Hydrophobic system of heme (II) derivative hexa-coordination compound as well as preparation method and application thereof

Abstract

The invention discloses a hydrophobic system of a heme (II) derivative hexa-coordination compound as well as a preparation method and application of the hydrophobic system. The structure of the hydrophobic system of the hexa-coordinated compound of the heme (II) derivative is # imgabs0. The hydrophobic system of the hexa-coordinated compound of the heme (II) derivative is prepared by carrying out coordination reaction on the heme (II) derivative, a ligand (strong ligand Ls) with strong central ion coordination capability and a ligand (weak ligand Lw) with weak coordination capability in a hydrophobic solvent; good biocompatibility is realized; oxygen can be efficiently and reversibly carried, the oxidation rate is slow, and an oxidation product is stable and cannot catalytically generate active oxygen; the artificial oxygen carrier can be used for treating ischemia and hypoxia diseases.

Description

Technical Field

[0001] The present invention relates to a hydrophobic system of a hexacoordinate compound of a heme (II) derivative, a preparation method and application thereof, belonging to the technical field of biomaterials. Background Art

[0002] Blood transfusion is a vital, life-saving clinical treatment. However, global blood supply is limited. According to data released by the World Health Organization (WHO), nearly 120 million units of blood are donated annually, yet this still falls short of global demand. Furthermore, with advances in medical technology and the widespread use of blood transfusions, the demand for blood is increasing. However, practical applications present numerous challenges, such as the short shelf life of blood, the requirement for cross-matching prior to transfusion, and the risk of transmitting hepatitis and acquired immunodeficiency syndrome. Consequently, efforts have been underway to find safe and effective artificial oxygen carriers to alleviate the pressure on blood supply.

[0003] Artificial oxygen carriers can be divided into three categories based on their raw materials: hemoglobin-based oxygen carriers (HBOCs), perfluorocarbon-based oxygen carriers (PFOCs), and porphyrin-based oxygen carriers (PBOCs). These are the most intensively studied oxygen carriers in this field. The deoxygenated forms of existing HBOCs and PBOCs are both iron(II) porphyrin pentacoordinates, with the sixth coordination site vacant for oxygen binding. In aqueous solution, iron(II) porphyrin is easily oxidized to iron(III) porphyrin. This product is unable to bind O2 and catalyzes the production of reactive oxygen species (ROS), triggering adverse reactions. This is an inherent toxicity that pentacoordinate iron(II) porphyrin cannot address. It is precisely this toxicity that prevents HBOCs and PBOCs from being administered in large doses, and they can only be used in small doses as oxygen therapy for various comorbidities. Summary of the Invention

[0004] The present invention provides a heme (II) derivative hexacoordination compound hydrophobic system, its preparation method and application. The heme (II) derivative hexacoordination compound hydrophobic system of the present invention can effectively carry oxygen and be used as an oxygen carrier for treating ischemic hypoxia diseases. During the preparation of the present invention, a solvent method is used to make the heme (II) derivatives react with the strong ligand L s , weak ligand L w The coordination reaction is carried out to generate a hexaligand of a heme (II) derivative with good biocompatibility. The hexaligand is dissolved in a hydrophobic solvent to obtain a heme (II) derivative hexaligand hydrophobic system.

[0005] In order to solve the above technical problems, the technical solutions adopted by the present invention are as follows:

[0006] A hydrophobic system of a hexacoordinate compound of a heme (II) derivative is characterized in that the hexacoordinate compound of the heme (II) derivative is soluble in a hydrophobic solvent.

[0007] The hydrophobic solvent is one or more of soybean oil for injection or medium chain triglyceride for injection.

[0008] The structure of the heme (II) derivative hexamer is:

[0009]

[0010] Including: heme (II) derivative parent component, the fifth coordination is a strong ligand L s The sixth coordination is a weak ligand L w .

[0011] The above-mentioned heme (II) derivative parent component has the structure:

[0012]

[0013] X1 represents an oxygen atom, a sulfur atom, a nitrogen atom, a hydrazine group (-NH-NH-), a diamine group One or more of wherein said n represents 0 to 20;

[0014] X2 represents an oxygen atom, a sulfur atom, a nitrogen atom, a hydrazine group (-NH-NH-), a diamine group One or more of wherein said n represents 0 to 20;

[0015] Wherein, the R1 represents C1~C 20 Alkyl, substituted C1~C 20 Alkyl, phenyl, substituted phenyl, C1~C 20 Acyl, substituted C1~C 20 One or more of acyl groups, polyether chains, substituted polyether chains, amides, polyamide chains, substituted polyamide chains, polyesters, and polyurethanes;

