Fluorine modified dihydroartemisinin as well as preparation method and application thereof

Abstract

The invention provides fluorine-modified dihydroartemisinin as well as a preparation method and application thereof, and belongs to the field of dihydroartemisinin derivatives. The fluorine-modified dihydroartemisinin provided by the invention is dihydroartemisinin of which the 12-site hydroxyl group is substituted by a fluorine atom. The dihydroartemisinin is subjected to fluorine modification, so that the combining capacity of the dihydroartemisinin and a target can be enhanced, the activity of the dihydroartemisinin is improved, and the curative effect of the dihydroartemisinin is further improved; a C-F bond formed after fluorine modification improves the metabolism resistance of dihydroartemisinin, the action time of dihydroartemisinin can be prolonged, and the treatment effect is improved. Compared with dihydroartemisinin, the fluorine modified dihydroartemisinin provided by the invention has a better treatment effect.

Description

Technical Field

[0001] This invention relates to the field of dihydroartemisinin derivatives, and more particularly to a fluorine-modified dihydroartemisinin, its preparation method, and its application. Background Technology

[0002] Dihydroartemisinin (DHA) is a sesquiterpene lactone compound with the molecular formula C6H2O. 15 H 24 O5. The structure of DHA contains a peroxy bridge (-OO-) and multiple hydroxyl (-OH) groups, which gives it unique advantages in biological activity. DHA's antimalarial activity is several times higher than artemisinin, and its antimalarial effect is 4–8 times that of artemisinin. In addition to its antimalarial activity, DHA and its derivatives have potential value in antitumor activity. For example, DHA has shown antitumor properties against esophageal cancer by regulating cell cycle-related targets. Furthermore, DHA also has antiviral, anti-inflammatory, and immunomodulatory effects.

[0003] Although DHA and its derivatives have broad application potential, their pharmaceutical activity needs further improvement. Therefore, providing a DHA derivative with higher activity, thereby enhancing the efficacy of DHA in disease treatment, has become a technical problem urgently needing to be solved by those skilled in the art. Summary of the Invention

[0004] The purpose of this invention is to provide a fluorinated dihydroartemisinin, its preparation method, and its application. The fluorinated dihydroartemisinin provided by this invention exhibits high pharmacological activity and can enhance the efficacy of DHA in disease treatment.

[0005] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0006] This invention provides a fluorine-modified dihydroartemisinin having the following chemical structure:

[0007]

[0008] This invention also provides a method for preparing fluorine-modified dihydroartemisinin as described in the above technical solution, comprising:

[0009] Under an inert atmosphere, a fluorinating reagent is mixed with a dihydroartemisinin solution to carry out a substitution reaction to obtain fluorinated dihydroartemisinin; the fluorinating reagent is added at a rate of 0.2–0.5 mL / min.

[0010] Preferably, the temperature of the reaction system is controlled at -60 to -20°C during the mixing of the fluorinating reagent and the dihydroartemisinin solution; after the fluorinating reagent is added, the reaction system is transferred to 20 to 30°C.

[0011] Preferably, the fluorinating agent includes diethylaminosulfur trifluoride, N,N-bis(2-methoxyethyl)trifluoromethanesulfonamide, 2-tert-butyl-5-methyl-3-trifluoromethylthiobenzene, or 2-fluoropyridine-1-oxide.

[0012] Preferably, the molar ratio of dihydroartemisinin to fluorination reagent in the dihydroartemisinin solution is 1:(1-2).

[0013] Preferably, the solvent in the dihydroartemisinin solution is one or more of dichloromethane, acetonitrile, chloroform, and methanol.

[0014] Preferably, the concentration of the dihydroartemisinin solution is 0.03–0.04 mol / L.

[0015] Preferably, after the substitution reaction is completed, a quenching agent is added for quenching, and the quenching agent is an alkaline solution.

[0016] Preferably, the quenching process further includes sequentially separating the organic phase, drying, and desolventizing the obtained quenching system to obtain fluorine-modified dihydroartemisinin.

[0017] The present invention also provides the application of the fluorine-modified dihydroartemisinin described above in the preparation of antimalarial, antitumor, antiviral, antibacterial or anti-inflammatory drugs.

[0018] This invention provides a fluorine-modified dihydroartemisinin having the following chemical structure:

[0019]

[0020] This invention enhances the binding ability of dihydroartemisinin to its target by fluorine modification, thereby increasing its activity and therapeutic efficacy. The CF bonds formed after fluorine modification improve the antimetabolite activity of dihydroartemisinin, prolonging its duration of action and enhancing therapeutic efficacy. Results from the examples show that the fluorine-modified dihydroartemisinin provided by this invention significantly inhibits cell viability in various leukemia cell lines, with superior effects compared to dihydroartemisinin alone. Apoptosis experiments show that in three leukemia cell lines (THP-1, KG-1, and L1210), both dihydroartemisinin and fluorine-modified dihydroartemisinin significantly promote apoptosis, but the effect of fluorine-modified dihydroartemisinin is more significant, especially in the mid-amplitude stage of apoptosis, exhibiting a good dose-dependent effect. Western blot results show that, compared to dihydroartemisinin, fluorine-modified dihydroartemisinin significantly upregulates the expression of Caspase 3 / 9 protein. Furthermore, at a concentration of 0.5 μg / mL, the effect was statistically significant (P<0.001), indicating that its anticancer activity was superior to that of dihydroartemisinin. In in vivo antimalarial activity experiments, the spleen index of mice treated with fluorinated dihydroartemisinin at low, medium, and high doses was closer to that of the uninfected group, suggesting that fluorinated dihydroartemisinin may have a stronger antimalarial effect. Moreover, treatment with fluorinated dihydroartemisinin significantly reduced the proliferation of malaria parasites and alleviated organ damage caused by infection, especially in the heart and spleen. This indicates that fluorinated dihydroartemisinin can not only effectively inhibit the proliferation of malaria parasites but also has a protective effect against tissue damage caused by infection, thus exhibiting better therapeutic effects.

[0021] The preparation method provided by this invention is simple. Furthermore, the product yield can be improved by controlling the reaction system temperature, the molar ratio of dihydroartemisinin to fluorinating reagent in the dihydroartemisinin solution, the type of fluorinating reagent, and the type of solvent in the dihydroartemisinin solution during the mixing process of the fluorinating reagent and the dihydroartemisinin solution. Attached Figure Description

[0022] Figure 1 The infrared spectrum of dihydroartemisinin (DHA) in Example 1 of this invention;

[0023] Figure 2 The infrared spectrum of fluorine-modified dihydroartemisinin (F-DHA) in Example 1 of this invention;

[0024] Figure 3 The fluorine-modified dihydroartemisinin (F-DHA) in Example 1 of this invention 1 H-NMR spectrum;

[0025] Figure 4 The fluorine-modified dihydroartemisinin (F-DHA) in Example 1 of this invention 13 C-NMR spectrum;

[0026] Figure 5 The fluorine-modified dihydroartemisinin (F-DHA) in Example 1 of this invention 19 F-NMR spectrum;

[0027] Figure 6 This is a bar chart showing the effect of DHA and F-DHA on the viability of human monocytic leukemia (THP-1) cells in Example 1 of the present invention.

