CN113234117A

CN113234117A - Hederagenin C-28 polyethylene glycol modified derivative and preparation method thereof

Info

Publication number: CN113234117A
Application number: CN202110705056.7A
Authority: CN
Inventors: 毕毅; 陈晓倩; 程浩然; 黄文涛; 王炳华; 曾文轩; 马梦馨; 梁凯程
Original assignee: Yantai University
Current assignee: Yantai University
Priority date: 2021-06-24
Filing date: 2021-06-24
Publication date: 2021-08-10

Abstract

The invention discloses a C-28 polyethylene glycol modified hederagenin derivative and a preparation method thereof. The derivatives of the invention includeN- (23-hydroxyolean-12-en-28-oylo [3, 2-)b]Pyrazine) -4-aminobutyric acid diethylene glycol ester;N- (23-hydroxyolean-12-en-28-oylo [3, 2-)b]Triethylene glycol 4-aminobutyric acid, and the like. The hederagenin polyethylene glycol modified derivative and the medically acceptable salt thereof have excellent tumor drug resistance reversal activity and higher water solubility, so that the drug resistance reversal activity and the water solubility of the obtained hederagenin derivative are remarkably improved, and the hederagenin derivative has a drug property.

Description

Hederagenin C-28 polyethylene glycol modified derivative and preparation method thereof

Technical Field

The invention relates to the field of organic synthesis and pharmaceutical chemistry, in particular to a C-28 polyethylene glycol modified hederagenin derivative with a novel structure and a preparation method thereof.

Technical Field

Recent cancer data from the international agency for research on cancer (IARC) show: in 2020, the number of newly added cancers is estimated to be 1930 ten thousand in total, and the number of cancer death cases is estimated to be 1000 ten thousand in total. Once Multidrug Resistance (MDR) has developed, the chemotherapeutic effect of the drug will be significantly reduced, and thus, tumor MDR has become one of the major obstacles to chemotherapy, and the development of tumor MDR reversal agents is imminent. Natural products and derivatives thereof with various structural types have been reported to have tumor drug resistance reversing activity and become an important source of tumor drug resistance reversing agents.

Hederagenin (H) is a pentacyclic triterpene compound separated from Hederagenin leaves and other plants. In the previous studies, alpha-Hederagenin (H) is found to have the biological Activity of resisting tumors, and the structural modification study of pentacyclic triterpene A ring splicing heterocycles is reported in the literature, so that a plurality of heterocyclic derivatives with better biological Activity are obtained (see Milan Urban, Jan Sarek, Miroslav Kvasica, et al. But the prior research simultaneously highlights the technical problems that: how to obtain the hederagenin derivative which has excellent tumor drug resistance reversion activity and higher water solubility simultaneously enables the drug resistance reversion activity and the water solubility of the obtained hederagenin derivative to be obviously improved, and the hederagenin derivative has the drug property.

Disclosure of Invention

The invention aims to provide a C-28 polyethylene glycol modified hederagenin derivative and a preparation method thereof. The invention aims to solve the technical problem of finding a compound which has a novel structure and good water solubility and has excellent tumor drug resistance reversal activity and a preparation method thereof.

In order to solve the technical problems, the invention provides the following technical scheme:

c-28 polyethylene glycol modified hederagenin derivative shown in general formula I and its pharmaceutically acceptable salt,

wherein the content of the first and second substances,

n =2-13, and n is a natural number.

Preferably, some of the compounds of the present invention are:

general formula I:

N- (23-hydroxyolean-12-en-28-oylo [3, 2-)b]Pyrazine) -4-aminobutyric acid diethylene glycol ester;

N- (23-hydroxyolean-12-en-28-oylo [3, 2-)b]Triethylene glycol 4-aminobutyric acid pyrazine;

N- (23-hydroxyolean-12-en-28-oylo [3, 2-)b]Pyrazine) -4-aminobutyric acid tetraethylene glycol ester;

N- (23-hydroxyolean-12-en-28-oylo [3, 2-)b]Pyrazine) -4-aminobutyric acid pentaethylene glycol ester;

N- (23-hydroxyolean-12-en-28-oylo [3, 2-)b]Pyrazine) -4-aminobutanoic acid hexaethylene glycol ester;

N- (23-hydroxyolean-12-en-28-oylo [3, 2-)b]Pyrazine) -4-aminobutyric acid heptaethylene glycol ester.

