CN110683932B

CN110683932B - Beta-elemene halide and preparation method thereof

Info

Publication number: CN110683932B
Application number: CN201910934274.0A
Authority: CN
Inventors: 谢恬; 叶向阳; 李震; 高园; 卓晓韬; 戚香
Original assignee: Hangzhou Normal University
Current assignee: Hangzhou Normal University
Priority date: 2019-09-29
Filing date: 2019-09-29
Publication date: 2022-05-27
Anticipated expiration: 2039-09-29
Also published as: CN114890866B; CN114890866A; CN110683932A

Abstract

The invention provides a beta-elemene halide and a preparation method and application thereof. Specifically, the invention provides a preparation method of a compound (each group is defined as the specification) with a structure shown as the following formula (I); the invention also provides a compound with a structure shown as the following formula (II) (each group is defined as the specification), a pharmaceutical composition containing the compound of the formula (II), the compound, and isotopic derivatives, chiral isomers, variants, different salts, prodrugs, preparations and the like of the compound. The compound can be used as intermediate for preparing elemene derivatives, and treating various diseases which can be treated by elemene, including lung cancer, breast cancer, liver cancer, etc.

Description

Beta-elemene halide and preparation method thereof

Technical Field

The invention belongs to the field of preparation of beta-elemene derivative intermediates, and particularly relates to a beta-elemene halide and a preparation method thereof.

Background

Elemene element is a sesquiterpene compound extracted and separated from radix curcumae, and compared with other plant anticancer drugs such as paclitaxel, hydroxycamptothecin and vinblastine, the elemene element has the advantages of minimum molecular weight, minimum toxicity, broad-spectrum antitumor effect and the like, and is approved as an antitumor plant drug with independent intellectual property rights in China by the national drug administration in 1994. The emulsion with elemene as the main component has been approved by the Ministry of health as a new national class II anti-cancer drug in 2008 to enter the second-phase clinical research, and the new anti-tumor natural product gradually shows a strong clinical treatment effect in various tumors. At present, the composition is mainly used for chemotherapy of malignant serosal cavity effusion, lung cancer, digestive tract tumor, brain tumor and other superficial tumors clinically, and also has certain curative effect on esophageal cancer, gastric cancer, breast cancer, liver cancer, bladder cancer and the like. Research shows that elemene hardly produces side effects, does not produce drug-induced toxicity on kidney and liver functions, and particularly has no inhibitory effect on spinal cord (zhanjia, zhongpeng, huaxin, yuan korean, chemical bulletin 2010, 6: 499). In addition, the elemene and other targeted small molecule drugs are used together, so that the effects of sensitization and synergy are also shown, and meanwhile, the drug resistance of the small molecule targeted drugs can be reversed or delayed. In recent years, scientists have gained fruitful results in the elemene field, and have successively obtained a plurality of national family prizes. Develops a new generation of anticancer new drug with better activity on the basis of elemene, and has very wide prospect.

Elemene contains various isomers including alpha-, beta-, gamma-, delta-and the like. The research shows that the beta-elemene is the main component and is also the isomer with the best anti-tumor effect, and the structure of the beta-elemene is shown as follows:

studies have also shown that three separate double bonds on β -elemene may be major contributors to antitumor efficacy (but there is no specific evidence). Aiming at the problems of low water solubility (belonging to volatile oil) and low bioavailability of beta-elemene, a plurality of research groups modify the chemical structure of the beta-elemene and have made certain progress (reference (1) Liu, G.; Kong, Z.; Shen, Y. Synthesis, chromatography, and in vitro antibacterial activity of novel beta-elemene monomeric derivatives [ J].Med.Chem.Res.2013,22,3536-3540.(2)Sun,Y.；Liu,G.；Zhang,Y.；Zhu,H.；Ren,Y.；Shen,Y.-M.Synthesis and in vitro anti-proliferative activity ofβ-elemene monosubstituted derivatives in HeLa cells mediated through arrest of cell cycle at the G1phase[J].Bioorganic&Medicinal Chemistry 2009,17,1118–1124.(3)Ren,Y.；Sun,Y.；Cheng,K.；Liu,G.；Shen,Y.Synthesis and radiolabelling of Re(CO)₃-β-elemene derivatives as potential therapeutic radiopharmaceuticals[J].J.Label Compd.Radiopharm 2009,52,139–145.(4)Chen,J.；Wang,R.；Wang,T.et al.Antioxidant Properties of Novel Dimers Derived from Naturalβ-Elemene through Inhibiting H₂O₂-Induced Apoptosis[J].ACS Medicinal Chemistry 2017,8(4),443-448.(5)Xu,L.；Tao,S.；Wang,X.et al.The synthesis and anti-proliferative effects ofβ-elemene derivatives with mTOR inhibition activity.Bioorganic&Medicinal Chemistry 2006,14, 5351-5356). The above documents share a common feature that the chlorination reaction is performed at the 13-position and/or 14-position, and then other derivatives, including bromides, are derived, and several β -elemene halides reported in the prior documents have the following structures:

in 1991, Giardian et al (Giardian, Yankeen, Lianhui et al, organic chemistry, 1991,11(6):608-₂With calcium hypochlorite in the CH of beta-elemene₂Cl₂-H₂The disubstituted chloride 4 of beta-elemene (yield: 36%) and the mixture of the monosubstituted chlorides 2 and 3 (total yield: 28%) are synthesized for the first time in the O mixed solution. Compounds 2 and 3 could not be separated according to the ratio¹δ H ═ 3.90 in H NMR: Δ H was obtained as a peak area ratio of 3.98, and was 8: 3. In addition, there are many documents that use elemene as a raw material and introduce chlorine into allylic position by using sodium hypochlorite and glacial acetic acid to obtain a mixture of 3 kinds of chlorinated products, 13-monochloro 2, 14-monochloro 3 and 13, 14-dichloro 4. Silica gel column chromatography can only separate 13, 14-dichloride 4 from 13-monochloride 2 and 14-monochloride 3, and 13-monochloride 2 and 14-monochloride 3 cannot be separated.

