A deferoxamine conjugate, its preparation method and application

Abstract

A deferoxamine conjugate as well as a preparation method and application thereof belong to the technical field of medicines and specifically relate to a 2-sulfo-9-fluorenylmethoxycarbonyl-deferoxamine conjugate as well as a preparation method and application thereof. On one hand, the 2-sulfo-9-fluorenylmethoxycarbonyl-deferoxamine conjugate solves the problems of high toxicity, short half-life and high frequency of administration of deferoxamine itself, thereby improving patient medication compliance and reducing drug-related toxic side effects. On the other hand, the present application provides a drug combination containing the 2-sulfo-9-fluorenylmethoxycarbonyl-deferoxamine conjugate and application thereof in preparation of a medicine for treating iron overload and a medicine for treating tumors.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of medicine, in particular to 2-sulfo-9-fluorenylmethoxycarbonyl-desferrioxamine conjugate and its application. BACKGROUND

[0002] Desferrioxamine (DFO) as a metal ion complexing agent can form a stable desferrioxamine-iron complex with free iron ions through hydroxamic acid groups, and finally be excreted by urine. Desferrioxamine can bind to free and unstable iron pools in the body, and treat acute or chronic iron overload by removing excess iron in the body. In recent years, desferrioxamine has also been applied to many clinical trials, such as subarachnoid hemorrhage, diabetic foot ulcer, triple-negative breast cancer, etc., and has great potential in future clinical applications.

[0003] Compared with other FDA-approved iron chelators, desferrioxamine is the only FDA-approved iron chelator for parenteral administration, and its administration method is suitable for more general clinical cases. Desferrioxamine still has many drawbacks in clinical administration. First, the half-life of desferrioxamine is very short (about 5.5 min) and the dosage is large (30-60 mg / kg / day), and patients need to be continuously injected subcutaneously for 8-12 hours at a time, and at least five days a week, which leads to poor patient compliance. Second, desferrioxamine itself has hepatotoxicity, and high-dose administration and high administration frequency can cause some toxic side effects to the respiratory system, auditory system and cardiovascular system of patients. Therefore, methods to reduce the toxicity of desferrioxamine itself, prolong the half-life of desferrioxamine, and reduce the administration frequency of desferrioxamine are expected to improve patient compliance and provide more valuable possibilities for clinical applications.

[0004] Most of the previous studies use high-molecular-weight water-soluble polymers to synthesize conjugates with desferrioxamine. This conjugate escapes renal clearance to reduce the excretion rate of the conjugate, thereby prolonging the half-life of desferrioxamine. However, iron overload patients in clinical practice often have impaired liver function, which hinders the hepatobiliary excretion of desferrioxamine-iron ion complexes. In addition, such high-molecular-weight conjugates are difficult to freeze-dry and fill in industrial production. Moreover, high-molecular-weight conjugates increase the viscosity of desferrioxamine clinical injection, which hinders the application of the drug in subcutaneous injection. Currently, high-molecular-weight desferrioxamine conjugates have no clinical trial applications. SUMMARY

[0005] The present application aims to provide a 2-sulfo-9-fluorenylmethoxycarbonyl-desferrioxamine conjugate and its application to solve the problems of high toxicity, short half-life, and high administration frequency of desferrioxamine mentioned in the background art, thereby improving the pharmacokinetics and pharmacodynamics of desferrioxamine and improving patient compliance and reducing drug-related side effects.

