organic phase was collected and dried over anhydrous sodium sulfate. The filtrate was dried by filtration and then purified by silica gel column chromatography (dichloromethane / methanol = 12:1).

[0040] Example 2: TZD and Cu + Investigation of Cu(DTC)2 generation after co-incubation

[0041] The kinetics of TZD and Cu + Cu(DTC)2 generation after co-incubation. TZD and CuCl were dissolved in acetonitrile, CuCl2 was dissolved in pure water, TZD and copper ions were fully mixed at a molar ratio of 1:2, and the samples were injected and analyzed at different time points to examine the changes in the characteristic absorption peak of Cu(DTC)2 at 269nm. At the same time, TZD solution was set as a negative control group and Cu(DTC)2 solution was set as a positive control group to assist in the analysis.

Claims

1. A copper dithiocarbamate complex prodrug compound responsive to monovalent copper, characterized in that: It is a compound shown in general formula I: General formula I: in, represents a dithiocarbamic acid group, R1 and R2 can be independently C1-C 10 Alkyl, C2-C 10 Alkenyl, C2-C 10 Alkynyl, C1-C6 alkyl optionally substituted by one or more hydroxyl, amino, halogen, cyano, nitro, sulfonyl, aromatic ring, aromatic heterocycle and condensed ring aromatic group, or may be combined to form C3-C7 cycloalkyl, C3-C7 heterocycloalkyl, C6-C 10 Aryl, 5-10 membered heteroaryl, condensed ring aromatic group, C6-C 10 Heterocyclic group; It is a self-fragmenting structure; R3 can be N, O, S; R4 can be -H or -NO2; It represents an organic ligand that recognizes metal ions, which is generally a multidentate ligand containing N, S, and O coordination groups, and X is C or N; R5 and R6 can be two identical or different coordination groups containing N, S, and O, or one can be a multidentate coordination group and the other can be hydrogen.

2. The prodrug compound of the monovalent copper-responsive copper dithiocarbamate complex according to claim 1, characterized in that: R1 and R2 in the general formula I are preferably the following groups: they are the same or different C1-C6 alkyl groups independently of each other, or they may be combined to form a C3-C7 cycloalkyl group.

3. The prodrug compound of the monovalent copper-responsive copper dithiocarbamate complex according to claim 1, characterized in that: The R5 and R6 may include but are not limited to the following structures: 、 、 、 。 4. The prodrug compound of the monovalent copper-responsive dithiocarbamate copper complex according to claim 2, characterized in that: The R1 and R2 are preferably: the same ethyl group which is independent of each other, or combined to form a C3-C7 cycloalkyl group.

5. The prodrug compound of the monovalent copper responsive dithiocarbamate copper complex according to claim 1, characterized in that: The R3 is preferably O; R4 is preferably H.

6. The prodrug compound of the monovalent copper-responsive copper dithiocarbamate complex according to claim 3, characterized in that: The R5 and R6 are preferably .

7. The prodrug compound of the monovalent copper-responsive copper dithiocarbamate complex according to claims 1-6, characterized in that: Cu + With organic ligand After coordination occurs, in the presence of O2, the molecular structure of Formula I undergoes oxidative cleavage, initiating a 1,6-elimination reaction of the self-fragmented structure, releasing free dithiocarbamate groups, accompanied by Cu + Cu 2+ The released dithiocarbamate competes for the Cu 2+ A metal coordination reaction occurs to form a copper dithiocarbamate complex that is highly cytotoxic.

8. The prodrug compound of the monovalent copper-responsive copper dithiocarbamate complex according to claims 1-6, characterized in that: The compound, its optical isomer and pharmaceutically acceptable salt or solvate, and application thereof in preventing and / or treating tumor diseases.

9. The prodrug compound of the monovalent copper-responsive copper dithiocarbamate complex according to claims 1-6, characterized in that: The invention relates to a pharmaceutical composition formed by using the compound, its optical isomers and pharmaceutically acceptable salts or solvates thereof as active drugs and pharmaceutically acceptable excipients, and the use thereof in preventing and / or treating tumor diseases.

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