FeCl for tiger et al (patent applications CN 1462745A, CN 1462746)₃The elemene bromide is prepared by combining the catalyst and NBS, but specific nuclear magnetic data are not reported in the specification. Xu jin Yi (ACS med.Lett.2017, 8(4):443-448) reported a method for synthesizing compound 5, which has many reaction steps. The specific process is as follows: firstly, carrying out chlorination reaction on elemene, then carrying out substitution reaction on the chloride and sodium acetate, hydrolyzing the obtained substituted acetate to obtain a mixture of 13-hydroxyl substituted elemene and 14-hydroxyl substituted elemene, separating the mixture by using a chromatographic column to obtain pure 13-pure hydroxyl substituted elemene, and converting the 13-hydroxyl elemene into 13-bromo elemene by using triphenylphosphine and NBS, wherein the specific synthetic route is as follows:

in summary, the synthesis method of the β -elemene halide reported in the prior art is complex, and many prepared compounds exist in the form of mixture, the mixture is difficult to separate and purify, and part of the mixture can not be separated at all.

Disclosure of Invention

The invention aims to provide a preparation method of elemene allylic halide, which greatly simplifies the synthesis steps, improves the product yield and has wide application prospect in elemene derivatization.

In a first aspect of the present invention, there is provided a process for preparing a β -elemene halide compound having a structure represented by the following formula (I):

in the formula (I), R¹And R²Independently selected from hydrogen, Cl, Br, I, and R¹And R²Not hydrogen at the same time;

the method comprises the following steps: dissolving a beta-elemene raw material in a solvent, and carrying out allylic halogenation reaction with a halogenated source reagent under the condition of adding or not adding a transition metal salt to obtain the beta-elemene compound; the transition metal salt is CeCl₃Or Yb (OTf)₃(ii) a The solvent is water, dichloromethane, trichloromethane, tetrahydrofuran and acetic acidOne or more of (a); the halogenated source reagent is NaClO, NCS, NaOBr, NBS, N-bromophthalimide or NIS.

The invention adopts the reaction of monochloro elemene (such as 13-chloro elemene, 14-chloro elemene) of elemene and NaI in a certain solvent (such as acetone) to prepare the corresponding elemene iodo-compound. Or, 13, 14-di-chloro elemene and NaI are reacted in a certain solvent (such as acetone) to prepare the corresponding 13, 14-di-iodo elemene.

Further, the structure of the beta-elemene halogenated compound is at least one of the following formulas:

the beta-elemene halide is a mixture of a compound of a formula 2 and a compound of a formula 3, a mixture of a compound of a formula 5 and a compound of a formula 11, or a mixture of a compound of a formula 12 and a compound of a formula 13, and the method comprises the following steps: dissolving beta-elemene raw material in solvent, adding CeCl₃The system is obtained by allylic halogenation reaction with a halogenated source reagent.

The separation method of the mixture comprises the following steps: dissolving the mixture in acetonitrile, adding into a circulating preparative liquid chromatograph, wherein the mobile phase is pure acetonitrile, the flow rate is 3.5mL/min, clicking a circulating button at 1h, and collecting a main peak after four cycles of raw materials and at the fifth cycle.

Preferably, the β -elemene halide is a mixture of a compound of formula 2 and a compound of formula 3, the method comprising: beta-elemene and NaClO are put in CeCl₃The allyl halogenation is carried out in the system. The separation method of the compound 2 comprises the following steps: and dissolving the mixture in acetonitrile, adding into a circulating preparative liquid chromatograph, wherein the mobile phase is pure acetonitrile, the flow rate is 3.5mL/min, clicking a circulating button at 1h, and collecting a main peak after the raw materials pass through four cycles and at the fifth cycle to obtain a compound 2.

The beta-elemene halide is a compound of formula 4, and the method comprises the following steps: dissolving a beta-elemene raw material in a solvent containing acetic acid, and then carrying out allylic halogenation reaction with NaClO to obtain the beta-elemene compound.

The beta-elemene halide is a compound shown in a formula 5, and the method comprises the following steps: dissolving a beta-elemene raw material in a solvent containing acetic acid, and then carrying out allylic halogenation reaction with NBS to obtain the beta-elemene.

In a second aspect of the present invention, there is provided a β -elemene halide of the following formula (II), or an optical isomer (including racemate, single enantiomer, and possible diastereoisomer), deuterated derivative, hydrate, or solvate thereof:

in the formula (II), R⁴Selected from hydrogen, Cl, Br, I, R³And R⁵Each independently selected from Cl or Br; the carbon labeled with "-" may be in the (R) configuration or the (S) configuration.