[0028] Figure 7 This is a bar chart showing the effects of DHA and F-DHA on the viability of human acute myeloid leukemia (KG-1) cells in Example 1 of the present invention.

[0029] Figure 8 This is a bar chart showing the effects of DHA and F-DHA on the viability of murine leukemia (L1210) cells in Example 1 of the present invention.

[0030] Figure 9 This is a statistical graph showing the apoptosis regulation results of the blank control group of this invention on the THP-1 leukemia cell line;

[0031] Figure 10 This is a statistical graph showing the effect of 0.1 μg / mLDHA on apoptosis regulation of the THP-1 leukemia cell line in Example 1 of the present invention.

[0032] Figure 11 This is a statistical graph showing the effect of 0.1 μg / mLF-DHA on apoptosis regulation of the THP-1 leukemia cell line in Example 1 of the present invention.

[0033] Figure 12 This is a statistical graph showing the apoptosis regulation effect of 0.5 μg / mLF-DHA on the THP-1 leukemia cell line in Example 1 of the present invention.

[0034] Figure 13 This is a statistical graph showing the regulatory effects of different concentrations of DHA and F-DHA on different apoptosis proteins in the THP-1 leukemia cell line in the blank group and Example 1 of this invention.

[0035] Figure 14 The bar chart shows the statistical ratio of different concentrations of DHA and F-DHA to Caspase3 protein in the blank group and Example 1 of this invention.

[0036] Figure 15 The following figures show the results of DHA and F-DHA inhibiting the tumor volume of L1210 leukemia cell line tumor-bearing mice in the model group, positive drug group, and Example 1 of this invention.

[0037] Figure 16 This is a bar chart showing the tumor weight of mice bearing L1210 leukemia cell lines inhibited by DHA and F-DHA in the model group, positive drug group, and Example 1 of this invention.

[0038] Figure 17 Line graph showing the statistical changes in tumor volume over time in the model group, positive drug group, and Example 1 of this invention, where DHA and F-DHA inhibited the tumor volume of L1210 leukemia cell line tumor-bearing mice.

[0039] Figure 18 These are smears of malaria parasite infection in mice treated with 30 mg / kg / day DHA and 30 mg / kg / day F-DHA in the blank group, model group, and Example 1 of this invention.

[0040] Figure 19 The results of H&E staining of organ pathological sections from mice infected with parasites during the treatment period are shown in the blank group, model group, and Example 1 of this invention, using 30 mg / kg / day DHA and 30 mg / kg / day F-DHA.

[0041] Figure 20 This is a bar chart comparing the spleen index of malaria-infected mice after treatment with different concentrations of DHA and F-DHA in the blank group, model group, and Example 1 of this invention. Detailed Implementation

[0042] This invention provides a fluorine-modified dihydroartemisinin having the following chemical structure:

[0043]

[0044] The fluorine-modified dihydroartemisinin provided by this invention has high activity and good anti-tumor activity; it can effectively inhibit the proliferation of Plasmodium and also has a protective effect against tissue damage caused by infection, thus having better therapeutic efficacy.

[0045] This invention enhances the binding ability of dihydroartemisinin to its target and improves its activity by fluorinating it, thereby improving its therapeutic effect. The CF bond formed after fluorination enhances the antimetabolite activity of dihydroartemisinin, prolonging its duration of action and improving the therapeutic effect.

[0046] This invention also provides a method for preparing fluorine-modified dihydroartemisinin as described in the above technical solution, comprising:

[0047] Under an inert atmosphere, a fluorinating reagent is mixed with a dihydroartemisinin solution to carry out a substitution reaction, yielding fluorinated dihydroartemisinin.

[0048] Unless otherwise specified, all reagents used in this invention are commercially available reagents in the field of drug synthesis and analysis that are well known to those skilled in the art.

[0049] In one embodiment of the present invention, the gas used in the inert atmosphere may be argon or nitrogen.

[0050] In this invention, the fluorinating agent preferably includes diethylaminosulfur trifluoride, N,N-bis(2-methoxyethyl)trifluoromethanesulfonamide, 2-tert-butyl-5-methyl-3-trifluoromethylthiobenzene, or 2-fluoropyridine-1-oxide, more preferably diethylaminosulfur trifluoride or N,N-bis(2-methoxyethyl)trifluoromethanesulfonamide, and even more preferably diethylaminosulfur trifluoride. Limiting the type of fluorinating agent to the above range in this invention can improve the yield of the prepared fluorinated dihydroartemisinin.

[0051] In this invention, the structural formula of the diethylaminosulfur trifluoride is:

[0052]

[0053] The structural formula of the N,N-bis(2-methoxyethyl)trifluoromethanesulfonamide is:

[0054]

[0055] The structural formula of the 2-tert-butyl-5-methyl-3-trifluoromethylthiobenzene is:

[0056]

[0057] The structural formula of the 2-fluoropyridine-1-oxide is:

[0058]

[0059] In an embodiment of the present invention, the dihydroartemisinin in the dihydroartemisinin solution is derived from Leyan, with CAS number 71939-50-9.

[0060] In this invention, the molar ratio of dihydroartemisinin to the fluorinating reagent in the dihydroartemisinin solution is preferably 1:(1-2), more preferably 1:(1.5-2). Limiting the molar ratio of dihydroartemisinin to the fluorinating reagent in the dihydroartemisinin solution to the above range in this invention is beneficial for improving the yield of fluorinated dihydroartemisinin.

[0061] In this invention, the concentration of the dihydroartemisinin solution is preferably 0.03–0.04 mol / L, more preferably 0.032–0.038 mol / L, and even more preferably 0.034–0.036 mol / L. Limiting the concentration of the dihydroartemisinin solution to the above range promotes the smooth progress of the reaction.

[0062] In this invention, the solvent in the dihydroartemisinin solution is preferably one or more of dichloromethane, acetonitrile, chloroform, and methanol, more preferably dichloromethane or chloroform, and even more preferably dichloromethane. Limiting the solvent in the dihydroartemisinin solution to the above-mentioned range can improve the yield of fluorine-modified dihydroartemisinin.