The preparation route of the hederagenin polyethylene glycol derivative provided by the invention is as follows:

the C-28 polyethylene glycol modified hederagenin derivative with the general formula I is synthesized and prepared by the following method:

a. taking hederagenin as a raw material, and protecting carboxyl by benzyl bromide in the presence of inorganic base;

b. protecting hydroxyl at the C-23 position by using tert-butyldimethylsilyl chloride;

c. the intermediate product protected by TBS is oxidized by chloropyridine chromate to obtain C-3 hydroxyl;

d. reacting with ethylenediamine under the catalysis of sulfur;

e. under the acidic condition, removing a C-23 protecting group;

f. under the catalysis of palladium-carbon, with H₂Reacting to remove C-28 benzyl;

g. reacting with oxalyl chloride to prepare acyl chloride, and reacting with methyl 4-aminobutyrate under the catalysis of triethylamine;

h. under the catalysis of DMAP and EDCI, reacting with diethylene glycol, triethylene glycol, tetraethylene glycol, pentaethylene glycol, hexaethylene glycol and heptaethylene glycol to obtain a crude product;

i. and purifying the crude product by using a dialysis method and a column chromatography method in sequence to obtain the target compound.

The C-28 polyethylene glycol modified hederagenin derivative has good water solubility and excellent tumor drug resistance reversal activity, and can be used for preparing tumor drug resistance reversal agents.

The C-28 polyethylene glycol modified hederagenin derivative and the optical isomer of the compound or the pharmaceutically acceptable solvate thereof.

The effective amount of the compound of the general formula I or the salt thereof and the pharmaceutically acceptable carrier are used for treating diseases or symptoms such as oral epithelial cancer, gastric cancer, lung cancer, cervical cancer, breast cancer or colon cancer and the like by combining with clinical common antitumor drugs.

The C-28 polyethylene glycol modified derivative of hederagenin of the general formula I and the application of the medical acceptable salt thereof are used for preparing tumor drug resistance reversal agents and/or medicinal carriers for treating mammals, preferably human diseases or symptoms.

The C-28 polyethylene glycol modified hederagenin derivative shown in the general formula I and the pharmaceutically acceptable salt thereof have excellent tumor drug resistance reversion activity and high water solubility, so that the drug resistance reversion activity and the water solubility of the obtained hederagenin derivative are remarkably improved, and the hederagenin derivative has drug properties.

Detailed Description

The invention is characterized in that the C-28 polyethylene glycol modified derivative of hederagenin in the general formula I and the medically acceptable salt thereof have the advantages that compared with the existing hederagenin derivative, the tumor drug resistance reversal activity is improved, the water solubility is also obviously improved, and the hederagenin derivative has good drug forming property.

The present invention will be described in further detail below by way of examples, but the present invention is not limited to only the following examples.

Example 1 N- (23-hydroxyolean-12-en-28-oylo [3, 2-)b]Synthesis and characterization of pyrazine) -4-aminobutyric acid diethylene glycol (H628 + D + P100)

Dissolving compound hederagenin (472.0 mg, 1.0 mmol) in N, N-dimethylformamide (15.0 mL), adding potassium carbonate (300.0 mg, 2.1 mmol) and benzyl bromide (0.2 mL, 1.3 mmol), and stirring at 50 deg.C for 6-10 h. The reaction mixture was diluted with ethyl acetate (25.0 mL), washed with water three times, washed with saturated brine two times, dried over anhydrous sodium sulfate, filtered, evaporated under reduced pressure to remove the solvent, and subjected to silica gel column chromatography (V)_{Petroleum ether}:V_{Ethyl acetate}= 10:1-5:1) to give a white solid (470.0 mg, 83.0%).

The above compound (460.0 mg, 0.8 mmol) was dissolved in 20.0 mL of dichloromethane, 4-dimethylaminopyridine (122.0 mg, 1.0 mmol) and tert-butyldimethylsilyl chloride (360.0 mg, 2.4 mmol) were added, and the mixture was stirred at room temperature for 4-8 h. The dichloromethane was evaporated, diluted with ethyl acetate (20.0 mL), washed acidic with 5% HCl, washed neutral with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and column chromatographed (V)_{Petroleum ether}:V_{Ethyl acetate}= 30:1-15:1) to give a white solid (383.0 mg, 70.0%).

