CN103159814B - A kind of morellic acid ester derivative and its production and use - Google Patents

Abstract

The invention relates to a gambogic acid ester derivative and its preparation method and application. The structural formula of the derivative is shown in formula (I); the preparation method comprises: under nitrogen protection, gambogic acid, gemcitabine and triphenyl The phosphine is dissolved in anhydrous tetrahydrofuran, injected into a THF solution containing diethyl azodicarboxylate DEAD, reacted for 12-24 hours, and the product is obtained after washing, drying, rotary evaporation and column chromatography. The derivatives can be used for preparing compositions, antineoplastic drugs, antifungal drugs or antiangiogenic drugs or used in combination with other anticancer active ingredients. The derivative of the present invention not only shows good activity to various tumor cells, but also shows good targeting to tumor cells, which will lay the foundation for the development of new drugs.

Description

A kind of gambogic acid ester derivative and its preparation method and application

technical field

The invention belongs to the field of natural product derivatives of traditional Chinese medicine, and in particular relates to a gambogic acid ester derivative and its preparation method and application.

Background technique

Malignant tumors are one of the major diseases that seriously threaten human life and quality of life. At present, most antineoplastic drugs are synthetic drugs, which have the disadvantages of large adverse reactions and immunosuppression. Therefore, finding new anticancer drugs and methods, reducing adverse reactions of chemotherapy, and improving the quality of life of patients has become a hot spot in medical research. Because of their ability to inhibit tumor growth and have less toxic and side effects, traditional Chinese medicines and herbal medicines have received widespread attention from medical workers.

Gambogic acid is a compound with high anti-tumor activity and good cell selectivity extracted from Garcinia Hanburryi. Experimental research results show that gambogic acid has strong anti-tumor activity on various tumors, and its toxic and side effects are relatively small within the effective dose range. There is no effect on the function, which is not available in current tumor chemotherapy drugs. This provides novel prospects for finding new anticancer drugs and may reveal new anticancer mechanisms.

本发明的反应与吉西他滨反应生成酯类化合物的类似（见参考文献：AndriesM.Bergman,AukeD.Adema&JanBalzarin,SkjalgBruheim,etal.Antiproliferativeactivity,mechanismofactionandoralantitumoractivityofCP-4126,afattyacidderivativeofgemcitabine,ininvitroandinvivotumormodels.InvestNewDrugs,2011,29:456–466. ).

Performance change: Adding a lipophilic group makes the present invention no longer need to pass through the hent1 transport enzyme compared with the substrate, making it easier to absorb.

Contents of the invention

The technical problem to be solved by the present invention is to provide gambogic acid ester derivatives and their preparation methods and applications. The derivatives not only show better inhibitory activity against various tumor cells, but also are easier to target than gambogic acid. Cell combination shows good targeting to tumor cells, which will lay the foundation for the development of anti-tumor drugs.

In the formula,

R is independently selected from 5'-gemcitabine;

X is independently O.

The structural formula of the derivative is:

In an embodiment of the present invention, the gambogic acid derivatives, each substituent group combination includes but is not limited to the following table:

Table 1

The present invention also provides a preparation method of gambogic acid ester derivatives, comprising:

Under nitrogen protection, dissolve gemcitabine gambogic acid and triphenylphosphine in anhydrous tetrahydrofuran, cool to 0°C, inject a THF solution containing diethyl azodicarboxylate DEAD with a syringe within 10 minutes, and return to room temperature naturally , continue to react for 12-24h; add ethyl acetate to the reactant to quench, then pour into saturated brine to wash, dry over anhydrous sodium sulfate, evaporate ethyl acetate; column chromatography, to obtain crude product; gambogic acid, The ratio of gemcitabine, triphenylphosphine, anhydrous tetrahydrofuran, DEAD, THF, ethyl acetate, and saturated saline is: 0.71mmol: 0.64mmol: 2.13mmol: 5.0mL: 2.13mmol: 5ml: 50ml: 30ml.

The inert solvent is anhydrous tetrahydrofuran or diethyl ether.

The saturated saline was washed 3 times, 10 ml each time.

The eluent is a mixture of methanol and dichloromethane in a volume ratio of 3:100.

The yield is 47.4%.

The principle of the preparation method of the present invention is: in an inert solvent, gambogic acid in formula (II) is reacted with RXH, thereby forming the compound of formula (I):

In the formula, the definitions of R and X are as shown in Table 1, and the salt of RXH can be an inorganic acid salt or an organic acid salt.

Preferably, in the preparation method of the above compound, the corresponding ester is prepared by reacting gambogic acid in formula (II) with RXH under the catalysis of a base, and the base is selected from NaOH, KOH, K ₂ CO ₃ , Na ₂ CO ₃ , Any of NaHCO ₃ , Cs ₂ CO ₃ or NaH.

