CN114533882B - Tocotrienol compositions for the treatment of cancer - Google Patents

Abstract

The invention relates to a composition containing tocotrienols for treating cancers and application thereof. The invention discloses the killing effect of different compositions formed by the 4 compound monomers based on 2 or 3 compound monomers in the 4 compound monomers on different cancer cell lines, which is obviously improved compared with the anti-cancer effect of the respective compound monomers of tocotrienol, metformin, curcumin and geranylgeraniol, and the synergistic effect of the compositions is reflected. In addition, the development of the existing hypoglycemic drug metformin in the new medical application field is also discussed. The application of the composition has potential to be developed into anticancer drugs and therapies.

Description

Tocotrienol compositions for the treatment of cancer

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a tocotrienol composition for treating cancers.

Background

Cancer, also known as malignant tumor, is a disease that is caused by genetic factors or causes mutation of genes induced by external factors and causes rapid proliferation of variant cells out of control, it can occur in any organ of the human body, it can invade adjacent parts of the body through blood or lymph fluid and spread to other organs, a process called cancer metastasis, and is a major cause of cancer death.

At present, cancer is one of the leading causes of death worldwide, and the world health organization international cancer research Institute (IARC) has issued the latest cancer burden data worldwide in 2020. Recent morbidity, mortality, and cancer development trends for 36 cancer types in 185 countries worldwide are estimated. The data show that 1929 ten thousand new cancer cases are global in 2020, up to 226 ten thousand new cases are global breast cancer, more than 220 ten thousand cases of lung cancer are global, and breast cancer replaces lung cancer and becomes the first cancer worldwide. 457 ten thousand new cancer cases in 2020, namely 300 ten thousand cancer death cases in 2020, 182 ten thousand men and 118 ten thousand women, occur on average every day.

The survival rate of the united states total cancer is 66%, the survival rate of the Chinese total cancer is 31%, and the survival rate is less than half of that of the united states. Therefore, the prevention of cancer is always a major topic to be solved in the medical community, especially in China, the urgent need is to develop new autonomous innovative anticancer drugs to increase the overall cure rate or 5-year survival rate of Chinese cancer patients, and bring benefit to people suffering from cancer in China and even worldwide.

The main modes of non-operative treatment of cancer at present are chemotherapy, targeted therapy, cellular immunotherapy and the like. But all have their respective drawbacks and limitations. The main expression is as follows:

1) The conventional chemotherapy can only relieve but not cure cancer due to the clinical application, and can produce serious adverse reactions in treatment, such as bone marrow function inhibition, nausea, diarrhea, vomiting and other gastrointestinal adverse reactions due to the fact that cancer cells and normal cells are killed at the same time; reduces the safety of the medicine and is easy to relapse after treatment due to the inability to kill cancer stem cells. The cost of conventional chemotherapy generally takes tens of thousands to hundreds of thousands of primordial notes for one treatment course;

2) Targeted therapy: there is a targeting therapy based on small molecules or antibodies, the curative effect is slightly better than that of conventional chemotherapy, but more than 90% of cancer death is caused by tumors transferred to important organs, but many genetically engineered targeting drugs still always lock targets on targets of primary cancers due to single acting targets, and drug resistance often occurs faster and cancerogenesis recurs. Because of the lack of specificity, sometimes the toxic side effects caused by targeted drugs are even more severe than the disease itself. These therapeutic bottlenecks indicate that anticancer agents targeting a single or two three signaling pathway molecules cannot prevent or destroy cancer. It is also difficult to cure cancer, which has the advantage that it can only extend the life of patients on average for several months over chemotherapy. In recent years, although immune drug therapies such as PD-1 (programmed cell death-1) and PD-L1 (programmed development-ligand 1) inhibitors have better therapeutic effects, they are limited to the population with high expression rate of PD-1, which is about 20% of cancer patients, and thus the application area is limited, and the current medical community is expanding the combination therapy of PD-1 inhibitors and other antitumor drugs to increase the therapeutic effects. In addition, the cost of targeted therapy is still high, and generally, one treatment course also takes ten to hundreds of thousands of yuan;

3) Chimeric antigen receptor T cell immunotherapy (CAR-T); at present, although the drug is very popular because of good short-term curative effect, the drug has the limitation that the drug can still be found to relapse if being subjected to long-term follow-up. Juno's JCAR015 primary clinical data shows that its complete remission rate in acute lymphoblastic leukemia humans is 87%, i.e., 87% of patients are not able to find cancer cells after treatment. However, about 60% of these 87% of patients relapse soon, with 59% of people who have passed 6 months. So even in the field of leukemia where CAR-T is the most powerful, it is currently not able to "attack" completely. Most CAR-T is directed against the antigen CD19, CD19 being expressed only on B lymphocytes, so it is able to kill B lymphocyte derived acute lymphoblastic leukemias and non-hodgkin lymphomas, but is completely ineffective against other tumors. The incidence rate of several kinds of hematological tumors in China is not increased even in the first ten times; in addition, the side effects of CAR-T are relatively large. A large number of T cells attack tumor cells, which release a large amount of cytokines in a short time, a process called "cytokine storm", and improper treatment is fatal; in addition, there is a disadvantage in terms of popularization, in that CAR-T is only reconstructed from immune cells derived from a patient, and CAR-T and other oncology drugs are different in that it is equivalent to a personalized therapy, and immune cells in each patient need to be separated and genetically reconstructed, and amplified and then returned to the patient, so that mass production cannot be performed, and the resulting high cost is not affordable to general people, and the price of each CAR-T treatment course in the united states is up to over 40 ten thousand dollars. At present, although the first type of Chinese CAR-T is marketed in China, the cost is extremely high, 120 ten thousand RMB is needed for one treatment course, and the treatment course is far from the common public of the salary level.