[0006] In order to achieve the above-mentioned purpose, the application provides a desferrioxamine conjugate which is prepared by connecting 2-sulfo-9-fluorenylmethoxycarbonyl-N-succinimidyl carbonate and desferrioxamine molecules through an amide bond; the 2-sulfo-9-fluorenylmethoxycarbonyl-N-succinimidyl carbonate is prepared by sulfonating 9-fluorenylmethoxycarbonyl-N-succinimidyl carbonate; and the structure of the desferrioxamine conjugate is as follows:

[0007]

[0008] The 9-fluorenylmethoxycarbonyl-N-succinimidyl carbonate has the following structure:

[0009]

[0010] The 2-sulfo-9-fluorenylmethoxycarbonyl-N-succinimidyl carbonate has the following structure:

[0011]

[0012] The desferrioxamine has the following structure:

[0013]

[0014] The application provides a synthesis method of the 2-sulfo-9-fluorenylmethoxycarbonyl-desferrioxamine conjugate, and the specific method is as follows: 9-fluorenylmethoxycarbonyl-N-succinimidyl carbonate and chlorosulfonic acid are dissolved in dichloromethane to react, and after the reaction is completed, purification is performed, and then freeze-drying is performed after purification to prepare 2-sulfo-9-fluorenylmethoxycarbonyl-N-succinimidyl carbonate; subsequently, 2-sulfo-9-fluorenylmethoxycarbonyl-N-succinimidyl carbonate and desferrioxamine are dissolved in a solvent to react under the action of DIPEA, and after the reaction is completed, purification is performed, and then freeze-drying is performed after purification to obtain the final product 2-sulfo-9-fluorenylmethoxycarbonyl-desferrioxamine conjugate, and the synthesis route is as shown in the following:

[0015]

[0016] Preferably, the reaction solvent is anhydrous N, N-dimethylformamide.

[0017] Preferably, the 2-sulfo-9-fluorenylmethoxycarbonyl-desferrioxamine conjugate obtained by the reaction is purified by preparative liquid phase to obtain a conjugate with a molecular weight of 861.37.

[0018] Preferably, the in-vitro iron chelating ability and cytotoxicity of the 2-sulfo-9-fluorenylmethoxycarbonyl-desferrioxamine conjugate and desferrioxamine are compared.

[0019] Preferably, the half-lives of the 2-sulfo-9-fluorenylmethoxycarbonyl-desferrioxamine conjugate and desferrioxamine in vivo are compared.

[0020] Preferably, the in vivo toxicity of the 2-sulfo-9-fluorenylmethyloxycarbonyl-desferrioxamine conjugate compared to desferrioxamine;

[0021] The present application provides the use of 2-sulfo-9-fluorenylmethyloxycarbonyl-desferrioxamine conjugate in the preparation of a medicament for treating iron overload diseases;

[0022] The present application provides the use of 2-sulfo-9-fluorenylmethyloxycarbonyl-desferrioxamine conjugate in the preparation of a medicament for treating tumors.

[0023] The present application has the following beneficial effects:

[0024] The present application provides a 2-sulfo-9-fluorenylmethyloxycarbonyl-desferrioxamine conjugate which has lower toxicity than desferrioxamine at the same concentration; has a longer half-life; has stronger ability to remove excess iron in the plasma; can treat iron overload at a low administration frequency; and has better attenuation effect. The present application provides a drug combination of 2-sulfo-9-fluorenylmethyloxycarbonyl-desferrioxamine conjugate and 5-aminolevulinic acid, which can improve the diagnosis and treatment effect of tumors. BRIEF DESCRIPTION OF DRAWINGS

[0025] Figure 1 NMR spectra of desferrioxamine, 2-sulfo-9-fluorenylmethyloxycarbonyl-N-succinimidyl carbonate, and 2-sulfo-9-fluorenylmethyloxycarbonyl-desferrioxamine conjugate in the examples;

[0026] Figure 2 Mass spectra of desferrioxamine, 2-sulfo-9-fluorenylmethyloxycarbonyl-N-succinimidyl carbonate, and 2-sulfo-9-fluorenylmethyloxycarbonyl-desferrioxamine conjugate in the examples;

[0027] Figure 3 Iron ion chelating ability graphs of 2-sulfo-9-fluorenylmethyloxycarbonyl-desferrioxamine conjugate (FMS-DFO) and DFO in the examples;