Preferably, in the preparation method of the above-mentioned compound, gambogic acid in the formula (II) prepares corresponding ester by condensation catalytic reagent and alcohol reaction, and above-mentioned condensation reagent can be independently selected from: diisopropyl azodicarboxylate ( DIAD), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI);

The gambogic acid ester derivative of the present invention can be used to prepare a composition, which contains a safe and effective amount of the gambogic acid ester derivative and a pharmaceutically acceptable carrier. The "safe and effective amount" refers to: the amount of the compound is sufficient to improve the condition without causing serious side effects. The safe and effective dose is determined according to the age, condition, course of treatment, etc. of the subject to be treated.

The gambogic acid ester derivative of the present invention can be used to prepare antitumor drugs, antifungal drugs or anti-angiogenesis drugs; the tumor is any one of breast cancer, lung cancer, liver cancer and colon cancer.

The compounds of the present invention have good antitumor activity, and they can be used to treat tumors, including esophagus, stomach, intestine, rectum, oral cavity, pharynx, larynx, lung, colon, breast, uterus, endometrium, ovary, prostate, testis , bladder, kidney, liver, pancreas, bone, connective tissue, skin, eye, brain, and central nervous system, as well as thyroid cancer, leukemia, Hodgkin's disease, lymphoma, and myeloma.

A gambogic acid ester derivative of the present invention can be used in combination with other one (or more) anticancer active ingredients, especially anticancer compounds, such as alkylating agents, such as alkyl sulfonate (buxiao An), dacarbazine, procarbazine, nitrogen mustard compounds (chlormethine, phenylalanine nitrogen mustard, tumor canine), cyclophosphamide or ifosfamide; nitrosoureas, such as card Mustin, lomustine, semustine, or streptozotocin; antineoplastic alkaloids, such as vincristine or vinblastine; taxanes, such as paclitaxel or taxotere, antineoplastic antibiotics Classes such as actinomycin; intercalating reagents, antineoplastic metabolites, folate antagonists, or methotrexate; purine synthesis inhibitors; purine analogs such as 6-mercaptoguanine; pyrimidine synthesis inhibitors, aromatase inhibitors drugs, capecitabine, or pyrimidine analogs such as fluorouracil, cytarabine, cytosine arabinoside; brequinar, topoisomerase inhibitors such as camptothecin or etoposide; anticancer hormones Agonists and antagonists, including tamoxifen; kinase inhibitors, imatinib mesylate (imatinib), growth factor inhibitors, anticancer metal complexes, anthracyclines, etc.

It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described in the following (such as embodiments) can be combined with each other to form new or preferred technical solutions.

Nucleoside drugs are very hydrophilic, while gambogic acid is very lipophilic. The products synthesized from them through ester bonds have hydrophilicity and lipophilicity just between the two, which is easier Combined with targeted cells, it has stronger anti-tumor properties.

Beneficial effect

(1) The present invention provides a class of novel Chinese medicine natural product derivatives based on gambogic acid.

(2) The present invention has broad-spectrum anti-tumor activity, and has significant growth-inhibiting effect on various solid tumors.

(3) Compared with gambogic acid, the present invention is easier to combine with targeted cells, has strong targeting, stable chemical structure, and is expected to be developed into an innovative anti-tumor drug.

Detailed ways

Below in conjunction with specific embodiment, further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. In addition, it should be understood that after reading the teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

Compound of the present invention can adopt gambogic acid to be raw material (the preparation of raw material sees references: Chen Baoren. Separation of gambogic acid and identification of structure, Jiangxi Medical College Journal, 1980, 7. and TimothyJ.R.Weakley, SuiX.Cai , Han-ZhongZhang, and JohnF.W.Keana.Crystalstructureofthepyridinesaltofgambogicacid.J.ChemCrystallogr,December2001,31:11-12), prepared by repeated recrystallization, the method is simple, the yield is high, and it is easy to realize industrialization.