Therefore, the accelerated research is urgently needed in China to develop various novel, efficient, multi-target, multi-mechanism, broad-spectrum, safe, low-cost and anticancer new drugs which can reduce the recurrence rate and the drug resistance, and the novel drugs have extremely important significance.

The invention relates to a key concept, namely 'drug synergistic effect', namely when a certain number of drug monomer components are used as single drugs, only weak anticancer drug effect is generated, so that cancers cannot be controlled and effectively treated, but when the several compound components are combined together, the drug effect generated is many times stronger than the drug effect generated by using the single compound components (namely, the multiplied relationship), rather than the simple addition of the drug effects generated by using the single compound components (namely, the added relationship), the drug effect of the composite preparation of two monomer combinations can be visually understood as 1+1>2, or the drug effect of the composite preparation of 3 monomer combinations is 1+1+1>3.

The application cases of 'drug synergy' in the medical field are all kinds of traditional Chinese medicines which are formed by combining a plurality of or up to ten traditional Chinese medicines throughout the history of 5000 years, for example, the traditional Chinese medicines are a class of traditional prescriptions, and the ancestry of Chinese people is shown to fully utilize 'drug synergy' to treat diseases.

See how modern medicine uses "drug synergy". As is well known, it is not possible to effectively control aids by means of one anti-aids drug alone, so that doctor He Dayi in the united states invented a combination therapy which utilizes the synergistic anti-aids effect of 3 anti-aids drugs which are already on the market and have different mechanisms of action to treat aids, and has achieved great success, while it is not possible to cure aids, it has made aids better controlled than in the past, becoming the standard therapy for the current treatment of aids. It is reported that the gelan smith announces recently that a single-chip compound preparation, named siameteka, with the new generation of integrase inhibitor teweika, for the treatment of HIV is formally marketed in china. This is the first complete therapeutic regimen of single-piece compound preparation in the field of HIV treatment in the country. For example, the medicine can be used for four-way therapy such as omeprazole capsules, colloidal bismuth pectin, amoxicillin sodium, potassium clavulanate, clarithromycin and the like to play roles in stopping acid, protecting gastric mucosa and killing helicobacter pylori. For another example, the national drug administration approved the new coronavirus therapeutic drug of the feijie, nemorvir tablet/ritonavir tablet combination package (i.e., paxlovid) for import registration under the condition of day 11, 2, 2022. The Paxlovid is a protease inhibitor anti-RNA virus compound therapy, and the components of Paxlovid are Nemacavir and ritonavir, the Nemacavir and ritonavir can block SARS-CoV-2-3CL protease activity, and the protease is an enzyme required by coronavirus replication, and can block the activity of the protease and further block the virus replication. Ritonavir is also a protease inhibitor, but low doses of ritonavir can also increase the blood level of nematavir by inhibiting liver metabolism. The data show that Paxlovid can reduce the risk of hospitalization or mortality of new coronavirus patients by 89%. Meanwhile, according to the latest experiments, the medicine is effective on the Omikovia variant virus.

These are all clinically useful combinations.

Cancers like AIDS are very refractory diseases, and treatment of difficult and complicated diseases often requires unusual means and innovative breakthrough methods, and most cancers cannot be effectively controlled or cured by 1 or 2 anticancer chemotherapeutics only depending on the traditional cancer treatment methods. The invention is supposed to achieve the highly synergistic anticancer effect by screening a large number of natural compounds layer by layer to obtain a method that 2 or 3 component monomers with definite anticancer efficacy and basically no toxicity are combined into different compositions, inhibit or kill cancer cells by inhibiting or promoting apoptosis by utilizing complementary different anticancer mechanisms and multiple target groups, so that the cancer cells are everywhere and are shaped, serious adverse reactions generated during conventional chemotherapy can be avoided, the resistance of a human body to anticancer drugs is reduced, the probability of cancer recurrence is reduced, and the control and even the probability of cancer cure are greatly improved. This would be a significant innovation in anticancer methodology.

Disclosure of Invention

In order to solve the problems in the prior art, the invention provides a tocotrienol compound preparation for treating cancers.

The technical scheme of the invention is as follows:

a composition of tocotrienols for the treatment of cancer, the raw material components comprising different compositions based on 2 or 3 substance monomers of the four types of tocotrienols, metformin, curcumin and geranylgeraniol;

the tocotrienol comprises tocotrienol and tocotrienol isomers: 4 isomers of alpha-tocotrienol, beta-tocotrienol, gamma-tocotrienol, and delta-tocotrienol, tocotrienol salts, tocotrienol co-crystals, tocotrienol precursors, or tocotrienol derivatives;

the metformin substances comprise any one of metformin, metformin isomers, metformin salts, metformin eutectic, metformin precursors or metformin derivatives;

the curcumin substance comprises any one of curcumin, curcumin isomer, curcumin salt, curcumin eutectic or curcumin derivative.

The geranylgeraniol substance is any one of geranylgeraniol, geranylgeraniol isomer, geranylgeraniol salt, geranylgeraniol eutectic, geranylgeraniol precursor and geranylgeraniol derivative.

