Horsfiquiinone B derivative as well as preparation method and application thereof

Abstract

The invention relates to the technical field of medicines, and discloses a Horsfiquione B derivative as well as a preparation method and application thereof, the applicant of the invention finds that Horsfiquione B hydroxylamine derivatives 1 and 2 are prepared by adopting a reaction between Horsfiquione B and hydroxylamine, and finds that the anti-tumor activity of the Horsfiquione B hydroxylamine derivatives is doubled, and the Horsfiquione B hydroxylamine derivatives have remarkable anti-breast cancer and leukemia tumor cell proliferation activity. The application prospect of tumor treatment or adjuvant treatment is realized.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and particularly to Horsfiequinone B derivatives, their preparation methods and applications. More specifically, it relates to derivatives of diarylpropane dimer Horsfiequinone B, pharmaceutical compositions containing the same as active ingredients, their preparation methods, and their applications in the preparation of anti-cancer drugs. Background Art

[0002] The statements in this section only provide background information related to the disclosure of the present application and may not constitute prior art.

[0003] At present, effective cancer treatment remains a major challenge. However, the anti-tumor drugs currently used clinically have problems such as poor prognosis and easy development of drug resistance. Therefore, it is of great significance to develop new and efficient anti-cancer candidate drugs.

[0004] Natural products are an important source of new anti-cancer drugs, and more and more new compounds and preparations based on natural products have been developed for cancer treatment. There are research reports indicating that the natural diarylpropane dimer compound Horsfiequinone B has anti-tumor activity. Nevertheless, its natural content is low, and its anti-tumor activity needs to be improved. Therefore, it is particularly important to study its analogs to increase the possibility of developing drugs from such compounds.

[0005] Derivatization is the main method used in the development of natural product drugs. However, the anti-tumor mechanism of the natural product Horsfiequinone B is not clear, which further increases the complexity of its structural optimization. In the absence of clear targets and action pathways, the design of its derivatives often relies on the trial-and-error method. Summary of the Invention

[0006] The purpose of the present invention is to provide Horsfiequinone B derivatives, their preparation methods and applications in view of the problem that the anti-tumor activity of the natural product Horsfiequinone B is weak at present. It is first discovered that the O-aminoxime condensation derivatives of the natural diarylpropane dimer Horsfiequinone B have significant anti-proliferative activity against a variety of tumor cell lines, significantly enhancing the anti-tumor activity of Horsfiequinone B.

[0007] The technical solution of the present invention is as follows:

[0008] On the one hand, the present invention provides a Horsfiequinone B derivative, the structural formula of which is as follows:

[0009]

[0010] On the other hand, the present invention provides a method for preparing a hydroxamic acid condensation derivative of Horsfiequinone B as described above, which comprises the following steps:

[0011] React Horsfiequinone B with hydroxylamine hydrochloride and pyridine. After the product is extracted with an organic solvent, it is washed successively with a saturated copper sulfate solution and a saturated saline solution, and then subjected to flash silica gel column chromatography to obtain the diarylpropane dimer Horsfiequinone B derivative 1 or 2.

[0012] According to a preferred embodiment, it comprises the following steps:

[0013] Step (1): React Horsfiequinone B with hydroxylamine hydrochloride (NH2OH·HCl) and pyridine. After the product is extracted with an organic solvent, it is washed successively with a saturated copper sulfate solution and a saturated saline solution, and then subjected to flash silica gel column chromatography to obtain the diarylpropane dimer Horsfiequinone B derivative 1;

[0014] According to a preferred embodiment, it comprises the following steps:

[0015] Step (1): React Horsfiequinone B with O-methylhydroxylamine hydrochloride (NH2OMe·HCl) and pyridine. After the product is extracted with an organic solvent, it is washed successively with a saturated copper sulfate solution and a saturated saline solution, and then subjected to flash silica gel column chromatography to obtain the diarylpropane dimer Horsfiequinone B derivative 2.

[0016] On the other hand, the present invention provides the use of a Horsfiequinone B derivative as described above in the preparation of a drug for treating or adjuvantly treating cancer.