[0028] Figure 4 24h cytotoxicity MTT experiment results of 2-sulfo-9-fluorenylmethyloxycarbonyl-desferrioxamine conjugate (FMS-DFO) and DFO in the examples;

[0029] Figure 5 48h cytotoxicity MTT experiment results of 2-sulfo-9-fluorenylmethyloxycarbonyl-desferrioxamine conjugate (FMS-DFO) and DFO in the examples;

[0030] Figure 6 72h cytotoxicity MTT experiment results of 2-sulfo-9-fluorenylmethyloxycarbonyl-desferrioxamine conjugate (FMS-DFO) and DFO in the examples;

[0031] Figure 7 Pharmacokinetic results of 2-sulfo-9-fluorenylmethoxycarbonyl-deferoxamine conjugate (FMS-DFO) in the examples;

[0032] Figure 8 The pharmacokinetic results of the 2-sulfo-9-fluorenylmethoxycarbonyl-deferoxamine conjugate (FMS-DFO) and DFO in the examples are compared;

[0033] Figure 9 The pharmacodynamic iron clearance test results of five administrations of 2-sulfo-9-fluorenylmethoxycarbonyl-deferoxamine conjugate (FMS-DFO) and DFO in the example;

[0034] Figure 10 The pharmacodynamic results of the serum ferritin concentration of the 2-sulfo-9-fluorenylmethoxycarbonyl-deferoxamine conjugate (FMS-DFO) in the example are as follows:

[0035] Figure 11 The pharmacodynamic results of serum iron content of 2-sulfo-9-fluorenylmethoxycarbonyl-deferoxamine conjugate (FMS-DFO) after three doses in the example are shown;

[0036] Figure 12 The pharmacodynamic antioxidant test results of five doses of 2-sulfo-9-fluorenylmethoxycarbonyl-deferoxamine conjugate (FMS-DFO) and DFO in the example;

[0037] Figure 13 The graph shows the experimental results of liver tissue pathological manifestations of 2-sulfo-9-fluorenylmethoxycarbonyl-deferoxamine conjugate (FMS-DFO) and DFO in the examples.

[0038] Figure 14 The figures show the experimental results of the embodiment of 2-sulfo-9-fluorenylmethoxycarbonyl-deferoxamine conjugate (FMS-DFO) and DFO for enhancing the intratumoral accumulation of protoporphyrin IX.

[0039] Figure 15 These are the experimental results of the photodynamic therapy for cancer using 2-sulfo-9-fluorenylmethoxycarbonyl-deferoxamine conjugate (FMS-DFO) and DFO-enhanced 5-aminolevulinic acid. DETAILED DESCRIPTION

[0040] The technical solutions of the present application will be clearly and completely described below in combination with the embodiments of the present application. Obviously, the described embodiments are only some of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative work fall within the scope of the present application.

[0041] Example 1: Synthesis of 2-sulfo-9-fluorenylmethoxycarbonyl-desferrioxamine conjugate

[0042] 9-Fluorenylmethoxycarbonyl-N-succinimidyl carbonate (337.4 mg, 1 mmol) was weighed into 4 mL of dichloromethane (CH2Cl2) and reduced to 0°C, 0.03% chlorosulfonic acid CH2Cl2solution was added dropwise, after 15 min the reaction system was transferred to 25°C for further reaction, after 1 h of reaction at room temperature, the precipitate was centrifuged. The precipitate was washed 4 times in a cyclohexane: dichloromethane = 1:1 mixed solution, the precipitate was dissolved in water and freeze-dried to obtain 2-sulfo-9-fluorenylmethoxycarbonyl-N-succinimidyl carbonate.