According to the tocotrienol composition for treating cancer, when the raw material components are two monomers of the tocotrienol monomer and the metformin monomer to form a composition, the mass ratio between the tocotrienol monomer and the metformin monomer is 1-60:1-100; or when the raw material components are two monomers of the tocotrienol monomer and the geranylgeraniol monomer to form a composition, the mass ratio of the tocotrienol monomer to the geranylgeraniol monomer to form the composition is 1-60:1-100.

According to the composition of tocotrienols for treating cancer, when the raw material components are three monomers of a tocotrienol monomer, a metformin monomer and a curcumin monomer to form a composition, the mass ratio of the tocotrienol monomer, the metformin monomer and the curcumin monomer is 1-60:1-100:1-100; when the raw material components are three monomers of a tocotrienol monomer, a curcumin monomer and a geranylgeraniol monomer to form a composition, the mass ratio of the tocotrienol monomer to the curcumin monomer to the geranylgeraniol monomer is 1-60:1-100:1-100.

The composition of tocotrienols for the treatment of cancer according to said composition of tocotrienols wherein tocotrienol is represented primarily by the most active delta-tocotrienol in its isomers and wherein when the starting materials comprise three monomers of delta-tocotrienol monomer, metformin monomer, and curcumin monomer in a composition, the preferred mass ratio of delta-tocotrienol monomer, metformin monomer, and curcumin monomer ranges from about 1-5:1-10:1-10; or when the starting components are three monomers of delta-tocotrienol monomer, curcumin monomer, and geranylgeraniol monomer to form a composition, the more desirable preferred ranges for the delta-tocotrienol monomer, curcumin monomer, and geranylgeraniol monomer mass ratios are about 1-5:1-10:1-10.

According to the tocotrienol composition for treating cancer, the tocotrienol monomer, the metformin monomer, the curcumin monomer and the geranylgeraniol monomer are extracted from natural plant compounds, or are derived from or are artificially synthesized.

The preparation form of the composition is any one of tablets, capsules, granules, suspension, solution, sublingual tablets, injection, freeze-dried powder, aerosol, nasal spray or drops and skin emplastrum.

The tocotrienol composition for treating cancer according to the present invention further comprises pharmaceutically acceptable excipients.

A drug delivery system comprising the composition of any one of nanoparticles, polymeric micelles, passive targeting vectors, and active targeting vectors.

According to the drug delivery system, the nanoparticle is any one of liposome, nanoliposome, nanoemulsiond particle, nanocrystal, nanosuspension, nanomicelle, polypeptide copolymer micelle, nanoschitosan, nanovesicle, carbon nanotube, or carbon nanodisk.

Use of any of the compositions in the manufacture of a medicament for the treatment of breast, pancreatic, colorectal, lung, liver, nasopharyngeal, oral, tongue, larynx, oesophageal, kidney, cardia, stomach, pylorus, bladder, prostate, cervical, uterine, ovary, lip, skin, bone, sarcoma, malignant glioma, ewing's tumor, hodgkin's lymphoma, non-hodgkin's lymphoma, multiple myeloma, and various leukemias in humans.

In order to facilitate understanding of the present invention, the crude drug in the present invention will now be further described.

1. Research on anticancer effects of tocotrienols

Chinese name: tocotrienols

Foreign language name: tocotrienols

The vitamin E family is divided into the 2 major sub-groups of the isomeric tocopherols and tocotrienols, namely tocotrienols are different names of the same substance from tocotrienols, the 2 sub-groups being distinguished by the fact that tocotrienols have 3 double bonds (also called olefinic bonds) in the figure, which are unsaturated in the side chains, compared to tocopherols, and are therefore known as tocotrienols.

Chemical structure: the following are listed below

Tocotrienol in the above figure is tocotrienol in english and tocophenol is tocopheryl in english.

Tocotrienols are one of the 2 major subclasses in the large vitamin E family.

Overview of vitamin E Structure

The 2 major subclasses of tocopherols and tocotrienols can be represented by functional groups (R ₁ And R is ₂ ) The positions of (a) are further divided into four isomers of alpha, beta, gamma and delta. Such as the following table lists the methyl functional groups (R ₁ And R is ₂ ) Quilt CH ₃ Or H, and forms alpha, beta, gamma, and delta tocotrienols with an activity ratio: tocotrienol isomers as shown in the following table:

isomer of tocotrienol	R 1	R 2	Ratio of Activity
				Alpha-tocotrienols	CH3	CH3	100
Beta tocotrienols	CH3	H	40
				Gamma-tocotrienol	H	CH3	10
Delta-tocotrienols	H	H	1

The above table shows that delta-tocotrienol activity is highest in all four tocotrienol isomers, so this patent will be presented primarily as delta-tocotrienol and related supportive testing will be conducted.