[0016] Wherein, the R2 represents C1~C 20 Alkyl, substituted C1~C 20 Alkyl, phenyl, substituted phenyl, C1~C 20 Acyl, substituted C1~C 20 One or more of acyl groups, polyether chains, substituted polyether chains, amides, polyamide chains, substituted polyamide chains, polyesters, and polyurethanes;

[0017] The strong ligand L S , its structure is: Fat-soluble imidazole derivatives Fat-soluble imidazole derivatives

[0018] or fat-soluble pyridine derivatives One or more of;

[0019] Wherein, the R3 is C1~C 20 one or more of alkyl or substituted alkyl;

[0020] Wherein, X3 represents an oxygen atom, a sulfur atom, a nitrogen atom, a hydrazine group (-NH-NH-), a diamine group One or more of wherein said n represents 0 to 20;

[0021] Wherein, the R4 represents C1~C 20 Alkyl, substituted C1~C 20 Alkyl, phenyl, substituted phenyl, C1~C 20 Acyl, substituted C1~C 20 One or more of acyl groups, polyether chains, substituted polyether chains, amides, polyamide chains, substituted polyamide chains, polyesters, and polyurethanes;

[0022] Wherein, the R5 represents C1~C 20 Alkyl, substituted C1~C 20 One or more of an alkyl group, a phenyl group, a substituted phenyl group, a polyether chain, or a substituted polyether chain;

[0023] X4 represents an oxygen atom, a sulfur atom, a nitrogen atom, a hydrazine group (-NH-NH-), a diamine group One or more of wherein said n represents 0 to 20;

[0024] The R6 represents C1~C 20 Alkyl, substituted C1~C 20 Alkyl, phenyl, substituted phenyl, C1~C 20 Acyl, substituted C1~C 20 One or more of acyl groups, polyether chains, substituted polyether chains, amides, polyamide chains, substituted polyamide chains, polyesters, and polyurethanes;

[0025] X5 represents an oxygen atom, a sulfur atom, a nitrogen atom, a hydrazine group (-NH-NH-), a diamine group One or more of wherein said n represents 0 to 20;

[0026] Wherein, the R7 represents C1~C 20 Alkyl, substituted C1~C20 Alkyl, phenyl, substituted phenyl, C1~C 20 Acyl, substituted C1~C 20 One or more of acyl groups, polyether chains, substituted polyether chains, amides, polyamide chains, substituted polyamide chains, polyesters, and polyurethanes;

[0027] The weak ligand L w , its structure is:

[0028] One or more of vitamin A, vitamin D, vitamin E, and cholesterol;

[0029] One or more of R5OH, wherein said R5 represents C 2-20 Alkyl, C 14 Alkyl, C 16 Alkyl, C 18 Alkyl or substituted alkyl, C6~C 15 Aryl, C6~C 15 One or more of the heterocyclic groups, preferably R5 is C 12 Alkyl, C 14 Alkyl, C 16 Alkyl, C 18 alkyl;

[0030] One or more of polyether, substituted polyether, polyester, and polyurethane;

[0031] The preparation method of the heme (II) derivative hexaligand is as follows: using hemin chloride as a raw material, a heme (II) derivative B is obtained by condensation reaction and reduction reaction, and the compound B is reacted with a strong ligand L s , weak ligand L w It is obtained by coordination reaction.

[0032] The hemin chloride and strong ligand L used in the present invention s , weak ligand L w All are biocompatible materials.

[0033] The preparation method of the above-mentioned hexameric heme (II) derivative is as follows:

[0034]

[0035] As a specific implementation scheme, the preparation method of the hexaligand of the heme (II) derivative is as follows: a solution of hemin chloride in an organic solvent is reacted with a nucleophilic reagent RXH under the action of a condensing agent to obtain compound A; compound A is reduced to compound B under the action of a reducing agent; compound B reacts with a strong ligand L s , weak ligand L wThe coordination reaction is carried out in a fat-soluble solvent to obtain the final product T.

[0036] The molar ratio of hemin chloride to RXH is 1:2.0-10.0, the reaction time is 6-120 h, and the reaction temperature is room temperature to 80° C., more preferably the molar ratio is 1:3, the reaction time is 72 h, and the reaction temperature is room temperature;

[0037] The organic solvent is one or more of N,N-dimethylformamide (DMF), acetonitrile or polyhalogenated alkane, preferably dichloromethane, chloroform or DMF;

[0038] The above heme (II) derivative B, and the strong ligand L s , weak ligand L w The molar ratio is 1:1.0-2.0:1.0-30.0, more preferably 1:1.1:8.0;

[0039] The fat-soluble solvent used is one or more of soybean oil for injection and medium-chain triglycerides for injection;

[0040] The hydrophobic system of the hexacoordinate compound of the heme (II) derivative can be used as an artificial oxygen carrier for treating ischemic and hypoxic diseases.

