Icaritin derivatives, and preparation method and use thereof

Abstract

The present invention belongs to the field of pharmaceutical chemistry and relates to icariin derivatives, such as formula (I), and their pharmacologically tolerable salts and processes for their preparation, and their use for the preparation of medicaments for the treatment of asthma and myelosuppression.

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry, specifically relating to icariin derivatives and their pharmacologically permissible salts, as well as their preparation methods and applications. Background Technology

[0002] Icaritin (ICT) is a polyhydroxyflavonoid monomer from Epimedium, a plant belonging to the genus Epimedium in the family Berberidaceae. Its structural formula is as follows:

[0003]

[0004] Icariin can be isolated from icariin or its in vivo metabolites (Sun Pengyue, Xu Ying, Wen Ye et al., Chemical constituents of Korean icariin, Chinese Journal of Phytochemistry, 1998, 8(2): 122-125; Liu Tiehan, Wang Yi, Wu Lijun et al., Study on intestinal bacterial metabolism of icariin I. Metabolic transformation of icariin by intestinal bacteria, 2000, 31(11): 834-837), or obtained by enzymatic hydrolysis of icariin (Ye Haiyong, Liu Jian, Lou Yijia, Preparation of icariin derivatives and its estrogen-like effects, Journal of Zhejiang University, 2005, 34(2): 131-136).

[0005] Literature reports that icariin has an anti-apoptotic effect on Aβ-induced apoptosis in primary cultured rat neurons (Zhang Xiangnan, Wang Huanhuan, Wang Zhiqiang, et al., Anti-apoptotic effect of icariin on Aβ-induced apoptosis in primary cultured rat neurons, Journal of Zhejiang University, 2007, 36(3): 224-226). Chinese patent CN101836976A discloses that icariin has an anti-tumor angiogenesis effect. Chinese patent CN101428015A discloses that icariin has an anti-endotoxemia effect. Chinese patent CN101284000A discloses that icariin has a role in preventing and treating obesity or fatty liver. Chinese patent CN1869204A discloses the use of icariin in inducing in vitro directed differentiation of stem cells.

[0006] Icariin possesses a wide range of pharmacological activities, but its solubility is poor. It is slightly soluble in dichloromethane and ethyl acetate, almost insoluble in methanol, anhydrous ethanol, and water, and almost insoluble or insoluble in various pH buffers. Icariin has low oral bioavailability. Summary of the Invention

[0007] The purpose of this invention is to provide a novel icariin derivative and its pharmacologically permissible salt.

[0008] In a first aspect of the invention, icariin derivatives and pharmacologically permissible salts thereof are provided, the structures of which are shown in formula (I).

[0009]

[0010] In formula (I), R1 and R3 are independently selected from hydrogen, C groups substituted with amino, hydroxyl, and carboxyl groups, respectively. 1-4 Alkyl, L, -(CH2) m OX, C 1-4 Alkyl-substituted aminoacyl group;

[0011] L is selected from hydrogen, glycine, alanine, valine, isoleucine, leucine, threonine, serine, glutamic acid, lysine, and -CO(CH2). m COOH;

[0012] The halogen is selected from F, Cl, Br or I;

[0013] R7 and R8 are independently selected from hydrogen, halogen, hydroxyl, and amino, respectively.

[0014] R1 and R3 are not simultaneously represented by H or C groups substituted with amino, hydroxyl, or carboxyl groups. 1-4 alkyl.

[0015] Furthermore, the C 1-4 Alkyl groups are methyl, ethyl, propyl, isopropyl, butyl, isobutyl, and tert-butyl.

[0016] Furthermore, equation (I) is:

[0017]

[0018] The preparation method of the icariin derivative of the present invention can be carried out by, but is not limited to, the following methods:

[0019] Method 1: When R1 and R3 have the same functional group, the steps are as follows:

[0020] 1)

[0021] Icariin (A) undergoes an addition reaction with R7-R8 to give compound 1;

[0022] 2)

[0023] Compound 1 is directly condensed with L or halogenated R1 under condensing agent conditions to generate compound (I); or icariin A is first substituted with a haloalcohol under the action of a base to generate an intermediate, which is then condensed with L under condensing agent conditions to generate compound (I).