In a third aspect of the present invention, there is provided a process for producing a compound represented by formula (II), comprising: dissolving a beta-elemene raw material in a solvent, and carrying out allylic addition reaction with a halogenated source reagent under the condition of adding or not adding a transition metal salt to obtain the beta-elemene compound; the transition metal salt is CeCl₃Or Yb (OTf)₃(ii) a The solvent is one or a mixture of water, dichloromethane, trichloromethane, tetrahydrofuran and acetic acid; the halogenated source reagent is NCS, NaOCl, NBS, NaOBr and Br₂Or NIS.

Preferably, the beta-elemene halide has a structure of at least one of the following formulas:

the structure of the beta-elemene halide is a compound shown in a formula 16, and the method comprises the following steps: dissolving a raw material of beta-elemene in a solvent containing acetic acid, and then carrying out allylic addition reaction with NaOBr to obtain the beta-elemene.

The beta-elemiThe alkene halide has the structure of formula 17, the process comprising: dissolving beta-elemene raw material in solvent, adding CeCl₃The system is obtained by allylic addition reaction with NBS.

The structure of the beta-elemene halide is a compound of a formula 18 or a compound of a formula 19, and the method comprises the following steps: dissolving the compound of the formula 17 in acetonitrile, adding the acetonitrile into a circulating preparation liquid chromatograph, wherein the mobile phase is pure acetonitrile, the flow rate is 3.5ml/min, clicking a circulating button at 1h, the raw materials pass through four cycles, collecting a main peak at the fifth cycle to obtain a mixture, respectively carrying out rotary evaporation on samples with different proportions, then repeating the circulating preparation operation, and respectively collecting effluents with the retention time of 15.167min and 15.338min to respectively obtain the compound of the formula 18 and the compound of the formula 19.

In the above formulae, the groups are as defined above. Reagents and conditions for each step may be selected from those conventional in the art for carrying out such preparation methods, and such selection may be made by those skilled in the art after the structure of the compound of the present invention is disclosed, according to the knowledge in the art.

More specifically, the compounds represented by the general formula I of the present invention can be prepared by the following method, however, the conditions of the method, such as reactants, solvent, amount of the compound used, reaction temperature, time required for the reaction, etc., are not limited to the following explanation. The compounds of the present invention may also be conveniently prepared by optionally combining various synthetic methods described in the present specification or known in the art, and such combinations may be readily carried out by those skilled in the art to which the present invention pertains.

In the preparation method, each reaction is usually carried out in an inert solvent, the reaction temperature is usually-10-55 ℃, and preferably 0-room temperature. The reaction time in each step is usually 0.5 to 48 hours, preferably 2 to 12 hours. Dissolving beta-elemene in a solvent, sequentially adding a halogenated source reagent, stirring for 30 min-72 h at 0-55 ℃ with or without a transition metal catalyst, adding saturated sodium thiosulfate or saturated sodium bisulfite to quench reaction, extracting, and carrying out column chromatography to obtain a structural compound shown in formula (I) or formula (II) (each group is defined as above).

Wherein the molar concentration of the solution of the beta-elemene is 0.1-0.5 mol/L; the molar ratio of the intermediate beta-elemene to the halogenated source reagent to the transition metal catalyst is 1 (0-10) to (0-4); preferably 1 (1-10) to 0.5-1.

In the fourth aspect of the invention, the invention also provides application of the compound shown in the formula (II) as an elemene derivatization reaction intermediate. The compound is obtained by addition reaction of one carbon-carbon double bond in beta-elemene and halogen. These compounds are key intermediates in elemene derivatization reactions, and they can be used to prepare various elemene derivatives.

Compared with the prior art, the invention has the following beneficial effects:

(1) the method of the invention utilizes the method of preparing beta-elemene halide by allylic halogenation reaction and addition reaction, and adopts the method of circularly preparing liquid chromatogram to separate out pure halogenated elemene, the method has the characteristics of simplicity, convenience, easy operation, high yield and the like, and can be widely applied to elemene derivatization in the future.

(2) The invention also relates to a double bond selective addition product of three carbon-carbon double bonds in beta-elemene and halogen, which belongs to a new compound, the compounds are important intermediates for derivatization of elemene, and the compounds have the same or slightly stronger activity of inhibiting proliferation of various tumor cell strains in vitro as beta-elemene.

Detailed Description

The inventor has found out the result of the invention through long-term research, namely, the allylic halogenation reaction of beta-elemene is carried out by using a specific research method. The specific study was carried out in CeCl₃Under the catalytic conditions of (2), a halogen source reagent (such as NaClO, NCS, NBS, NIS and the like) is used for carrying out a halogenation reaction on the allyl position of the beta-elemene. Most of the products are monohalides, except that a small amount of the double halide in the 13, 14-position is obtained. The monohalides of beta-elemene include monochloro (compounds 2 and 3), monobromoSubstitutes (compounds 5 and 11), monoiodo (compounds 12 and 13). They have no way to achieve separation by conventional silica gel column chromatography or conventional liquid chromatography. The compounds are useful intermediates for the preparation of other elemene derivatives. The invention has great significance for further research on the derivatization and the anti-tumor activity of the beta-elemene, and even research on the action mechanism and the cell pathway of the beta-elemene. On the other hand, the invention also includes that the elemene allyl is a halogenated compound and shows similar or better inhibiting effect than beta-elemene in the proliferation experiment of tumor cell strains. The beta-elemene allylic halogenated compounds are expected to be applied to the research and development of anti-tumor drugs. Wherein the compounds 2, 3, 4 and 5 are compounds which have been reported in the literature.