[0041] The hydrophobic system of the hexacoordinate compound of the heme (II) derivative can effectively carry oxygen and has a slow oxidation rate, and its oxidation product μ-oxo dimer does not catalyze the generation of reactive oxygen species.

[0042] The technologies not mentioned in this invention are all referred to the prior art.

[0043] The advantages of the present invention are:

[0044] In the hydrophobic system based on hexaligands of heme (II) derivatives of the present invention, the parent component is a heme (II) derivative, the fifth coordination is a strong ligand, and the sixth coordination is a weak ligand. The hexaligand can carry oxygen efficiently; its oxidation rate is slow, and its oxidation product is μ-oxo dimer, which does not catalyze the production of reactive oxygen species, fundamentally solving the inherent oxidative toxicity of the pentaligand; oxygen and weak ligands compete for central ions to achieve oxygenation and oxygen release, and the oxygenation curve is adjustable, with a wide range of applications. The hexaligand oxygen-carrying system of heme (II) derivatives in a hydrophobic environment has not been reported in the literature. This hydrophobic system can be prepared into different preparations for the treatment of ischemic and hypoxic diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] Figure 1 The UV spectrum of heme (II) didodecanol ester of the present invention;

[0046] Figure 2 The infrared spectrum of heme (II) didodecanol ester of the present invention;

[0047] Figure 3 LC-MS chart of heme (II) didodecanol ester of the present invention;

[0048] Figure 4 Oxygenation curve of heme (II) didodecanol ester hexaligand of the present invention;

[0049] Figure 5 The oxygen saturation of the hemoglobin (II) didodecanol ester hexaligand of the present invention changes in medium-chain triglycerides at 37° C. and air conditions for 0-6 hours. DETAILED DESCRIPTION

[0050] In order to better understand the present invention, the content of the present invention is further illustrated below in conjunction with the examples, but the content of the present invention is not limited to the following examples.

[0051] Example 1

[0052] Preparation of heme(II)-didodecanol ester hexaligand and its oxygenation and oxidation properties:

[0053] Hemin (400 mg, 0.610 mmol), dodecanol (457 mg, 2.440 mmol), and EDC·HCl (515 mg, 2.680 mmol) were mixed in 8 mL of dry dichloromethane and poured into a round-bottom flask. DMAP (16 mg, 0.130 mmol) was added under an ice-water bath. The reaction mixture was allowed to stand at room temperature for 24 h. After completion of the reaction, the mixture was washed with 0.1 M HCl, saturated NaHCO₃, and water. Drying was performed over MgSO₄, and the solvent was removed under reduced pressure. Separation was performed by column chromatography.

[0054] Dissolve 0.51 g of hemoglobin didodecanol ester (III) in 40 mL of deoxygenated dichloromethane. Add a saturated sodium dithionite solution and stir under nitrogen for 30 minutes. Wash the organic phase with water and separate again. Dry the organic layer over anhydrous sodium sulfate and evaporate it by rotary evaporation to obtain hemoglobin didodecanol ester (II).

[0055] At 37 ° C, hemoglobin didodecanol ester (II) (0.15 g, 1.55 × 10 -4mol) and 1-hexadecyl imidazole and 1-dodecanol are dissolved in medium-chain triglycerides and stirred. Repeatedly evacuate and pass nitrogen. Subsequently, adjust the system pressure to one atmosphere and record the reading as V3. Then, read the value every 5 minutes until the reading stabilizes, and record it as V4. The difference between the two readings ΔV2 is the total amount of gas absorbed by the complex solution. ΔV2 is composed of ΔV and the amount of gas dissolved in the DMF solvent ΔV1. Therefore, the oxygen binding capacity ΔV of hemoglobin didodecanol ester (II)-hexadecyl imidazole-dodecanol hexaplex is calculated by formula (1). According to the ideal gas equation, 1.55×10 -4 The maximum oxygen saturation of mol hemoglobin didodecanol ester (II)-hexadecyl imidazole-dodecanol hexaplex is 3.46mL.

[0056] ΔV=ΔV2-ΔV1 (1)

[0057] Although the present invention has been described in detail above using general descriptions and specific implementation plans, some modifications or improvements can be made on the basis of the present invention. These modifications or improvements made without departing from the core of the present invention are all within the scope of protection claimed by the present invention.

Claims

1. A hydrophobic system of a hexacoordinate compound of a heme (II) derivative, characterized by: The hexaligand of the heme(II) derivative is soluble in a hydrophobic solvent.

2. The hydrophobic solvent according to claim 1, wherein: Soybean oil for injection, one or more of medium chain triglycerides for injection.

