Application of mangosteen extract or its monomer components in the preparation of β-glucuronidase inhibitors

Abstract

The invention discloses an application of a mangosteen extract or its monomer components in the preparation of a β-glucuronidase inhibitor, and belongs to the field of biomedicine technology. The invention discloses an application of a mangosteen extract or its monomer components in the preparation of a β-glucuronidase inhibitor, and through in vitro activity assays, it is found that the mangosteen extract or its monomer components have a high affinity and strong inhibitory activity to EcGUS, IC 50 The values ​​ranged from 0.23 to 20.66 μM, and the inhibition kinetic constant K i The value range is 0.24-11.68μM. In vivo experiments have found that mangosteen extract can alleviate irinotecan-induced weight loss and reduce irinotecan-induced disease activity index. Mangosteen extract or its monomer components have high biological activity and good safety, and have broad application prospects in the treatment of irinotecan-induced drug-induced enterotoxicity.

Description

Technical Field

[0001] The invention relates to the technical field of biomedicine, and in particular to application of a mangosteen extract or a monomer component thereof in the preparation of a beta-glucuronidase inhibitor. Background Art

[0002] Irinotecan (CPT-11) is a first-line drug for the clinical treatment of solid tumors such as colorectal cancer. However, the incidence of intestinal toxic side effects, mainly grade III-IV delayed diarrhea, is as high as 40% during its use, resulting in the suspension of chemotherapy, affecting the efficacy, and even posing a risk of death, which greatly limits the clinical application of CPT-11. Studies have shown that the intestinal toxicity of CPT-11 is closely related to its metabolism in the "gut flora-gut-liver" axis. The metabolic process of CPT-11 mainly includes three steps: (1) Carboxylesterase 2 (CES2) first rapidly hydrolyzes CPT-11 into an active and toxic product, 7-ethyl-10-hydroxycamptothecin (SN-38). (2) Liver uridine diphosphate glucuronyltransferase 1A1 (UGT1A1) catalyzes SN-38 to produce a non-toxic and inactive metabolite, SN-38-O-glucuronide (SN-38G). (3) SN-38G is excreted into the intestinal cavity with bile. The β-glucuronidase (GUS) secreted by the intestinal flora in the intestinal cavity can convert SN-38G into active and toxic SN-38 again. The SN-38 accumulated in the intestine can be absorbed by the intestinal wall and enter the liver, resulting in an enterohepatic circulation phenomenon. Among them, the intestinal bacterial GUS enzyme is the key "toxic enzyme" that converts CPT-11 into SN-38 in the intestine and causes intestinal toxicity (intestinal mucosal damage and delayed diarrhea). Its activity plays a decisive role in the intestinal concentration of SN-38.

[0003] β-Glucuronidase (GUS) is an important type of hydrolase produced by intestinal bacteria, mainly including Clostridium perfringens GUS, Streptococcus agalactiae GUS and Escherichia coli GUS, which catalyzes the hydrolysis of glucuronide metabolites of various drugs (irinotecan, indomethacin, diclofenac, etc.) and endogenous substances (bilirubin, testosterone, estradiol, etc.) in the intestine to generate aglycones that accumulate in the intestine. The use of safe and efficient inhibitors of intestinal bacterial GUS activity can inhibit the production of SN-38 in the intestine, reduce the exposure of intestinal SN-38, shorten the contact time of SN-38 with the intestine, and to a certain extent alleviate gastrointestinal adverse reactions such as delayed diarrhea caused by CPT-11. Therefore, GUS has become an important target for alleviating the intestinal adverse reactions of CPT-11. The development of efficient and safe inhibitors of intestinal bacterial GUS is of great significance for alleviating fatal diarrhea caused by CPT-11. Currently, compounds with intestinal bacterial β-GUS inhibitory activity include marketed drugs (amoxapine, crizotinib, nicotinamide, etc.), synthetic compounds (azacoumarins, indolyl thiadiazole analogs, piperazine / piperidines, etc.) and natural products (flavonolignans, flavonoid glucuronides, flavonoid aglycones, etc.). Although there are many intestinal bacterial GUS inhibitors, there are problems such as low activity, poor selectivity, low safety, and poor pharmacokinetic properties. Therefore, it is urgent to expand the intestinal bacterial GUS inhibitors with new skeleton types, improve the intestinal bacterial GUS inhibitory activity, and subsequently develop new therapeutic preparations.