[0043] DFO (26.2716 mg, 0.04 mmol) was weighed into 7 mL of N,N-dimethylformamide (DMF), heated and stirred at 55°C until DFO was completely dissolved, then 2-sulfo-9-fluorenylmethoxycarbonyl-N-succinimidyl carbonate (34.4 mg, 0.04 mmol) was dissolved in 1 mL of DMF, added to the DFO solution, and finally 20 μL of DIPEA was added to adjust the reaction system to weak alkaline, after 4 h of reaction at 25°C, the reaction solution was rotary evaporated under reduced pressure. After determining the generation of the new compound by high performance liquid chromatography (HPLC), the reaction mixture was separated and purified by preparative liquid chromatography, and finally freeze-dried to obtain 2-sulfo-9-fluorenylmethoxycarbonyl-desferrioxamine conjugate. First, nuclear magnetic resonance hydrogen spectrum was used to confirm the structure of the compound in Example 1, and the results are shown in Figure 1 The H at 3, 12, 17, 19; 5, 9; 4, 11, 16 in 2-sulfo-9-fluorenylmethoxycarbonyl-desferrioxamine conjugate correspond to the H at bdqr; ip; cjq in the desferrioxamine molecule, respectively, in addition, the H at 23-26 corresponds to the H at E-H in the 2-sulfo-9-fluorenylmethoxycarbonyl-succinimidyl structure, respectively. The H at A, B only exists in the 2-sulfo-9-fluorenylmethoxycarbonyl-succinimidyl structure and does not exist in the 2-sulfo-9-fluorenylmethoxycarbonyl-desferrioxamine conjugate, proving that the succinimidyl structure and the amino group of desferrioxamine are successfully synthesized in the experiment. Thereafter, as shown in Figure 2As shown, the conjugate with a molecular weight of 861.37 was tested by mass spectrometry, confirming that the 2-sulfo-9-fluorenylmethyloxycarbonyl-desferrioxamine conjugate was successfully synthesized.

[0044] Example 2: Test of iron chelating ability of 2-sulfo-9-fluorenylmethyloxycarbonyl-desferrioxamine conjugate

[0045] (6.56 mg 0.01 mmol) of desferrioxamine and (8.61 mg 0.01 mmol) of 2-sulfo-9-fluorenylmethyloxycarbonyl-desferrioxamine conjugate (the conjugate and the free drug of desferrioxamine have the same amount of desferrioxamine) were respectively dissolved in 10 ml of distilled water, and the two solutions were mixed with the same amount of 10 mM ferrous ammonium sulfate solution and incubated for 30 min. After dilution four times, the absorbance was measured at a wavelength of 430 nm by UV-visible spectrophotometry to investigate the iron ion chelating ability. The results are shown in Figure 3 As shown, the characteristic absorption peak of desferrioxamine chelating iron ions is at 430 nm, and the absorption peak of 2-sulfo-9-fluorenylmethyloxycarbonyl-desferrioxamine conjugate chelating iron ions is also at 430 nm. Therefore, the 2-sulfo-9-fluorenylmethyloxycarbonyl-desferrioxamine conjugate synthesized in the present application has the same iron chelating ability as the same molar concentration of desferrioxamine.

[0046] Example 3: Cytotoxicity test of 2-sulfo-9-fluorenylmethyloxycarbonyl-desferrioxamine conjugate

[0047] The cytotoxicity was detected by MTT method, and the samples used for detection were desferrioxamine and 2-sulfo-9-fluorenylmethyloxycarbonyl-desferrioxamine conjugate. The above samples were respectively dissolved in DMEM medium to prepare a solution with an equivalent concentration of 1 mM of desferrioxamine, which was used in the following cytotoxicity experiment. When used, the DMEM complete medium was diluted to a solution with an equivalent concentration of 750 μM, 500 μM, 250 μM, 200 μM, 125 μM, 100 μM, 50 μM, and 10 μM of desferrioxamine. The control group used the same volume of DMEM medium. RAW246.7 mouse mononuclear macrophages were inoculated in a 96-well plate at a density of 5 x 10 3 Each well, and the culture medium was discarded after 12 h of culture and replaced with the above DMEM complete medium containing different concentrations of desferrioxamine and 2-sulfo-9-fluorenylmethyloxycarbonyl-desferrioxamine conjugate. At 24 h, 48 h, and 72 h of culture, the cytotoxicity was detected by MTT method. The results of cytotoxicity MTT test are shown in Figure 4 、 Figure 5 and Figure 6 As can be seen from the figure, in all time ranges, the toxicity of 2-sulfo-9-fluorenylmethyloxycarbonyl-desferrioxamine conjugate is significantly lower than that of desferrioxamine at the same concentration. This indicates that after forming the 2-sulfo-9-fluorenylmethyloxycarbonyl-desferrioxamine conjugate, the toxicity of desferrioxamine is significantly reduced.