[0024] Method 2: When R1 and R3 take different functional groups, the steps are as follows:

[0025] 1) Starting material B undergoes an addition reaction with R7-R8 to give compound 2;

[0026]

[0027] 2) Compound 2 is directly condensed with L or haloR3 under condensing agent conditions to generate compound 3; or B is first substituted with haloalcohol under the action of base to generate an intermediate, which is then condensed with L under condensing agent conditions to generate compound 3.

[0028] 3) 3. Remove Boc protection under acidic conditions; then react with L or halo-R1 under alkaline conditions to generate (I).

[0029] The condensing agent is selected from one of the following: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride / 4-dimethylaminopyridine EDCI / DMAP, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride / 1-hydroxybenzotriazole / N,N-diisopropylethylamine EDCI / HOBT / DIPEA, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate / N,N-diisopropylethylamine HATU / DIPEA, and dicyclohexylcarbodiimide / 4-dimethylaminopyridine DCC / DMAP.

[0030] The amount of the condensing agent used is 0.1-25 equivalents.

[0031] Another object of the present invention is to provide a pharmaceutical composition comprising the epimedium aglycone derivatives described herein or a pharmacologically permissible salt thereof.

[0032] Another object of the present invention is to provide the use of icariin derivatives and their pharmacologically permissible salts in the preparation of medicaments for the treatment of asthma or myelosuppression.

[0033] The icariin derivatives of this invention exhibit significantly better therapeutic effects than icariin itself for asthma. Furthermore, the icariin derivatives of this invention show significantly better therapeutic effects than icariin itself for bone marrow suppression induced by radiotherapy or chemotherapy. Finally, the bioavailability of the icariin derivatives of this invention is significantly better than that of icariin itself, indicating that the compounds of this invention possess significant technical advantages in terms of drug-likeness.

[0034] The icariin derivative of this invention exhibits significantly superior therapeutic effects on asthma compared to the compound of Formula I. The icariin derivative of this invention also shows significantly superior therapeutic effects on bone marrow suppression induced by radiotherapy or chemotherapy compared to the compound of Formula I. Furthermore, the bioavailability of the icariin derivative of this invention is significantly better than that of the compound of Formula I, indicating a clear technical advantage in terms of drug-likeness. More importantly, long-term toxicity studies revealed that the compound of Formula I exhibits certain hepatotoxicity and nephrotoxicity, and causes mammary hyperplasia in female rats, while the compound of this invention shows no significant toxicity, demonstrating higher clinical safety compared to the compound of Formula I.

[0035] Compound I is compound 5 from the literature (Dell'Agli M, Galli GV, Dal CE, et al. Potent inhibition of human phosphodiesterase-5 by icariin derivatives.[J]. Journal of Natural Products, 2008, 71(9):1513-1517.). The structural formula of compound I is as follows:

[0036] Detailed Implementation

[0037] The following detailed embodiments illustrate the present invention. However, these embodiments should not be construed as limiting the invention. Various substitutions or modifications made based on common technical knowledge and conventional methods in the art without departing from the aforementioned spirit of the invention are all included within the scope of this invention.

[0038] Example 1

[0039]

[0040] 500 mg (1.4 mmol) of icariin (ICT) was dissolved in 20 ml of acetone, and 188 mg (1.4 mmol) of potassium carbonate and 0.1 ml (1.4 mmol) of 2-bromoethanol were added. The mixture was refluxed until the reaction was complete. The solvent was evaporated under reduced pressure, dissolved in ethyl acetate, and purified by column chromatography to give 1,378 mg of a yellow solid intermediate, with a yield of 61%.

[0041] 1H NMR (300MHz, CDCl3): 12.44 (s, 1H), 8.13 (d, J = 9.00Hz, 2H), 7.08 (d, J = 9.06Hz, 2H), 6.43 (s, 1H), 5.20 (q, J = 15.53Hz, 1H), 4.20 (t, J=8.91Hz,2H),4.02(m,4H),3.93(s,3H),3.83(m,2H),3.56(d,J=6.72Hz,2H),1.82(s,3H),1.72(s,3H).ESI-MS(m / z):479[M+Na] + .