[0004] Mangosteen, also known as mangosteen, is the fruit of the evergreen tree mangosteen of the genus Garcinia in the family Garciniaceae. Mangosteen fruit contains vitamin B, multiple amino acids, proteins and rich minerals, has high edible value, and is known as the "Queen of Tropical Fruits". Mangosteen shell is a traditional medicinal material in Thailand, Myanmar, India and other countries, and is widely used to treat inflammation, trauma, skin and wound infections, etc. Mangosteen contains a variety of secondary metabolites and is increasingly valued by the medical community. However, there has been no report on the inhibitory effect of mangosteen and its chemical components on intestinal bacteria GUS, nor has there been any report on the use of mangosteen and its chemical components to alleviate CPT-11 intestinal toxicity. Summary of the invention

[0005] The purpose of the present invention is to provide an application of a mangosteen extract or its monomer components in the preparation of a β-glucuronidase inhibitor, so as to solve the above problems existing in the prior art.

[0006] To achieve the above object, the present invention provides the following solutions:

[0007] One of the technical solutions of the present invention is the use of a mangosteen extract in the preparation of a β-glucuronidase inhibitor.

[0008] Technical solution 2 of the present invention: Application of a mangosteen extract in the preparation of a drug for treating irinotecan-induced drug-induced enterotoxicity

[0009] Furthermore, the active ingredients in the mangosteen extract include one or more of α-mangostin, β-mangostin, γ-mangostin, gambogic acid C, gambogic acid D, 8-deoxy-gambogic acid glycoside, gambogic acid glycoside and 3-isomangostin.

[0010] Furthermore, the mangosteen extract is obtained by taking mangosteen peel as raw material and extracting with ethanol aqueous solution as extraction solvent.

[0011] Furthermore, the concentration of the ethanol aqueous solution is 90wt%; the usage ratio of the mangosteen peel to the ethanol aqueous solution is 1g:10mL.

[0012] Furthermore, the preparation of the mangosteen extract specifically includes: mixing mangosteen peel and ethanol aqueous solution, soaking for 4 hours, ultrasonicating for 2 hours, filtering, and rotary evaporating to obtain the mangosteen extract.

[0013] Furthermore, the β-glucuronidase is derived from intestinal bacteria, preferably Escherichia coli.

[0014] Furthermore, the irinotecan-induced drug-induced enterotoxicity is specifically irinotecan-induced drug-induced diarrhea; the drug for treating irinotecan-induced drug-induced enterotoxicity is used in combination with irinotecan to inhibit the hydrolysis of irinotecan metabolite SN-38G in the intestine, thereby reducing the local exposure of irinotecan active metabolite SN-38 in the intestinal system, and mitigating the serious adverse reaction of delayed diarrhea caused by it.

[0015] The third technical solution of the present invention: an application of a monomer component in mangosteen in the preparation of a β-glucuronidase inhibitor, wherein the monomer component is α-mangostin, β-mangostin, γ-mangostin, gambogic acid C, gambogic acid D, 8-deoxygarcin, gambogic acid or 3-isomangostin.

[0016] Technical solution four of the present invention: Use of a monomer component in mangosteen in the preparation of a drug for treating irinotecan-induced drug-induced enterotoxicity, wherein the monomer component is α-mangostin, β-mangostin, γ-mangostin, gambogic acid C, gambogic acid D, 8-deoxy-gambogic acid glycoside, gambogic acid glycoside or 3-isomangostin.

[0017] Technical solution five of the present invention: a β-glucuronidase inhibitor, the active ingredient of which includes mangosteen extract, or one of α-mangostin, β-mangostin, γ-mangostin, gamboge C, gamboge D, 8-deoxygarcin, gamboge glycosides, and 3-isomangostin, or a composition composed of two or more of α-mangostin, β-mangostin, γ-mangostin, gamboge C, gamboge D, 8-deoxygarcin, gamboge glycosides, and 3-isomangostin.

[0018] That is, the β-glucuronidase inhibitor can be used in the form of a mangosteen extract, or in the form of a single component, or in the form of a combination.

[0019] Technical solution six of the present invention: A drug for treating irinotecan-induced drug-induced enterotoxicity, the active ingredient of which includes mangosteen extract, or one of α-mangostin, β-mangostin, γ-mangostin, gamboge C, gamboge D, 8-deoxygarcin, gamboge glycoside, and 3-isomangostin, or a composition composed of two or more of α-mangostin, β-mangostin, γ-mangostin, gamboge C, gamboge D, 8-deoxygarcin, gamboge glycoside, and 3-isomangostin.

