CN111419853B

CN111419853B - Cucurbitacin and ibrutinib composition for treating breast cancer

Info

Publication number: CN111419853B
Application number: CN202010459184.3A
Authority: CN
Inventors: 吴莹; 邓意辉; 邓吉林
Original assignee: Deliwei Beijing biological Technology Co ltd
Current assignee: Deliwei Beijing biological Technology Co ltd
Priority date: 2020-05-27
Filing date: 2020-05-27
Publication date: 2022-07-19
Anticipated expiration: 2040-05-27
Also published as: CN111419853A

Abstract

The invention relates to a pharmaceutical composition for treating breast cancer, in particular triple negative breast cancer, which comprises cucurbitacin and ibrutinib. The invention also relates to application of the combination of cucurbitacin and ibrutinib in preparation of a medicine for treating triple-negative breast cancer. The cucurbitacin and ibrutinib combined drug has a remarkable synergistic effect on treatment of breast cancer, particularly triple negative breast cancer, brings a new hope for patients with the breast cancer, provides a scientific basis for research and development of novel tumor drugs, and has a wide application prospect.

Description

Cucurbitacin and ibrutinib composition for treating breast cancer

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a pharmaceutical composition containing cucurbitacin and ibrutinib and application thereof in treating breast cancer, particularly triple negative breast cancer.

Background

The breast cancer is the first mortality rate of female malignant tumors in China. Triple Negative Breast Cancer (TNBC) is a breast cancer in which an Estrogen Receptor (ER), a Progestogen Receptor (PR), and a human epidermal growth factor receptor2 (HER-2) are negative, and accounts for 15% of the incidence rate of breast cancer. TNBC is a special subtype of breast cancer, and tumor cells lack effective targets for ER, PR and HER-2, and no better treatment scheme exists so far. The triple negative breast cancer is generally treated according to the conventional targeting standard of the breast cancer, but the treatment prognosis is poor, invasion and metastasis easily occur, the life of a patient is seriously threatened, and the mortality rate is high.

Cucurbitacins (Cucurbitacins) belong to tetracyclic triterpene compounds with 19-methyl group appearing on C-9 position, are mainly distributed in cucurbitaceae plants, and are also found in higher plants of cruciferae, scrophulariaceae, begoniaceae, Elaeagnaceae, Tetradactylaceae and the like and some macrofungi. At present, Cucurbitacin (Cucurbitacin) which is approved to be produced in China is an extract of pedicellus melo (also known as pedicellus melo) which is a traditional Chinese medicine and mainly contains components such as Cucurbitacin B, E, and the Cucurbitacin B content in the existing extract is more than 60%. The cucurbitacin medicament approved by China to be on the market is used for the auxiliary treatment of persistent hepatitis, chronic hepatitis and primary liver cancer caused by excessive damp-heat toxicity, and the cucurbitacin medicament is available in two types, namely cucurbitacin tablets and cucurbitacin capsules.

The application of cucurbitacin in treating breast cancer has been disclosed. Journal literature "research on growth inhibition of cucurbitacin B on breast cancer cells in vitro" (zhangmeiman et al, practical drug and clinical, 2010, stage 01) discloses that cucurbitacin B has an inhibition effect on proliferation of human breast cancer cells and induces apoptosis of breast cancer cells through in vitro experiments, thereby exerting an anti-tumor effect. WO2008071968A discloses that cucurbitacins, in particular cucurbitacin B, inhibit the growth of cancer cells, such as cells in breast cancer, by modulating signal transduction pathways. Patent document CN106074567A discloses the use of cucurbitacin B inhibitor protein phosphatase 2A cancer suppressor factor (CIP2A) to activate its substrate, oncostatin PP2A, for the treatment of gastric cancer, breast cancer, lung cancer, and glioma. To date, no report on the treatment of triple negative breast cancer by cucurbitacin B is available.