[0048] Example 4: Pharmacokinetics test of 2-sulfo-9-fluorenylmethyloxycarbonyl- desferrioxamine conjugate

[0049] Take 10 male healthy rats, weighing 180-200g, randomly divided into 2 groups, 5 in each group, according to the equivalent dose of desferrioxamine 50mg / kg, desferrioxamine and 2-sulfo-9-fluorenylmethyloxycarbonyl-desferrioxamine conjugate were weighed and dissolved in appropriate amount of normal saline, through the tail vein injection, according to the time of orbital blood, centrifugal to obtain plasma, by high performance liquid chromatograph to determine the concentration of drug in plasma. The pharmacokinetics test results are shown in Figure 7 and Figure 8 The pharmacokinetics research parameters are shown in Table 1; from Figure 7 and Figure 8 It can be seen that within 30min after the tail vein injection, free desferrioxamine has been completely metabolized, while 2-sulfo-9-fluorenylmethyloxycarbonyl-desferrioxamine conjugate can be rapidly combined with HSA after entering the body, and can be completely cleared in the plasma after 8h circulation, at the same time, the amide bond will be broken, gradually releasing free DFO for 24h, which shows that 2-sulfo-9-fluorenylmethyloxycarbonyl-desferrioxamine conjugate obtained by structural modification of DFO can successfully prolong the half-life of DFO in vivo.

[0050] Table 1 Pharmacokinetics parameters

[0051]

[0052]

[0053] Example 5: Test of 2-sulfo-9-fluorenylmethyloxycarbonyl-desferrioxamine conjugate in treating iron overload

[0054] Healthy BALB / c female mice were selected and housed in separate cages. The mice were weighed the day before modeling. A dextran iron solution (Dextran / Fe, normal saline) with an iron content of 35% was prepared. After filtering and sterilizing the solution, the mice were injected into the tail vein at a dose of 150 mg / kg to establish a mouse iron overload model. On the eighth day after the injection of dextran iron, the mice were randomly divided into three groups, with 5 mice in each group, namely the control group (Control group), the DFO group and the FMS-DFO group. A group of healthy mice was also set up as a blank group (Blank group). The corresponding doses for the DFO group and the FMS-DFO group were 100 mg / kg and 153.63 mg / kg, respectively. The Control group and the Blank group were given equal volumes of normal saline. Each group of mice was given the drug once every two days by tail vein injection, for a total of five times. After the start of drug administration, the mice were fed an iron-deficient diet. Seven days after the last administration, the eyeballs of all mice were removed and blood was collected. The blood was placed in a heparin-coated blood collection tube and centrifuged at 13,000 rpm / min for 5 minutes. The supernatant was collected for later use. The ferritin content in mouse plasma was determined by competitive enzyme-linked immunosorbent assay (ELISA). The results are as follows: Figure 9 As shown in the figure, from the comparison results between the Blank group and the Control group, the plasma FE content in the Control group was significantly increased, indicating that the mouse iron overload model was successfully established; the results of the other two groups showed that both DFO and FMS-DFO could reduce the plasma ferritin content in mice, but the ferritin content in the FMS-DFO group was lower, and there was a significant difference compared with the DFO group, indicating that the ability of 2-sulfo-9-fluorenylmethoxycarbonyl-deferoxamine conjugate to clear excess iron in plasma is significantly improved than deferoxamine.