Delta-tocotrienol:

english name: delta-tocotrienol

Molecular weight: 396.605

CAS accession number: 25612-59-3

The function is as follows: antioxidant, anticancer, etc

Delta-tocotrienol can inhibit tumor cell proliferation, and SHAH and other researches show that tocotrienol has growth inhibition effect on human breast cancer MDA-MB-435 cells, and the inhibition effect is enhanced with the increase of drug concentration and action time. Horse leaps and other research results show that under the action of delta-tocotrienol, the expression level of beta-catenin and wnt-1 in colon cancer SW620 cells is reduced, which indicates that the activities of the beta-catenin and wnt-1 are inhibited or degraded in the cells, and the proliferation of the SW620 cells is influenced. Studies show that tocotrienol can induce HepG2 cells, HT-29 cells, SGC-7901 cells and the like to undergo apoptosis, such as nuclear chromatin condensation, nuclear shrinkage, membrane foaming germination, organelle swelling, cell membrane sinking, encapsulation of nuclear fragments and organelles, formation of apoptosis minibodies and DNA trapezoid strips and the like. Zhang Zhongquan and other studies show that delta-tocotrienol incubated human liver cancer HepG2 cells obviously reduce the nuclei of the HepG2 cells through 48h, reduce the mitochondrial membrane potential, concentrate and marginalize chromatin and form apoptotic bodies. Delta-tocotrienol also has a regulatory effect on Bcl-2 family protein expression, induces up-regulation of pro-apoptotic factor Bax expression and down-regulation of anti-apoptotic factor Bcl-2 expression in a concentration-dependent manner, and causes Bid protein activation. Meanwhile, delta-tocotrienol can release cytochrome C from mitochondria into cytoplasm by increasing the permeability of mitochondrial membrane, activate caspase-3, caspase-8 and caspase-9, and finally cause apoptosis of HepG2 cells. In addition, studies on human breast cancer cells and human colon cancer cells have also found that tocotrienols can alter Bcl-2 to Bax ratios, while the presence of ruptured mitochondria in the cells, a decrease in mitochondrial membrane potential, release of cytochrome C from mitochondria, and caspase-9 activation have been found.

Delta-tocotrienol can inhibit tumor neovascularization. Neovascularization is the process of creating new blood vessels from pre-existing blood vessels, such as capillaries and postcapillary venules, precisely regulated by angiogenic and anti-angiogenic factors. Including endothelial cell activation, basement membrane rupture, migration, proliferation, and vascular endothelial cell tube formation. Since the vascular endothelial growth factor receptor VEGFR-2 is a key protein in endothelial cell growth factor signaling. Thus, modulation of VEGFR-2 activation is a potential molecular target for anti-angiogenic compounds. Weng et al have found that tocotrienols can inhibit VEGF-activated tubular formation. Furthermore, the administration of TRF treatment to BALB/C mice significantly reduced serum VEGF levels. MIYAZWA et al showed that implantation of human colon cancer DLD-1 cells into dorsal air sac model mice, daily dietary supplementation with palm oil enriched with tocotrienols 10mg inhibited tumor neovascularization.

2. Research on anticancer effect of metformin

Metformin, an organic compound of the formula C ₄ H ₁₁ N ₅ For the treatment ofType 2 diabetes mellitusParticularly for patients with overweight.

Drug name: metformin hydrochloride

Foreign language name: metformin

Alias name: 1, 1-dimethylbiguanide

The function is as follows: reducing blood sugar, resisting cancer, etc

The chemical formula: c (C) ₄ H ₁₁ N ₅

Molecular weight: 129.164

CAS accession number: 657-24-9

Chemical structure:

metformin was approved by the FDA in 1994 as an oral drug for the treatment of type 2 diabetes. With the continued intensive research into metformin, researchers have found that metformin has an anticancer effect in addition to lowering blood sugar.

As one study showed, metformin has reduced TOR pathway signaling that can promote breast cancer.

Another study reported that metformin has an antihypoxic effect. The results of this study show that metformin therapy reduces the positive areas of hypoxia and reduces HIF-1 a and pro-angiogenic factor levels. Metformin can inhibit HIF-1 a accumulation in hepatocellular carcinoma by resisting hypoxia. Scharping et al report that metformin can relieve the hypoxic state in tumors by reducing oxygen consumption, thereby improving T cell immune function of Tumor Microenvironment (TME), so that the combined use of metformin and an immune checkpoint inhibitor targeting PD-1 has a remarkable synergistic anti-tumor effect.

In 2019, cancer Cell journal published an article, and scientific researchers found that taking metformin in a fasted state can significantly inhibit tumor growth, and simultaneously proposed that the PP2A-GSK3 beta-MCL-1 signaling pathway could be a new target for tumor treatment.

3. Research on anticancer effect of curcumin

Curcumin is an active ingredient from turmeric rhizome and is used as a pharmaceutical in both india and china, which is also commonly used as a flavouring in india. The multi-target, multi-way and bi-directional regulation effect of the curcumin is a bright spot, and the curcumin can be extracted from plants or synthesized artificially.

Chinese name curcumin

Foreign language name Curcumin

The function is as follows: antioxidant, antitumor, senile dementia resisting, blood lipid reducing, etc

CAS registry number 458-37-7

Molecular formula C ₂₁ H ₂₀ O ₆

Molecular weight 368.39

The chemical structural formula: the following are listed below

The anti-tumor effect of curcumin was first proposed by indian scholars Kuttan in 1985 and has been widely paid attention to domestic and foreign scholars. Curcumin has been listed as a 3 rd generation cancer chemopreventive drug by the national tumor institute since it can inhibit the growth of various tumor cell lines, prevent the formation of various tumors in chemically and radioactively induced experimental animals, significantly reduce the number of tumors, and reduce tumor volume.