[0063] In this invention, the reaction system temperature is controlled at -60 to -20°C, preferably -40 to -20°C, and more preferably -40 to -30°C during the mixing of the fluorinating reagent and the dihydroartemisinin solution. Controlling the reaction system temperature within this range during the mixing process ensures that the initial reaction is not too vigorous, preventing the formation of a large number of byproducts. This allows the F group to replace the hydroxyl group at position 12 of dihydroartemisinin, thereby increasing the yield of fluorinated dihydroartemisinin.

[0064] In this invention, the addition rate is preferably 0.2–0.5 mL / min, more preferably 0.25–0.35 mL / min. Limiting the addition rate to the above range ensures the complete progress of the reaction.

[0065] In this invention, after the fluorinating reagent is added, the reaction system is transferred to 20–30°C. This method of transferring the reaction system to 20–30°C after the addition of the fluorinating reagent ensures efficient reaction. As one embodiment of this invention, the reaction system can be stirred after being transferred to 20–30°C.

[0066] The present invention does not have a special limitation on the stirring speed; stirring speeds commonly used by those skilled in the art can be used to promote the reaction of the reactants.

[0067] The present invention does not have a specific time limit for the substitution reaction; it is sufficient to detect the absence of dihydroartemisinin in the reaction system using TLC.

[0068] In one embodiment of the present invention, after the substitution reaction is completed, a quenching agent is added for quenching, and the quenching agent is an alkaline solution. In an embodiment of the present invention, the quenching agent can be a saturated sodium bicarbonate solution. In an embodiment of the present invention, the specific operation of quenching the substitution reaction with saturated sodium bicarbonate solution is as follows: the system after the substitution reaction is transferred to a 500mL separatory funnel, 200mL of saturated sodium bicarbonate solution is slowly added, and the reaction system is vigorously shaken until no bubbles are generated.

[0069] In this invention, the quenching process preferably further includes sequentially separating the organic phase, drying, and desolventizing the obtained quenching system to obtain fluorine-modified dihydroartemisinin.

[0070] This invention does not impose any particular limitation on the operation of separating the organic phase; any operation well-known to those skilled in the art can be used. This invention enables the separation of the organic phase from the quenching system.

[0071] In one embodiment of the present invention, the obtained organic phase can be washed with water after separation. The present invention does not specifically limit the number of washes, as long as the resulting aqueous phase is neutral. In an embodiment of the present invention, the number of washes is 3, and the amount of water used for each wash is 50 mL.

[0072] In one embodiment of the present invention, the desiccant used for drying can be anhydrous sodium sulfate.

[0073] In one embodiment of the present invention, the solvent removal can be performed under vacuum conditions. The present invention does not impose any particular limitation on the solvent removal operation; any solvent removal operation commonly used by those skilled in the art can be employed.

[0074] This invention ensures a good yield of the synthesized product by adjusting the temperature of the reaction system during the mixing of the fluorinating reagent and the dihydroartemisinin solution. Furthermore, the product yield is improved by controlling the temperature of the reaction system, the molar ratio of dihydroartemisinin to the fluorinating reagent in the dihydroartemisinin solution, the type of fluorinating reagent, and the type of solvent in the dihydroartemisinin solution during the mixing of the fluorinating reagent and the dihydroartemisinin solution.

[0075] The present invention also provides the application of the fluorine-modified dihydroartemisinin described above in the preparation of antimalarial, antitumor, antiviral, antibacterial or anti-inflammatory drugs.

[0076] The technical solutions of this invention will be clearly and completely described below with reference to the embodiments thereof. Obviously, the described embodiments are only a part of the embodiments of this invention, and not all of them. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this invention.

[0077] Example 1

[0078] Under an argon atmosphere, a fluorinating reagent (diethylaminotrifluoride sulfur, DAST) was added to a dihydroartemisinin solution (the solvent in the dihydroartemisinin solution was dichloromethane, and the concentration of the dihydroartemisinin solution was 0.036 mol / L) at a rate of 0.3 mL / min (the temperature of the reaction system was controlled at -40℃ during the mixing of the fluorinating reagent and the dihydroartemisinin solution). After the addition of the fluorinating reagent was completed, the reaction system was transferred to room temperature (25℃), stirred, and the reaction was monitored by TLC to confirm completeness. After 5 hours, the system after the substitution reaction was transferred to a 500 mL separatory funnel, and 200 mL of saturated sodium bicarbonate solution was slowly added. The reaction system was shaken vigorously until no bubbles were generated. The organic phase was separated and washed with water (3 × 50 mL) until the aqueous phase was neutral. The organic phase was dried with anhydrous sodium sulfate and then desolvated under vacuum to obtain fluorinated dihydroartemisinin (F-DHA). The molar ratio of dihydroartemisinin to fluorinating reagent in the dihydroartemisinin solution was 1:1.5.

[0079] The yield of fluorinated dihydroartemisinin (F-DHA) obtained in Example 1 was 90%.

[0080] The raw material dihydroartemisinin (DHA) and the prepared fluorine-modified dihydroartemisinin (F-DHA) in Example 1 were tested using an infrared spectrometer. The infrared spectra of dihydroartemisinin (DHA) are shown below. Figure 1 As shown, the infrared spectrum of fluorine-modified dihydroartemisinin (F-DHA) is as follows. Figure 2 As shown. From Figure 1 As can be seen from this, 3372cm -1 The absorption peak at 2936 cm⁻¹ is the signal of the stretching vibration of the hydroxyl group (OH). -1 and 2853cm -1 The absorption peak at 1446 cm⁻¹ is the signal of saturated CH stretching vibration. -1 and 1377cm -1 The absorption peak at 1092 cm⁻¹ is the signal of saturated CH bending vibration. -1 1014cm -1 and 983cm -1 The absorption peak at that point represents the CO stretching vibration signal. From... Figure 2 As can be seen, compared to dihydroartemisinin, the hydroxyl (OH) stretching vibration signal of fluorinated dihydroartemisinin disappeared, indicating that the hydroxyl group was substituted. Other signals, 2952 cm⁻¹ -1 and 2873cm -1 The absorption peak at 1455 cm⁻¹ is the signal of saturated CH stretching vibration. -1 and 1379cm -1 The absorption peak at 1112 cm⁻¹ is the signal of saturated CH bending vibration. -1 The absorption peak at that point represents the CO stretching vibration signal.