[0020] The present invention discloses the following technical effects:

[0021] The invention discloses the application of a mangosteen extract in the preparation of a beta-glucuronidase inhibitor, enriches the types of the beta-glucuronidase inhibitor, and broadens the application field of the mangosteen extract.

[0022] The present invention finds through in vitro activity assay that mangosteen extract or its monomer components α-mangostin, β-mangostin, γ-mangostin, gambogelin C, gambogelin D, 8-deoxygarcin, gambogelin, and 3-isomangostin have high affinity and strong inhibitory activity to EcGUS, inhibiting the hydrolysis reaction of DDAOG glucuronide mediated by EcGUS, IC 50 The values ​​ranged from 0.23 to 4.54 μM, and the inhibition kinetic constant K i The value range is 0.24-2.44 μM; inhibiting EcGUS-mediated SN-38G glucuronide hydrolysis reaction, IC 50 The values ​​ranged from 0.87 to 20.66 μM, and the inhibition kinetic constant K i The value range is 0.64-11.68μM; in vivo experiments found that mangosteen extract can alleviate irinotecan-induced weight loss and reduce irinotecan-induced disease activity index. Mangosteen extract or its monomer components have high biological activity and good safety, and have good therapeutic effects, application prospects and economic benefits in the treatment of irinotecan-induced drug-induced diarrhea. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0024] Figure 1 is the inhibition curve of mangosteen extract on EcGUS;

[0025] Figure 2 The chemical fingerprint of mangosteen extract (A), the inhibitory effect spectrum of mangosteen extract fractions on EcGUS (B) and the chemical structural formula of active substances after identification (C);

[0026] Figure 3 The results of the test on the effect of mangosteen extract on alleviating CPT-11-induced intestinal toxicity, where (a) is the change of weight loss of Bablc female mice over time; (b) is the change of disease activity index over time; (c) is the change of diarrhea incidence rate over time; (d) is the β-glucuronidase activity in the feces of Bablc female mice; (e) is the SN-38 content in the serum of Bablc female mice; (f) is the SN-38G content in the serum of Bablc female mice. DETAILED DESCRIPTION

[0027] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0028] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0029] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0030] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0031] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0032] The extraction method of the mangosteen extract used in the following examples is as follows: 50 g of mangosteen peel is added with 500 mL of 90 wt% ethanol aqueous solution, sealed with plastic wrap, soaked at room temperature for 4 h, and then ultrasonicated for 2 h (ultrasonic frequency is 100 KHz, ultrasonic temperature is 37 ° C), filtered to remove the residue, rotary steamed in a large bottle to dryness, directly scraped and transferred to a white plastic bottle, cooled at room temperature, tightened the bottle cap and marked for storage at -20 ° C.

[0033] The GUS used in the following examples is EcGUS (β-glucuronidase secreted by Escherichia coli), purchased from Sigma-Aldrich (Shanghai) Trading Co., Ltd.

[0034] Example 1 Determination of the inhibitory activity of mangosteen extract on EcGUS

[0035] The hydrolysis and metabolism reaction of 7-ethyl-10-hydroxycamptothecin-O-glucuronide (SN38G) was used as the probe reaction, and the inhibitory activity of mangosteen extract on GUS was tested by a multifunctional ELISA plate assay method with the help of an in vitro GUS enzyme incubation system. The specific experimental process is as follows:

[0036] (1) In vitro GUS metabolic reaction system, including PBS buffer (100 mM) at pH 6.5, EcGUS (0.2 U / mL), and mangosteen extract (0, 0.25, 0.5, 1, 2, 4, 6, and 8 μg / mL), was pre-incubated at 37°C with shaking for 5 min. The concentration of mangosteen extract in the metabolic reaction system was changed, and multiple single-factor experiments were performed;

[0037] (2) Add SN38G (final concentration 1.0 μM) to the reaction system to start the reaction;

[0038] (3) After 30 min, add 200 μL of glacial acetonitrile and shake vigorously to terminate the reaction;

[0039] (4) After high-speed centrifugation at 4°C and 20,000 × g for 10 min, the supernatant was collected and subjected to mass spectrometry. SN-38G was detected in positive ion mode with a parent ion of 393.100 and a daughter ion of 349.100.

