CN115611796B - Menthol monoterpene dimer cyhalothanol B and application thereof in preparation of antitumor drugs - Google Patents

Abstract

The present invention belongs to the technical field of anti-tumor drug research and development, and relates to a novel plant metabolic component, pileadimenthenol B, and its use in pharmaceutical manufacturing. Specifically, it relates to menthol monoterpene dimer pileadimenthenol B and its use in preparing anti-tumor drugs. The present invention prepares a monoterpene dimer pileadimenthenol B from the folk medicine Pilea brevis. The molecular structure of this type of component is novel, and it is the first peroxide-bridged menthane-type monoterpene dimer. In vitro experiments show that the pileadimenthenol B can significantly inhibit the growth of human lung cancer cell line A549. Further, the pileadimenthenol B can be used to prepare anti-tumor drugs.

Description

Menthol monoterpene dimer Pieris ethyl and its use in the preparation of anti-tumor drugs

Technical Field

The present invention belongs to the technical field of anti-tumor drug research and development, and relates to a novel plant metabolic component, Pilea ethanol, and its use in pharmaceutical manufacturing, and specifically relates to Pilea ethanol, a menthol monoterpene dimer, and its use in preparing anti-tumor drugs.

Background technique

It is reported that malignant tumors have become one of the major diseases that seriously threaten human life and health. Studies have reported that the high incidence of tumors is related to a variety of factors such as population aging and environmental pollution. While social development and medical progress have significantly extended human life, they have also brought some new medical challenges. Due to the high incidence of cancer, the difficulty of treatment, and the serious threat to the life and health of patients, research on it has always been an important field of life sciences.

Practice shows that the treatment of tumors has made great progress in terms of methods, efficacy and patient tolerance. However, the low drug efficiency, large toxic side effects, tumor metastasis and tumor resistance are still serious. Finding more effective drugs and methods to fight cancer is an important task for today's medical workers. In clinical practice, anti-tumor treatment methods mainly include surgical treatment, chemotherapy, radiotherapy, immunotherapy, traditional Chinese medicine treatment, etc. Among them, chemotherapy is currently the main method for treating malignant tumors. Therefore, the research and development of new anti-tumor drugs is still a difficult and important part of conquering tumor diseases.

Research practice shows that natural products, especially medicinal plants, are an important source of anti-tumor drugs; for example, camptothecin (CPT) and 10-hydroxycamptothecin (HCPT) are isolated from the bark of Camptotheca acuminata Decne., a plant of the Davidiaceae family that is unique to China. These ingredients show significant cytotoxic effects by inhibiting DNA topoisomerase I (TOPO I). After years of research, camptothecins represented by CPT and HCPT have developed into a mature class of anti-tumor drugs that can be used to treat bladder cancer, liver cancer, gastric cancer, and intestinal cancer. Paclitaxel is another anti-tumor star molecule. It was originally discovered in the bark of Pacific yew and is a new type of diterpene alkaloid. Due to its strong anti-tumor activity and novel mechanism of action, paclitaxel is known as one of the three major achievements of international anti-cancer drugs in the 20th century. According to incomplete statistics, dozens of anti-tumor drugs used in clinical practice are derived from natural products. Therefore, natural products have important significance and great potential in the field of anti-tumor.

Pilea aquarum subsp.brevicornuta is a plant of the genus Pilea. It is used as a folk medicine as "four-wheel grass" to clear away heat and detoxify, and treat boils. According to the research team of this application, the plant contains a variety of chemical components with novel structural types, such as sesquiterpenes, triterpenes, and the first discovered peroxide-bridged menthane monoterpene dimers. Literature reports that peroxidized natural products have a variety of biological activities, especially showing good application prospects in antimalarial, antiviral, and antitumor aspects.

Based on the foundation and current status of the prior art, the inventor of this application intends to provide a new plant metabolite, Pilea ethanol, and its use in pharmaceutical preparation, specifically Pilea ethanol, a menthol monoterpene dimer, and its use in the preparation of anti-tumor drugs.

Summary of the invention

The purpose of the present invention is to provide a new chemical entity with anti-tumor activity based on the basis and current status of the prior art, and relates to a new plant metabolic component, pyrethroid alcohol ethyl and its use in pharmaceutical manufacturing, and specifically to menthol monoterpene dimer pyrethroid alcohol ethyl and its use in the preparation of anti-tumor drugs.