[0081] The fluorinated dihydroartemisinin (F-DHA) prepared in Example 1 was tested using nuclear magnetic resonance spectroscopy. 1 H-NMR spectrum as shown Figure 3 As shown, fluorine-modified dihydroartemisinin (F-DHA) 13 C-NMR spectrum as shown Figure 4 As shown, fluorine-modified dihydroartemisinin (F-DHA) 19 F-NMR spectrum as shown Figure 5 As shown. From Figures 3-5 It can be seen that the infrared spectrum of the product after the reaction shows that the stretching vibration signal of the hydroxyl group (OH) of dihydroartemisinin has disappeared, and the fluorine spectrum shows that there is a signal of a fluorine atom at -135.2 ppm. The carbon atom adjacent to the fluorine atom has also split, indicating that the hydroxyl group at position 12 of dihydroartemisinin has been replaced by a fluorine atom.

[0082] The NMR data of the fluorine-modified dihydroartemisinin prepared in Example 1 are as follows: 1 HNMR (600MHz, CDCl3): δ0.89-0.91(m,1H),0.94(d,J=6.0Hz,3H),0.97(d,J=6.0Hz,3H),1.21-1 .26(m,1H),1.30-1.37(m,1H),1.38(s,3H),1.37-1.49(m,2H),1.50-1.60(m,2H),1.65(dd,J=12 .0,6.0Hz,1H),1.81(dd,J=12.0,6.0Hz,1H),1.85-1.90(m,1H),2.02(dt,J=18.0,6.0Hz,1H),2 .34(td,J=12.0,6.0Hz,1H),2.59(dt,J=36.0,12.0Hz,1H),5.52(s,1H),5.55(d,J=60.0Hz,1H);

[0083] 13 C NMR (150MHz, CDCl3): δ12.3,20.2,24.2(d, 2 J=6.0Hz),24.6,25.8,30.7(d,J=24.0Hz),34.5,36.2,37.4,43.4(d,J=1.5Hz),52.2,80.4,88.7,104.4,110.7(d, 1 J = 222Hz);

[0084] 19F NMR(MHz,CDCl3):δ-135.2(s,1F).HRMS(ESI)calcd.for C15H22FO3 - [MO] - :269.1553; found:269.1367.IR(KBr):2951,2872,2001,1456,1380,1179,1112,1038,872,540cm -1 .

[0085] Example 2

[0086] The only difference between Example 2 and Example 1 is that the molar ratio of dihydroartemisinin to fluorination reagent in the dihydroartemisinin solution is 1:1. Otherwise, they are the same as in Example 1.

[0087] The yield of fluorinated dihydroartemisinin (F-DHA) obtained in Example 2 was 76%.

[0088] Example 3

[0089] The only difference between Example 3 and Example 1 is that the molar ratio of dihydroartemisinin to fluorination reagent in the dihydroartemisinin solution is 1:2. Otherwise, they are the same as in Example 1.

[0090] The yield of fluorinated dihydroartemisinin (F-DHA) obtained in Example 3 was 91%.

[0091] Example 4

[0092] The only difference between Example 4 and Example 1 is that the temperature of the reaction system is controlled at -20°C during the mixing of the fluorinating reagent and the dihydroartemisinin solution. Otherwise, they are the same as in Example 1.

[0093] The yield of fluorinated dihydroartemisinin (F-DHA) obtained in Example 4 was 62%.

[0094] Example 5

[0095] The only difference between Example 5 and Example 1 is that the solvent in the dihydroartemisinin solution is acetonitrile; otherwise, they are the same as in Example 1.

[0096] The yield of fluorinated dihydroartemisinin (F-DHA) obtained in Example 5 was 64%.

[0097] Example 6

[0098] The only difference between Example 6 and Example 1 is that the solvent in the dihydroartemisinin solution is chloroform; otherwise, they are the same as in Example 1.

[0099] The yield of fluorinated dihydroartemisinin (F-DHA) obtained in Example 6 was 86%.

[0100] Example 7

[0101] The only difference between Example 7 and Example 1 is that the solvent in the dihydroartemisinin solution is methanol; otherwise, they are the same as in Example 1.

[0102] The yield of fluorinated dihydroartemisinin (F-DHA) obtained in Example 7 was 45%.

[0103] Example 8

[0104] The only difference between Example 8 and Example 1 is that the fluorinating agent is N,N-bis(2-methoxyethyl)trifluoromethanesulfonamide (BAST), otherwise it is the same as Example 1.

[0105] The yield of fluorinated dihydroartemisinin (F-DHA) obtained in Example 8 was 77%.

[0106] Example 9

[0107] The only difference between Example 9 and Example 1 is that the fluorinating agent is 2-tert-butyl-5-methyl-3-trifluoromethylthiobenzene (Fluolead), otherwise it is the same as Example 1.

[0108] The yield of fluorinated dihydroartemisinin (F-DHA) obtained in Example 9 was 52%.

[0109] Example 10

[0110] The only difference between Example 10 and Example 1 is that the fluorinating agent is 2-fluoropyridine-1-oxide (PyFluo), otherwise it is the same as Example 1.

[0111] The yield of fluorinated dihydroartemisinin (F-DHA) obtained in Example 10 was 38%.

[0112] Comparative Example 1

[0113] The only difference between Comparative Example 1 and Example 1 is that the temperature of the reaction system was controlled at room temperature (25°C) during the mixing of the fluorinating reagent and the dihydroartemisinin solution. Otherwise, they are the same as in Example 1.

[0114] The yield of fluorine-modified dihydroartemisinin (F-DHA) obtained in Comparative Example 1 was 5%.

[0115] Comparative Example 2

[0116] The only difference between Comparative Example 2 and Example 1 is that the temperature of the reaction system was controlled at 0°C during the mixing of the fluorinating reagent and the dihydroartemisinin solution; otherwise, they were the same as in Example 1.

[0117] The yield of fluorinated dihydroartemisinin (F-DHA) obtained in Comparative Example 2 was 35%.

[0118] Test example

[0119] (1) Anti-leukemia activity test was carried out on the raw material dihydroartemisinin (DHA) in Example 1 and the prepared fluorine-modified dihydroartemisinin (F-DHA).

[0120] The cells used included: human acute myeloid leukemia (KG-1) cells, human monocytic leukemia (THP-1) cells, and murine leukemia (L1210) cells, all purchased from the National Cell Bank of Biomedical Experiments.

[0121] Experimental animals: SPF-grade Balb / C Nude mice (male, 6-8 weeks old, body weight 18-22 g), purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. License numbers: SCXK(Beijing)2021-0011, SCXK(Beijing)2021-0006. Animal ethics number: 2025B014. The animals were housed in the South Animal Building of the Institute of Chinese Materia Medica, in a barrier environment. Experimental facility license SYXK(Beijing)2015-0041.