[0040] (5) A curve was drawn with the concentration of mangosteen extract as the horizontal axis and the residual activity of EcGUS as the vertical axis to obtain the inhibition curve of mangosteen extract on EcGUS, such as Figure 1 As shown by Figure 1 It can be seen that mangosteen extract showed strong inhibitory activity against EcGUS. The residual activity of EcGUS in the reaction system with a mangosteen extract concentration of 1 μg / mL was 28.11%, and the residual activity of EcGUS in the reaction system with a mangosteen extract concentration of 8 μg / mL was 4.10%. The IC 50 The value is 0.60μg / mL.

[0041] Example 2 Activity-guided tracking of EcGUS inhibitors in mangosteen

[0042] With the help of high performance liquid chromatography, the fingerprint of mangosteen extract was established, the eluent of the spectrum was collected, and the hydrolysis and metabolic reaction of 1,3-dichloro-7-hydroxy-9,9-dimethyl-2(9H)-acridone-O-glucuronide (DDAOG) was used as a probe reaction to determine the inhibitory activity of all eluents on EcGUS. The fingerprint and inhibition spectra were compared, and the main monomer components of mangosteen that had an inhibitory effect on EcGUS were identified by comparing the retention time, ultraviolet spectrum, and high-resolution mass spectrum with those of the standard. The specific experimental process is as follows:

[0043] (1) Fingerprint establishment: The fingerprint of mangosteen extract was established using Waters high performance liquid chromatography, Acquity UPLC HSS T3C18 (2.1×100 mm, 1.8 μm) column. The mobile phase composition was acetonitrile (A) and water (B), the flow rate was 0.8 mL / min, and the detection wavelength was 285 nm. The effluent was directly collected into a black 96-well plate every 60 seconds and used for EcGUS inhibition experiments after vacuum drying.

[0044] (2) Establishing an inhibition profile: Using the GUS-specific fluorescent probe substrate DDAOG, the inhibitory effect of the monomeric compounds in the mangosteen extract on the EcGUS activity was evaluated. The specific steps are as follows:

[0045] 1) The incubation system contained EcGUS (0.2 U / mL), 100 mM PBS (pH 6.5), and the chromatographic elution fraction of mangosteen extract, and was pre-incubated at 37°C for 5 min;

[0046] 2) Add DDAOG (10.0 μM) to the reaction system to start the reaction;

[0047] 3) After 30 min, add 200 μL of glacial acetonitrile and shake vigorously to terminate the reaction;

[0048] 4) After high-speed centrifugation at 4°C and 20,000×g for 10 min, the supernatant was collected and subjected to fluorescence detection (Ex=600nm, Em=660nm);

[0049] 5) A curve was drawn with the acquisition time of the chromatographic elution fraction of the mangosteen extract as the abscissa and the residual activity of EcGUS in the fraction as the ordinate to obtain an inhibition spectrum.

[0050] Figure 2 The chemical fingerprint of mangosteen extract (A), the spectrum of the inhibitory effect of mangosteen extract fractions on EcGUS (B) and the chemical structural formula after active substance identification (C). It can be seen from (A) and (B) that there are 8 components in mangosteen extract that show a strong inhibitory effect on EcGUS. By comparing with the retention time, ultraviolet spectrum and high-resolution mass spectrum of the standard, they are identified as gamboge C, gamboge D, γ-mangosteen, 8-deoxygamboge glycoside, gamboge glycoside, α-mangosteen, 3-isomangosteen and β-mangosteen. The comparison process is shown in Table 1. The chemical structural formula after active substance identification is shown in Figure 2 (C).

[0051] Table 1 Compounds in mangosteen extract that have strong inhibitory ability against EcGUS

[0052]

[0053] Example 3 In vitro inhibition test to quantitatively evaluate the inhibitory ability of mangosteen extract series compounds on EcGUS

[0054] The IC values ​​of different concentrations of mangosteen extract monomer components (α-mangostin, β-mangostin, γ-mangostin, gambogic acid C, gambogic acid D, 8-deoxygambogic acid glycoside, gambogic acid glycoside, and 3-isomangostin) on EcGUS inhibition were evaluated by high-throughput assay using the in vitro GUS enzyme incubation system and a multifunctional ELISA plate assay. 50 and K i The specific experimental process is as follows:

[0055] (1) In vitro GUS metabolic reaction system, including PBS buffer (100 mM) at pH 6.5, EcGUS (0.2 U / mL), and different concentrations of monomer components of mangosteen extract (final concentration 0.005-40 μM), pre-incubated at 37°C with shaking for 3 min;