The above-mentioned compound (380.0 mg, 0.6 mmol) was dissolved in 15.0 mL of dichloromethane, and fresh pyridinium chlorochromate (300.0 mg, 1.3 mmol) was added thereto and stirred at room temperature for 6 to 10 hours. Evaporating to remove dichloromethane, diluting with ethyl acetate (20.0 mL), washing with water, washing with saturated saline to neutrality, drying with anhydrous sodium sulfate, filtering, concentrating, and performing column chromatography (V)_{Petroleum ether}:V_{Ethyl acetate}= 35:1-20:1) to give a white solid (319.0 mg, 84.0%).

Dissolving the above compound (500.0 mg, 0.7 mmol) in morpholine (25.0 mL), adding sulfur (0.3 g, 10.0 mmol) and ethylenediamine (0.3 g, 4.5 mmol), refluxing for 6-10 h, diluting with ethyl acetate (30.0 mL), washing with water three times, washing with saturated salt water twice, drying over anhydrous sodium sulfate, filtering, concentrating, and performing column chromatography (V)_{Petroleum ether}:V_{Ethyl acetate}30:1-10:1) to give a white solid (357.0 mg, 68.0%).

Dissolving the above product (300.0 mg, 0.4 mmol) in acetone (20.0 mL), adding 10% HCl (2.0 mL), stirring at room temperature for 3-5 h, diluting with ethyl acetate, washing with water to neutrality, washing with saturated salt water, drying with anhydrous sodium sulfate, filtering, concentrating, and performing column chromatography (V)_{Petroleum ether}:V_{Ethyl acetate}5:1-8:1) to give H6 (225.0 mg, 89.0%) as a white solid.

Dissolve H6 (1.0 g, 1.7 mmol) in methanol: tetrahydrofuran =2:3 (20 mL) mixed solution, 10% Pd/C (802.0 mg, 7.5 mmol) was added, and H was introduced into the reaction system₂And stirring and reacting for 5-8 h at room temperature. After the reaction is finished, vacuum filtration is carried out to obtain filtrate, and the filtrate is concentrated to obtain crude product H6-Bn which can be directly used for the next reaction without further purification.

Dissolving H6-Bn (1.0 g, 1.98 mmol) in anhydrous dichloromethane (15 mL), placing in an ice bath for 10 min, adding oxalyl chloride (2.68 mL), stirring at room temperature for 30 min, evaporating to dryness, dissolving with anhydrous dichloromethane (15 mL), adding methyl 4-aminobutyric acid hydrochloride (1.52 g, 9.88 mmol) and triethylamine (1.01 mL), stirring at room temperature for 3H, concentrating, diluting with dichloromethane, washing with water once in sequence, washing with saturated salt water twice, drying with anhydrous sodium sulfate, and concentrating to obtain white solid H628+ D-OCH3 (0.96 g, 93.77%).

Dissolving H628+ D-OCH3 (800 mg, 1.32 mmol) in a mixed solution (40 ml) of tetrahydrofuran and methanol (3: 2), adding 10% NaOH solution (8 ml) dropwise, stirring at room temperature for 1H, concentrating, diluting with EA, adjusting pH to acidity with 5% HCl, washing with water and saturated brine in turn three times, drying over anhydrous sodium sulfate, and concentrating to obtain H628+ D as a white solid (632 mg, 80.92%).