Ibrutinib (also called Ibrutinib) is the first globally marketed selective and covalent inhibitor of Bruton Tyrosine Kinase (BTK) developed by Johnson and pharmaceuticals, with 3 approved indications: (1) mantle cell lymphoma; (2) chronic lymphocytic leukemia; (3) chronic lymphocytic leukemia with 17p deletion. Ibrutinib has been approved as a capsule on the market. Journal articles "Ibrutinib as a potential therapeutic option for HER2 overexpression Breast cancer-the role of STAT3 and p 21" (Prabaharan Chandra Bose et al, "Investigational new drugs" 2019) disclose Ibrutinib as a Tyrosine Kinase Inhibitor (TKI), which has proven to be an effective treatment option for HER2 overexpressing breast cancers. However, reports on the treatment of triple negative breast cancer by ibrutinib are not found so far.

The development of combination drugs has become a new trend in the development of modern drugs. The combination of cucurbitacin B and ibrutinib for breast cancer, particularly triple negative breast cancer, is not researched at present, and the development of the composition of the cucurbitacin B and the ibrutinib can provide a new treatment method for patients with the breast cancer and a new research strategy for the combination of other medicaments.

Disclosure of Invention

In order to overcome the defects and shortcomings of the prior art, the invention provides a pharmaceutical composition of cucurbitacin and ibrutinib and application thereof. The composition combines cucurbitacin and ibrutinib, has remarkable synergistic effect on the treatment of breast cancer, especially has synergistic effect on the treatment of triple negative breast cancer, and has unexpected technical effect.

The invention aims at providing a pharmaceutical composition which comprises cucurbitacin or a pharmaceutically acceptable salt thereof and ibrutinib or a pharmaceutically acceptable salt thereof.

The cucurbitacin or a pharmaceutically acceptable salt thereof and ibrutinib or a pharmaceutically acceptable salt thereof can be respectively present in free or solvate forms. For example, may exist in the form of a hydrate by absorbing moisture in the air. The solvate is not particularly limited as long as it is pharmaceutically acceptable. Specifically, the solvate is preferably a hydrate, an ethanol solvate, or the like. The pharmaceutically acceptable salts include salts with suitable organic or inorganic acids/bases.

In one embodiment of the present invention, the cucurbitacin or a pharmaceutically acceptable salt thereof and ibrutinib or a pharmaceutically acceptable salt thereof may be contained in the same formulation as an active ingredient. In another embodiment, the cucurbitacin or a pharmaceutically acceptable salt thereof and ibrutinib or a pharmaceutically acceptable salt thereof are contained in different formulations and administered simultaneously or at different times.

The cucurbitacin comprises cucurbitacin B, and the purity of the cucurbitacin B in the cucurbitacin is 1-100% (for example, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%). Preferably, the purity of cucurbitacin B in the cucurbitacin is more than 90%. More preferably, the purity of cucurbitacin B in the cucurbitacin is 100%.

In the pharmaceutical composition, the weight ratio of cucurbitacin (calculated as cucurbitacin B) to ibrutinib is 1: 1-1: 100. Preferably, in the pharmaceutical composition, the weight ratio of cucurbitacin (calculated as cucurbitacin B) to ibrutinib is 1: 5-1: 50. More preferably, in the pharmaceutical composition, the weight ratio of cucurbitacin (calculated as cucurbitacin B) to ibrutinib is 1: 10-1: 40.

The pharmaceutical composition can be prepared into any dosage form suitable for human or animal with pharmaceutically acceptable carriers. For example, the pharmaceutical composition may be directly mixed to form a formulation; or mixing cucurbitacin or pharmaceutically acceptable salt thereof and ibrutinib or pharmaceutically acceptable salt thereof in the pharmaceutical composition with corresponding carriers respectively to prepare preparations, and then packaging or combining the preparations in a conventional manner; or the cucurbitacin or pharmaceutically acceptable salt thereof and ibrutinib or pharmaceutically acceptable salt thereof in the pharmaceutical composition are respectively mixed with corresponding carriers and then mixed to prepare the preparation. The pharmaceutically acceptable carrier refers to various organic carrier substances or inorganic carrier substances used as preparation materials. Pharmaceutically acceptable carriers include, for example, but are not limited to, excipients, lubricants, binders, and disintegrants in solid formulations, or solvents, solubilizers, suspending agents, isotonic agents, buffers, pH adjusting agents, and the like in liquid formulations. In addition, additives such as preservatives, antioxidants, coloring agents, and sweeteners can be used as necessary. Any dosage form suitable for human or animal use may for example be a liquid dosage form, a solid dosage form or a semi-solid dosage form. Liquid dosage forms include, but are not limited to, solutions, suspensions, injections (including water injections, powder injections, and infusions); the solid dosage forms include but are not limited to tablets (including common tablets, enteric-coated tablets, buccal tablets, dispersible tablets, chewable tablets, effervescent tablets, orally disintegrating tablets), capsules (including hard capsules, soft capsules and enteric-coated capsules), granules, powder, pellets, dropping pills and the like; semisolid dosage forms include, but are not limited to, ointments, gels, pastes, and the like.

