TR201721581A2 - Synthesis of a New Benzimidazolium Salt with High Anticancer Activity - Google Patents

Abstract

Cancer, which is one of the important health problems of our country as well as developed and developing countries and which develops due to genetic changes and environmental effects, is one of the most common diseases threatening human life. Therefore, in order to protect human health and to improve the quality of life continuously, the innovative aspect of national and international health science needs to be continuously improved. Due to the increasing cancer-related mortality rates, the idea that chemotherapy drugs should be developed has been developed and to date, the complex has been synthesized by researchers in a complex structure that exhibits a large number of anticancer drug potentials. However, due to the high cost of preparation and low solubility in water, the search for new drug candidates is continuing unabated. Therefore, in this study, unlike the other researchers, a new compound which could be a candidate for anticancer drug in salt structure instead of complex was developed in order to reduce the cost and increase the water solubility. This compound was tested using the MTT assay method against breast cancer, which is the most common cause of death among women in the world and the second most common cause of death, and colon cancer cells, which are also common in men. The compound SA-73 gave excellent results. For example ; IC50 values of cisplatin used as a positive control drug against cancer colon and breast cells and busulfan used in solid tumors, respectively; (4.38 = 0.05 MM, 5.77 = 0.4 Umm), (173.2 MM,> 200 )m), while the compound SA-73 (2.761 = 0.43 MM, 2.682 = 0.24 MM). This shows that the unsymmetric substituted salt structure, which can be produced at low prices (about 150 TL) and has no water solubility problem, is a much more effective and promising compound than positive control drugs.

Description

DESCRIPTION' A NEW BENZIMIDAZOLIUM WITH HIGH ANTICANCANCER ACTIVITY SYNTHESIS OF SALT Technical Area An effective and novel chemotherapeutic drug candidate for the treatment of breast and colon cancer. development State of the Art Malignant malignancy that occurs when cells in an organ or tissue divide and multiply irregularly. Tumors are called cancer. Uncontrolled proliferation of cells in various parts of our body There are more than 100 types of cancer caused by cancer. The common feature of these cancer types It begins with the out-of-control proliferation of abnormal cells. Serious if untreated can cause illness and even death.

Various drugs such as cisplatin, oxaliplatin, carboplatin, nedaplatin, busulfan for cancer treatment Anticancer drugs are used clinically. However, death rates from cancer are very high. high and these drugs have a high toxic effect on healthy cells. Therefore, there is a great need to develop more effective anticancer drugs.

Therefore, the research and development of new anticancer drugs is being done by scientists. it is done in an intense way.

Technical Problems That the Invention Aims to Solve It is very common both in our country and in the world and can cause death. Breast and colon cancer pose a great risk to humans. So this cancer It is much better than cisplatin and busulfan, which are known and used as clinical drugs. A new organic-based drug candidate has been developed that gives results.

Description of the Developed Compound General methods Structure control of the developed compound Bruker 400 MHz Ultra Shield Nuclear Magnetic Resonance (NMR) spectroscopy, High Resolution Mass Spectrometer (HRMS), and Shimadzu Fourier (FT-IR) 8400 spectrophotometer. If cell counting is done, Invitrogen Countess automatically was done with a cell counting device. 1-[2-(4-nitrophenyl)ethyl-3-(4-methylbenzyl)-1H-benzo[d]imidazole-S-ium bromide (SA-73) Benzimidazole (1 mmol) and potassium hydroxide (1 mmol) were dissolved in ethyl alcohol (20 mL).

After stirring the mixture for one hour at room temperature, 1-(chloromethyl)-4-methylbenzene (1 mmol) added to the medium and reIluxed for 6 hours. Then the reaction mixture was returned to room temperature. The potassium chloride formed by cooling was filtered and the obtained 1-(4-methylbenzyl)benzimidazole ethyl It was purified by crystallization in alcohol.

Synthesized 1-(4-methylbenzyl)benzimidazole (0.59 g, 1 mmol), 1-(2-bromoethyl)-4-nitrobenzene (influencing in the targeted main product (452.34 g/mol) was synthesized. 24 h for the reaction to occur Stirring was continued at 80 °C. DMF in the environment after the reaction is complete removed by applying vacuum. The developed product was crystallized in ethyl alcohol and purified.