3. The hexaplex of heme (II) derivative according to claim 1, characterized in that Its structure is: a heme (II) derivative parent component, a fifth coordination ligand (strong ligand L) with a strong coordination effect with Fe (II) ions s ), the sixth coordination is a ligand with weak coordination effect (weak ligand L w ).

4. The heme (II) derivative precursor component according to claim 3, characterized in that: Its structure is: Wherein, X1 represents an oxygen atom, a sulfur atom, a nitrogen atom, a hydrazine group (-NH-NH-), a diamine group One or more of wherein said n represents 0 to 20; Wherein, X2 represents an oxygen atom, a sulfur atom, a nitrogen atom, a hydrazine group (-NH-NH-), a diamine group One or more of wherein said n represents 0 to 20; Wherein, the R1 represents C1~C 20 Alkyl, substituted C1~C 20 Alkyl, phenyl, substituted phenyl, C1~C 20 Acyl, substituted C1~C 20 One or more of acyl groups, polyether chains, substituted polyether chains, amides, polyamide chains, substituted polyamide chains, polyesters, and polyurethanes; Wherein, the R2 represents C1~C 20 Alkyl, substituted C1~C 20 Alkyl, phenyl, substituted phenyl, C1~C 20 Acyl, substituted C1~C 20 One or more of acyl groups, polyether chains, substituted polyether chains, amides, polyamide chains, substituted polyamide chains, polyesters, and polyurethanes; 5. The strong ligand L as claimed in claim 3 S , characterized by: Its structure is: fat-soluble imidazole derivative or fat-soluble pyridine derivatives One or more of; Wherein, the R3 is C1~C 20 one or more of alkyl or substituted alkyl; Wherein, X3 represents an oxygen atom, a sulfur atom, a nitrogen atom, a hydrazine group (-NH-NH-), a diamine group One or more of wherein said n represents 0 to 20; Wherein, the R4 represents C1~C 20 Alkyl, substituted C1~C 20 Alkyl, phenyl, substituted phenyl, C1~C 20 Acyl, substituted C1~C 20 One or more of acyl groups, polyether chains, substituted polyether chains, amides, polyamide chains, substituted polyamide chains, polyesters, and polyurethanes; Wherein, the R5 represents C1~C 20 Alkyl, substituted C1~C 20 One or more of an alkyl group, a phenyl group, a substituted phenyl group, a polyether chain, or a substituted polyether chain; Wherein, X4 represents an oxygen atom, a sulfur atom, a nitrogen atom, a hydrazine group (-NH-NH-), a diamine group One or more of wherein said n represents 0 to 20; Wherein, the R6 represents C1~C 20 Alkyl, substituted C1~C 20 Alkyl, phenyl, substituted phenyl, C1~C 20 Acyl, substituted C1~C 20 One or more of acyl groups, polyether chains, substituted polyether chains, amides, polyamide chains, substituted polyamide chains, polyesters, and polyurethanes; Wherein, X5 represents an oxygen atom, a sulfur atom, a nitrogen atom, a hydrazine group (-NH-NH-), a diamine group One or more of wherein said n represents 0 to 20; Wherein, the R7 represents C1~C 20 Alkyl, substituted C1~C 20 Alkyl, phenyl, substituted phenyl, C1~C 20 Acyl, substituted C1~C 20 One or more of acyl groups, polyether chains, substituted polyether chains, amides, polyamide chains, substituted polyamide chains, polyesters, and polyurethanes; 6. The weak ligand L according to claim 3 w ,characterized in that, Its structure is: One or more of vitamin A, vitamin D, vitamin E, and cholesterol; One or more of R5OH, wherein said R5 represents C 12 Alkyl, C 14 Alkyl, C 16 Alkyl, C 18 Alkyl or substituted alkyl, C6~C 15 Aryl, C6~C 15 One or more heterocyclic groups; One or more of polyether, substituted polyether, polyester, and polyurethane; 7. The hexaplex of a heme (II) derivative according to claim 3, wherein: Heme (II) derivative parent component, strong ligand L s , weak ligand L w The molar ratio is 1:1.0~2.0:1.0~30.0; 8. Use of the hydrophobic system of a hexacoordinate compound of a heme (II) derivative as claimed in claim 1, characterized in that: Artificial oxygen carriers for the treatment of ischemic and hypoxic diseases; 9. Use of the hydrophobic system of the hexacoordinate compound of the heme (II) derivative according to claim 8, characterized in that: Oxidation rate is slow; 10. Use of the hydrophobic system of the hexacoordinate compound of the heme (II) derivative according to claim 8, characterized in that: Its oxidation product, μ-oxo dimer, does not catalyze the generation of reactive oxygen species (ROS).

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