The invention separates a natural component from the folk medicine Pilea aquarum subsp. brevicornuta (commonly known as four-wheel grass): monoterpene dimer Pileadimenthenol B. The molecular structure of this component is novel and it is the first peroxide-bridged menthane-type monoterpene dimer. In vitro experiments show that the Pileadimenthenol B can significantly inhibit the growth of human lung cancer cell line A549. Furthermore, the Pileadimenthenol B can be used to prepare anti-tumor drugs.

Specifically, the present invention provides a menthol monoterpene dimer, pyrethrin, having the following structure:

The invention provides an extraction and preparation process of euphorbia cerifera alcohol 1, and uses the traditional extraction and purification process of natural products to achieve the purpose of purifying the target compound.

In the present invention, the menthol monoterpene dimer Pilea ethanol B is obtained by extracting the above-ground part of Pilea breviscapus according to the following method and steps:

(1) The whole herb of Brachypodium album was extracted by solvent method.

The solvent is ethanol, methanol, ethyl acetate or acetone and water, and a single solvent or a mixture of two or more solvents is used for immersion extraction at room temperature or reflux extraction under heating conditions, and the extract is distilled under reduced pressure to obtain a crude extract;

(2) adding an appropriate amount of water to the crude extract in step (1) to form a suspension solution, extracting with an equal volume of petroleum ether, cyclohexane or chloroform, a low-polarity organic solvent, and concentrating the extract under reduced pressure to obtain an extract of a low-polarity part of the crude extract;

(3) The low polarity extract obtained in step (2) is repeatedly separated by a silica gel column and a dextran gel Sephadex LH-20 column to obtain the menthol monoterpene dimer pyrethroid ethyl.

In the present invention, the novel peroxymenthane monoterpene dimer - pyrethrin, two menthane units are connected by a peroxide bond; the compound is the first monoterpene derivative of this type found in nature; the X-ray single crystal structure of the compound (as shown in FIG1 ) and the spectrum data are as follows:

HRESIMS m/z 361.2352[M+Na] ⁺ (calcd.for C ₂₀ H ₃₄ NaO ₄ ,361.2349, Δ=0.6ppm). ¹ H NMR (600MHz, CDCl ₃ ) δ5.88 (brd, J=5.5Hz, 1H),5.74(brs,1H),4.33(brd,J＝8.0Hz,1H),4.13(dd,J＝3.4,5.5Hz,1H),3.96(dd,J＝3.4,3.8Hz,1H), 2.30(ddd,J＝13.0,2.0,2.2Hz),1.94(pd,J＝6.9,4.3Hz,1H),1.82(s, 7H),1.74(m,1H),1.67(m,1H),1.60(overlapped,1H),1.45(ddd,J＝13.0,10.0,3.5Hz,1H),1.39(m,1H),1.02(d ,J＝6.6Hz,3H),1.01(d,J＝6.6Hz,3H),0.96(d,J＝6.9Hz,3H),0.89(d,J＝6.9Hz,3H). ¹³ CNMR (151MHz, CDCl ₃ ) δ18.3,20.5,20.9,21.0,21.4,25.2,26.9,28.2,30.6,37.7,40.1,64.7,68.0,80.1,80.2,125.2,130.9,134.2,139.3.

Table 1.Crystal data and structure refinement for pileadimenthenol B

In the present invention, the prepared menthol monoterpene dimer pyrethrol can be derivatized, including: all or part of the 3-OH and 6'-OH in the molecule are alkylated, acetylated or carbonylated.

The results of in vitro experiments of the present invention show that the physalis alcohol B has a significant inhibitory effect on the human lung cancer cell line A549.

Furthermore, the present invention provides the use of the menthol monoterpene dimer Pilea alcohol ethyl in the preparation of anti-tumor drugs; the menthol monoterpene dimer Pilea alcohol ethyl can be used directly or after derivatization.

Furthermore, the present invention also provides a pharmaceutical composition prepared by using the menthol monoterpene dimer, Pilea ethanol or its derivatives, and other drugs for use together.