Figure-1: 1-[2-(4-nitrophenyl)ethyl]-3-(4-methylbenzyl)-1H-benzo[d]imidazole-3-ium bromide synthesis of salt Yield: 55%, mp: ; NCH; 5.82 (t, J: 8.0 Hz, 2H, Cytotoxic activity studies of the developed compound Human colon cancer cell (DLD-1) and human breast cancer cell (MDA-MB-231), 10% Cultured in DMEM medium containing PES. Cells were plated 1x103 in sterile 96-well plates. seeded at a cell/well density and a moist soil containing 5% COZ for 24 hours. It was incubated at 37 °C under atmosphere. Cells were then treated with nine different concentrations of drug, ranging from 0.5 µM to 72 hours. exposed to the candidate. Stock solutions of the developed compound Phosphate buffered saline (PBS) was also prepared. As a positive control, the anti-cancer agents cisplatin and busulfan (50 µL) was added to each well and incubated in the incubator for 4 hours. Then each added to the well and the plate was placed on the shaker for half an hour. was abandoned. Absorbance values with standard ELISA microplate device (Biorad 6800) at 595 mn read. IC50 values were calculated with the GraphPad Prism software program.

Table 1. IC50 results of the developed compound against DLD-1 and MDA-MB-231 cell lines.

Compounds IC 50 (iiM) DLD-l MDA-MB-23l The product developed according to Table 1 gives much better results than the well-known cisplatin and busulfan gave. Low IC50 value of compound SA-73 against both colon and breast cancer cells appears to be a suitable drug candidate.

Explanation of Parts Shown with Reference Numbers in Figure-1 selected as follows: may contain one of the groups H, F, Cl, Br, I, NH2, NO2, OH, CH3, ON=O.

Substituted or unsubstituted C1-C5 alkyl, Substituted or unsubstituted C1-C5 alkenyl, Substituted or unsubstituted C6-C8 alkynyl, Substituted or unsubstituted C6-C9 alkyl, Substituted or unsubstituted C6-C9 alkenyl, Substituted or unsubstituted C6-C9 alkynyl, Substituted or unsubstituted C 10-C14 alkyl, Substituted or unsubstituted Cio-CM alkenyl, It may contain a substituted or unsubstituted C10-C14 alkynyl group.

"C1-C5 alkyl" as used herein, either alone or in compound terms The term straight-chain or branched saturated hydrocarbon having 1 to 5 carbon atoms. refers to groups. Limited with methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl Suitable alkyl groups are present. "C1-C5 alkyl" optionally one or more may contain more substituents. Substituents "Cr-C5 alkyl" in the carbon atom chain any carbon atom or one or more hydrogens on the carbon atom can replace the atom.

The term "Cl-C5 alkenyl" as used herein means 1 to 5 carbons, either alone or in combined terms. straight chain or branched atoms containing at least one carbon-carbon double bond refers to unsaturated hydrocarbon groups. Limited with ethenyl, propenyl or butenyl may contain suitable alkenyl groups, but not necessarily Carbon-carbon double bond, any two adjacent between carbon atoms. "C1-C5 alkenyl" optional with one or more substituents can be substituted. Substituents in the "Ci-C5 alkenyl" carbon atom chain any carbon atom or one or more hydrogens on the carbon atom can replace the atom.

Here; R6, R7, R8 and R9 are independently H, F, ci, Br, i, NH2, OH, ON=O may contain groups. Substituted or unsubstituted C6-C9 alkyl, substituted or as unsubstituted C6-C9 alkenyl, substituted or unsubstituted C6-C9 alkynyl can be used.

R3 is CH3; R12 contains No2 group.

Organic solvent is used to synthesize this compound. Preferably polar as solvent aprotic solvents are used. More ethyl acetate, tetrahydrofuran (THF), dichloromethane (CHZClz), acetone, acetonitrile, dimethylformamide (DMF), DMSO or mixtures thereof is preferred. But mostly DMF is used as solvent. preferably from formic acid, acetic acid, n-butanol, isopropanol, n-propanol, ethanol or methanol the resulting solvents are selected. Preferably with an apolar solvent as most crystallization agent. A mixture of polar aprotic solvent is used. Ethyl alcohol synthesized compound used for crystallization.

The temperature of the reaction mixture may range from approximately 60 °C to 100 °C. However, approx. It is preferred that it be between 70 °C and 90 °C. In the invention, it is preferred that the temperature be 80 °C. compound is prepared. Preferably colon cancer or breast cancer in a cancer treatment Much better results than the well-known cisplatin when using this product developed against has been taken.