[0017] Preferably, the cancer is human breast cancer or leukemia.

[0018] On the other hand, the present invention provides a pharmaceutical composition for treating or adjuvantly treating cancer, which comprises a Horsfiequinone B derivative as described above as an active pharmaceutical ingredient.

[0019] Preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient or carrier.

[0020] Preferably, the excipient includes a bulking agent.

[0021] The present invention has no special limitation on the pharmaceutical carrier or the bulking agent, and a pharmaceutically acceptable carrier or bulking agent well-known in the art can be used, specifically one or more of a solid, semi-solid or liquid diluent, a filler or a pharmaceutical product adjuvant.

[0022] In the present invention, in the pharmaceutical composition for treating or adjuvantly treating cancer, the mass ratio of the Horsfiequinone B derivative to the excipient or carrier is 1:1 - 1:10.

[0023] Preferably, the pharmaceutical composition is prepared as a liquid preparation, a solid preparation, a spray or an aerosol. The liquid preparation preferably includes an injection, a suspension, an emulsion, a solution or a syrup; the solid preparation preferably includes a tablet, a capsule, a granule or an infusion. In the present invention, there is no specific limitation on the preparation method of the pharmaceutical composition of the oxime condensation derivative of Horsfiequinone B, and those skilled in the art can routinely adjust the dosage and the like.

[0024] In the present invention, the administration method of the pharmaceutical composition for treating or adjuvantly treating cancer is preferably injection, oral administration or sublingual administration; the injection preferably includes intravenous injection, intravenous drip, intramuscular injection, intraperitoneal injection or subcutaneous injection.

[0025] In the present invention, the pharmaceutical composition for treating or adjuvantly treating cancer is preferably used in the form of the dosage per unit body weight. The dosage per unit body weight is preferably 0.5 - 20 mg / kg.

[0026] Compared with the existing technology, the beneficial effects of the present invention are as follows:

[0027] 1. After the structure of Horsfiequinone B is improved and optimized in the present application, the anti-tumor activity of the prepared Horsfiequinone B derivative is significantly improved; specifically, the oxime condensation derivative of Horsfiequinone B has good anti-proliferation activity against tumor cell lines including human triple-negative breast cancer cell line MDA-MB-468, HER2-positive breast cancer cell line JIMT-1, and human leukemia tumor cell line HL-60, and its anti-tumor activity is about 2 times that of the original Horsfiequinone B.

[0028] 2. A class of new oxime condensation derivatives of diarylpropane dimer Horsfiequinone B is provided;

[0029] 3. The application of the oxime condensation derivative of the diarylpropane dimer Horsfiequinone B or the pharmaceutical composition of the oxime condensation derivative of the diarylpropane dimer Horsfiequinone B in the preparation of a medicament for treating cancer is also provided. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 It is a schematic diagram of the preparation method of Horsfiequinone B derivative 1;

[0031] Figure 2 Schematic diagram of the preparation method of Horsfiequinone B derivative 2;

[0032] Figure 3 Structural formulas of Horsfiequinone B derivatives 1 and 2;

[0033] Figure 4 Structural formulas of products 3, 4, 5 and 6 of the ester condensation of Horsfiequinone B;

[0034] Figure 5 IC of different cells treated with Horsfiequinone B 50 IC of different cells treated with compounds 1 and 2 respectively 50 Comparison (Note: The IC50 of different cells treated with Horsfiequinone B is compared with the IC of different cells treated with compounds 1 and 2 respectively 50 Comparison (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001)). Detailed implementation manners

[0035] The specific examples listed in the present invention are only examples of the present invention, and the present invention is not limited to the specific examples described below. For those skilled in the art, any equivalent modifications and substitutions to the examples described below are also within the scope of the present invention. Therefore, equivalent transformations and modifications made without departing from the spirit and scope of the present invention should be covered within the scope of the present invention. For those conditions not specified in the examples, they are carried out according to conventional conditions or the conditions recommended by the manufacturer. All reagents or instruments not indicating the manufacturer can be obtained through commercially available conventional products. To better illustrate the present invention, numerous specific details are given in the following detailed implementation manners. Those skilled in the art should understand that the present invention can be implemented without some specific details. In other examples, methods, means, equipment and steps well-known to those skilled in the art are not described in detail to highlight the gist of the present invention.