[0055] Example 6: Trial of 2-sulfo-9-fluorenylmethoxycarbonyl-deferoxamine conjugate to reduce the frequency of administration for the treatment of iron overload

[0056] Healthy BALB / c female mice were selected and housed in cages. The mice were weighed one day before modeling. A dextran / Fe solution (35% iron content) was prepared and filtered to remove bacteria. The mice were injected with the solution via the tail vein at a dose of 150 mg / kg to establish an iron overload model. Eight days after the injection, the mice were randomly divided into three groups of five mice each: a control group (Positive group), a Single group, and a Three-Times group. A group of healthy mice was also established as a blank group (Negative group). The doses of DFO and FMS-DFO were 100 mg / kg and 153.63 mg / kg, respectively. The Positive and Negative groups were given an equal volume of saline. The mice in each group were injected with the drugs via the tail vein every two days at the specified dose. The Single group was injected once, and the Three-Time group was injected three times. After the start of the administration, the mice were fed with iron-deficient feed. Seven days after the last administration, the mice were enucleated to collect blood, which was placed in a heparin-coated blood collection tube. The blood was centrifuged at 13,000 rpm / min for 5 min, and the supernatant was collected for use. The ferritin content in the mouse plasma was determined by competitive enzyme-linked immunosorbent assay (ELISA). The results are shown in FIG. 2. Figure 10 and Figure 11 As shown in FIG. 2, the iron content in the plasma of the Positive group was significantly higher than that of the Negative group, indicating that the mouse iron overload model was successfully established. The results of the other two groups showed that single administration and multiple administration could both reduce the ferritin content in the mouse plasma. However, the ferritin content in the FMS-DFO group was not significantly different from the healthy level, and there was a significant difference compared with the Positive group. This indicates that when the administration frequency is reduced to three times, the 2-sulfo-9-fluorenylmethoxycarbonyl-desferrioxamine conjugate still has the ability to remove excess iron in the plasma to a healthy level.

[0057] Example 7: Toxicity test of 2-sulfo-9-fluorenylmethoxycarbonyl-desferrioxamine conjugate

[0058] The iron-overloaded mice were administered DFO or FMS-DFO five times. Seven days after the last administration, the mice were enucleated to collect blood, which was placed in a heparin-coated blood collection tube. The blood was centrifuged at 13,000 rpm / min for 5 min, and the supernatant was collected for use. The malondialdehyde (MDA) content in the serum was determined by 2-thiobarbituric acid colorimetry. The experimental results are shown in FIG. 3. Figure 12 As shown in FIG. 3, both free DFO and FMS-DFO have the ability to reduce oxidative stress caused by iron overload. At the same molar dose, the FMS-DFO group has a significantly higher ability to down-regulate oxidative stress, indicating a higher antioxidant capacity.

[0059] Example 8: Liver histopathology of 2-sulfo-9-fluorenylmethoxycarbonyl-desferrioxamine conjugate in mice

[0060] After five doses of DFO or FMS-DFO, the liver tissues were dehydrated and embedded in paraffin, the embedded samples were cut into 4 μm slices using a tissue sample slicer, the samples were stained using hematoxylin and eosin, and finally the stained slice samples were observed under a microscope, and the results are shown in Figure 13 As shown in the figure, the slice of the positive control group showed extensive hemosiderin (arrow 1) and balloon swelling (arrow 2), and the cytoplasm of the hepatocytes disappeared, and the nucleus was located in the center. After DFO was given, the hemosiderin decreased, but a large range of balloon swelling (arrow 2) still appeared. FMS-DFO conjugate did not cause inflammatory infiltration, and only a small amount of balloon swelling (arrow 2) was found, which showed that both desferrioxamine and 2-sulfo-9-fluorenylmethoxycarbonyl-desferrioxamine conjugate can reduce liver toxicity caused by iron overload, but the attenuation effect of 2-sulfo-9-fluorenylmethoxycarbonyl-desferrioxamine conjugate is obviously better.