The invention is for use in the manufacture of a new and more effective drug for the treatment of cancer. has been made.

The synthesis method for the manufacture of the drug is also given in detail in the invention.

Disclosure of the Invention Industrial Application of the Invention The present invention is based on the discovery of the SA-73 compound and its anticancer properties. The compound of the invention has different types such as mouth, cheek, nose, vaginal, rectal, parenteral, lung, liver. It can also be formulated against cancer types. The compound synthesized within the scope of the invention is tablets, pills, lozenge, aqueous or oily suspension, dispersible powder or granule, emulsion, hard or soft It can be given to the patient as a capsule or syrup.

The appropriate daily dosage level is the kg body weight that can be administered in a single dose or in multiple doses. will be approximately 0.01 to 500 mg per weight. Preferably, the dosage level per day about 0.0] to 250 mg/kg; more preferably about 0.5 to about 100 mg/kg. a suitable It can be between mg/kg. Compositions preferably for oral administration 1.0 to 1000 milligrams active may contain milligrams. The compound may be administered 1 to 4 times a day, preferably once or twice a day.

However, the specific dose level and frequency of dosing may vary for a particular patient. and the activity, metabolic stability and duration of action of the specific compound to be used. It will be understood that it will depend on various factors, including For example; age, body weight, general health, gender, diet, method and time of administration, excretion rate, drug combination, special like the seriousness of the situation and the host entering therapy.

In addition, the pharmaceutical composition may contain other therapeutically active compounds.

Selection of suitable substances to be used in combination therapy, conventional pharmaceutical according to the principles it can also be done by people with ordinary experience in the technical field.

Combination of therapeutic agents to treat the conditions described here can act synergistically. With this approach, each substance is administered at lower dosages. may gain therapeutic efficacy and thus the possibility of side effects may be reduced.

Pharmaceutical formulation application; binding agents, gelatins, fillers, well-known as lubricants, disintegrants, surfactants and colorants may also include conventional additives. Tablet, capsule, lozenge, dragee, gum or other solid formulation types in various dosages can be used. capsules; It can be in powder, liquid or gel form. The solid formulation can be swallowed or may be of the absorbable or chewable type. Available invention, bottle or blister such as tube Dosing unit other than packages can be presented in packages.

Compositions intended for oral use are flavoring, coloring and/or preservative. may contain one or more components such as agents. Tablets, suitable for the manufacture of tablets active ingredient in mixture with physiologically acceptable excipients may contain. Such excipients include, for example, calcium carbonate, sodium carbonate, lactose, inert diluents such as calcium phosphate or sodium phosphate, corn starch or alginic acid granulating and dispersing agents such as starch, binding agents such as gelatin or acacia, and lubricating agents such as magnesium stearate or stearic acid may be present. tablets may be uncoated or delay its breakdown and absorption in the gastrointestinal tract. and thus to provide a sustained effect over a longer period of time, with known techniques they can be covered. For example, a time delayer such as glyceryl monosterate or glyceryl distearate material can be used.

Formulations for oral use, active gelatin, calcium carbonate, calcium phosphate or It can be mixed with an inert solid diluent such as kaolin or into soft gelatinous capsules. may also be offered as gelatin capsules. The active ingredient here is also in the water or oil environment (ground peanut oil, liquid paraffin or olive oil).

Aqueous suspensions, active ingredient(s) suitable for the production of aqueous suspensions Contains in mixture with excipients. These excipients are sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, tragacanth gum and suspending agents such as acacia gum and naturally occurring phosphatids (e.g. lecithin) dispersing or wetting agents such as fatty acids such as polyoxyethylene stearate of an alkylene oxide, ethylene oxide with long-chain aliphatic alcohols such as heptadecaethyleneoxycetanol Condensed products, partial esters derived from fatty acids with ethylene oxide and condensation products of hexitol, such as polyoxyethylene sorbitol mono-oleate, or ethylene oxide condensation products partial esters derived from fatty acids and polyethylene sorbitan hexitol anhydrides such as monooleate. Aqueous suspensions are also or more preservatives, such as ethyl or n-propyl p-hydroxybenzoate, one or more a coloring agent, one or more flavoring agents, and one or more such substances as sucrose or saccharin. It may also contain more sweetening agent.