[0042] Example 2: Synthesis of compound W-4.

[0043]

[0044] 1,456 mg (1 mmol) of intermediate was dissolved in 20 mL of dichloromethane, and 217 mg (1 mmol) of BOC-valine, 192 mg (1 mmol) of carbodiimide (EDCI), and 122 mg (1 mmol) of 4-dimethylaminopyridine (DMAP) were added. The mixture was stirred at room temperature for 2 h. After the reaction was complete, the product was purified by column chromatography to obtain 632 mg of yellow solid product, with a yield of 89%.

[0045] The obtained yellow solid 632 mg product was dissolved in 10 ml of dichloromethane, and 139 mg (1.39 mmol) of concentrated hydrochloric acid was added under stirring. After the addition was completed, the mixture was stirred at room temperature for 20 min, and then filtered to obtain yellow solid W-4 520 mg, with a total yield of 70%.

[0046] 1 H NMR (300MHz, CDCl3): 12.67 (s, 1H), 8.58 (s, 6H), 8.15 (d, J = 9Hz, 2H), 7.13 (d, J = 9Hz, 2H), 6.64 (s, 1H), 4.52-4.42 (m, 8H), 3.87 (s,1H),2.95(d,J=9.24Hz,2H),2.21-2.11(m,2H),1.99-1.89(m,2H),1.65(s,6H),1.00-0.91(m,6H).ESI-MS(m / z):691[M+H] + .

[0047] Example 3

[0048]

[0049] 500 mg (1.1 mmol) of Boc-protected icariin (Boc-ICT) was weighed into a 100 mL round-bottom flask. 15 mL of dichloromethane was added as solvent, and 0.14 mL (1.65 mmol) of isopropyl isocyanate and 32 μl (0.22 mmol) of triethylamine were added under stirring at room temperature. The mixture was refluxed. After the reaction was complete, the product was purified by column chromatography (petroleum ether: ethyl acetate = 4:1) to give 450 mg of a yellow solid. This solid was then placed in a 100 mL round-bottom flask, and 15 mL of dichloromethane was added as solvent. The mixture was cooled in an ice bath, and 5 mL of trifluoroacetic acid was added. After the reaction was complete, the product was separated by column chromatography (petroleum ether: ethyl acetate = 4:1) to give 340 mg of a yellow solid, with a yield of 91%.

[0050] 340 mg (0.75 mmol) of the obtained yellow solid product was placed in a 100 mL round-bottom flask, 20 mL of acetone was added as solvent, along with 0.1 mL (1.5 mmol) of 2-bromoethanol and 207 mg (1.5 mmol) of potassium carbonate. The reaction was carried out at 60 °C. After the reaction was complete, the mixture was filtered, the filtrate was evaporated to dryness, and separated by column chromatography (petroleum ether: ethyl acetate = 4:1) to give 2290 mg of the yellow solid intermediate, with a yield of 78%.

[0051] Example 4

[0052]

[0053] Weigh 2,450 mg of the intermediate into a 100 ml round-bottom flask, add 15 ml of dichloromethane as solvent, cool in an ice bath, add 5 ml of trifluoroacetic acid, and after the reaction is complete, separate by column chromatography to obtain a yellow solid. Then, place the solid into a 100 ml round-bottom flask, add 20 ml of acetone as solvent, add 0.1 ml of 2-bromoethanol and 207 mg of potassium carbonate, and react at 60 °C. After the reaction is complete, filter, evaporate the filtrate to dryness, separate by column chromatography to obtain a yellow solid, then dissolve it in 10 ml of dichloromethane, add 139 mg of concentrated hydrochloric acid under stirring, and stir at room temperature for 20 min after the addition is complete. Filter to obtain 363 mg of yellow solid W-4-1, with an overall yield of 69%.

[0054] MS: 534.3.

[0055] Example 5: Effect of the compounds of the present invention on chemotherapy-induced bone marrow suppression in tumor-bearing mice.