Curcumin can inhibit the function and expression of P-glycoprotein, activate caspase-3, and reverse the multi-drug resistant human gastric cancer cell line SGC 7901/VCR. The growth of tumor requires the establishment of a vascular supply system for continuous oxygen and nutrient supply. Hypoxia-inducible factor-1 (HIF-1) is the primary regulator of angiogenesis induced by malignant tumors, where HIF-alpha protein determines HIF-1 activity. Sun Jun and the like, through experimental researches on the action of curcumin on a human liver cancer cell strain BEL-7402, the curcumin can reduce the expression of HIF-lα protein of human liver cancer cells through a proteasome way. And according to the pharmacological characteristics of curcumin and the pharmacokinetic characteristics of various dosage forms, a learner proposes to mix a large dose of curcumin with iodized oil to perform interventional therapy on liver tumor. The Bcl-2 regulatory protein family plays an important role in regulating apoptosis and Bcl-2 family members can be divided into two classes according to different functions: bcl-2 is an apoptosis inhibitor, and Bax is an apoptosis promoter. Experiments prove that curcumin can up-regulate the levels of D53 serine phosphorylation and Bax and simultaneously down-regulate the levels of Bc-2, caspase precursor-3 (pro-caspase-3) and cysteine precursor-9 (pro-caspase-9), so that apoptosis of colon cancer cells HT-29 is induced, and the curcumin can be used as an effective medicament for treating colon cancer. Curcumin can also inhibit proliferation of cisplatin-resistant human ovarian cancer cells by inducing cell G2/M arrest, apoptosis, and peroxidation. In addition, the curcumin derivatives, namely maleic acyl L angiotensin (NVC) and maleic acyl glycine (NGC), can effectively improve the activity of apoptosis protease caspase-3 in human bladder cancer cell lines. In addition, curcumin can also play a certain role in treating prostate cancer, cancers of blood systems and the like.

The medical doctor Su Ni group of university of arizona medical school in the united states teaches at 42 th annual meeting of the american cancer control society held in 2014 that a natural product, curcumin, can modulate tens of antitumor targets. Through medical personnel research experiments, curcumin is proved to be capable of regulating and controlling cancer signal pathway molecules by group targets and reversing tumor progress. The achievement is hopeful to realize research breakthrough of natural targeted drugs and greatly reduce the treatment cost of the targeted drugs.

Researchers Su Ni, in the united states, have demonstrated by global multinational research specialists in the charge of the professor aloweil, the center for cancer, m.d., that 86 anti-cancer targets for curcumin-mediated intervention have been reached by 2018, with 23 anti-cancer targets approved as drugs by the united states Food and Drug Administration (FDA) and 63 anti-cancer targets not yet approved as new drugs. Curcumin can regulate and control a plurality of target groups such as inflammatory factors, tumor metastasis factors, growth factors, protein kinases, oncoproteins and the like. Su Ni it is introduced that in clinic, curcumin has been applied to more than 1000 cancer patients, and most of these patients have poor traditional treatment effects such as surgery and radiotherapy and chemotherapy or have recurrent metastasis after traditional treatment, including pancreatic cancer, intestinal cancer, lung cancer, breast cancer, liver cancer and other multiple diseases, and the disease condition of the patients is reversed by adopting curcumin combined with nutritional diet support, psychological intervention, home support system construction and other modes. Su Ni discloses that curcumin has completed a first and second phase clinical trial at multiple research centers worldwide and has been approved by the FDA in the united states to enter a third phase clinical trial.

The toxicology test of curcumin shows that the oral acute toxicity test of curcumin rats or mice belongs to an actual nontoxic substance; no potential mutagenic, micronuclear and teratogenic effects are seen; the feeding test of the rats for 30 days does not have obvious toxic reaction, has no obvious influence on the growth and development of the rats, blood chemical indexes, blood biochemistry, dirty body ratio and other indexes, and has no obvious adverse effect on the general anatomy and histological observation results of the rats. Experimental results show that the curcumin food has high safety and basically has no toxic or side effect. Therefore, the curcumin has very broad prospect in the application and development of the fields of foods, medicines and the like.

Shalma et al studied the pharmacokinetics of curcumin by orally administering turmeric extract to 15 patients with advanced colon cancer. The patient orally takes 180mg of curcumin, and the HPLC measurement of blood plasma and urine hardly detects curcumin and metabolites thereof, but the curcumin is detected in the patient's feces, which indicates that the patient orally takes 180mg of curcumin every day, is safe and well tolerated, but is not easy to absorb after oral administration and has low bioavailability. Lao et al studied the maximum tolerability and safety of curcumin by dose-scale-up, 24 healthy subjects orally administered curcumin extract powder, with doses increasing from 500mg to 12000 mg/time in sequence, with only 7 cases showing slightly non-dose related toxic effects, so curcumin oral single dose tolerability was considered good, up to 8000mg. Cheng et al performed a prospective clinical study on 25 patients with 5 malignant diseases, and evaluated the safety of curcumin. The initial dose was 500mg/d, and if no consecutive grade 2 or more toxic reactions occurred, the doses were sequentially increased to 1000, 2000, 4000, 8000 and 12000 mg/day. The results show that the continuous oral dose of 8000 mg/d curcumin is 3 months, and the body does not have toxic reaction related to curcumin; after 3 months, the patient's diseased tissue was examined and found to be significantly improved. Curcumin has been found to target a number of oncogenic proteins such as NF-kB, STAT3, AP-1 and the like.