Oily suspensions of active ingredients in peanut oil, olive oil, sesame oil or in a vegetable oil such as coconut oil or in a mineral oil such as liquid paraffin It can be formulated by suspending. Oily suspensions wax, hard may contain a thickening agent such as paraffin or cetyl alcohol. refillable mouth flavoring agents and/or fragrances such as those described above to provide the preparations donor agents can be added. These suspensions are supplemented with an antioxidant such as ascorbic acid. can be protected.

Dispersible powders and granules suitable for the preparation of an aqueous suspension by adding water, active ingredient, a dispersing or wetting agent, suspending agent and one or more It can be supplied in combination with a preservative. Sweetener, flavoring and coloring matter Additional excipients such as substances may also be present.

The pharmaceutical compositions may also be in the form of oil-in-water emulsions.

Oily phase, a vegetable oil such as olive or peanut oil, a mineral such as liquid paraffin oil or a mixture of these. Suitable emulsifying agents, gum acacia or naturally occurring gums such as tragacanth gum, naturally occurring gums such as soybean lecithin phosphatides and fatty acids and esters or partial esters derived from hexitol, sorbitan anhydrides such as monoleates and condensation products of partial esters from fatty acids and may be derived from hexitol with ethylene oxide, such as polyoxyethylene sorbitan monoleate. A The emulsion may also contain one or more sweetening and/or flavoring agents.

Syrups and elixirs are mixed with flavoring agents such as glycerol, propylene glycol, sorbitol or sucrose. can be formulated. Such formulations contain one or more preservatives, flavorings and / or may contain coloring matter. Suitable selections of additional vehicle ingredients include alcohols (eg, ethanol, isopropyl alcohol or butyric solvents such as glycerin), glycols such as butylene, isoprene or propylene glycol, lanolin aliphatic alcohols, such as water and a mixture of organic solvents, and lipid-based materials such as glycerin, fatty acids, acylglycerols, including oils such as mineral oil, and natural or synthetic origin Protein-based materials such as oils, phosphoglycerides and waxes, collagen and gelatin, silicone based materials (both volatile and non-volatile) and hydrocarbons such as polymer matrices basic materials are available.

Composition, stabilizing agents, emulsifying agents, viscosity adjusters, gelling agents, preservatives, antioxidants, skin penetration enhancers, stability or efficacy of the applied formulation, such as humectants and sustained release. It may contain one or more components adapted to increase Examples of these components are Martindale-Extra Pharmacopeia (Pharmaceutical Press, London 1993) and Remington's It is described in Pharmaceutical Sciences. Formulations, hydroxymethylcellulose or gelatin microcapsules, liposomes, albumin microspheres, microemulsions, nanoparticles or may include microcapsules such as nanocapsules. Product formulation; including paste, cream, foam, lotion, gel, powder, emulsion and spray It can be prepared in various physical forms. The physical appearance of such forms and emulsifier(s) and viscosity adjuster(s) whose viscosity is in the formulation can be adjusted by its presence and amount. Solids are generally solid and do not spill. Generally They are formulated in stick or particle form. Solids can be opaque or transparent and can be optionally solvent, emulsifier, moisturizer, softener, perfume, dye/colorant, may contain preservatives and other active ingredients that increase the efficacy of the final product. Creams and lotions are mostly similar to each other and differ in viscosity by weight.

Both lotions and creams can be opaque, translucent or clear and are moisturizing, softener, perfume, dye/colourant, preservative and final substance (product) emulsifier, solvent and viscosity adjuster with other active ingredients that increase its effectiveness contain agents. Gels range from thick or high Viscosity to thin or low viscosity. Can be prepared with a range of viscosity. Solvent, such as formulations, lotions and creams, emulsifier, humectant, emollient, perfume, dye/colourant, preservative and final It may contain other active ingredients that increase the effectiveness of the product. Pimples from cream, lotion or gel it is thinner and usually does not contain emulsifiers. Liquid products are mostly solvents, emulsifier, moisturizer, emollient, perfume, dye/colorant, preservative and final product They contain other active ingredients that increase their effectiveness.

Formulations; emulsifiers, ionic emulsifiers, polyoxyethylene oleyl ether, PEG-40 non-ionic such as stearate, cetearet-12, cetearet-20, cetearet-30, cetearete alcohol, PEG May contain emulsifiers. Viscosity adjusting agents, hydroxyethylcellulose, xanthan gum, magnesium aluminum silicate, silica, beeswax, paraffin and cetyl palmitate may contain protective colloids or nonionic gums. A gel composition, chitosan, methyl cellulose, ethyl cellulose, polyvinyl alcohol, polyquatemiums, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, carbomer or ammoniacal glycyrrhine It can be formed by adding a gellestinne agent such as Suitable surfactants, may contain nonionic, amphoteric, ionic and anionic surfactants. For example formulations include polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, lauramide DEA, cocamide DEA and cocamide MEA, oleyl betaine, cocamidopropyl phosphatidyl PG-diammonium One or more of chloride and ammonium laurethsulfate may be used.