Term(s) for

Unless otherwise specified, the term "pharmaceutically acceptable salt" refers to salts that are suitable for contact with the tissues of a subject (e.g., a human) without undue side effects. In some embodiments, pharmaceutically acceptable salts of a certain compound of the invention include salts of a compound of the invention having an acidic group (e.g., potassium, sodium, magnesium, calcium) or a basic group (e.g., sulfate, hydrochloride, phosphate, nitrate, carbonate).

Use of

The invention provides a novel preparation method of a compound shown in a formula (I). In particular, CeCl is used₃And a halogen source (such as NaClO, NCS, NaOBr, NBS, NIS) to perform a halogenation reaction on the allyl position of the beta-elemene.

The present invention provides a cyclic preparative liquid chromatography technique for separating the following mixtures which cannot be separated by conventional silica gel column chromatography and conventional high pressure liquid chromatography techniques to obtain pure single monohalogenated compounds.

The invention provides a compound of formula (I), or a deuterated derivative, a salt, an isomer (enantiomer or diastereoisomer if existing), a hydrate, a pharmaceutically acceptable carrier or an excipient thereof, and application of the compound of formula (I) or the deuterated derivative, the salt, the isomer (enantiomer or diastereoisomer if existing), the hydrate, the pharmaceutically acceptable carrier or the excipient thereof in inhibiting proliferation of in vitro tumor cell strains. The compounds belong to elemene derivatives, and have similar or stronger proliferation activity to beta-elemene in vitro inhibition of various tumor cell strains.

The compound has the same or stronger activity of inhibiting various tumor cell strains in vitro as that of elemene, and the elemene is an anti-tumor medicament approved by the national drug administration. The diseases include liver cancer, rectal cancer, bladder cancer, throat cancer, non-small cell lung cancer, lung adenocarcinoma, lung squamous carcinoma, breast cancer, prostate cancer, glioma, ovarian cancer, head and neck squamous carcinoma, cervical cancer, esophageal cancer, renal cancer, pancreatic cancer, colon cancer, skin cancer, lymphoma, stomach cancer, multiple myeloma cancer, solid tumor and the like.

The compounds of the invention can be used with biologicals such as PD-1 inhibitors

And

can be used as a combined medicine for treating various cancers and related diseases.

The compounds of the present invention and deuterated derivatives thereof, as well as pharmaceutically acceptable salts or isomers thereof (if present) or hydrates thereof and/or compositions can be formulated together with pharmaceutically acceptable excipients or carriers and the resulting compositions can be administered to mammals, such as men, women and animals, in vivo for the treatment of conditions, symptoms and diseases. The composition may be: tablets, pills, suspensions, solutions, emulsions, capsules, aerosols, sterile injections. Sterile powders, and the like. In some embodiments, pharmaceutically acceptable excipients include microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, dibasic calcium phosphate, mannitol, hydroxypropyl- β -cyclodextrin, β -cyclodextrin (plus), glycine, disintegrants (such as starch, croscarmellose sodium, complex silicates, and polymeric polyethylene glycols), granulation binders (such as polyvinylpyrrolidone, sucrose, gelatin, and acacia), and lubricants (such as magnesium stearate, glycerol, and talc). In a preferred embodiment, the pharmaceutical composition is in a dosage form suitable for oral administration, including but not limited to tablets, solutions, suspensions, capsules, granules, powders. The amount of a compound or pharmaceutical composition of the present invention administered to a patient is not fixed and is generally administered in a pharmaceutically effective amount. Also, the amount of the compound actually administered can be determined by a physician, in the light of the actual circumstances, including the condition being treated, the chosen route of administration, the actual compound administered, the individual condition of the patient, and the like. The dosage of the compounds of the invention will depend on the particular use being treated, the mode of administration, the state of the patient, and the judgment of the physician. The proportion or concentration of the compounds of the invention in the pharmaceutical composition will depend on a variety of factors including dosage, physicochemical properties, route of administration and the like.

It is to be understood that within the scope of the present invention, the above-described features of the present invention and those specifically described below (e.g., in the examples) may be combined with each other to form new or preferred embodiments.

Pharmaceutical compositions and methods of administration

The compound has the same or stronger proliferation activity of inhibiting various tumor cell strains as that of elemene, so that the compound and various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates thereof and a pharmaceutical composition containing the compound as a main active ingredient can be used for treating, preventing and relieving various diseases which can be treated by elemene, including various cancers.

The pharmaceutical composition of the present invention comprises the compound of the present invention or a pharmacologically acceptable salt thereof in a safe and effective amount range and a pharmacologically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-2000mg of a compound of the invention per dose, more preferably, 5-1000mg of a compound of the invention per dose. Preferably, said "dose" is a capsule or tablet.

"pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances which are suitable for human use and must be of sufficient purity and sufficiently low toxicity. By "compatible" is meant herein that the components of the composition are capable of intermixing with and with the compounds of the present invention without significantly diminishing the efficacy of the compounds. Examples of pharmaceutically acceptable carrier moieties are cellulose and its derivatives (e.g. sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (e.g. stearic acid, magnesium stearate), calcium sulfate, vegetable oils (e.g. soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (e.g. propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (e.g. tween, etc.)

) Wetting agents (e.g., sodium lauryl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, and the like.