[0056] 1. Model preparation and grouping: Seventy-two Balb / C mice were selected, weighed, and randomly divided into six groups: normal group, blank control group, icariin group, Formula I group, and each group treated with the compound of this invention, with 12 mice in each group. Mice were intraperitoneally injected with 50 mg / kg cyclophosphamide daily for one week, which significantly reduced platelet count. One week before model establishment, each treatment group received the following treatment drugs:

[0057] Normal group: Administered an equal volume of sodium carboxymethyl cellulose;

[0058] Blank control group: Administered an equal volume of sodium carboxymethyl cellulose;

[0059] Epimedium aglycone group: 5 mg / kg epimedium aglycone was administered by gavage;

[0060] Formula I group: 5 mg / kg of Formula I compound was administered by gavage;

[0061] Group W-4: 0.5 mg / kg of compound W-4 was administered by gavage;

[0062] Group W-4-1: 0.5 mg / kg of compound W-4-1 was administered by gavage;

[0063] Each treatment group was administered the drug once daily and fed normally. After 3 weeks of continuous administration, mice were anesthetized, blood was collected, and white blood cell and platelet counts were measured to investigate the effects of the compound of this invention on white blood cells and platelets.

[0064] 2. Experimental Results

[0065] Through the effects of the compounds of the present invention on cyclophosphamide-induced white blood cells and platelets in mice in this embodiment (Table 1), it was found that the compounds of the present invention have an unexpected effect in alleviating chemotherapy-induced bone marrow suppression.

[0066] Compared with the model control group, the total number of white blood cells and the total number of platelets in each compound group of the present invention were increased, and the differences were extremely significant.

[0067] Compared with the icariin group, the total number of white blood cells and platelets in each compound group of the present invention increased significantly.

[0068] Compared with Formula I, the total number of white blood cells and the total number of platelets in each compound group of the present invention are significantly increased.

[0069] Table 1. Effects of each compound on cyclophosphamide-induced leukopenia and thrombocytopenia.

[0070]

[0071] Compared with the blank control group, $ p<0.05, $$ p<0.01;

[0072] Compared with the icariin group, # p<0.05, ## p<0.01;

[0073] Compared with Equation I, & p<0.05,&& p<0.01.

[0074] Example 6: Compounds of the present invention 60 Effect of Co-irradiated mice on blood cell count

[0075] 1. Model preparation and grouping for drug administration: Ninety Kunming mice were selected. Except for the normal control group (n=10), the other groups of mice were irradiated with 4 Gy. 60 Co radiation was administered via a single whole-body irradiation session with an absorbed dose of 4 Gy at a rate of 0.88 Gy / min. Whole blood cell counts were determined by collecting blood samples from the orbital vein on days 3, 7, and 10 post-irradiation. A white blood cell count below 3.0 × 10⁻⁶ was considered acceptable. 9 / L or platelet count less than 500×10 9 Mice with a weight of / L were removed, and the remaining mice were used for experiments.

[0076] Mice that met the experimental requirements after irradiation were randomly divided into the following groups: model group, icariin group, formula I group, W-4-1 group, and W-4 group, with 10 groups in each group and half male and half female. Each group was treated or administered drugs as follows.

[0077] Normal group: Administered an equal volume of sodium carboxymethyl cellulose;

[0078] Blank control group: Administered an equal volume of sodium carboxymethyl cellulose;

[0079] Icariin group: intraperitoneal injection of 1 mg / kg icariin;

[0080] Formula I group: Intraperitoneal injection of 1 mg / kg of Formula I compound;

[0081] Group W-4: Intraperitoneal injection of 0.1 mg / kg of compound W-4;

[0082] Group W-4-1: Intraperitoneal injection of 0.1 mg / kg of compound W-4-1;

[0083] Each treatment group was administered the drug once daily and fed normally. After 10 consecutive days of administration, mice were anesthetized, blood was collected, and the numbers of white blood cells and platelets were measured to investigate the effect of the compound of this invention on white blood cells and platelets.

[0084] 2. Experimental Results

[0085] Through this embodiment, the various compounds of the present invention are used to accept... 60 The effects of Co radiation on white blood cells and platelets in mice (Table 2) revealed that the compounds of this invention have an unexpected effect in alleviating radiotherapy-induced bone marrow suppression.

[0086] Compared with the model control group, the total number of white blood cells and the total number of platelets in each compound group of the present invention were increased, and the differences were significant.