4. Research on anticancer effect of geranylgeraniol

Chinese name: geranylgeraniol

English name; geranyl geraniol

Molecular formula C ₂₀ H ₃₄ O

Molecular weight: 290.48

CAS login number:24034-73-9

structural formula:

the function is as follows: geranylgeraniol itself has a wide range of physiological activities, such as killing viruses, resisting tumors, and the like;

geranylgeraniol is a biochemical precursor of a variety of biochemically significant products, such as terpenes, carotenes, steroids, cholesterol, and paclitaxel. Meanwhile, geranylgeraniol is also an important organic synthesis intermediate of multiple drugs, such as coenzyme Qn (I), coenzyme Q10, vitamin K2 and the like, and the research on geranylgeraniol by the scientific community is currently in a relatively early stage, but is in continuous penetration. Researchers gradually excavate and discover that geranylgeraniol has certain anticancer effects, HMG-CoA reductase is prenylated and Ras-and polyphenol-mediated glycosylation of growth factor receptors of nuclear laminin. Diterpene geranylgeraniol down regulates HMG-CoA reductase levels, thereby inhibiting human liver, lung growth, ovarian, pancreatic, colon, gastric and hematological tumors. For example, in one study it was found that geraniol inhibits the HMG-CoA reductase level of the rate-limiting enzyme of the mevalonate pathway and thereby reduces the viability of human prostate cancer cells DU 145. Geranylgeraniol-induced dose-dependent inhibition of human DU145 prostate cancer cell viability (ic50=80±18 μmol/L, n=5) after 72 hours incubation in 96-well plates. The arrest of the cell cycle in the G1 phase is accompanied by a decrease in cyclin D1 protein. Flow cytometry analysis to detect geranylgeraniol-induced apoptosis using immunohistochemistry, fluorescent microscopy to observe the brominated double staining, caspase-3 activation. Geranylgeraniol-induced inhibition of activity is accompanied by a decrease in the concentration of HMG-CoA reductase levels. The results of this study suggest that geranylgeraniol may have potential in the chemoprevention and/or treatment of prostate cancer.

The beneficial effects of the invention are as follows:

the composition for treating cancer provided by the invention has the advantages that the raw material components comprise two or three of tocotrienols, metformin, curcumin and geranylgeraniol, the raw material components can complement through different anticancer mechanisms and inhibit or promote apoptosis to kill different cancer cells through multiple targets, as an exemplary experiment, the composition has obvious inhibition effect on panc-1 human pancreatic cancer cell strains and MDA-MB-231 human breast cancer cell strains, and the number of anticancer targets and the anticancer spectrum are overlapped after combination so as to obviously improve the anticancer effect, so that the significant anticancer synergistic effect exists among the raw materials of the monomer compounds in the composition, meaningful attempts are made for exploring an innovative method for treating cancer, and the composition has further development potential and prospect.

Detailed description of the in vitro cytological assays

In order to make the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be described in detail below. It will be apparent that the described embodiments are only some, but not all, embodiments of the invention. Based on the examples herein, it will be appreciated by those of ordinary skill in the art that all other embodiments that could be made without the inventive effort are within the scope of the invention.

The following experimental examples are for illustrative purposes only and are not intended to limit the invention.

In the examples below, 1 part by weight represents l. Mu.g. And the composition of any of the embodiments can be used to perform experiments directed to different cancer cell lines, such as inhibition experiments directed to the panc-1 human pancreatic cancer cell line, and the MDA-MB-231 human breast cancer cell line. In all examples, the tocotrienols will be represented by delta-tocotrienols, the metformin will be represented by metformin, the curcuminoids will be represented by curcumin, and the geranylgeraniols will be represented by geranylgeraniols, respectively, in combination for experimental implementation.

Example 1

The embodiment provides a tocotrienol composition for treating cancers, which comprises the following raw materials in parts by weight: 20 parts by weight of delta-tocotrienol, and 40 parts by weight of metformin.

Example 2

The embodiment provides a tocotrienol composition for treating cancers, which comprises the following raw materials in parts by weight: 20 parts by weight of delta-tocotrienol, 40 parts by weight of metformin, and 40 parts by weight of curcumin.

Example 3

The embodiment provides a tocotrienol composition for treating cancers, which comprises the following raw materials in parts by weight: 25 parts of delta-tocotrienol, 60 parts of geranylgeraniol.

Example 4

The embodiment provides a tocotrienol composition for treating cancers, which comprises the following raw materials in parts by weight: 25 parts by weight of delta-tocotrienol, 60 parts by weight of geranylgeraniol, and 50 parts by weight of curcumin.

Comparative example 1

The comparative example provides a monomer for treating cancers, which comprises the following raw materials in parts by weight: 50 parts by weight of delta-tocotrienol.

Comparative example 2

The comparative example provides a monomer for treating cancers, which comprises the following raw materials in parts by weight: 40 parts of metformin.

Comparative example 3

The comparative example provides a monomer for treating cancers, which comprises the following raw materials in parts by weight: 50 parts by weight of curcumin.

Comparative example 4

The comparative example provides a monomer for treating cancers, which comprises the following raw materials in parts by weight: 100 parts by weight of geranylgeraniol.

Comparative example 5

The comparative example provides a monomer for treating cancers, which comprises the following raw materials in parts by weight: 100 parts of doxorubicin.

Test example: the invention takes the form of in vitro anti-tumor activity test and research

1. Test purpose: the compositions for treating cancer according to the present invention were tested for cytotoxicity to cancer cells by MTT assay.