Another object of the present invention is to provide a use of the pharmaceutical composition for preparing a medicament for treating breast cancer diseases. Preferably, the invention provides an application of the pharmaceutical composition in preparing a medicament for treating triple negative breast cancer diseases. The pharmaceutical composition comprises cucurbitacin or a pharmaceutically acceptable salt thereof and ibrutinib or a pharmaceutically acceptable salt thereof.

Still other objects of the present invention include providing a method of treating breast cancer diseases, particularly triple negative breast cancer. The method comprises administering to the patient an effective amount of the pharmaceutical composition. The pharmaceutical composition comprises cucurbitacin or a pharmaceutically acceptable salt thereof and ibrutinib or a pharmaceutically acceptable salt thereof.

The pharmaceutical composition or medicament provided in the present invention can be administered in an amount appropriately determined to achieve an effective amount according to sex, age, body weight, symptoms and the like of the patient. An effective amount refers to the amount of a pharmaceutical composition or drug necessary to achieve a desired result, which is the amount necessary to achieve a delay, inhibition, prevention, reversal or cure in the progression of the state of treatment or management.

The dosage of the pharmaceutical composition of the invention is usually 0.1-1 mg/kg, preferably 0.2-0.8 mg/kg, most preferably 0.3-0.6 mg/kg of cucurbitacin or pharmaceutically acceptable salt thereof (calculated as cucurbitacin B); the dose of ibrutinib or pharmaceutically acceptable salt thereof is 1-50 mg/kg, preferably 5-20 mg/kg, and most preferably 10 mg/kg. The administration may be once or in several portions a day. When the pharmaceutical composition is administered in multiple doses, the pharmaceutical composition is preferably administered 1 to 4 times a day. The pharmaceutical composition may be administered at various administration frequencies such as daily, every other day, once a week, every other week, and once a month. Preferably, the frequency of administration is readily determined by a physician or the like. In addition, an amount outside the above range may be used, as necessary.

The pharmaceutical composition according to the present invention may also be used in combination with other anticancer agents. The combination means that the pharmaceutical composition and the anticancer agent are administered to the same patient over a prescribed period. When the drug composition and the anticancer agent are used in combination, they are preferably administered at the same time in terms of time, but may be administered separately before and after the time when the effect of one remains. In the combined use, the administration routes of the pharmaceutical composition and the anticancer agent may be the same or different.

The anticancer agent to be used in combination is not particularly limited as long as it is effective for breast cancer, and examples of the anticancer agent include alkylating agents, antimetabolites, anticancer antibiotics, plant alkaloids, platinum agents (platinum compound agents), biotherapeutics, hormonal agents, and molecular targeting drugs. Specific examples of the anticancer agent include cisplatin, carboplatin, oxaliplatin, cyclophosphamide, methotrexate, fluorouracil, doxorubicin, irinotecan, gemcitabine hydrochloride, tegafur, trastuzumab, pertuzumab, lapatinib, neratinib, sorafenib, gefitinib, sunitinib, erlotinib, temsirolimus, everolimus, pipindoxeclare, and docetaxel.

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

(1) the cucurbitacin (especially cucurbitacin B) and ibrutinib are combined for use, so that the cucurbitacin B has a remarkable synergistic effect on treatment of breast cancer, especially triple negative breast cancer, brings a new hope for patients with the breast cancer, provides a scientific basis for research and development of novel tumor medicaments, and has a wide application prospect.

(2) According to the invention, cucurbitacin (especially cucurbitacin B) and ibrutinib are combined, so that on one hand, growth and proliferation of breast cancer cells can be regulated and inhibited in multiple targets and multiple paths; on the other hand, the combination of the cucurbitacin (especially cucurbitacin B) and the ibrutinib composition has small toxic and side effects on cells and is also ensured in the aspect of safety.