[0087] Compared with the icariin group, the total number of white blood cells and platelets in each compound group of the present invention increased significantly.

[0088] Compared with Formula I, the total number of white blood cells and the total number of platelets in each compound group of the present invention are significantly increased.

[0089] Table 2. Pairs of each compound 60 Effect of Co-irradiated mice on blood cell count

[0090]

[0091] Compared with the blank control group, $ p<0.05, $$ p<0.01;

[0092] Compared with the icariin group, # p<0.05, ## p<0.01;

[0093] Compared with Equation I, & p<0.05, && p<0.01.

[0094] Example 7: Inhibitory effect of the compounds of the present invention on airway smooth muscle

[0095] Airway remodeling is an important pathological feature of bronchial asthma, and airway smooth muscle cells (ASMCs) are the main effector cells causing airway remodeling. Compared with normal individuals, patients with severe asthma have significantly increased ASMC levels, a phenomenon mainly caused by smooth muscle cell proliferation.

[0096] 1.1 Materials

[0097] 150–200 g SD rats; DM EM medium (Gibico); fetal bovine serum (Hangzhou Sijiqing Co., Ltd.); trypsin (Sigma).

[0098] 1.2 Experimental Methods

[0099] 1.2.1 Culture of rat ASM C

[0100] Following the method described in relevant literature: Under aseptic conditions, the trachea was longitudinally dissected, the outer membrane was carefully peeled off, and the inner membrane was gently scraped away. The tracheal segment was carefully cut into small tissue blocks (1mm × 1mm × 1mm) using ophthalmic scissors. These blocks were then placed on the bottom of a 5cm × 5cm culture flask, arranged at equal intervals. 2mL of DM EM (high glucose) medium containing 25% fetal bovine serum was added, ensuring the culture medium did not contact the tissue blocks. The culture flask was placed bottom-up and incubated at 37°C with 5% carbon dioxide for approximately 3 hours to allow the tissue blocks to nearly dry. The culture flask was then gently inverted so that the culture medium just covered the surface of the tissue blocks. After 3 days of semi-open, absolutely static culture, the culture medium was added to 5mL. The medium was completely changed after 6 days, and thereafter changed every 3 days. After approximately 7 days, the cells reached confluence and were passaged. Cells from passages 4-5 were used in the experiment. The cultured rat ASM C cells were identified morphologically.

[0101] 1.2.2 Detection of ASMC proliferation in rats using the CCK-8 assay

[0102] Fourth-generation rat ASMCs were cultured and prepared into a single-cell suspension, at a concentration of 1×10⁻⁶ cells / cells. 4 Cells were seeded per well in a 96-well plate and cultured at 37°C with 5% carbon dioxide for 24 hours. When the cells reached a confluent state, culture medium containing 0.1% fetal bovine serum was added (to arrest cell growth in the G0 phase) and the cells were cultured for another 24 hours.

[0103] The culture medium was replaced with one containing 1% fetal bovine serum, and the cells were randomly divided into:

[0104] Control group: DMEM was added to each well only;

[0105] The icariin group, with an icariin concentration of 1×10⁻⁶. -6 mol / L;

[0106] Formula I group: The concentration of compound I is 1×10 -6 mol / L;

[0107] Group W-4-1: The concentration of compound W-4-1 is 1×10 -6 mol / L;

[0108] Group W-4: The concentration of compound W-4 was 1×10⁻⁶ -6 mol / L;

[0109] Each group was configured with 5 replicates, and a control group (containing no cells, but with the same DMEM concentration as the corresponding group) was also included in each group. After culturing at 37℃ and 5% CO2 for 48 h, 10 μL of CCK-8 reagent was added to each well, and the cells were cultured for another 4 h. The OD value of each well was measured at 450 nm. The cell growth inhibition rate of each group was calculated using the formula: Cell growth inhibition rate = 1 - [(OD value of each drug-treated group - OD value of the corresponding control group) / (OD value of the control group - OD value of the corresponding control group)] × 100%.