Methylparaben, propylparaben, sorbic acid, benzoic acid, formaldehyde, vitamin E, sodium ascorbate/ascorbic acid, propyl gallate and antioxidants are among the preservatives. takes. Lactic acid, other hydroxy acids and their salts, glycerin, propylene glycol and butylene glycol is among the suitable humectants. Lanolin alcohol, lanolin, lanolin derivatives, cholesterol, petrolatum, isostearyl neopentanoate and mineral oils are suitable are softeners. Among the available fragrances and colors, FD & C Red No. 40 and FD&C Yellow no. There are 5 Other suitable additional additives that may be included in the formulation, abrasives, absorbents, sabotage agents, chamomile extract, binders/excipients, buffering agents, chelating agents, film-forming agents, thickening agents, propellants, opacifying agents, pH adjusters and preservatives alcohol and herbal extracts.

The prepared composition is applied with a physical applicator such as a cloth, tissue, stick or brush. applicable. Also, mist, aerosol or foam spray, dropper application, sprinkling, may include spraying methods such as soaking and rinsing. Controlled salivenne tools too can be used.

Pharmaceutical compositions are slowly released after administration of the modulator. It can be formulated as sustained-release formulations such as a release capsule.

Such formulations can generally be prepared using well-known technology and by oral, rectal or subcutaneous implantation or implanted in the desired target area can be applied. Carriers for use in such formulations are biological is compatible and also biodegradable. Preferably, the formulation is relatively stable. provides a level of modulator release. Modulator in a sustained-release formulation The amount depends, for example, on the site of implantation, the rate of release and expected duration, and depends on the nature of the disorder to be treated or prevented.

The invention relates to a method for treating a subject. This method is considered pharmaceutical a pharmaceutically effective amount of the compound SA-73 in an acceptable form includes its implementation.

A "subject" as used herein; human, monkey, horse, cow, sheep, goat, dog, cat, refers to an animal such as a mammal, mouse, rat, chicken, or bird species, including a guinea pig.

The activity of the specific compound used; age, body weight, general health, gender, diet, the time of administration will depend on several factors, including the route of administration, and excretion rate, combination of drugs (ie other drugs used to treat the patient), and the severity of the particular disorder undergoing therapy should not exceed the specific dose level for any patient. will be effective in determining

Claims (1)

REQUESTS It is a benzimidazolium salt synthesis and its feature is; o 1 ininol beiiziinidazoluii 1 mmol potassium hydroxide is taken into its solution in 20 mL of ethyl alcohol and mixed, o 1 mmol 1-(chloromethyl1)-4-methylbenzene is added to this mixture after one hour, o refluxing the mixture under reflux for 6 hours, o The reaction mixture is mixed in room Removal of potassium chloride formed by cooling to temperature, o Purification of the obtained 1-(4-methylbenzyl)benzimidazole by crystallization in ethyl alcohol, o 1-(4-methylbenzyl1)benzimidazole (0.59 g, 1 mmol) solution in dried DMF. )-4-nitrobenzene (0.61 g, 1 mmol) and stirring, o Continuation of the reaction at 80 °C under argon gas for 24 h, o After the reaction is completed, DMF in the environment is removed by applying vacuum, o Developed product (SA-73 ) is characterized by the steps of crystallization in ethyl alcohol and purification. The formula indicated by SA-73 is a compound containing a pharmaceutically acceptable carrier, diluent or excipient; the independently selected H group is that 0 R3 contains the CH3 group, and 0 Rlz contains the NO2 group. It is a compound according to claim 2, its feature is; The synthesized compound is organic and easily soluble in hot water. The product mentioned in claim 1 is pharmaceutically acceptable and its feature is; use in a pharmaceutical composition for the treatment of a disease. According to claim 4, the disease mentioned here is cancer and its feature is; colon cancer or breast cancer. It is a compound synthesis and application method as in any of the claims 1-5 and its feature is; a process for the manufacture of a drug for use in the treatment of cancer.