The mode of administration of the compounds or pharmaceutical compositions of the present invention is not particularly limited, and representative modes of administration include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia; (c) humectants, for example, glycerol; (d) disintegrating agents, for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) slow solvents, such as paraffin; (f) absorption accelerators, e.g., quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glycerol monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage forms may also comprise buffering agents.

Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shells such as enteric coatings and other materials well known in the art. They may contain opacifying agents and the release of the active compound or compounds in such compositions may be delayed in release in a certain part of the digestive tract. Examples of embedding components which can be used are polymeric substances and wax-like substances. If desired, the active compound may also be in microencapsulated form with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly employed in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1, 3-butylene glycol, dimethylformamide and oils, in particular, cottonseed, groundnut, corn germ, olive, castor and sesame oils or mixtures of such materials and the like.

In addition to these inert diluents, the compositions can also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

Suspensions, in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances, and the like.

Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols and suitable mixtures thereof.

Dosage forms of the compounds of the present invention for topical administration include ointments, powders, patches, sprays, and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants which may be required if necessary.

The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds.

When the pharmaceutical composition is used, a safe and effective amount of the compound of the present invention is suitable for mammals (such as human beings) to be treated, wherein the administration dose is a pharmaceutically-considered effective administration dose, and for a human body with a weight of 60kg, the daily administration dose is usually 1 to 5000mg, preferably 5 to 2000 mg. Of course, the particular dosage will depend upon such factors as the route of administration, the health of the patient, and the like, and is within the skill of the skilled practitioner.

Example 1: preparation of a mixture of 13-chloro-beta-elemene (2) and 14-chloro-beta-elemene (3) (sodium hypochlorite and cerium chloride systems)

To a solution of β -elemene starting material (218mg,1.069mmol) in dichloromethane (5mL) and water (5mL) under ice-bath was added tetrabutylammonium iodide (197mg, 0.53mmol) and cerium trichloride heptahydrate (1.153g, 3.09 mmol). Sodium hypochlorite (1.2mL, 8-13% strength, 3.207mmol) was then slowly added dropwise to the above mixture. The reaction solution was kept ice-cooled, stirred for 0.5 hour and reacted at room temperature overnight. The starting material was checked for complete reaction by thin plate chromatography and the mixture was extracted with ethyl acetate (3X5 mL). The combined organic phases were washed successively with water (2X 5mL) and saturated brine (2X 5mL) and dried over anhydrous sodium sulfate. The drying agent was removed by filtration, the filtrate was concentrated under reduced pressure, and the resulting crude product was purified by silica gel column chromatography (eluting with pure petroleum ether) to give a mixture of compounds 2 and 3 (188.5mg, yield 74.1%, colorless liquid).

GC-MS conditions: MS Agilent Technologies 5977A MSD; GC systems: agilent Technologies 7890B; agilent gas chromatograph and gas workstation, FID detector, capillary chromatography (Agilent 19091S-433UI, HP-5ms Ultra insert, 60 ℃ -325 ℃ (350 ℃), 30m × 250 μm × 0.25 μm); sample inlet temperature: 250 ℃, detector temperature: the temperature rising program is as follows: the initial temperature is 50 deg.C, maintained for 2min, increased to 80 deg.C at a rate of 20 deg.C per minute, maintained for 2min, increased to 150 deg.C at a rate of 30 deg.C per minute, and maintained for 5 min; carrier gas: helium, helium flow rate: 24.2mL/min, column flow: 1.2mL/min, pressure: 9.8psi, tail gas blowing flow: 3mL/min, sample size: 1 μ L, split ratio: 100: 1. GC-MS confirmed that Compound 2 and 3 had retention times of 11.606min and 11.339min, respectively; the peak area ratio was 5: 2.

Example 2: preparation of a mixture of 13-chloro-beta-elemene (2) and 14-chloro-beta-elemene (3) (NCS and cerium chloride system)

To a solution of β -elemene starting material (201.1mg,1.03mmol) in acetic acid (3mL) under ice-cooling (10-15 deg.C) was added NCS (136.7mg, 1.00 mmol). The reaction was cooled in an ice bath (10-15 deg.C), stirred for 8 hours, thin plate chromatography checked that the starting material did not react completely, reacted overnight at room temperature (25-28 deg.C. the reaction was quenched by slowly adding saturated sodium bisulfite (7.5mL) and water (8mL) dropwise, the mixture was extracted with petroleum ether (3X5mL), the combined organic phases were washed successively with saturated sodium bisulfite (3X5mL) and water (3X5mL) and saturated brine (3X5mL), drying with anhydrous sodium sulfate, filtering to remove desiccant, concentrating the filtrate under reduced pressure, and purifying the obtained crude product by silica gel column chromatography (eluting with pure petroleum ether) to obtain colorless liquid compound A007-028-02(71.7mg, yield 38.6%). GC-MS confirms that the retention times of compounds 2 and 3 are 11.610min and 11.338min, respectively; the peak area ratio was 3: 1.