[0110] 2 Results

[0111] 2.1 ASMC Identification

[0112] Observation with an inverted phase contrast microscope revealed that ASMCs were spindle-shaped before merging, and after merging, cells in some areas were arranged in bundles, exhibiting a typical "peak and valley" shape.

[0113] 2.2 Effects on ASMC proliferation

[0114] After 48 hours of treatment with ASMCs, the OD values ​​of the cells in each compound group of the present invention were significantly lower than those in the icariin group, with statistically significant differences.

[0115] Compared with the icariin group of cells, the inhibition rate of all groups of cells in this invention was significantly increased, and the differences were statistically significant.

[0116] Compared with the cells in Formula I, the inhibition rates of all groups of cells in this invention were significantly increased, with statistically significant differences (see Tables 3 and 4).

[0117] Table 3. OD values ​​of each compound after 48 hours of treatment with ASMC

[0118]

[0119] Compared with the normal control group, $ p<0.05, $$ p<0.01;

[0120] Compared with the icariin group, # p<0.05, ## p<0.01;

[0121] Compared with Equation I, & p<0.05, && p<0.01.

[0122] Table 4. Inhibition rate of each compound on ASMC after 48 hours.

[0123]

[0124] Compared with the icariin aglycone group, $ p < 0.05, $$ p < 0.01;

[0125] Compared with the group of Formula I, & p < 0.05, && p < 0.01.

[0126] Example 8 Determination of the Bioavailability of the Compounds of the Present Invention

[0127] 1. Animal Grouping and Administration

[0128] Forty-eight Wistar rats (270 ± 30) g, half male and half female, were provided by the Experimental Animal Center of Shandong Xinsdailai Pharmaceutical Co., Ltd., with the production license number: SCXK (Lu) 20060019. They were raised under the conditions of temperature 20 - 22°C, relative humidity 45% - 65%, and light / dark 12 h / 12 h, with free access to food and water.

[0129] Intragastric Administration Group of the Compounds of the Present Invention: Twenty-four healthy Wistar rats that had fasted for 12 hours and had free access to water, half male and half female, were divided into 4 groups: the icariin aglycone group (intragastrically administered icariin aglycone), the group of Formula I (intragastrically administered the compound of Formula I), the W-4-1 group (intragastrically administered the compound W-4-1), and the W-4 group (intragastrically administered the compound W-4). Each group was administered intragastrically once, with the administration dose of 3 mg / kg. They fasted for 12 h before administration and had free access to water. Approximately 300 μL of blood was taken from the retroorbital venous plexus at 0 h (before administration), 0.083, 0.25, 0.5, 1, 1.5, 2, 3, 4, 6, 8, 12, and 24 h after administration. The blood was anticoagulated with heparin and centrifuged at 12000 rpm at 4°C for 5 min to separate plasma, which was stored in a -20°C low-temperature refrigerator. During the experiment, they had free access to water and ate 2 h after intragastric administration.

[0130] Intravenous Administration Group of the Compounds of the Present Invention: Twenty-four healthy Wistar rats that had fasted for 12 hours and had free access to water, half male and half female, were divided into 4 groups: the icariin aglycone group (injected with icariin aglycone), the group of Formula I (injected with the compound of Formula I), the W-4-1 group (injected with the compound W-4-1), and the W-4 group (injected with the compound W-4). Each group was administered by tail vein injection, with the administration dose of 3 mg / kg. Approximately 300 μL of blood was taken from the retroorbital venous plexus at 0 h (before administration), 0.033, 0.083, 0.25, 0.5, 1, 1.5, 2, 3, 4, 6, 8, 12, and 24 h after administration. The blood was anticoagulated with heparin and centrifuged at 12000 rpm at 4°C for 5 min to separate plasma, which was stored in a -20°C low-temperature refrigerator. During the experiment, they had free access to food and water.

[0131] [[ID=二十九]]2. Determination of Plasma Samples

[0132] All processed plasma samples were subjected to UPLC-MS / MS quantitative analysis to determine plasma drug concentration.

[0133] 3. Calculation of bioavailability

[0134] The measured blood drug concentration-time data were used to calculate pharmacokinetic parameters using DAS software (Drug and Statistics, compiled by Sun Ruiyuan et al., Chinese Society for Mathematical Pharmacology). The absolute bioavailability of each compound was calculated according to the formula, where t is the sampling time at which the final measurable drug concentration was obtained.