Dissolving H628+ D (120 mg, 0.20 mmol) in anhydrous DCM (6 mL), adding diethylene glycol (319. mu.L, 4 mol), adding DMAP (471 mg, 3.8 mmol) and EDCI (466 mg, 2.4 mmol), stirring at room temperature for 9H, concentrating and redissolving the reaction solution in methanol (4 mL) after the reaction is finished, dialyzing the solution in deionized water (MWCO 500 Da) for 28H, and replacing dialysis every 7HAnd (4) liquid. Concentrating the solution under reduced pressure, and performing silica gel column chromatography (V)_DCM:V_MA=90: 1) to give H628+ D + P100 as a white solid (117 mg, 84.92%).¹H NMR (400 MHz, Chloroform-d) δ 8.38 (d, J = 2.2 Hz, 1H, H-pyrazine), 8.34 (d, J = 2.4 Hz, 1H, H-pyrazine), 6.11 (t, J = 5.5 Hz, 1H, NH), 5.48 (t, J = 3.3 Hz, 1H, H-12), 4.30-4.21 (m, 2H, CH₂O PEG group), 3.80 (d, J = 10.5 Hz, 1H, H-23a), 3.75-3.69 (m, 4H, CH₂O PEG group), 3.62-3.58 (m, 2H, CH₂O PEG group), 3.50 (d, J = 10.5 Hz, 1H, H-23b), 3.41 (dq, J = 13.6, 7.2 Hz, 1H, N-CH₂a), 3.08 (dq, J = 13.4, 6.9 Hz, 1H, N-CH₂b), 3.00 (d, J = 16.5 Hz, 1H, H-1a), 2.58 (dd, J = 12.8, 3.5 Hz, 1H, H-18), 2.52 (d, J = 16.6 Hz, 1H, H-1b), 2.39 (t, J = 7.2 Hz, 2H, CH₂), 2.10 (dd, J = 8.8, 3.3 Hz, 2H CH₂), 1.31 (s, 3H, CH₃), 1.21 (s, 3H, CH₃), 0.95 (s, 3H, CH₃), 0.93 (s, 3H, CH₃), 0.92 (s, 3H, CH₃), 0.86 (s, 3H, CH₃).

Example 2 N- (23-hydroxyolean-12-en-28-oylo [3, 2-)b]Synthesis and characterization of triethylene glycol 4-aminobutyric acid (H628 + D + P150)

Referring to the synthesis of example 1, H628+ D was reacted with triethylene glycol, dialyzed (MWCO 500 DA), and subjected to silica gel column chromatography (V)_{Methylene dichloride}: V_Methanol= 85: 1) to give compound H628+ D + P150 as a white solid (120 mg, 81.81%).¹H NMR (400 MHz, Chloroform-d)δ 8.38 (s, 1H, H-pyrazine), 8.34 (s, 1H, H-pyrazine), 6.15 (t, J = 5.3 Hz, 1H, NH), 5.48 (t, J = 3.3 Hz, 1H, H-12), 4.29-4.21 (m, 2H, CH₂O PEG group), 3.80 (d, J = 10.6 Hz, 1H, H-23a), 3.71 (dt, J = 9.2, 4.6 Hz, 4H, CH₂O PEG group), 3.68-3.64 (m, 4H, CH₂O PEG group), 3.63-3.55 (m, 2H, CH₂O PEG group), 3.50 (d, J = 10.5 Hz, 1H, H-23b), 3.40 (dq, J = 13.5, 7.0 Hz, 1H, N-CH₂a), 3.08 (dt, J = 13.2, 6.4 Hz, 1H, N-CH₂b), 3.00 (d , J = 16.5 Hz, 1H, H-1a), 2.60 (d, J = 9.8 Hz, 1H, H-18), 2.52 (d, J= 16.5 Hz, 1H, H-1b), 2.39 (t, J = 7.2 Hz, 2H, CH₂), 2.10 (dd, J = 8.6, 2.9 Hz, 2H, CH₂), 1.31 (s, 3H, CH₃), 1.21 (s, 3H, CH₃), 0.95 (s, 3H, CH₃), 0.93 (s, 3H, CH₃), 0.92 (s, 3H, CH₃), 0.86 (s, 3H, CH₃).

Example 3N- (23-hydroxyolean-12-en-28-oylo [3, 2-)b]Synthesis and characterization of tetraethylene glycol 4-aminobutyrate (H628 + D + P200)