2. Sample to be tested:

(A) Delta-tocotrienol (standard, purity > 98%);

(B) Metformin (standard, purity > 98%);

(C) Curcumin (standard, purity > 98%);

(D) Geranylgeraniol (standard, 93.5% purity);

(E) Doxorubicin, as a positive control (standard, purity > 99%).

3. Cell lines:

panc-1 human pancreatic cancer cell line;

MDA-MB-231 human breast cancer cell line;

all the cell lines were stored and passaged in liquid nitrogen in the laboratory.

4. Other materials:

DMEM medium (GIBCO product), inlet fetal bovine serum (PAA product), cell digests (trypsin+edta), PBS (-), MTT (5 mg/ml), cell lysates, and the like.

5. Instrument:

full-wavelength multifunctional enzyme labeling instrument, model: varioskan Flash, manufacturer: thermo scientific.

6. The test method comprises the following steps:

collecting cancer cell strain in logarithmic growth phase, stopping after digestion, centrifuging, collecting, making into cell suspension, and regulating cell count to 5×10 ⁴ 1000. Mu.L, i.e., 1mL, of each well was placed in a 96-well plate at 37℃with 5% CO ₂ Culturing in an incubator for 24 hours. The test samples are dissolved by DMSO, diluted by PBS (-) to the required concentration and added into each well, double wells are arranged at the same concentration, and 0.1% DMSO is additionally arranged as a control group. After further culturing for 72 hours, 200. Mu.L of MTT solution (5 mg/ml) was added to each well, and after 4 hours in the incubator, 100. Mu.L of the solution was added to each well, and the incubator was left overnight. OD at 570nm was measured with a microplate reader, and IC50 (half inhibition concentration in. Mu.g/ml) was calculated by calculating the cell growth inhibition rate. Cell growth inhibition = (OD control group-OD test group)/OD control group x 100%, and cell growth inhibition is plotted on the abscissa with drug concentration as the abscissa, and IC50 value, i.e. drug concentration value with cell growth inhibition of 50%. The lower the IC50 value, the lower the concentration of the compound required to achieve inhibition of half of the cancer cells, thus reflecting the greater the potency of the compound.

7. The test results are shown in Table 1

TABLE 1 results of cancer cell killing test (when the IC50 value of a composition is lower, the stronger the efficacy of the composition is shown)

8. Conclusion (note: the concentration values in brackets below were all omitted in units of. Mu.g/ml')

Group 1 experiments on human pancreatic cancer panc-1 cell lines:

in the test results of 1a (2 drug monomer combination), the composition of the 2 component monomer combination of delta-tocotrienol and metformin (20:40 ratio) achieves a half inhibition rate IC50 value (4.79) of human pancreatic cancer cell line panc-1 which is about 6.19 times stronger than the IC50 value (29.69) of delta-tocotrienol monomer (50) on the half inhibition rate of human pancreatic cancer cell line panc-1, reflecting the synergistic effect of delta-tocotrienol and metformin (20:40 ratio) 2 components on the human pancreatic cancer cell line (panc-1).

In the test results of 1b (3 drug monomer combination), the IC50 value (0.001) of the half inhibition rate of the composition composed of delta-tocotrienol and 3 component monomers of metformin and curcumin (20:40:40 ratio) on human pancreatic cancer cell line (panc-1) was about 29690 times lower than the half inhibition rate IC50 value (29.69) of delta-tocotrienol monomer on human pancreatic cancer cell line (panc-1), namely, about 29690 times stronger; simultaneously, compared with a compound preparation of 2 component monomer combination of delta-tocotrienol and metformin (20:40 ratio), the half inhibition rate IC50 value (4.79) of human pancreatic cancer cells panc-1 is 4790 times stronger; in addition, the efficacy is also stronger than the half inhibition rate IC50 value (1.19) of the positive control drug doxorubicin monomer 100 on the human pancreatic cancer cell strain (panc-1), thereby reflecting that the 3 component monomers of delta-tocotrienol, metformin and curcumin (20:40:40 ratio) have strong synergistic effect on resisting the human pancreatic cancer cell strain (panc-1).

1c (2 drug monomer combination) in the test results, the composition of the 2 component monomer combination of geranylgeraniol and delta-tocotrienol (60:25 ratio) achieved a half-inhibition IC50 value (< 6E-06) for human pancreatic cancer cell line panc-1 that was about 6E+04 times greater than the half-inhibition IC50 (about 100) for geranylgeraniol monomer for human pancreatic cancer cell line panc-1; in addition, the efficacy is far better than the half inhibition rate IC50 value (1.19) of the positive control drug doxorubicin monomer 100 on the human pancreatic cancer cell strain (panc-1), which reflects the synergistic effect of geranylgeraniol and delta-tocotrienol (60:25 ratio) 2 components on the human pancreatic cancer cell strain (panc-1).

In the test results of 1d (3 drug monomer combination), the half inhibition rate IC50 value (< 6E-06) of the composition of geranylgeraniol combined with 3 component monomers of delta-tocotrienol and curcumin (60:25:50 ratio) on human pancreatic cancer cell line panc-1 was about 6E+04 times stronger than the half inhibition rate IC50 (about 100) of geranylgeraniol monomer on human pancreatic cancer cell line panc-1; thus reflecting the strong synergy between geranylgeraniol and 3 component monomers of delta-tocotrienol and curcumin (20:40:40 ratio) against human pancreatic cancer cell line (panc-1).