[0056] (2) Add DDAOG or SN-38G (IC 50 The final concentration of DDAOG in the assay was 10 μM, and the final concentration of SN-38G was 1 μM; K iThe final concentrations of DDAOG in the assay were 5, 10, 20, and 40 μM, and the final concentrations of SN-38G were 0.5, 1, 2, and 4 μM) to initiate the reaction;

[0057] (3) After 30 min, add 200 μL of glacial acetonitrile and shake vigorously to terminate the reaction;

[0058] (4) After high-speed centrifugation at 4°C and 20,000 × g for 20 min, the supernatant was collected for fluorescence detection and mass spectrometry. The maximum excitation and emission wavelengths of DDAOG were 600 nm and 660 nm, respectively. SN-38G was detected in positive ion mode with a parent ion of 393.100 and a daughter ion of 349.100.

[0059] (5) Calculate IC using GraphadPrism9.0 software 50 and K i Numeric value, used to determine the suppression type.

[0060] The results are shown in Table 2. The eight monomer components in the mangosteen extract showed dose-dependent inhibition on the EcGUS-mediated hydrolysis of DDAOG and SN-38G glucuronides. The IC values ​​of α-mangostin, β-mangostin, γ-mangostin, gambogelin C, gambogelin D, 3-isomangostin, gambogelin, and 8-deoxygambogelin on the hydrolysis of DDAOG glucuronides mediated by EcGUS were 0. 50 The values ​​were 1.90, 1.42, 0.46, 1.10, 1.44, 1.56, 0.23 and 4.54 μM, respectively, and K i The IC values ​​of SN-38G glucuronide hydrolysis mediated by EcGUS were 1.15, 1.41, 0.28, 0.53, 0.80, 1.87, 0.24 and 2.44 μM, respectively. 50 The values ​​were 2.59, 3.04, 0.87, 3.14, 1.99, 5.78, 20.7 and 8.31 μM, respectively, and K i They are 1.42, 2.59, 0.64, 2.36, 1.25, 3.23, 11.7 and 9.15 μM respectively.

[0061] Table 2 IC values ​​of monomer components in mangosteen extracts for inhibiting EcGUS-mediated hydrolysis of DDAOG glucuronide and hydrolysis of SN-38G glucuronide 50 and K i Numeric

[0062]

[0063] Example 4 Study on the effect of mangosteen extract on alleviating CPT-11-induced intestinal toxicity

[0064] Healthy female Balb / c mice (18-22 g) were selected to evaluate the alleviating effect of mangosteen extract on CPT-11-induced intestinal toxicity. The specific experimental process is as follows:

[0065] (1) Healthy female Balb / c mice were selected and randomly divided into a blank control group (Group 1), a CPT-11 model group (Group 2), and a CPT-11 and mangosteen extract combined group (Group 3), with 6 mice in each group;

[0066] (2) The blank control group (Group 1) was intravenously injected with an equal volume of saline for 4 consecutive days and intragastrically administered with an equal volume of saline for 4 consecutive days. The CPT-11 model group (Group 2) was intraperitoneally injected with CPT-11 (50 mg / kg) for 4 consecutive days and intragastrically administered with an equal volume of saline for 4 consecutive days. The CPT-11 combined with mangosteen extract group (Group 3) was intravenously injected with CPT-11 (50 mg / kg) and intragastrically administered with mangosteen extract (200 mg / kg). The drug dosage and administration schedule were the same as those of Group 2.

[0067] (3) Record the incidence, severity, and pathological morphology of diarrhea in mice, as well as the weight and survival time of mice (starting from the first day of administration) to evaluate the intestinal toxicity of CPT-11;

[0068] (4) LC-MS / MS technology was used to detect the levels of CPT-11, SN-38, and SN-38G in serum (serum was collected on the first day after drug administration was stopped, i.e., the fifth day of the experiment). About 5 mg of freeze-dried mouse feces (mouse feces were feces excreted on the third day of drug administration) was weighed and placed in a 2 mL EP tube. About 0.5 mL of PBS was added to make a 10 mg / mL solution, which was ultrasonically disrupted and centrifuged at 4°C, 13,000 × g for 5 min in a high-speed refrigerated centrifuge. The solution was diluted 10 times and then analyzed by LC-MS / MS.