[0061] Example 9: 2-sulfo-9-fluorenylmethoxycarbonyl-desferrioxamine conjugate combined with 5-aminolevulinic acid for tumor diagnosis

[0062] In the present application, BALB / c mice inoculated subcutaneously with 4T1 cells were used as a tumor model, and when the tumor volume reached ~ 700 mm 3 When the tumor volume reached ~ 700 mm Figure 14 ), 2-sulfo-9-fluorenylmethoxycarbonyl-desferrioxamine conjugate can enhance the accumulation of protoporphyrin IX, which is internalized by 5-aminolevulinic acid, at the tumor site, and has a statistically significant advantage compared with desferrioxamine itself, and 2-sulfo-9-fluorenylmethoxycarbonyl-desferrioxamine conjugate has no significant effect on liver accumulation. The results show that 2-sulfo-9-fluorenylmethoxycarbonyl-desferrioxamine conjugate can enhance the fluorescence localization ability of protoporphyrin IX at the tumor site, and improve the tumor diagnosis effect of 5-aminolevulinic acid.

[0063] Example 10: 2-sulfo-9-fluorenylmethoxycarbonyl-desferrioxamine conjugate combined with 5-aminolevulinic acid for tumor treatment

[0064] The 4T1 cell subcutaneously tumor-bearing BALB / c mice were randomly divided into four groups, one of which was used as a positive control group, intravenous injection and oral D-PBS(-) were performed. The other three groups were respectively injected intravenously with D-PBS(-), DFO or FMS-DFO, and the doses of DFO and FMS-DFO were 560 μg / each and 861 μg / each, respectively, and then 100 mg / kg 5-aminolevulinic acid was orally taken. The tumors of the groups were irradiated for 10 min at 3 h after administration by using a 635 nm laser (200 mW / cm 2 , 10 min). The tumor sizes of the mice treated differently were measured every 2 days after administration until the 8th day. As shown in Figure 15 compared with the use of 5-aminolevulinic acid alone and the use of 5-aminolevulinic acid in combination with deferoxamine, the 2-sulfo-9-fluorenylmethyloxycarbonyl-deferoxamine conjugate in combination with 5-aminolevulinic acid has a significant advantage in treating tumors, which indicates that the 2-sulfo-9-fluorenylmethyloxycarbonyl-deferoxamine conjugate in combination with 5-aminolevulinic acid has a higher tumor treatment effect.

[0065] Although the embodiments of the present application have been shown and described, it can be understood by those of ordinary skill in the art that various changes, modifications, substitutions and variations can be made to the embodiments without departing from the principles and spirit of the present application, and the scope of the present application is defined by the appended claims and their equivalents.

Claims

1. A deferoxamine conjugate, characterized in that: The structural formula is as follows: ； The synthetic route is as follows: ； The specific steps are: Step (1): dissolving 9-fluorenylmethoxycarbonyl-N-succinimidyl carbonate in dichloromethane and reacting with chlorosulfonic acid to prepare 2-sulfo-9-fluorenylmethoxycarbonyl-N-succinimidyl carbonate; Step (2): 2-sulfo-9-fluorenylmethoxycarbonyl-N-succinimidyl carbonate and deferoxamine are dissolved in N,N-dimethylformamide and reacted under the action of DIPEA. After the reaction is complete, purification is performed to obtain a deferoxamine conjugate.

2. Use of the deferoxamine conjugate or a pharmaceutically acceptable salt thereof according to claim 1 in the preparation of a medicament for treating iron overload.

3. A pharmaceutical composition comprising the deferoxamine conjugate or a pharmaceutically acceptable salt thereof according to claim 1, and a pharmaceutically acceptable excipient.

4. Use of the pharmaceutical composition according to claim 3 in the preparation of a drug for treating iron overload.

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