Group 2 experiments on human breast cancer MDA-MB-231 cell lines:

in the test results of 2a cases (2 drug monomer combination), the half inhibition rate IC50 value (6.63) of the composition of the 2 component monomer combination of delta-tocotrienol and metformin (20:40 ratio) on the human breast cancer MDA-MB-231 cell strain is 18.24 times stronger than the half inhibition rate IC50 value (120.94) of the delta-tocotrienol monomer (50) on the human breast cancer MDA-MB-231 cell strain, which shows that the composition of the 2 component monomer combination of delta-tocotrienol and metformin (20:40 ratio) has the synergistic effect of resisting the human breast cancer MDA-MB-231 cell strain.

In the test results of 2b cases (3 drug monomer combination), the IC50 value (0.03) of the composition of the delta-tocotrienol combined with 3 component monomers of metformin and curcumin (20:40:40) on the human breast cancer MDA-MB-231 cell line is 4031 times stronger than the IC50 value (120.94) of the delta-tocotrienol monomer (50) on the human breast cancer MDA-MB-231 cell line, and 221 times stronger than the half inhibition rate IC50 value (6.63) of the composition of the delta-tocotrienol combined with 2 component monomers of metformin (20:40:40) on the human breast cancer MDA-MB-231 cell line; in addition, the drug effect is also stronger than the half inhibition rate IC50 value (0.061) of the positive control drug doxorubicin monomer (100) on the human breast cancer cell strain MDA-MB-231, and the half inhibition rate IC50 value shows that the delta-tocotrienol has the synergistic effect of the human breast cancer cell strain MDA-MB-231 with 3 component monomers of metformin and curcumin (20:40:40 ratio).

In the test results of 2c (2 drug monomer combination), the half inhibition rate IC50 value (< 6E-06) of the composition of the 2 component monomer combination of geranylgeraniol and delta-tocotrienol (60:25 ratio) on the human breast cancer MDA-MB-231 cell strain is about 6E+04 times stronger than the half inhibition rate IC50 (> 100) of geranylgeraniol monomer on the human breast cancer MDA-MB-231 cell strain; in addition, the drug effect is far stronger than the half inhibition rate IC50 value (0.061) of the positive control drug doxorubicin monomer (100) on the human breast cancer cell strain MDA-MB-231, and the half inhibition rate IC50 value reflects the strong synergistic effect of geranylgeraniol and delta-tocotrienol (60:25 ratio) 2 component monomers on the human breast cancer cell strain MDA-MB-231.

In the test results of 2d (3 drug monomer combination), the IC50 value (< 6E-06) of the composition of geranylgeraniol combined with 3 component monomers of delta-tocotrienol and curcumin (60:25:50 ratio) was about 6E+04 times stronger than the half-maximal inhibition rate IC50 (> 100) of geranylgeraniol monomer on human breast cancer MDA-MB-231 cell line; thus reflecting the strong synergy between geranylgeraniol and 3 component monomers of delta-tocotrienol and curcumin (60:25:50 ratio) against human breast cancer MDA-MB-231 cell line.

In summary, the compositions in all examples show better anticancer synergistic effect against different kinds of human cancer cell lines, and the implementation of the exemplary anticancer test is carried out on the panc-1 human pancreatic cancer cell line, the MDA-MB-231 human breast cancer cell line, the synergy or superposition of the anticancer effects are better than 1+1>2 or 1+1+1>3, especially the 1c, 1d, 2c and 2d cases in the implementation of the cytology test, and the anticancer effects of the tested compositions are very remarkable, which suggests that the different compositions formed by two or three of the raw material components including delta-tocotrienol, metformin, curcumin and geranylgeraniol have potential to be developed into innovative anticancer drugs for replacing traditional chemotherapeutics.

The present invention is a composition that the inventors have been diligent and diligent in over 10 years to screen from hundreds of natural and synthetic compounds through layer-by-layer exploratory testing. The above experiments are only exemplary embodiments of the present invention, but the scope of the present invention is not limited thereto, and any other corresponding composition embodiments obtained by adding or subtracting 1 to 2 chemical groups to or from the chemical structure of the above compound monomers can be easily considered by those skilled in the art within the scope of the present invention, and thus all other composition embodiments are within the scope of the present invention.

Claims (4)

1. A composition of tocotrienols for the treatment of cancer, characterized by the following raw material components: metformin or geranylgeraniol, tocotrienol, curcumin;

the tocotrienol is tocotrienol;

the metformin substance is metformin;

the curcumin substance is curcumin;

the geranylgeraniol is geranylgeraniol;

when the raw material components are tocotrienols, metformin and curcumin, the mass ratio of the tocotrienols, the metformin and the curcumin is 20:40:40;

when the raw material components are tocotrienols, curcumin and geranylgeraniol to form a composition, the mass ratio of the tocotrienols, the curcumin and the geranylgeraniol is 25:50:60;

the cancer is human breast cancer or pancreatic cancer;

the tocotrienol is delta-tocotrienol.

2. The composition of matter of claim 1, wherein said tocotrienols, metformin, curcumin and geranylgeraniol are extracted from natural plant compounds or are synthetically produced.

3. The tocotrienol composition for the treatment of cancer according to claim 1, wherein the dosage form of said composition is any one of a tablet, a capsule, a granule, a suspension, a solution, an injection, a lyophilized powder, an aerosol and a skin patch.

4. The tocotrienol composition for the treatment of cancer according to claim 1, wherein said composition further comprises a pharmaceutically acceptable adjuvant.