[0122] The experimental reagents used and their sources are shown in Table 1

[0123] Table 1 Reagents and manufacturers used in the in vitro anti-leukemia activity experiment

[0124] Reagent Name factory CAS / Item Number Dihydroartemisinin (DHA) Leyan 71939-50-9 Fluorinated dihydroartemisinin (F-DHA) Preparation in Example 1 RPMI1640 medium Gibco C11875500BT Fetal bovine serum (FBS) Sigma-Aldrich 1943609-65-1 CCK-8 reagent kit biosharp BS350B PMSF protease inhibitors Solarbio Biotechnology Co., Ltd. P0100 penicillin-streptomycin solution Gibco 15640055 RIPA pyrolysis fluid Solarbio Biotechnology Co., Ltd. 2312013 Universal antibody diluent Coolaber SL351221100 Trizol Xavier 144701-48-4 daunorubicin Leyan 20830-81-7

[0125] The experimental instruments used, as well as their manufacturers and models, are shown in Table 2.

[0126] Table 2 Experimental instruments, manufacturers and models

[0127]

[0128]

[0129] The experimental data of this invention were analyzed by the One-Way ANOVA method.

[0130] (1) In vitro anti-leukemia activity of F-DHA

[0131] (1.1) The CCK-8 method was used to detect the activity levels of DHA and F-DHA against tumor cells

[0132] The cells cultured the previous day were digested, centrifuged, and counted. Three groups of cells (200 μL / well for 3000-7000 cells / well) were seeded into 96-well plates: human monocytic leukemia (THP-1) cells, human acute myeloid leukemia (KG-1) cells, and murine leukemia (L1210) cells, respectively. Different concentrations of DHA and F-DHA solutions were prepared for each group of cells (the concentrations of DHA and F-DHA solutions for human monocytic leukemia cells and murine leukemia cells were 0.5 μg / mL, 1 μg / mL, 2.5 μg / mL, and 5 μg / mL, respectively). The concentrations of DHA and F-DHA solutions were 2.5 μg / mL, 5 μg / mL, 10 μg / mL, 25 μg / mL, 50 μg / mL, 100 μg / mL, and 200 μg / mL for detecting human acute myeloid leukemia cells. Three replicates (three parallel experiments) of each concentration were added to 96-well plates and cultured at 37°C, 5% CO2, and 90% humidity for 48 h. CCK-8 was thawed and centrifuged at room temperature before use. Cell growth was assessed using an inverted microscope, and 20 μL of CCK-8 solution was added to each well. Cells were cultured at 37°C, 5% CO2, and 90% humidity for 4 h. Absorbance was measured at 450 nm using a microplate reader.

[0133] The effects of DHA and F-DHA on the viability of human monocytic leukemia (THP-1) cells in Example 1 are shown in Table 3, and the bar chart is as follows. Figure 6 As shown in the figure; the data on the effects of DHA and F-DHA on the viability of human acute myeloid leukemia (KG-1) cells in Example 1 are shown in Table 4, and the bar chart is shown in the figure. Figure 7 As shown in the figure; the data on the effects of DHA and F-DHA on the viability of mouse leukemia (L1210) cells in Example 1 are shown in Table 5, and the bar chart is shown in the figure. Figure 8 As shown.

[0134] Table 3. Data on the effects of DHA and F-DHA on the viability (cell survival rate) of human monocytic leukemia (THP-1) cells in Example 1.

[0135] unit(%)

[0136]

[0137] Table 4. Effects of DHA and F-DHA on the viability (cell survival rate) of human acute myeloid leukemia (KG-1) cells in Example 1.

[0138] unit(%)

[0139]

[0140] Table 5. Data on the effects of DHA and F-DHA on the viability (cell survival rate) of mouse leukemia (L1210) cells in Example 1.

[0141] unit(%)

[0142]

[0143] In statistics, P represents a probability value, used to determine statistical significance. Figures 6-8 ** indicates P < 0.01 (highly significant), and *** indicates P < 0.001 (extremely significant). From Tables 3-5 and... Figures 6-8 It can be seen that F-DHA, compared with DHA, can significantly inhibit cell viability in different leukemia cell lines.

[0144] (1.2) Flow cytometry was used to detect the effects of DHA and F-DHA on tumor cell apoptosis.

[0145] THP-1 cells at 2×10 5 mL -1 Cells were seeded at different concentrations in 6-well plates and cultured for 48 h with different concentrations of sample solutions (blank control group, 0.1 μg / mL LDH, 0.1 μg / mL LF-DHA, and 0.5 μg / mL LF-DHA). Cells from each group were collected, the supernatant was discarded, and the cell density was adjusted to 1 × 10⁶ cells / well. 6 mL -1 1000r·min -1 Centrifuge for 5 min, discard the supernatant, resuspend in 195 μL Annexin V-FITC binding buffer in the dark, add 5 μL Annexin V-FITC, add 10 μL propidium iodide staining solution, mix well, incubate at room temperature in the dark for 15 min, and then perform flow cytometry analysis.

[0146] The statistical graph of the apoptosis regulation results of the blank control group on the THP-1 leukemia cell line is shown in the figure below. Figure 9 As shown in the figure, the statistical graph of the apoptosis regulation effect of 0.1 μg / mLDHA on the THP-1 leukemia cell line in Example 1 is as follows. Figure 10 As shown in the figure, the statistical graph of the apoptosis regulation effect of 0.1 μg / mLF-DHA on the THP-1 leukemia cell line in Example 1 is as follows. Figure 11 As shown in the figure, the statistical results of the apoptosis regulation of the THP-1 leukemia cell line by 0.5 μg / mLF-DHA in Example 1 are as follows: Figure 12 As shown, region Q1 contains mechanically damaged necrotic cells, region Q2 contains late-stage apoptotic or necrotic cells, region Q3 contains early-stage apoptotic cells, and region Q4 contains normal cells. From... Figures 9-12It can be seen that in the THP-1 leukemia cell line, both DHA and F-DHA can significantly promote cell apoptosis. Compared with DHA, F-DHA has a greater effect on the mid-stage of cell apoptosis and shows a good dose-dependent effect.

[0147] (1.3) Western blotting was used to detect the regulatory levels of DHA and F-DHA on tumor cell apoptosis proteins.