Detailed Description

The present invention will be described in more detail with reference to examples. It is to be understood that the practice of the invention is not limited to the following examples, and that any variations and/or modifications may be made thereto without departing from the scope of the invention.

EXAMPLE 1 injection solution

Prescription:

ibrutinib 40mg

F68 1000mg

Sterile water for injection is added to 5ml

The preparation method comprises the following steps: placing Ibrutinib and F68 in a beaker, adding appropriate amount of ethanol, heating for dissolving, injecting sterilized water for injection under magnetic stirring, and adjusting volume to 5 ml.

EXAMPLE 2 injection

Prescription:

cucurbitacin B1 mg

F68 1000mg

Sterile water for injection is added to 5ml

The preparation method comprises the following steps: placing cucurbitacin B and F68 with the purity of more than 99% in a beaker according to the prescription amount, adding a proper amount of ethanol, heating for dissolving, injecting sterilized water for injection under magnetic stirring, and adjusting the volume to 5ml to obtain the cucurbitacin B and F68 oral liquid.

EXAMPLE 3 injection

Prescription:

the preparation method comprises the following steps: placing Ibrutinib, cucurbitacin B with purity of more than 99% and F68 in a beaker, adding appropriate amount of ethanol, heating for dissolving, injecting sterilized water for injection under magnetic stirring, and adjusting volume to 5 ml.

EXAMPLE 4 injection solution

Prescription:

the preparation method refers to example 3.

EXAMPLE 5 injection solution

Prescription:

the preparation method refers to example 3.

EXAMPLE 6 injection

Prescription:

the preparation method is referred to example 3.

EXAMPLE 7 tablets

Prescription:

the preparation method comprises the following steps: weighing cucurbitacin B, ibrutinib, lactose and F68 according to the formula, uniformly mixing, adding 10% starch slurry prepared from 1000g of starch, performing spray granulation, adding PVPP100g, magnesium stearate 100g, sodium bisulfite and citric acid, uniformly mixing, and pressing into 10000 tablets.

Experimental example 1 anti-mouse 4T1 Breast cancer cell study

Experimental materials: mouse breast cancer 4T1 cells (shanghai cell bank in chinese academy of sciences) were selected as a three-negative breast cancer model.

The experimental method comprises the following steps: and (3) taking the preserved mouse breast cancer 4T1 cells out of the liquid nitrogen, freezing and storing the tubes, and rapidly putting the tubes into water at 37 ℃ for resuscitation. The recovered 4T1 cell suspension is cultured in vitro, counted under an inverted microscope, and diluted into cell suspension by adding physiological saline when the activity of the tumor cells is more than 95 percent and the dilution ratio is adjusted. Sterilizing with 75% alcohol, inoculating 4T1 cell suspension on Balb/c mouse breast pad, inoculating 35 mice, and randomly dividing the mice into three groups7 groups of 5. The tumor volume of each group of mice reaches 100mm³The administration was started later (dose as given in table 1), 1 time every 3 days, 5 times in total (8, 11, 14, 17 and 20 days after inoculation), and the control group was given 0.9% sodium chloride injection. Data for tumor volume, body mass, death events, etc. were recorded throughout the duration of the pharmacodynamic test. Tumor inhibition (%) - (average mass of control tumor-average mass of experimental group tumor)/average mass of control tumor × 100%. The results are shown in Table 1.

TABLE 1 results of experiments on the inhibition of mouse 4T1 by cucurbitacin B and ibrutinib

The experimental results are as follows: as can be seen from Table 1, the preparation in example 1 (ibrutinib alone) has almost no effect on triple negative breast cancer, and the preparations in examples 4-6 have obvious synergistic effect on triple negative breast cancer.

Experimental example 2 different administration doses of cucurbitacin B

Selecting 1: 20 (mass ratio of cucurbitacin B to ibrutinib) different cucurbitacin B doses were studied.