Example 3: preparation of 13-chloro beta-elemene (2) (circulation preparation liquid phase chromatographic separation method)

Beta-elemene chloride (product of example 1, ratio of compounds 2 and 3 5:2, 480mg,2.02mmol) was dissolved in acetonitrile (10mL) and then slowly pumped into a circulating preparative liquid phase apparatus (Japan Analytical Industry, LC-9130 NEXT.) the mobile phase was pure acetonitrile and the flow rate was set at 3.5 mL/min. The column model was GEL-ODS-AP-L, SP-120-15(Serial No.051206728), and the cycle button was clicked at 1h, and the starting material was subjected to four cycles, and the main peak was collected at the fifth cycle to obtain pure compound 2(212mg, yield 44%).¹H NMR(500MHz,Chloroform-d)δ5.83(ddd,J＝17.9,10.5,2.4Hz,1H),5.16(d,J＝2.2Hz,1H),5.04(d,J＝2.2Hz,1H),4.97–4.91(m,1H),4.90(d,J＝2.4Hz,1H),4.87–4.81(m,1H),4.59(s,1H),4.10(d,J＝2.4Hz,2H),2.21(td,J＝11.6,3.3Hz,1H),2.05(dt,J＝12.6,3.0Hz,1H),1.85–1.39(m,9H),1.02(d,J＝2.4Hz,3H)。

Example 4: preparation of 13, 14-bischloro beta-elemene (4) (sodium hypochlorite and acetic acid system)

To a solution of β -elemene (3.8g,18.6mmoL) in dichloromethane (25mL) and glacial acetic acid (22mL) was added TBAF (0.06mL, 1.0M THF solution, 0.06mmoL) with ice-bath cooling, followed by slow dropwise addition of NaClO (28mL,84mmoL) in water (isobaric dropping funnel, complete over 5 hours). After the addition, the stirring was continued at 0 ℃ for 1 hour. The reaction was quenched successively with 10% aqueous sodium sulfite (30mL) and saturated aqueous sodium bicarbonate (20 mL). Extract with ethyl acetate (3 × 50 mL). The combined organic phases were washed with saturated brine (30mL) and dried over anhydrous sodium sulfate. The drying agent was removed by filtration, the filtrate was concentrated under reduced pressure, and the resulting crude product was purified by silica gel column chromatography (100% petroleum ether) to give compound 4(1.84g, yield 37%) as a colorless liquid.¹H NMR(400MHz,Chloroform-d)δ5.80(dd,J＝17.1,11.1Hz,1H),5.29(d,J＝1.0Hz,1H),5.18(d,J＝1.0Hz,1H),5.05(s,1H),4.98–4.90(m,3H),4.14–4.08(dd,J＝11.6,0.8Hz,1H),4.11(s,2H),3.98(dd,J＝11.6,0.8Hz,1H),2.37–2.22(m,2H),1.79–1.63(m,2H),1.59–1.43(m,4H),0.99(s,3H)。

Example 5: preparation of 13-bromo-beta-elemene (5) (NBS and acetic acid system)

NBS (183mg,1.03mmol) was added to a solution of beta-elemene (210mg,1.03mmol) in acetic acid (3mL) under ice-bath (10-15 ℃), the reaction solution was cooled under ice-bath (10-15 ℃) and stirred for 6 hours, and the starting material was not completely reacted by thin-plate chromatography. The reaction was warmed to room temperature and stirred overnight (25-28 ℃). The reaction was quenched by slowly adding saturated sodium bisulfite (5mL) and water (5mL) dropwise, and the mixture was extracted with ethyl acetate (3X5 mL). The combined organic phases were washed successively with water (3X5mL) and saturated brine (3X5mL) and dried over anhydrous sodium sulfate. The drying agent was removed by filtration, the filtrate was concentrated under reduced pressure, and the resulting crude product was purified by silica gel column chromatography (eluting with pure petroleum ether) to give compound 5(72mg, yield 25.4%) as a colorless liquid.¹H NMR (400MHz, Chloroform-d) δ 5.89-5.76 (m,1H),5.21(s,1H),5.04(t, J ═ 1.1Hz,1H), 4.97-4.81 (m,3H),4.59(dt, J ═ 1.9,0.9Hz,1H),4.04(d, J ═ 0.7Hz,2H), 2.33-2.17 (m,1H),2.06(dd, J ═ 12.6,3.5Hz,1H), 1.74-1.70 (m,3H), 1.69-1.39 (m,6H),1.01(s, 3H). The conditions for GC-MS were as above. Mass spectrum peak 283.0[ M ]]。

Example 6: preparation of 13-bromo-beta-elemene (5) (N-bromophthalimide and acetic acid system)

Under ice bath (10-15 ℃), adding cerous chloride heptahydrate (107mg,0.285mmol) into acetic acid (1mL) solution of beta-elemene (117mg,0.57mmol), keeping the reaction solution at ice bath cooling (10-15 ℃), adding N-bromophthalimide (126mg, 0.57mmol), stirring for 8 hours, and detecting that the raw materials are not completely reacted by thin plate chromatography. The reaction was quenched by slowly adding saturated sodium bicarbonate (5mL) and water (5mL) dropwise. The mixture was extracted with ethyl acetate (3 × 5 mL).The combined organic phases were washed successively with water (3X5mL) and saturated brine (3X5mL) and dried over anhydrous sodium sulfate. The drying agent was removed by filtration, the filtrate was concentrated under reduced pressure, and the resulting crude product was purified by silica gel column chromatography (eluting with pure petroleum ether) to give compound 5(59mg, yield 36.6%) as a colorless liquid.¹H NMR(400MHz,Chloroform-d)δ5.83(dd,J＝17.8,10.5Hz,1H),5.21(s,1H),5.04(d,J＝1.1Hz,1H),4.97–4.80(m,3H),4.59(dt,J＝1.9,0.9Hz,1H),4.06–4.00(m,2H),2.25(dd,J＝9.7,6.1Hz,1H),2.06(dd,J＝12.6,3.5Hz,1H),1.72(t,J＝1.2Hz,3H),1.57(s,4H),1.56–1.41(m,2H),1.01(s,3H)。