[0148] THP-1 cells at 2×10 5 mL -1 Cells were seeded at different concentrations in 6-well plates and cultured for 48 h with different concentrations of sample solutions (blank control group, 1 μg / mL LDH, 5 μg / mL LDH, 0.5 μg / mL LF-DHA, and 1 μg / mL LF-DHA). Cells from each group were collected, the supernatant was discarded, and the cell density was adjusted to 1 × 10⁶ cells / well. 6 mL -1 Centrifuge at 10000 rpm for 5 min, discard the supernatant, add 100 μL LRIPA lysis buffer (containing PMSF) to each well, and set up three parallel experiments for each concentration. Incubate at 4℃ on a shaker for 30 min, centrifuge at 10000 rpm for 10 min, collect the supernatant, and add 5× loading buffer after BCA protein quantification. Boil the sample at 95℃. Prepare the appropriate lower gel according to the protein size, add freshly prepared electrophoresis buffer to the inner layer, and add recycled electrophoresis buffer to the outer layer. Run the gel at 90V for 20 min, then switch to 120V for 90 min. Transfer the membrane at 90mA for 100 min. Remove the membrane and place it in 5% skim milk powder. Shake for 1 h, add the corresponding antibody, and incubate at 4℃ overnight. Remove the membrane and wash it three times with TBST for 15 min each time. Add secondary antibody and shake at room temperature for 1 h. Remove the membrane and wash it three times with TBST for 15 min each time. Add developing solution and perform chemiluminescence detection, and take pictures for grayscale analysis.

[0149] The statistical results of the regulation of different apoptosis proteins in the THP-1 leukemia cell line by different concentrations of DHA and F-DHA in the blank group and Example 1 are shown in the figure below. Figure 13 As shown in Table 6, the ratios of different concentrations of DHA and F-DHA to Caspase3 protein in the blank group and Example 1 are presented in the bar chart. Figure 14 As shown in Table 6. Table 6 shows the ratios of different concentrations of DHA and F-DHA to Caspase 3 protein in the blank group and Example 1.

[0150]

[0151] Figure 14 The asterisk (*) in the table indicates P < 0.05, as shown in Table 6 and... Figures 13-14It can be seen that DHA did not show statistically significant differences in the regulation of apoptosis-related proteins in the THP-1 cell line. In contrast, F-DHA had a significant upregulation effect on Caspase3 / 9 proteins in the apoptosis pathway, and the difference was statistically significant at a concentration of 0.5 μg / mL (P<0.05). This experiment can prove that the anticancer activity of fluorinated dihydroartemisinin products is superior to that of dihydroartemisinin.

[0152] (2) F-DHA in vivo anti-leukemia activity

[0153] Experimental methods:

[0154] (2.1) Dosage:

[0155] The F-DHA dosage used in the mouse antitumor experiment in this test case was based on the following: Clinical dose conversion: Referring to the adult clinical dose of dihydroartemisinin (DHA) (60 mg / day, equivalent to 1 mg / kg / day), the equivalent dose for mice was calculated to be 12.3 mg / kg / day using the body surface area normalization method (conversion factor 12.3). The equivalent dose of F-DHA was set at 15 mg / kg / day.

[0156] Dosage range expansion: To assess the dose-response relationship, a high-dose group (30 mg / kg / day) was added.

[0157] (2.2) Route of administration

[0158] Both DHA and F-DHA are administered via gavage (orally, via gavage).

[0159] Daunorubicin is administered via intraperitoneal injection (ia).

[0160] (2.3) Animal grouping

[0161] Balb / C Nude mice were randomly divided into 5 groups (6 mice in each group) according to their body weight.

[0162] Model group (Untreated group): L1210 cells were administered, along with a solvent.

[0163] Positive drug group (Daunorubicin): 2 mg / kg / week.

[0164] DHA treatment group: 30mg / kg / 3 days.

[0165] F-DHA treatment group: 15mg / kg / 3 days, 30mg / kg / 3 days.

[0166] (2.4) Subcutaneous seed tumor

[0167] Collect cultured L1210 cells by centrifugation (800×g, 10 min), and adjust the concentration to 1×10⁻⁶ cells using serum-containing RPMI 1640 medium. 8 pcs·mL -1 Each group of mice received a subcutaneous injection of 0.2 mL of cell suspension into the axilla of the right forelimb.

[0168] (2.5) Administration method

[0169] Five days after subcutaneous injection, tumors were visible under the skin of mice. Drug administration was then initiated, administered by gavage every three days at a fixed time, for a total of five doses before discontinuation. The positive control drug, daunorubicin, was administered via intraperitoneal injection once weekly.

[0170] (2.6) Observation indicators

[0171] Body weight changes: The body weight of mice was recorded every 3 days, and the weight of six mice was recorded.

[0172] Tumor volume changes: Tumor volume of mice was recorded every 3 days. Tumor volumes of four mice were recorded in the model group, DHA treatment group and 15 mg / kg / 3 days F-DHA treatment group. Tumor volumes of five mice were recorded in the positive control group (daunorubicin) and 30 mg / kg / 3 days F-DHA treatment group.

[0173] (2.7) Organ collection

[0174] After euthanizing the mice, the heart, liver, spleen, kidneys, and tumor tissues were quickly removed, preserved in tissue fixative, and then stained with hematoxylin and eosin (HE) for pathological observation.

[0175] (2.8) Data Processing and Statistical Analysis

[0176] Data are expressed as mean ± standard deviation (Mean ± SD). One-way ANOVA was performed using GraphPadPrism 9.0. Tukey's test was used for comparisons between groups, and P < 0.05 was considered statistically significant.

[0177] The results of DHA and F-DHA inhibiting the tumor volume of L1210 leukemia cell line in the model group, positive drug group, and Example 1 are shown in the figure. Figure 15 As shown in Table 7, the tumor weight statistics are presented in the bar chart below. Figure 16 As shown in Table 8, the statistical data on tumor volume changes over time are presented in Table 8, and the line graph of tumor volume changes over time is shown in the figure. Figure 17 As shown.

[0178] Table 7. Statistical data on tumor weight in L1210 leukemia cell line-bearing mice in the model group, positive drug group, and Example 1, showing the inhibition of tumor weight by DHA and F-DHA.

[0179]

[0180] Table 8 Statistical data on the changes in the tumor volume of tumor-bearing mice in the L1210 leukemia cell line in the model group, positive drug group, and DHA and F-DHA in Example 1 over time

[0181] Unit (mm 2 )

[0182]

[0183]

[0184] Figure 16 and 17 The ** in [] indicates P < 0.01 (highly significant), and *** indicates P < 0.001 (extremely significant). From Tables 7 and 8 and Figures 15-17 it can be seen that both DHA and F-DHA have obvious inhibitory effects on the anti-tumor effect of L1210-1 tumor-bearing mice at the same administration dose. The statistical data of the DHA treatment group show (**(P < 0.01)), and the high and low doses of F-DHA (***(P < 0.001)) have better effects in inhibiting tumor growth and are positively correlated with the administration dose. It shows that F-DHA can produce a more effective effect in inhibiting tumor growth.