[0036] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art. Without special instructions, the units used in this specification are all international standard units, and the numerical values and numerical ranges appearing in the present invention should be understood to include the inevitable systematic errors in industrial production.

[0037] The compound Horsfiequinone B of the present invention can be prepared according to the method described in the literature (Lumin Li; Ruiling Liu; Rui Yang; Hanwei Li; Ya Liu; Rui Zhan; Wenjing Wang; Lidong Shao; Dashan Li, Biomimetic Total Synthesis of Horsfiequinone B Enabled by a Friedel-Crafts Alkylation-Type Non-homodimerization, Organic Chemistry Frontiers, 2024, 11(19), 5358-5362).

[0038] The features and properties of the present invention will be further described in detail below in conjunction with the examples.

[0039] Example 1 Preparation of the diarylpropane dimer Horsfiequinone B derivative 1

[0040] Adopt Figure 1 the preparation method shown for preparation, which specifically includes the following preparation steps:

[0041] Dissolve the raw material Horsfiequinone B (30 mg, 0.05 mmol) in anhydrous ethanol (2 mL). At room temperature, successively add hydroxylamine hydrochloride NH2OH·HCl (35 mg, 0.5 mmol) and pyridine (60 μL, 0.75 mmol), transfer to a sealed tube and react at 70 °C for 3 h. Monitor by TLC until the reaction is completed. Dilute with 5 mL of water and extract with ethyl acetate three times (10 mL×3). Combine the organic phases, wash the organic phases twice with saturated copper sulfate solution (5 mL×2), then wash twice with saturated brine (5 mL×2), dry over anhydrous sodium sulfate, filter, concentrate under reduced pressure in vacuo, and subject the residue to flash silica gel column chromatography (eluted with dichloromethane:methanol = 10:1) to obtain the diarylpropane dimer Horsfiequinone B derivative 1 (23 mg) in a yield of 75%, which is a yellow amorphous powder.

[0042] Structure identification:

[0043] 11H NMR (500 MHz, acetone-d6) δ 7.40 (s, 1H), 7.08 (s, 1H), 6.69 (m, 2H), 6.63 (s, 1H), 6.60 (dd, J = 7.9, 1.5 Hz, 1H), 6.49 (s, 1H), 6.43 (s, 1H), 5.91 (s, 2H), 5.83 (s, 2H), 5.72 (s, 1H), 4.44 (t, J = 7.6 Hz, 1H), 3.78 (s, 3H), 3.67 (s, 3H), 2.66–2.59 (m, 4H), 2.55–2.43 (m, 2H), 2.13–2.08 (m, 2H), 1.91–1.84 (m, 2H); 13 13C NMR (200 MHz, acetone-d6) δ 186.9, 162.9, 157.3, 155.0, 150.1, 148.5, 146.6, 146.5, 145.64, 141.2, 137.2, 130.7, 122.0, 121.5, 121.4, 120.9, 119.0, 110.2, 109.6, 108.7, 105.0, 101.6, 101.4, 100.3, 98.4, 56.16, 55.8, 37.1, 36.5, 34.8, 31.6, 30.5. HR-ESI-MS (m / z): calculated for C 34 H 32 NO 10 [M-H] - : 614.2032, found: 614.2029.