Example 7: preparation of 13-bromo-beta-elemene (14) (NBS and Yb (OTf))₃)

NBS (169.9mg,0.95mmol) was added to a solution of β -elemene as a starting material (194.7mg,0.95mmol) in dichloromethane (1.6mL) and tetrahydrofuran (0.4mL) under ice-cooling, the reaction mixture was cooled in ice-cooling, and Yb (OTf) was added thereto after stirring for 5min₃(60.3mg,0.095mmol), TMSCl (12. mu.L, 0.095 mmol). And (3) cooling the reaction solution in an ice bath, stirring for 3 hours, adding NBS (0.5eq), reacting in the ice bath for 5 hours, and detecting that the raw materials are not completely reacted by thin plate chromatography. The reaction was quenched by slowly adding saturated sodium bisulfite and ice water dropwise. The mixture was extracted with petroleum ether (3X5 mL). The combined organic phases were washed successively with saturated sodium bisulfite solution (3 × 5mL), water (3 × 5mL) and dried over anhydrous sodium sulfate. The drying agent was removed by filtration, the filtrate was concentrated under reduced pressure, and the resulting crude product was purified by silica gel column chromatography (eluting with pure petroleum ether) to give compound 14(32.1mg, yield 18.6%) as a colorless liquid. Further, a mixture of compounds 5 and 11 (51.2mg, yield 29.7%) was obtained.

Compound 14:¹H NMR(400MHz,Chloroform-d)δ5.79(dd,J＝17.3,10.9Hz,1H),5.32(s,1H),5.22(s,1H),5.05(t,J＝1.0Hz,1H),4.98–4.89(m,3H),4.08–3.89(m,4H),2.42–2.28(m,2H),1.80–1.59(m,3H),1.57–1.40(m,3H),0.99(s,3H)。

mixing of Compounds 5 and 11An object:¹H NMR(400MHz,Chloroform-d)δ5.83(dd,J＝17.8,10.6Hz,1H),5.21(s,1H),5.05(d,J＝1.0Hz,1H),4.98–4.82(m,3H),4.60(dd,J＝1.9,0.9Hz,1H),4.04(s,2H),2.31–1.74(m,2H),1.73(t,J＝1.1Hz,3H),1.69(s,1H),1.59–1.42(m,3H),1.26(s,2H),1.02(s,3H)。

example 8: preparation of Compound 16 (liquid bromine and acetic acid System)

Under ice bath (10-15 ℃), adding liquid elementary bromine (28 muL, 0.55mmol) into acetic acid (1mL) solution of beta-elemene raw material (112mg,0.55mmol), dissolving in acetonitrile (150 muL), keeping the reaction liquid at ice bath for cooling (10-15 ℃), stirring for 5 hours, detecting that the raw material is not completely reacted by thin plate chromatography, and reacting at room temperature overnight (25-28 ℃). The reaction was quenched by slowly adding saturated sodium bisulfite (5mL) and water (5mL) dropwise, and the mixture was extracted with ethyl acetate (3X5 mL). The combined organic phases were washed successively with water (3X5mL) and saturated brine (3X5mL) and dried over anhydrous sodium sulfate. The drying agent was removed by filtration, the filtrate was concentrated under reduced pressure, and the resulting crude product was purified by silica gel column chromatography (eluting with pure petroleum ether) to give compound 16(15.2mg, yield 11.1%) as a colorless liquid. The retention time of compound 16 was 16.723min and the molecular weight was 364.02 as determined by GC-MS. The purity was 98.4%.

Example 9: preparation of Compound 16 (liquid bromine and acetic acid System)

Liquid bromine (4g) was slowly added dropwise to a solution of sodium hydroxide (4g) in water (12mL) under ice-bath (5-10 ℃ C.), and the reaction solution was kept cooled in ice-bath and stirred for 0.5 hour to obtain an aqueous solution of sodium hypobromite as a yellow liquid.

Slowly dropwise adding sodium hypobromite (0.38mL,0.52mmol, prepared fresh) into acetic acid (1mL) solution of beta-elemene (106mg,0.52mmol) under ice bath (10-15 deg.C), cooling the reaction solution with ice bath (10-15 deg.C), stirringAfter 1 hour, the reaction of the raw materials was detected by thin plate chromatography. The reaction was quenched by slowly adding saturated sodium bisulfite (5mL) and water (5mL) dropwise, and the mixture was extracted with ethyl acetate (3X5 mL). The combined organic phases were washed successively with water (3X5mL) and saturated brine (3X5mL) and dried over anhydrous sodium sulfate. The drying agent was removed by filtration, the filtrate was concentrated under reduced pressure, and the resulting crude product was purified by silica gel column chromatography (eluting with pure petroleum ether) to give compound 16(6.6mg, yield 3%) as a colorless liquid.¹H NMR(500MHz,Chloroform-d)δ5.77(dd,J＝17.7,10.5Hz,1H),5.21–5.14(m,2H),5.04(d,J＝1.1Hz,1H),4.97–4.90(m,2H),4.84(s,1H),4.09(d,J＝0.9Hz,2H),4.09–3.94(m,2H),2.28–2.17(m,1H),2.07(dd,J＝12.3,3.9Hz,1H),1.77–1.69(m,1H),1.69–1.57(m,1H),1.57–1.44(m,4H),1.01(s,3H)。