Referring to the synthesis of example 1, H628+ D was reacted with tetraethylene glycol, dialyzed (MWCO 500 DA) and subjected to silica gel column chromatography (V)_{Methylene dichloride}: V_Methanol= 80: 1) to give compound H628+ D + P200 as a yellow clear oil (125 mg, 80.32%).¹H NMR (400 MHz, Chloroform-d) δ 8.40 (d, J = 1.8 Hz, 1H, H-pyrazine), 8.36 (d, J = 1.8 Hz, 1H, H-pyrazine), 6.13 (t, J = 5.6 Hz, 1H, NH), 5.47 (t, J = 3.3 Hz, 1H, H-12), 4.28-4.20 (m, 2H, CH₂O PEG group), 3.80 (d, J = 10.6 Hz, 1H, H-23a), 3.74-3.68 (m, 4H, CH₂O PEG group), 3.67-3.64 (m, 8H, CH₂O PEG group), 3.62-3.59 (m, 2H, CH₂O PEG group), 3.50 (d, J = 10.6 Hz, 1H, H-23b), 3.41 (dt, J = 13.5, 6.8 Hz, 1H, N-CH₂a), 3.12-3.05 (m, 1H, N-CH₂b), 3.02 (d, J= 16.6 Hz, 1H, H-1a), 2.60 (d, J = 8.4 Hz, 1H, H-18), 2.53 (d, J = 16.6 Hz, 1H, H-1b), 2.39 (t, J = 7.6 Hz, 2H, CH₂), 2.13-2.07 (m, 2H, CH₂), 1.33 (s, 3H, CH₃), 1.21 (s, 3H, CH₃), 0.96 (s, 3H, CH₃), 0.93 (s, 3H, CH₃), 0.92 (s, 3H, CH₃), 0.86 (s, 3H, CH₃).

Example 4 N- (23-hydroxyolean-12-en-28-oylo [3, 2-)b]Pyrazine) -4-aminobutanesSynthesis and characterization of Pentaethylene glycol acid ester (H628 + D + P240)

Referring to the synthesis of example 1, H628+ D was reacted with pentaethylene glycol, dialyzed (MWCO 500 DA) and subjected to silica gel column chromatography (V)_{Methylene dichloride}: V_Methanol= 75: 1) to give compound H628+ D + P240 as a yellow clear oil (130 mg, 79.01%).¹H NMR (400 MHz, Chloroform-d) δ8.38 (s, 1H, H-pyrazine), 8.35 (s, 1H, H-pyrazine), 6.15-6.08 (m, 1H, NH), 5.48 (t, J = 3.1 Hz, 1H, H-12), 4.28-4.20 (m, 2H, CH₂O PEG group), 3.80 (d, J = 10.6 Hz, 1H, H-23a), 3.71 (dd, J = 9.2, 4.4 Hz, 4H, CH₂O PEG group), 3.67-3.65 (m, 12H, CH₂O PEG group), 3.60 (dd, J = 5.2, 3.8 Hz, 2H, CH₂O PEG group), 3.50 (d, J = 10.6 Hz, 1H, H-23b), 3.40 (dq, J = 13.7, 7.1 Hz, 1H, N-CH₂a), 3.09 (dd, J = 11.7, 6.8 Hz, 1H, N-CH₂b), 3.01 (d, J= 16.6 Hz, 1H, H-1a), 2.60 (dd, J = 13.1, 2.4 Hz, 1H, H-18), 2.52 (d, J = 16.7 Hz, 1H, H-1b), 2.39 (t, J = 7.1 Hz, 2H, CH₂), 2.10 (dd, J = 8.8, 3.2 Hz, 2H, CH₂), 1.32 (s, 3H, CH₃), 1.21 (s, 3H, CH₃), 0.95 (s, 3H, CH₃), 0.93 (s, 3H, CH₃), 0.92 (s, 3H, CH₃), 0.86 (s, 3H, CH₃).

Example 5 N- (23-hydroxyolean-12-en-28-oylo [3, 2-)b]Synthesis and characterization of pyrazine) -4-aminobutanoic acid hexa-ethylene glycol ester (H628 + D + P280)