4T1 tumor model according to Experimental example 1, the cell suspension was inoculated on Balb/c mouse breast pads for a total of 30 mice, and the mice were randomly divided into 5 groups of 6 mice each. The tumor volume of each group of mice reaches 100mm³The administration (i.p.) was started 1 time every 3 days for 5 times (8, 11, 14, 17 and 20 days after inoculation) and the control group was given 0.9% sodium chloride injection. Data were recorded for tumor volume, body mass, death events, etc. throughout the pharmacodynamic test. Tumor inhibition (%) (average mass of control group tumor-average mass of experimental group tumor)/average mass of control group tumor × 100%. The results are shown in Table 2.

TABLE 2 results of experiments with different cucurbitacin B doses against mouse 4T1

As can be seen from the results in Table 2, although the tumor inhibition rate was the highest at the dose of cucurbitacin B of 1.0mg/kg, there was a problem that 2 mice died, and therefore, the highest dose of cucurbitacin B did not exceed 1.0mg/kg during the combination therapy.

Claims

1. A pharmaceutical composition for treating triple negative breast cancer, comprising cucurbitacin B or a pharmaceutically acceptable salt thereof and ibrutinib or a pharmaceutically acceptable salt thereof; wherein the weight ratio of the cucurbitacin B or the pharmaceutically acceptable salt thereof to the ibrutinib or the pharmaceutically acceptable salt thereof is 1: 40.

2. The pharmaceutical composition of claim 1, wherein the cucurbitacin B is more than 90% pure.

3. The pharmaceutical composition according to claim 1 or2, wherein the purity of cucurbitacin B is 100%.

4. The pharmaceutical composition according to any one of claims 1 to 3, wherein cucurbitacin B or a pharmaceutically acceptable salt thereof and ibrutinib or a pharmaceutically acceptable salt thereof are contained as active ingredients in the same formulation or in different formulations.

5. A pharmaceutical composition according to any one of claims 1 to 3, which can be formulated with a pharmaceutically acceptable carrier into a dosage form suitable for human or animal use.

6. The pharmaceutical composition of claim 5, wherein the dosage form comprises a liquid dosage form, a solid dosage form, or a semi-solid dosage form.

7. The pharmaceutical composition of claim 6, wherein the liquid dosage form comprises a solution, a suspension, an injection; the solid dosage forms include tablet, capsule, granule, powder, pellet, and dripping pill; semisolid dosage forms include ointments, gels, pastes.

8. The pharmaceutical composition of any one of claims 5-7, wherein the dosage form comprises water injection, powder injection, plain tablets, enteric-coated tablets, buccal tablets, dispersible tablets, chewable tablets, effervescent tablets, orally disintegrating tablets, hard capsules, soft capsules, enteric-coated capsules.

9. Use of a pharmaceutical composition comprising cucurbitacin B or a pharmaceutically acceptable salt thereof and ibrutinib or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of triple negative breast cancer; wherein the weight ratio of the cucurbitacin B or the pharmaceutically acceptable salt thereof to the ibrutinib or the pharmaceutically acceptable salt thereof is 1: 40.

10. Use according to claim 9, wherein the purity of cucurbitacin B is greater than 90%.

11. Use according to claim 9 or 10, wherein the purity of cucurbitacin B is 100%.

12. The use according to any one of claims 9 to 10, wherein the cucurbitacin B or a pharmaceutically acceptable salt thereof is administered in a dose of 0.1 to 1mg/kg based on cucurbitacin B.

13. The use according to any one of claims 9 to 10, wherein the cucurbitacin B or a pharmaceutically acceptable salt thereof is administered in an amount of 0.2 to 0.8mg/kg of cucurbitacin B

14. The use according to any one of claims 9 to 10, wherein the cucurbitacin B or pharmaceutically acceptable salt thereof is administered in a dose of 0.3 to 0.6mg/kg, based on cucurbitacin B.

15. The use of any one of claims 9-14, wherein ibrutinib or a pharmaceutically acceptable salt thereof is administered in an amount of 1-50 mg/kg.

16. The use of any one of claims 9-14, wherein the ibrutinib or a pharmaceutically acceptable salt thereof is administered in an amount of 5-20 mg/kg.

17. The use of any one of claims 9-14, wherein said ibrutinib or a pharmaceutically acceptable salt thereof is administered in an amount of 10 mg/kg.