[0185] (2) In vivo antimalarial activity test was carried out on dihydroartemisinin (DHA) in Example 1 and fluorine-modified dihydroartemisinin (F-DHA) prepared

[0186] Experimental animals: SPF-grade C57BL / 6 mice (male, 6 - 8 weeks old, body weight 18 - 22 g), purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. License numbers: SCXK (Beijing) 2021 - 0011, SCXK (Beijing) 2021 - 0006. Animal ethics number: 2024B228. The animals were housed in the South Animal Building of the Institute of Chinese Materia Medica in a barrier environment. Experimental facility license SYXK (Beijing) 2015 - 0041.

[0187] Plasmodium strain: Artemisinin-sensitive strain of Plasmodium berghei (Plasmodium berghei K173, PbK173-S), gifted by the Artemisinin Center of the Institute of Chinese Materia Medica, Chinese Academy of Chinese Medical Sciences.

[0188] The experimental reagents used and their sources are shown in Table 9

[0189] Table 9 Reagents and manufacturers used in the in vitro anti-leukemia activity experiment

[0190]

[0191]

[0192] The experimental instruments used, as well as their manufacturers and models, are shown in Table 10.

[0193] Table 10 Experimental Instruments, Manufacturers, and Models

[0194] name factory model Low temperature centrifuge Eppendorf 5424R optical microscope Olympus CX23 Analytical balance MettlerToledo XS205 Automated Blood Analyzer for Veterinary Use Mindray BC-2800vet

[0195] The experimental data of this invention were analyzed using the One-Way ANOVA method.

[0196] Experimental methods:

[0197] (1) Drug dosage

[0198] The F-DHA dosage used in the antimalarial mouse experiment in this test case was based on the following criteria:

[0199] Clinical dosage conversion was based on the adult clinical dose of dihydroartemisinin (DHA) (60 mg / day, equivalent to 1 mg / kg / day). Using body surface area normalization (conversion factor 12.3), the equivalent dose in mice was calculated to be 12.3 mg / kg / day. Combined with preliminary experimental data, the equivalent dose of F-DHA was set at 15 mg / kg / day.

[0200] Dosage range expansion: To evaluate the dose-response relationship, a low-dose group (5 mg / kg / day), a medium-dose group (15 mg / kg / day), and a high-dose group (30 mg / kg / day) were added.

[0201] (2) Drug preparation

[0202] Preparation of DHA and F-DHA solutions:

[0203] DHA and F-DHA powders were dissolved in 0.5% sodium carboxymethyl cellulose (CMC-Na) aqueous solution, vortexed and sonicated (30 min, 25 °C) to ensure uniform suspension.

[0204] According to the experimental dosage requirements (5, 15, 30 mg / kg / day), DHA solutions with concentrations of 0.5 mg / mL, 1.5 mg / mL, and 3.0 mg / mL, as well as F-DHA solutions of the same concentration, were prepared.

[0205] Solvent control group: The blank group and the model group were given an equal volume of 0.5% CMC-Na solution (without drug).

[0206] (3) Experimental Grouping

[0207] Eighty C57BL / 6 mice were randomly divided into eight groups (n=10 per group) according to their body weight.

[0208] Control group: Mice not infected with Plasmodium were given only an equal volume of 0.5% CMC-Na solution (without drugs).

[0209] Model group (untreated group after infection): Mice infected with Plasmodium were given an equal volume of 0.5% CMC-Na solution (without drugs).

[0210] DHA treatment group: Mice infected with Plasmodium were given DHA solutions at concentrations of 0.5 mg / mL, 1.5 mg / mL, and 3.0 mg / mL, respectively, in low, medium, and high dose groups.

[0211] F-DHA treatment group: Mice infected with Plasmodium were given F-DHA solution at concentrations of 0.5 mg / mL, 1.5 mg / mL, and 3.0 mg / mL, respectively, in low, medium, and high dose groups.

[0212] (4) Model building

[0213] Anticoagulated whole blood was collected from PbK173-infected mice, centrifuged (1500×g, 10 min) to collect infected red blood cells (pRBCs), and the concentration was adjusted to 1×10⁻⁶ with PBS. 8 pRBCs·mL -1 Mice in each group (except the control group) were intraperitoneally injected with 0.2 mL of pRBCs suspension (containing 1×10⁻⁶ pRBCs). 7 pRBCs).

[0214] (5) Model Validation

[0215] Seventy-two hours post-infection, blood was collected from the tail vein to prepare thin-layer blood smears, which were then stained with Giemsa stain and examined under a microscope. The criterion for successful infection was: a parasite-infected red blood cell percentage ≥20% (infection rate = parasite-infected red blood cell count / total red blood cell count × 100%).

[0216] (6) Administration method

[0217] The dosage volume should be dynamically adjusted according to the mouse's body weight, calculated based on a standard gavage volume of 10 mL / kg.

[0218] Example: If the mouse weighs 20g, the volume of each administration is 0.2mL (i.e., 20g × 10mL / kg ÷ 1000 = 0.2mL).

[0219] Start time: The first dose should be administered 24 hours after Plasmodium inoculation (day 1 post-infection).

[0220] Treatment course: Administer medication continuously for 5 days (from day 1 to day 5 post-infection).

[0221] (7) Observation indicators

[0222] Changes in body weight: Record the body weight of mice at the beginning and end of the experiment.

[0223] Hematological parameters: 24 hours after the last administration, blood was collected from the tail vein, anticoagulated with EDTA, and the red blood cell count (RBC), hemoglobin (HGB), hematocrit (HCT), white blood cell count (WBC), and platelet count (PLT) were measured.

[0224] Blood smear: to observe the density of Plasmodium infection.

[0225] (8) Organ harvesting

[0226] After euthanizing the mice, the heart, liver, spleen, and kidneys were quickly removed, preserved in tissue fixative, and then stained with hematoxylin and eosin (HE) for pathological observation. The heart, liver, and spleen tissues were blotted dry with filter paper, weighed, and their organ indices were calculated before fixation.

[0227] Organ index (%) = organ mass (g) / mouse body mass (g) × 100%.

[0228] (9) Data Processing and Statistical Analysis

[0229] Data are expressed as mean ± standard deviation (Mean ± SD). One-way ANOVA was performed using GraphPadPrism 9.0. Tukey's test was used for comparisons between groups, and P < 0.05 was considered statistically significant.

[0230] The control group, model group, and smears of 30 mg / kg / day DHA and 30 mg / kg / day F-DHA in mice infected with Plasmodium during the treatment period in Example 1 are shown below. Figure 18 As shown, from Figure 18 As can be seen, the model group exhibited typical Plasmodium proliferation characteristics, while the control group showed no signs of infection, ensuring the reliability of the experiment. Typical Plasmodium proliferation characteristics (such as multinucleated schizonts) were observed in the model group. The number of parasites was significantly reduced in the DHA and F-DHA groups, accompanied by schizont rupture and pigment granule dispersion.