Referring to the synthesis of example 1, H628+ D was reacted with hexaethylene glycol, dialyzed (MWCO 500 DA) and subjected to silica gel column chromatography (V)_{Methylene dichloride}: V_Methanol= 70: 1) to give compound H628+ D + P280 as a yellow clear oil (125 mg, 72.23%).¹H NMR (400 MHz, Chloroform-d) δ 8.39 (s, 1H, H-pyrazine), 8.38-8.33 (m, 1H, H-pyrazine), 6.10 (t, J = 5.4 Hz, 1H, NH), 5.48 (t, J = 3.4 Hz, 1H, H-12), 4.28-4.20 (m, 2H, CH₂O PEG group), 3.80 (d, J = 10.6 Hz, 1H, H-23a), 3.71 (m, 4H, CH₂O PEG group), 3.68-3.63 (m, 16H, CH₂O PEG group), 3.63-3.58 (m, 2H, CH₂O PEG group), 3.50 (d, J = 10.6 Hz, 1H, H-23b), 3.39 (dt, J = 13.6, 6.8 Hz, 1H, N-CH₂a), 3.12-3.04 (m, 1H, N-CH₂b), 3.01 (d, J = 16.4 Hz, 1H, H-1a), 2.59 (dd, J = 12.9, 4.0 Hz, 1H, H-18), 2.52 (d, J = 16.5 Hz, 1H, H-1b), 2.39 (t, J = 7.5 Hz, 2H, CH₂), 2.10 (dd, J = 8.8, 3.3 Hz, 2H, CH₂), 1.32 (s, 3H, CH₃), 1.21 (s, 3H, CH₃), 0.96 (s, 3H, CH₃), 0.93 (s, 3H, CH₃), 0.92 (s, 3H, CH₃), 0.86 (s, 3H, CH₃).

Example 6 N- (23-hydroxyolean-12-en-28-oylo [3, 2-)b]Synthesis and characterization of heptaethylene glycol pyrazine) -4-aminobutyric acid (H628 + D + P330)

Referring to the synthesis of example 1, H628+ D was reacted with heptaglycol, dialyzed (MWCO 500 DA) and subjected to silica gel column chromatography (V)_{Methylene dichloride}: V_Methanol= 60: 1) to give compound H628+ D + P330 as a yellow clear oil (106 mg, 58.11%).¹H NMR (400 MHz, Chloroform-d) δ 8.40 (s, 2H, H-pyrazine), 6.15 – 6.09 (m, 1H, NH), 5.47 (t, J = 3.5 Hz, 1H, H-12), 4.30-4.19 (m, 2H, CH₂O PEG group), 3.80 (d, J= 10.5 Hz, 1H, H-23a), 3.78-3.71 (m, 4H, CH₂O PEG group), 3.71-3.66 (m, 20H, CH₂O PEG group), 3.65-3.62 (m, 2H, CH₂O PEG group), 3.50 (d, J = 11.4 Hz, 1H, H-23b), 3.40 (dt, J = 13.6, 6.8 Hz, 1H, N-CH₂a), 3.12-3.04 (m, 1H, N-CH₂b), 2.59 (dd, J = 13.4, 4.5 Hz, 1H, H-18), 2.53 (d, J = 16.9 Hz, 1H, H-1b), 2.39 (t, J = 7.0 Hz, 2H, CH₂), 2.11 (dd, J = 8.6, 2.9 Hz, 2H, CH₂), 1.33 (s, 3H, CH₃), 1.21 (s, 3H, CH₃), 0.96 (s, 3H, CH₃), 0.93 (s, 3H, CH₃), 0.92 (s, 3H, CH₃), 0.86 (s, 3H, CH₃).

The following are the pharmacological tests and data for some of the compounds of the invention.

The experimental method comprises the following steps: examples 1-6 detection of the survival of the antitumor drug paclitaxel in KBV resistant strain cells

KBV cells in logarithmic phase are digested with 0.25% pancreatin to prepare single cell suspension with certain concentration. Based on the difference in cell growth rate, the cells were seeded at 4000 wells in a 96-well plate, and 100. mu.L of cell suspension was added to each well. After 24 h, complete medium was added at various concentrations of compound and 100 nM of paclitaxel and corresponding solvent control. mu.L of DMSO (final DMSO concentration < 0.1%) was added to each well, 3 wells were placed in each group, incubation was continued at 37 ℃ for 72 h, and the supernatant was discarded. After adding 100. mu.L of complete medium containing 0.5 mg/mL MTT per well, culturing was continued for 4 hours, the supernatant was discarded, 150. mu.L of DMSO was added per well to dissolve MTT formazan precipitate, the mixture was shaken and mixed by a micro shaker, and then an Optical Density (OD) was measured with a microplate reader at a reference wavelength of 450 nm and a detection wavelength of 570 nm. Using tumor cells treated by solvent control as a control group, and calculating the survival rate of different tumor cells under the action of each compound by using the following formula; tumor cells treated by solvent control are used as a control group, the inhibition rate of the compound on the tumor cells is calculated by the following formula, and the IC is calculated according to the middle effect equation₅₀。

Cell survival rate (%) = average OD value of administration group/average OD value of control group × 100%

IC₅₀= (control mean OD value-administration mean OD value)/control mean OD value × 100%

2, experimental results:

examples 1-6 cell viability when administered alone and in combination is shown in table 1.