[0069] (5) Using the GUS-specific fluorescent probe substrate DDAOG, the activity of β-glucuronidase in mouse feces was evaluated. The specific steps are as follows:

[0070] 1) The incubation system contained mouse fecal supernatant (5 μg / mL) and 100 mM PBS (pH 6.5), and was pre-incubated at 37°C for 5 min;

[0071] 2) Add DDAOG (10.0 μM) to the reaction system to start the reaction;

[0072] 3) After 30 min, add 200 μL of glacial acetonitrile and shake vigorously to terminate the reaction;

[0073] 4) After high-speed centrifugation at 4°C and 20,000×g for 10 min, the supernatant was collected and subjected to fluorescence detection (Ex=600nm, Em=660nm).

[0074] Figure 3 The results of the test on the effect of mangosteen extract on alleviating CPT-11-induced intestinal toxicity, where (a) is the change of weight loss of Bablc female mice over time; (b) is the change of disease activity index over time; (c) is the change of diarrhea incidence rate over time (the vertical axis Diarrhea-free represents the rate without diarrhea, and the side axis reflects the diarrhea incidence rate); (d) is the β-glucuronidase activity in the feces of Bablc female mice; (e) is the SN-38 content in the serum of Bablc female mice; (f) is the SN-38G content in the serum of Bablc female mice; Day 1 in the horizontal axis of (a)-(c) is the first day of administration, (a)-(c) are the average values ​​of 6 female mice in each group, (d)-(f) are the individual values ​​of 6 female mice in each group, and GM in the figure represents mangosteen extract. Figure 3 It can be seen that mangosteen extract can alleviate the weight loss of Bablc female mice induced by CPT-11, reduce the disease activity index of Bablc female mice induced by CPT-11, and reduce the incidence rate of diarrhea induced by CPT-11 in Bablc female mice. At the same time, mangosteen extract inhibits the activity of β-glucuronidase in the feces of Bablc female mice, reduces the SN-38 content in the serum of Bablc female mice, and increases the SN-38G content in the serum of Bablc female mice.

[0075] Comparative Example 1

[0076] Comparison of inhibitory activity of different plant extracts on EcGUS

[0077] Using DDAOG glucuronide hydrolysis and metabolism reaction as the probe reaction, with the help of in vitro GUS enzyme incubation system, the inhibitory activity of different plant extracts on GUS was tested by multifunctional ELISA plate assay method. The specific experimental process is as follows:

[0078] (1) In vitro GUS metabolic reaction system, including PBS buffer (100 mM) at pH 6.5, EcGUS (0.2 U / mL), and plant extract (10 μg / mL), was pre-incubated at 37°C with shaking for 5 min;

[0079] (2) Add DDAOG (final concentration 10.0 μM) to the reaction system to start the reaction;

[0080] (3) After 30 min, add 200 μL of glacial acetonitrile and shake vigorously to terminate the reaction;

[0081] (4) After high-speed centrifugation at 4°C and 20,000×g for 10 min, the supernatant was collected and subjected to fluorescence detection (Ex=600 nm, Em=660 nm).

[0082] In this comparative example, 6 different plant extracts were used for performance comparison. The extraction methods of various plant extracts were the same as the extraction method of mangosteen extract used in the present invention. The test results are shown in Table 3:

[0083] Table 3 Inhibitory activity of different plant extracts on EcGUS

[0084]

[0085] As shown in Table 3, the inhibitory activity of other plant extracts on β-glucuronidase is far inferior to that of mangosteen extract.

[0086] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A use of a mangosteen extract in the preparation of a drug for treating irinotecan-induced drug-induced enterotoxicity, characterized in that: The mangosteen extract is obtained by taking mangosteen peel as raw material and using ethanol aqueous solution as extraction solvent; The concentration of the ethanol aqueous solution is 90wt%; the usage ratio of the mangosteen peel to the ethanol aqueous solution is 1g:10mL.

2. The use according to claim 1, characterized in that The preparation of the mangosteen extract specifically comprises: mixing mangosteen peel and ethanol aqueous solution, soaking for 4 hours, ultrasonicating for 2 hours, filtering, and rotary evaporating to obtain the mangosteen extract.

3. The use according to claim 1, characterized in that The active ingredients in the mangosteen extract include one or more of α-mangostin, β-mangostin, γ-mangostin, gambogelin C, gambogelin D, 8-deoxygambogelin, gambogelin and 3-isomangostin.

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