Example 10: preparation of Compound 17 (NBS and CeCl)₃System)

To a solution of β -elemene starting material (204mg,1.0mmol) in dichloromethane (5mL) and water (5mL) under ice-bath was added tetrabutylammonium iodide (185mg, 0.5mmol) and cerium trichloride heptahydrate (1.117g, 3mmol), and finally NBS (267mg,1.5 mmol). The reaction solution was kept in an ice bath and cooled, and stirred for 2 hours. Detecting the complete reaction of the raw materials by thin-plate chromatography, and slowly dropwise adding saturated sodium bisulfite and ice water into the reaction solution for quenching. The mixture was extracted with ethyl acetate (3 × 5 mL). The combined organic phases were washed successively with water (2X 5mL) and saturated brine (2X 5mL) and dried over anhydrous sodium sulfate. The drying agent was removed by filtration, the filtrate was concentrated under reduced pressure, and the resulting crude product was purified by silica gel column chromatography (eluting with pure petroleum ether) to give compound 17(250mg, yield 88%) as a colorless liquid. GC-MS (conditions above) showed two peaks with retention times of 15.167min and 15.338min, respectively, and the molecular weights of both peaks were 318.07. The peak area ratio thereof was 1.2: 1. These two peaks are the two diastereomers of compound 17.

Example 11: preparation of Compounds 18 and 19

Beta-elemene bromide 17(297mg,1.05mmol) was dissolved in acetonitrile (10mL) and then slowly injected into a circulating preparative liquid phase apparatus (LC-9130 NEXT). The mobile phase was pure acetonitrile and the flow rate was set at 3.5 ml/min. The column model is GEL-ODS-AP-L, SP-120-15(Serial No.051206728), the cycle button is clicked at 1h, the starting material is subjected to four cycles, the main peak is collected at the fifth cycle, the mixture is obtained, then samples of different proportions are respectively rotary evaporated and the preparation operation is repeated in the cycle, and effluents of different retention periods are collected to obtain two components, compounds 18 and 19 respectively. Compounds 18 and 19 are a pair of diastereomers, the absolute stereoconfigurations of which are not determined. Their nuclear magnetic spectra are very close.

Compound 18:¹h NMR (500MHz, Chloroform-d) δ 5.85-5.75 (m,1H), 4.94-4.91 (m,1H),4.90(s,1H),4.85(p, J ═ 1.6Hz,1H), 4.64-4.60 (m,1H),3.82(d, J ═ 10.4Hz,1H),3.64(d, J ═ 10.4Hz,1H),2.01(dd, J ═ 12.0,4.3Hz,1H), 1.97-1.84 (m,1H), 1.76-1.70 (m,3H),1.68(s,3H), 1.67-1.54 (m,4H), 1.54-1.46 (m,2H),0.99(s, 3H). The molecular weight obtained by GC-MS measurement was 318.07.

Compound 19:¹h NMR (500MHz, Chloroform-d) δ 5.84-5.76 (m,1H), 4.95-4.91 (m,1H),4.90(s,1H),4.85(p, J ═ 1.7Hz,1H),4.62(d, J ═ 1.8Hz,1H),3.82(d, J ═ 10.2Hz,1H),3.64(d, J ═ 10.4Hz,1H),2.01(dd, J ═ 12.8,3.5Hz,1H), 1.97-1.83 (m,1H), 1.74-1.72 (m,3H),1.69(s,3H), 1.66-1.45 (m,6H),1.00(s, 3H). The molecular weight obtained by GC-MS measurement was 318.07.

Claims

1. A preparation method of a beta-elemene halide is characterized in that the structure of the beta-elemene halide is shown as the following formula (I):

the method comprises the following steps: dissolving a beta-elemene raw material in a solvent, and carrying out allylic halogenation reaction with a halogenated source reagent under the condition of adding a transition metal salt to obtain the beta-elemene compound;

the solvent is one or a mixture of water, dichloromethane, tetrahydrofuran and acetic acid;

when the transition metal salt is CeCl₃When the halogen source reagent is NaClO or N-bromophthalein imine; when the transition metal salt is Yb (OTf)₃When the current is over; the halo source reagent is NBS.

2. The method as claimed in claim 1, wherein the β -elemene halide has at least one of the following structures:

3. the method for preparing a β -elemene halide according to claim 2, wherein the β -elemene halide is a mixture of a compound of formula 2 and a compound of formula 3, a mixture of a compound of formula 5 and a compound of formula 11, or a mixture of a compound of formula 12 and a compound of formula 13;

the method comprises the following steps: dissolving beta-elemene raw material in solvent, adding CeCl₃The system is obtained by allylic halogenation reaction of the system and a halogenated source reagent.

4. A process for the preparation of a β -elemene halide as claimed in claim 3, wherein the separation of the mixture comprises: dissolving the mixture in acetonitrile, adding into a circulating preparative liquid chromatograph, wherein the mobile phase is pure acetonitrile, the flow rate is 3.5mL/min, clicking a circulating button at 1h, and collecting a main peak after four cycles of raw materials and at the fifth cycle.