[0135]

[0136] 4. Absolute bioavailability of each compound

[0137] Table 5 Bioavailability of each compound

[0138]

[0139] As can be seen from the table above, the bioavailability of each compound in this invention is significantly higher than that of icariin and compound I, indicating that the compounds in this invention have significant technical advantages in terms of drug-likeness.

[0140] Example 9: Toxicity study of the compound of the present invention and compound of formula I in rats after repeated injection.

[0141] SD rats were divided into four groups: normal control group, compound I group (intravenous injection of compound I 100 mg / kg / d), W-4 group (intravenous injection of W-4 ​​100 mg / kg / d), and W-4-1 group (intravenous injection of W-4-1 100 mg / kg / d).

[0142] Animals in each group were administered the drug continuously for 28 days, once daily, followed by a 4-week recovery period after drug withdrawal. At the end of the treatment period, blood samples were collected from the abdominal main vein for hematological, coagulation time, blood biochemistry, and electrolyte analysis. Subsequently, a systematic dissection was performed to observe the morphology of each organ and tissue. The brain, spleen, thymus, heart, kidneys (bilateral), liver, adrenal glands (bilateral), mammary glands (female rats), and prostate (male rats) were weighed, and organ coefficients were calculated.

[0143] Experiments revealed elevated levels of creatinine, alkaline phosphatase, and alanine aminotransferase in rats treated with compound I. Histopathological examination showed varying degrees of renal tubular damage, indicating significant nephrotoxicity. Mammary hyperplasia was observed in female rats. Compound I exhibited certain toxic side effects.

[0144] Under the conditions of this experiment, no abnormal changes related to W-4 toxicity were found. W-4 injection administration showed no significant toxicity, and its clinical use is highly safe.

[0145] Under the conditions of this experiment, no abnormal changes related to W-4-1 toxicity were found. W-4-1 injection administration showed no significant toxicity, and its clinical use is highly safe.

Claims

1. An icariin derivative, the structure of which is shown in formula (I): , or a pharmaceutically acceptable salt of a compound of formula (I); In formula (I), R1 is replaced by an amino or hydroxyl group. 1-4 Alkyl groups, R3 with C 1-4 Alkyl-substituted aminoacyl group; R7 is selected from halogens, and R8 is selected from hydrogen; the halogens are selected from F, Cl, Br or I.

2. The icariin derivative as described in claim 1, characterized in that, The compound of formula (I) is specifically: .

3. An icariin derivative, the structure of which is shown in formula W-4: 。 4. The method for synthesizing W-4 as described in claim 3, characterized in that... The synthesis steps are as follows: 1） ， The raw material icariin was reacted with 2-bromoethanol to obtain intermediate 1; 2） ， Intermediate 1 was reacted with BOC-valine to give W-4.

5. The method for synthesizing the icariin derivative as described in claim 1, characterized in that, The synthetic steps of compound (I) are as follows:

3. Remove Boc protection under acidic conditions; then react with halo-R1 under alkaline conditions to generate (I).

6. The synthesis method as described in claim 5, characterized in that, The synthesis steps of compound 3 are as follows: Compound 2 and halo-R3 directly condense under condensing agent conditions to form compound 3.

7. The synthesis method according to claim 6, characterized in that, The synthesis steps of compound 2 are as follows: Starting material B undergoes an addition reaction with R7-R8 to give compound 2; 。 8. The method for synthesizing the icariin derivative as described in claim 6 or 7, characterized in that, in, The condensing agent is selected from one of the following: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride / 4-dimethylaminopyridine EDCI / DMAP, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride / 1-hydroxybenzotriazole / N,N-diisopropylethylamine EDCI / HOBT / DIPEA, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate / N,N-diisopropylethylamine HATU / DIPEA, and dicyclohexylcarbodiimide / 4-dimethylaminopyridine DCC / DMAP.

9. A pharmaceutical composition comprising the icariin derivative of any one of claims 1-3 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

10. Use of the pharmaceutical composition of claim 9 in the preparation of a medicament for treating asthma or myelosuppression.

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