Example 1

Synthesis of SZY-182

Under nitrogen protection, gambogic acid (500mg, 0.71mmol), gemcitabine (169mg, 0.64mmol) and triphenylphosphine (555mg, 2.13mmol) were dissolved in anhydrous tetrahydrofuran (5.0mL), cooled to 0°C for 10 minutes Inject a solution of DEAD (321mg, 2.13mmol) in 5ml THF with a syringe, return to room temperature naturally, and continue the reaction for 12h. Ethyl acetate (50ml) was added to the reactant to quench, then poured into saturated brine (3×10ml) for washing, dried over anhydrous sodium sulfate, and evaporated to remove ethyl acetate. Column chromatography [V (methanol): V (dichloromethane) = 3: 100] gave 294 mg of an orange-yellow amorphous powder with a yield of 47.4%. ¹ HNMR (CDCl ₃ , 400MHz) δ: 12.87(s, 1H, 6-OH), 7.58(d, J=6.88Hz, 1H, 10-H), 6.68(d, J=10.12Hz, 1H, 4- H), 6.38(m, 1H, 1”-H), 6.14～6.18(m, 1H, 27-H), 5.87(d, J=6.96, 2H, 5’-H, 6’-H), 5.40 (d, J=10Hz, 1H, 3-H), 5.06～5.08(m, 1H, 32-H, 37-H), 4.18(m, 1H, 4”-H), 4.08(m, 1H, 5 "-H), 3.50～3.51(m, 1H, 11-H), 3.32～3.37(m, 1H, 31-Ha), 3.16～3.19(m, 1H, 31-Hb), 2.95-2.99(m, 1H,32-H),2.54(d,J＝9.24Hz,1H,22-H),2.33～2.35(m,1H,21-Ha),2.04～2.08(m,2H,36-H),1.76 (s,6H,34-H),1.75(s,3H,25-H),1.72(s,3H,29-H),1.63(s,3H,39-H),1.67(s,2H,20 -H), 1.58(s, 3H, 35-H), 1.46(s, 3H, 40-H), 1.28(s, 3H, 19-H), 1.31(m, 1H, 21b-H); ESI- MSm/z:874[M+H] ⁺ .

Example 2

In vitro anti-tumor activity test

1. Experimental cell line:

The tumor cell lines used in this experiment are: A549 (human lung adenocarcinoma cells), HCT116 (human colon cancer cells), PANC-1 (human pancreatic cancer cells) and HepG2 (human liver cancer cells) (provided by Shanghai Institute of Pharmaceutical Industry pharmacology laboratory cryopreservation and passage).

2. Sample preparation:

After dissolving with DMSO (Merck), add PBS(-) to make a 1000μg/ml solution or a homogeneous suspension, and then dilute with DMSO-containing PBS(-). The positive control drug was gambogic acid.

3. Test method

MTT method: 100 μl of cell suspension with a concentration of 4-5×10 ⁴ cells/ml was added to each well of a 96-well plate, and placed in a 37°C, 5% CO ₂ incubator. After 24 hours, add the sample solution, 10 μl/well, set up duplicate holes, and act at 37°C, 5% CO ₂ for 72 hours. Add 20 μl of 5 mg/ml MTT solution to each well, add the dissolving solution after 4 hours of action, 100 μl/well, place in the incubator, and measure the 570nmOD value with MK-2 automatic microplate reader after dissolution. (See Table 2 for antitumor activity in vitro)

Inhibitory effect of some samples in table 2 on the proliferation of human tumor cells in vitro

It can be seen from Table 2 that the gambogic acid ester derivatives of the present invention are similar to gambogic acid, and have good broad-spectrum anti-tumor activity on various human tumor cells such as lung cancer, colon cancer, breast cancer and liver cancer. Literature shows that gambogic acid also has good antitumor activity against gastric cancer, cervical cancer and melanoma, therefore, the gambogic acid derivatives of the present invention may also have good activity against these tumors. It is particularly worth pointing out that the compounds of the present invention can also significantly improve their water solubility by forming salts, which helps to increase the bioavailability of such compounds. All these show that the compound of the present invention has a good development prospect.

Claims (7)

1. a gambogic acid ester derivative, structural formula is as follows: 2. the preparation method of a kind of gambogic acid ester derivative according to claim 1, comprising: Under nitrogen protection, dissolve gambogic acid, gemcitabine and triphenylphosphine in anhydrous tetrahydrofuran, cool to 0°C, inject a THF solution containing diethyl azodicarboxylate with a syringe within 10 minutes, and naturally return to Continue to react at room temperature for 12-24h; add ethyl acetate to the reactant to quench, then pour into saturated brine to wash, dry over anhydrous sodium sulfate, and evaporate ethyl acetate; column chromatography to obtain the product; gambogic acid, The ratio of gemcitabine, triphenylphosphine, anhydrous tetrahydrofuran, diethyl azodicarboxylate, THF, ethyl acetate, and saturated saline is: 0.71mmol: 0.64mmol: 2.13mmol: 5.0mL: 2.13mmol: 5ml: 50ml: 30ml. 3. the preparation method of a kind of gambogic acid ester derivative according to claim 2, is characterized in that: described saturated saline is washed 3 times, each 10ml. 4. the application of a kind of gambogic acid ester derivatives according to claim 1, is characterized in that: be used for preparing a kind of composition, contain the gambogic acid ester derivatives of safe and effective dose and pharmaceutically acceptable carrier. 5. the application of a kind of gambogic acid ester derivative according to claim 4, is characterized in that: be used for preparing antitumor drug; Described tumor is any one in breast cancer, lung cancer, liver cancer, colon cancer kind. 6. The application of a gambogic acid ester derivative according to claim 5, characterized in that it is used in combination with one or more other anticancer active ingredients. 7. The application of a gambogic acid ester derivative according to claim 6, characterized in that it is used in combination with one or more other anticancer compounds.