[0044] Example 2 Preparation of the diarylpropane dimer Horsfiequinone B derivative 2

[0045] According to Figure 2 the preparation method shown, the synthesis is carried out specifically as follows:

[0046] Dissolve the raw material Horsfiequinone B (30 mg, 0.05 mmol) in absolute ethanol (2 mL). At room temperature, sequentially add O-methylhydroxylamine hydrochloride NH2OMe·HCl (42 mg, 0.5 mmol) and pyridine (60 μL, 0.75 mmol). Transfer the mixture to a sealed tube and react at 70 °C for 3 h. Monitor the reaction by TLC until the reaction is complete. Dilute with 5 mL of water and extract with ethyl acetate three times (10 mL × 3). Combine the organic phases, wash the organic phase twice with saturated copper sulfate solution (5 mL × 2), then wash twice with saturated brine (5 mL × 2). Dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure in vacuo. The residue is purified by flash silica gel column chromatography (eluted with dichloromethane:methanol = 10:1) to obtain compound 2 (18 mg) in a yield of 56%. It is a mixture of Z and E isomers that cannot be separated by column chromatography and high performance liquid chromatography, a yellow amorphous powder.

[0047] Structure identification:

[0048] 1 H NMR (600 MHz, CDCl3) δ 7.30 (s, 0.5H), 7.01 (d, J = 9.4 Hz, 1H), 6.74 (s, 0.5H), 6.68 (dd, J = 8.0, 2.5 Hz, 1H), 6.63 (s, 1H), 6.61 (d, J = 6.3 Hz, 1H), 6.57 (d, J = 7.9 Hz, 1H), 6.39 (d, J = 6.8 Hz, 1H), 6.31 (d, J = 5.8 Hz, 1H), 5.90 (s, 2.5H), 5.85 (s, 2.5H), 5.77 (s, 0.5H), 5.73 (s, 0.5H), 4.36–4.31 (m, 1H), 4.14 (s, 3H), 3.79 (s, 1.6H), 3.75 (s, 1.4H), 3.684 (s, 1.6H), 3.677 (s, 1.4H), 2.61 (t, J = 7.2 Hz, 2H), 2.53 (t, J = 7.8 Hz, 2H), 2.49–2.46 (m, 2H), 2.14–2.09 (m, 2H), 1.92 - 1.85 (m, 2H); 1313C NMR (150 MHz, CDCl3) δ 187.2, 186.6, 161.5, 158.2, 156.8 (C×2), 152.9, 152.8, 148.3, 148.2, 147.5 (C×2), 146.1, 146.0, 145.6 (C×3), 145.1, 143.9, 142.5, 141.4 (C×2), 136.5, 136.5 (C×2), 132.5, 130.9, 130.7, 122.1, 121.8, 121.3 (C×2), 120.3, 120.2, 119.1, 119.0, 118.8, 109.1 (C×3), 108.2 (C×2), 107.1, 105.1, 101.0, 100.9, 100.8 (C×2), 99.8, 99.7, 98.6, 65.4, 64.3, 56.3, 56.1, 55.7 (C×2), 37.6, 37.1, 35.4, 35.3, 34.2, 34.1, 30.0, 29.0, 28.9, 28.8, 28.7. HR-ESI-MS (m / z): calculated for C 35 H 34 NO 10 [M-H] - : 628.2188, found: 628.2189。

[0049] Example 3 Antitumor Proliferation Activities of Diarylpropane Dimer Horsfiequinone B Derivatives 1 and 2

[0050] The O - hydroxyamine condensation derivatives 1 and 2 of the compound diarylpropane dimer Horsfiequinone B of the present invention have significant anti - proliferation activities against tumor cell lines including human triple - negative breast cancer cell line MDA - MB - 468, HER2 - positive breast cancer cell line JIMT - 1, and human leukemia tumor cell line HL - 60.

[0051] The experimental methods and results are as follows:

[0052] I. Experimental Methods

[0053] 1. The samples of the O - hydroxyamine condensation derivatives of the diarylpropane dimer Horsfiequinone B to be tested were separately dissolved in DMSO to prepare solutions with a concentration of 20 μM and stored for later use.