The advantages of the present invention also lie in that the menthol peroxide monoterpene dimer is a novel structure. Chemical structure is the basis of pharmacological action, and novel chemical structure corresponds to novel biological activity. Since the chemical structure of menthol peroxide is different from known anti-tumor drugs, the clinical application transformation of this type of compound is worth looking forward to. In addition, as a natural product, menthol peroxide is derived from a herb, which is easier to achieve large-scale planting and harvesting, and the problem of drug resources is easy to solve.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is an X-ray single crystal structure diagram of the compound pyrethroid alcohol.

Detailed ways

The following examples further illustrate the present invention, but these examples are by no means limiting the present invention. Any changes made by those skilled in the art in the practice of the present invention under the guidance of this specification will fall within the scope of the claims.

Embodiment 1:

100 g of the dried aerial part of the short-horned wet cold water flower was crushed and soaked in 6 times the amount of 75% ethanol at room temperature for 48 hours, and the extract was collected. The residue was extracted 3-5 times in the same way. The extracts were combined and concentrated under reduced pressure at 60°C to obtain 10 g of extract. The extract was suspended with 1 times the amount of water (volume/mass, V/W) and extracted three times with petroleum ether of the same volume as the suspension. After the petroleum ether extract was concentrated under reduced pressure, 3.2 g of extract was obtained. The petroleum ether extract extract was first roughly divided by a silica gel column, with petroleum ether: ethyl acetate as the two-phase solvent as the fluidity, and gradient elution was performed at a volume ratio of 15:1 to 0:1 to obtain five elution components. Its component 5 was repeatedly separated by Sephadex LH-20 column chromatography (35×1200mm, MeOH) to obtain a compound, which was identified as cold water flower alcohol ethyl, about 2.3 mg.

Example 2: Cytotoxic effect of phytolamine B on human lung cancer cell line A549,

The cytotoxicity was determined by MTT method. 200 μL of cell culture medium (DMEM culture medium containing 10% FBS) was added to each well of a 96-well culture plate, and the number of cells was controlled at 3.5×10 ^4. The cells were cultured in a 5% CO ₂ incubator at 37°C for 24 hours. DMSO solutions of different concentrations of paclitaxel were added to make the final concentrations of the samples 0.04, 0.2, 1.0, 5.0, and 25 μM, respectively. Three replicate wells were set for each concentration, and blank controls and positive controls (paclitaxel) were set. After 24 hours, the survival of A549 cells was determined by MTT method. Data were processed by graphad prism software, and its IC ₅₀ value was 4.45 μM.

Example 3: Etherification reaction of ethanol hydroxyl group of Pleurotus eryngii

Example 4: Acylation reaction of ethanol hydroxyl group of Pleurotus eryngii

Claims (5)

1. Menthol monoterpene dimer pyrethrin ethyl with the following structure: . 2. The menthol monoterpene dimer Pilea ethanol according to claim 1, characterized in that the menthol monoterpene dimer Pilea ethanol is obtained by extracting from the aerial part of Pilea breviscapus by the following method and steps: (1) The whole herb of Shorthorn Wetwater Flower is extracted by solvent method. The solvent is ethanol, methanol, ethyl acetate or acetone and water, and a single solvent or a mixture of two or more solvents is used for immersion extraction at room temperature or reflux extraction under heating conditions, and the extract is distilled under reduced pressure to obtain a crude extract; (2) adding an appropriate amount of water to the crude extract in step (1) to form a suspension solution, extracting with an equal volume of petroleum ether, cyclohexane or chloroform, a low-polarity organic solvent, and concentrating the extract under reduced pressure to obtain an extract of a low-polarity part of the crude extract; (3) The low polarity extract obtained in step (2) is repeatedly separated by a silica gel column and a dextran gel Sephadex LH-20 column to obtain the menthol monoterpene dimer pyrethroid ethyl. 3. Use of the menthol monoterpene dimer Pilea alcohol ethyl described in claim 1 in the preparation of anti-tumor drugs; the menthol monoterpene dimer Pilea alcohol ethyl can be used directly. 4. The use according to claim 3, characterized in that the menthol monoterpene dimer, pyrethroid alcohol, significantly inhibits the growth of human lung cancer cell line A549. 5. A pharmaceutical composition, characterized in that it comprises the compound having the structure as described in claim 1 and other drugs for use in combination.