TABLE 1 cell viability in examples 1-6 when administered alone and in combination

Examples 1-6 analysis of the viability of anti-tumor drug paclitaxel in KBV resistant strain cells.

The survival rate evaluation results of KBV drug-resistant strain cells of the derivatives show that examples 1-6 have better tumor drug resistance reversal activity and can obviously increase the sensitivity of the drug-resistant KBV cells to paclitaxel, wherein the activity of examples 3-6 is better than that of an equal-dose positive control drug verapamil, and the action strength of the examples 5 and 6 with the best activity is better than that of a leader H6.

The following are solubility determination tests and data for some of the compounds of the present invention.

The experimental method comprises the following steps: solubility tests were performed on examples 1-6

The excess compound was added to 4 mL of deionized water, respectively, and the suspension was placed in a constant temperature shaker, shaken at 37 ℃ for 24 hours, and then centrifuged at 14,000 r/min in a centrifuge for 5 minutes. The supernatant was filtered through a 0.45 μm microporous membrane, diluted appropriately, and the concentration of the compound was measured by UV-visible spectrophotometry to obtain the solubility of the compound in water.

The experimental results are as follows:

examples 1-6 solubility data are shown in table 2.

TABLE 2 examples 1-6 solubility

The solubility measurement results of examples 1 to 6 show that the water solubility of the C-28 polyethylene glycol modified hederagenin derivative is remarkably improved compared with that of the lead compound H6, and the water solubility of the C-28 polyethylene glycol modified hederagenin derivative is improved by about 90 times in example 6.

Pharmacological tests and solubility determination tests prove that the C-28 polyethylene glycol modified hederagenin derivative provided by the invention improves water solubility while maintaining or improving tumor drug resistance reversal activity, and the water solubility is also obviously improved while improving the tumor drug resistance reversal activity in examples 5 and 6, so that the derivative can be used for preparing a tumor drug resistance reversal agent, is used together with common antitumor drugs, and exerts good antitumor activity.

The experiments show that the polyethylene glycol modification method is introduced into the C-28 structural modification of H6, and the polyethylene glycol is connected to H6 through the connecting chain 4-aminobutyric acid, so that the hederagenin C-28 polyethylene glycol modified derivative with improved tumor drug resistance reversal activity and remarkably improved water solubility and a novel structure is obtained, namely the structural modification of the hederagenin C-28 polyethylene glycol modified derivative with the general formula I and the medically acceptable salt thereof is successful, the water solubility is remarkably improved on the premise of having good tumor drug resistance reversal effect and the activity superior to the primer H6, and the hederagenin polyethylene glycol derivative with the general formula I and the medically acceptable salt thereof have good drug forming properties.

Although the preferred embodiments of the present invention have been described in detail, the present invention is not limited to the details of the embodiments, and various equivalent modifications can be made within the technical spirit of the present invention, and the scope of the present invention is also within the scope of the present invention. It should be noted that the various features described in the above embodiments may be combined in any suitable manner without departing from the scope of the invention. The invention is not described in detail in order to avoid unnecessary repetition. In addition, any combination of the various embodiments of the present invention is also possible, and the same should be considered as the disclosure of the present invention as long as it does not depart from the spirit of the present invention.

Claims

1. A C-28 polyethylene glycol modified derivative of hederagenin and a medically acceptable salt thereof are characterized in that the C-28 polyethylene glycol modified derivative of the hederagenin is:

2. The preparation method of the C-28 polyethylene glycol modified hederagenin derivative and the medically acceptable salt thereof according to claim 1, is characterized in that,

d. reacting with ethylenediamine under the catalysis of sulfur;

e. under the acidic condition, removing a C-23 protecting group;