[0054] 2. The working solution concentration starts at 20 μM and is diluted two-fold into 5 concentration gradients. The triple-negative breast cancer cell line MDA-MB-468 (3650 cells / well), HER2-positive breast cancer cell line JIMT-1 (4300 cells / well), and human leukemia tumor cell line HL-60 (8000 cells / well) in the logarithmic growth phase are seeded in 96-well plates and cultured overnight for 24 h. Gradient-diluted compounds are added for treatment, with three replicates for each concentration. At the same time, a control group and a blank group are set up. The morphological changes of the cells are observed after 24 h, 48 h, and 72 h. After 72 h, the viability of HL-60 cells is detected using CCK-8, and the viability of MDA-MB-468, JIMT-1, and HL-60 cells is detected using MTT.

[0055] Cell viability = (OD of the drug treatment group - OD of the blank) / (OD of the DMSO group - OD of the blank).

[0056] 3. IC 50 Determination

[0057] HL-60 (20 μM, 72 h) is selected for primary screening, and compounds 1 and 2 with an inhibition rate greater than 50% are obtained. The triple-negative breast cancer cell line MDA-MB-468 (3650 cells / well), HER2-positive breast cancer cell line JIMT-1 (4300 cells / well), and leukemia tumor cell line HL-60

[0058] (8000 cells / well) are seeded in 96-well plates and cultured overnight. After 24 h, the above compounds are added for a new round of rescreening (starting at a concentration of 20 μM and diluted two-fold in gradients) and treated for 72 h. Three replicates are set for each concentration. At the same time, a control group (vincristine, starting at a concentration of 20 μM and diluted two-fold in 5 concentration gradients) and a blank group (treated with PBS solution) are set up. For HL-60, cck-8 is used for staining at 37 °C for 1 h and the absorbance is detected at 450 nm. For the remaining cells, 5 mg / mL MTT is used for staining at 37 °C for 2.5 h. The culture medium is removed and 100 μL of DMSO is added, and the absorbance is detected at 490 nm. Nonlinear regression analysis is performed using GraphPad Prism 10 software to determine the IC 50 value.

[0059] II. Experimental Results

[0060] See Table 1. The present invention relates to two hydroxylamine condensation derivatives 1 and 2 of the diarylpropane dimer Horsfiequinone B and the ester condensation products 3, 4, 5, and 6 of Horsfiequinone B (structures as Figure 3As shown). In vitro anti-proliferative activity evaluation of glioma was carried out. The results showed that the O-aminohydroxyl condensation derivatives 1 and 2 of Horsfiequinone B had obvious inhibitory effects on the growth of human triple-negative breast cancer cell line MDA-MB-468, HER2-positive breast cancer cell line JIMT-1, and leukemia tumor cell line HL-60. Their anti-tumor activities were superior to those of the natural diarylpropane dimer compound Horsfiequinone B, indicating that the tested compounds have anti-tumor uses. In addition, the derivatives 3, 4, 5, and 6 of Horsfiequinone B with similar compound skeleton structures had no anti-tumor activities.

[0061] Table 1 Anti-proliferative activities of diarylpropane dimer Horsfiequinone B derivatives against various tumor cells a

[0062]

[0063] a The experimental results are the average values of three independent experiments. b Positive control.

[0064] Example 4 A pharmaceutical composition for treating or adjuvantly treating cancer

[0065] A pharmaceutical composition for treating or adjuvantly treating cancer, including the O-aminohydroxyl condensation derivatives 1 and 2 of the diarylpropane dimer Horsfiequinone B of the present invention as Figure 3 shown. The excipient is added in a weight ratio of 1:1 with the O-aminohydroxyl condensation derivatives 1 and 2 of the diarylpropane dimer Horsfiequinone B of the present invention, and granulated and tabletted. The excipients are starch paste, magnesium stearate, talc powder, polyethylene glycol (PEG), etc.

[0066] Example 5 A pharmaceutical composition for treating or adjuvantly treating cancer

[0067] A pharmaceutical composition for treating or adjuvantly treating cancer, the O-aminohydroxyl condensation derivatives 1 and 2 of the diarylpropane dimer Horsfiequinone B of the present invention, are made into capsules by a conventional capsule preparation method.