[0231] The results of H&E staining of organ pathological sections from mice infected with parasites during the treatment period in the blank group, model group, and Example 1 (30 mg / kg / day DHA and 30 mg / kg / day F-DHA) are as follows: Figure 19 As shown, from Figure 19It can be seen that, compared with the uninfected group, the perivascular spaces of the cardiac tissue in the infected group were widened, and focal myocardial fiber rupture was occasionally observed. In the kidney tissue, capillary congestion and basement membrane thickening were observed, with occasional immune complex deposition. Macrophages proliferated in the hepatic sinusoids and engulfed a large number of cells containing malarial pigment; a large number of erythrocytes were retained in the splenic sinusoids, and macrophages proliferated with malarial pigment deposition; brownish-brown malarial pigment granules (residues of hemoglobin digested by Plasmodium) were visible in the macrophage cytoplasm. The pathological severity of DHA and F-DHA in the treatment group was reduced compared with the infected group, and the treatment effect of the F-DHA group was more significant than that of the DHA group.

[0232] On day 1 after the end of the drug administration period, the mice were weighed, peripheral blood was collected into EDTA anticoagulant tubes, and the spleen, heart, and liver were harvested, weighed, and their organ indices were calculated. Organ index = organ weight (g) / mouse body weight (g) × 100%.

[0233] Table 11 shows the spleen index data of mice treated with different concentrations of DHA and F-DHA in the blank group, model group, and Example 1 (5 groups of effective spleen index data). The bar chart is shown below. Figure 20 As shown.

[0234] Table 11 Spleen index data of different concentrations of DHA and F-DHA in the blank group, model group, and Example 1 after treatment in malaria-infected mice.

[0235]

[0236] Figure 20 In the table, ### indicates P < 0.001 (extremely significant), *** indicates P < 0.001 (extremely significant), and **** indicates P < 0.0001 (highly significant). From Table 11 and... Figure 20 As can be seen, when comparing the spleen index of mice during the treatment period, there was a significant statistical difference between the untreated group and the control group (###(P<0.001)). Compared with the infected group, there were statistically significant differences in the low, medium, and high dose groups of DHA (***(P<0.001)), and similarly, there were statistically significant differences in the low, medium, and high dose groups of F-DHA (****(P<0.0001)). However, the spleen index of the F-DHA group was closer to that of the untreated group, indicating that the anti-malarial activity of F-DHA was superior to that of DHA.

[0237] The CCK-8 assay results of this invention showed that F-DHA significantly inhibited cell viability in various leukemia cell lines, with a superior effect compared to DHA. This indicates that fluorination modification may enhance the antitumor activity of the drug, enabling it to more effectively target and inhibit the proliferation of leukemia cells. Further apoptosis experiments showed that in three leukemia cell lines—THP-1, KG-1, and L1210—both DHA and F-DHA significantly promoted apoptosis, but the effect of F-DHA was more significant, especially in mid-apoptosis, where its effect showed a good dose-dependent relationship. This result demonstrates that F-DHA has a higher efficiency in inducing apoptosis in leukemia cells, possibly by more effectively activating the apoptosis pathway. Western blotting results further revealed the regulatory capacity of F-DHA in the apoptosis pathway. Compared to DHA, F-DHA significantly upregulated the expression of Caspase3 / 9 proteins, and this upregulation was statistically significant at a concentration of 0.5 μg / mL (P < 0.001). This indicates that fluorination enhances the interaction between the drug and key proteins in the apoptosis pathway, thereby triggering apoptosis more effectively.

[0238] F-DHA exhibited more significant antitumor activity than DHA in the treatment of leukemia. Its fluorination modification not only enhanced its ability to inhibit the viability of leukemia cells, but also significantly improved the efficiency of inducing apoptosis, especially in the regulation of apoptosis-related proteins.

[0239] This invention compares the spleen index of mice during the drug administration period to investigate the significant effects of F-DHA at different doses on PbK strain-infected mice. Compared with the untreated infected group, the DHA and F-DHA-treated groups showed statistically significant differences at all doses, indicating that these compounds can effectively inhibit the pathological changes caused by Plasmodium infection. In particular, the spleen index of mice treated with F-DHA at low, medium, and high doses was closer to that of the uninfected group, suggesting that F-DHA may have a stronger anti-malarial effect. Furthermore, observation of blood smears and organ pathology results revealed that F-DHA treatment significantly reduced the proliferation of Plasmodium and alleviated infection-induced organ damage, especially in the heart and spleen. These results indicate that F-DHA not only effectively inhibits the proliferation of Plasmodium but may also have a protective effect against infection-induced tissue damage.

[0240] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A fluorine-modified dihydroartemisinin having the following chemical structure:

2. The method for preparing fluorine-modified dihydroartemisinin according to claim 1, comprising: Under an inert atmosphere, a fluorinating reagent was mixed with a dihydroartemisinin solution to carry out a substitution reaction, yielding fluorinated dihydroartemisinin. The fluorinating agent is added at a rate of 0.2–0.5 mL / min.

3. The preparation method according to claim 2, characterized in that, During the mixing of the fluorinating reagent and the dihydroartemisinin solution, the temperature of the reaction system is controlled at -60 to -20°C; after the fluorinating reagent is added, the reaction system is transferred to 20 to 30°C.

4. The preparation method according to claim 2, characterized in that, The fluorinating agent includes diethylaminosulfur trifluoride, N,N-bis(2-methoxyethyl)trifluoromethanesulfonamide, 2-tert-butyl-5-methyl-3-trifluoromethylthiobenzene, or 2-fluoropyridine-1-oxide.

5. The preparation method according to claim 2 or 4, characterized in that, The molar ratio of dihydroartemisinin to fluorination reagent in the dihydroartemisinin solution is 1:(1-2).

6. The preparation method according to claim 2, characterized in that, The solvent in the dihydroartemisinin solution is one or more of dichloromethane, acetonitrile, chloroform, and methanol.

7. The preparation method according to claim 2 or 6, characterized in that, The concentration of the dihydroartemisinin solution is 0.03–0.04 mol / L.

8. The preparation method according to claim 2, characterized in that, After the substitution reaction is completed, a quenching agent is added to quench the reaction. The quenching agent is an alkaline solution.

9. The preparation method according to claim 8, characterized in that, The quenching process further includes sequentially separating the organic phase, drying, and removing the solvent from the obtained quenching system to obtain fluorine-modified dihydroartemisinin.

10. The use of the fluorine-modified dihydroartemisinin of claim 1 in the preparation of antimalarial, antitumor, antiviral, antibacterial or anti-inflammatory drugs.

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