[0068] Example 6 A pharmaceutical composition for treating or adjuvantly treating cancer

[0069] A pharmaceutical composition for treating or adjuvantly treating cancer, 10 mg in total of the O-aminohydroxyl condensation derivatives 1 and / or 2 of Horsfiequinone B, also includes appropriate amounts of starch, corn syrup, and magnesium stearate; the O-aminohydroxyl condensation derivatives 1 and / or 2 of Horsfiequinone B are mixed with the auxiliary agents starch, corn syrup, and magnesium stearate, granulated and tabletted to obtain tablets.

[0070] Example 7. A pharmaceutical composition for treating or adjuvantly treating cancer

[0071] A pharmaceutical composition for treating or adjuvantly treating cancer, comprising a total of 10 mg of the oxime condensation derivatives 1 and / or 2 of Horsfiequinone B, and also comprising appropriate amounts of starch and magnesium stearate; mixing the oxime condensation derivative of Horsfiequinone B or its pharmaceutical composition with the auxiliary agents starch and magnesium stearate, sieving, uniformly mixing in a suitable container, and filling the obtained mixture into hard gelatin capsules to obtain capsule preparations.

[0072] Example 8. A pharmaceutical composition for treating or adjuvantly treating cancer

[0073] A total of 2 mg of the oxime condensation derivatives 1 and / or 2 of Horsfiequinone B and 10 mg of sodium chloride are dissolved in an appropriate amount of water for injection, the obtained solution is filtered, and filled into ampoules under aseptic conditions. Ampoule preparations are obtained.

[0074] The above-described embodiments merely represent the specific embodiments of the present application, and their descriptions are relatively specific and detailed, but should not be construed as limiting the protection scope of the present application. It should be noted that for those of ordinary skill in the art, without departing from the concept of the technical solution of the present application, several modifications and improvements can still be made, and these all fall within the protection scope of the present application.

Claims

1. A Horsfiequinone B condensation derivative, characterized in that: It is a hydroxylamine condensation derivative of Horsfiequinone B.

2. A Horsfiequinone B derivative according to claim 1, characterized in that: Including derivatives 1 and derivatives 2 with the following structural formula:

3. A method for preparing a Horsfiequinone B hydroxylamine condensation derivative, characterized in that: The steps include: Horsfiequinone B is reacted with hydroxylamine hydrochloride and pyridine, the product is extracted with an organic solvent, and then washed with a saturated copper sulfate solution and a saturated saline solution in sequence, and subjected to rapid silica gel column chromatography to obtain a diarylpropane dimer Horsfiequinone B derivative 1 or 2.

4. The method for preparing the Horsfiequinone B hydroxylamine condensation derivative according to claim 3, characterized in that: When preparing the diarylpropane dimer Horsfiequinone B derivative 1, the hydroxylamine hydrochloride is NH2OH . HCl.

5. The method for preparing the Horsfiequinone B hydroxylamine condensation derivative according to claim 3, characterized in that: When preparing the diarylpropane dimer Horsfiequinone B derivative 2, the hydroxylamine hydrochloride is NH2OMe . HCl.

6. Use of a Horsfiequinone B derivative in the preparation of a drug for treating or assisting in the treatment of cancer, characterized in that: The Horsfiequinone B derivative is the Horsfiequinone B derivative as claimed in claim 1 or 2.

7. Use of a Horsfiequinone B derivative according to claim 6 in the preparation of a drug for treating or assisting in the treatment of cancer, characterized in that: The cancer is breast cancer or leukemia.

8. A pharmaceutical composition for treating or assisting in the treatment of cancer, characterized in that: The invention comprises the Horsfiequinone B derivative according to claim 1 or 2 and a pharmaceutically acceptable auxiliary material or carrier, wherein the mass ratio of the Horsfiequinone B derivative to the auxiliary material or carrier is 1:1-1:

10.

9. The pharmaceutical composition for treating or assisting in treating cancer according to claim 8, characterized in that: The cancer is breast cancer or leukemia.

10. The pharmaceutical composition for treating or assisting in treating cancer according to claim 8, characterized in that: The single dosage of the Horsfiequinone B derivative is 0.5 to 20 mg / kg.

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