TWI573589B - Novel lipophilic n-substituted norcantharimide derivatives and uses thereo - Google Patents

Description

Novel lipophilic N-substituted demethylcantharidin derivatives and their use way

The present disclosure is directed to a novel N-substituted desmethylcantharidin derivative and its use for the treatment of cancer, particularly leukemia.

Cantharidin (CA) is an active compound isolated from Mylabris , known as an inhibitor of serine/proline protease 1 (PP1) and protease 2A (PP2A), and has been It has been shown to have the ability to kill cancer cells, but unfortunately it also has nephrotoxicity and hepatotoxicity. Therefore, past research has focused on finding a new cantharidin derivative or analog that still retains the ability to kill cancer cells without causing damage to normal tissues. Accordingly, the researchers also identified two cantharidin derivatives, norcantharidin and norcantharimide. Demethylcantharidin is a form of demethylation of cantharidin with low renal toxicity, but demethylation also relatively reduces the biological activity of cantharidin. As for the methyl spot quinone imine, it has been found that the longer the alkyl chain carried on the N-, due to the lipophilic nature of the alkyl group, helps the cells to absorb the demethylated smectin.

Therefore, the related art requires a derivative of cantharidin and Or an analog that exhibits excellent selective toxicity to a variety of cancer cells without affecting normal cells.

The present disclosure is based, at least in part, on the discovery that N-substituted demethylcantharidin derivatives are not cytotoxic and can inhibit the growth of cancer cells. The above findings suggest that the N-substituted demethylcantharidin derivatives of the present disclosure are useful as lead compounds for the therapeutic agents for the treatment of cancer.

Accordingly, a first object of the present disclosure is directed to a compound of formula (I), Wherein R is farnesyl or farnesyloxy.

In one example, the compound of formula (I) is N-farneoxy-7-oxabicyclo[2,2,1]heptane-2,3-dimethylimine (NOC15). In another example, the compound of formula (I) is N-farnesyl-7-oxabicyclo[2,2,1]heptane-2,3-dimethylimine (NC15).

Accordingly, a second object of the present disclosure is to provide a pharmaceutical or pharmaceutical composition that can be used to treat cancer. The pharmaceutical or pharmaceutical composition comprises a therapeutically effective amount of a compound of formula (I), and a pharmaceutically acceptable adjuvant.

A cancer which can be treated by the pharmaceutical or pharmaceutical composition of the present disclosure is selected from the group consisting of leukemia, liver cancer, bladder cancer, rectal cancer, kidney cancer, ovarian cancer, cervical cancer, endometrial cancer, prostate Cancer, pancreatic cancer, lung cancer, breast cancer, melanoma, and squamous cell cancer (SCC). In one example, the cancer is leukemia; in another example, the cancer is liver cancer.

The compound of the formula (I) of the present invention comprises from about 0.1% to about 99% by weight based on the total weight of the pharmaceutical composition, based on the total weight of the pharmaceutical composition. In certain embodiments, the amount of the compound of formula (I) of the present invention is at least about 1% by weight based on the total weight of the pharmaceutical composition. In a particular embodiment, the amount of the compound of formula (I) of the present invention is at least about 5% by weight based on the total weight of the pharmaceutical composition. In other embodiments, the amount of the compound of formula (I) of the present invention is at least about 10% by weight based on the total weight of the pharmaceutical composition. In a further embodiment, the amount of the compound of formula (I) of the present invention is at least about 25% by weight based on the total weight of the pharmaceutical composition.

In certain embodiments, the pharmaceutical compositions of the present invention further comprise another therapeutic agent known to improve the therapeutic effect of cancer to help delay the growth of cancer cells. Preferably, the disclosed pharmaceutical or pharmaceutical composition further comprises a chemotherapeutic agent.

Accordingly, a third object of the present disclosure is directed to a method of treating a subject having cancer with respect to the disclosed compound of formula (I) or sodium of a pharmaceutical composition. The method comprises administering to the individual in need of treatment an effective amount of a compound of formula (I) or a pharmaceutical composition of the invention to inhibit growth of cancer cells in the subject. Cancers that can be treated by the disclosed methods are selected from the group consisting of leukemia, liver cancer, bladder cancer, rectal cancer, kidney cancer, Ovarian cancer, cervical cancer, endometrial cancer, prostate cancer, pancreatic cancer, lung cancer, breast cancer, melanoma, and squamous cell cancer (SCC). In one example, the individual has leukemia; in another example, the individual has liver cancer. The individual is preferably a mammal, preferably a human.

In certain embodiments, the method of the present invention further comprises administering to the individual another therapeutic agent known to improve the therapeutic effect of the cancer prior to, concurrently with, and/or after administration of the pharmaceutical composition of the present invention to help inhibit the treatment. Growth or metastasis of cancer cells in an individual. Preferably, the pharmaceutical or pharmaceutical composition of the invention is administered with another chemotherapeutic agent.

These and other features of the present disclosure will be more apparent from the following detailed description and appended claims. The above summary and the following detailed description are merely illustrative, and are not intended to limit the scope of the disclosure.

The above and other objects, features, advantages and embodiments of the present invention will become more <RTIgt; <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; Photographs of nuclear changes exhibited by HepG2 liver cancer cells after staining with Hearst staining, respectively; Figure 2 depicts images of HepG2 liver cancer cells treated with Compounds 2, 9 or 18, respectively, according to an embodiment of the present invention. , the results of cell cycle analysis; 3 is a view showing the results of analysis of apoptosis after treatment of HepG2 liver cancer cells with Compounds 2, 9 or 18, respectively, according to an embodiment of the present invention; and FIG. 4 is a view showing crotyl alcohol according to an embodiment of the present invention. In the Jurkat T cell mode activated by -12-tetradecanoate-13-acetate (PMA) plus ionomycin, (A) Compound 2 and (B) NOC15 treated human leukemia cells (Jurkat T, respectively) Analysis of cell survival after cells and human normal lymphocytes; Figures 5A-5C respectively depict treatment of human leukemia cells (Jurkat T cells) (A)0 with NOC15, respectively, according to an embodiment of the present invention, B) the results of cell cycle analysis after 48 hours of 24 or (C); the 5D plot is a quantitative analysis of the results of 5A-5C; the histogram of Fig. 6 depicts an embodiment according to the present invention, NC15 treats the results of cell survival analysis of human leukemia cells (Jurkat T cells) and human normal lymphocytes after about 48 hours (A) 24 or (B), respectively; and Figures 7A-7C respectively illustrate an embodiment according to the present invention. By way of NC15, human leukemia cells (Jurkat T cells) (A) 0, (B) 24 or (C) 4 were treated separately. After 8 hours, the results of cell cycle analysis; Figure 7D is a quantitative analysis of the results of Figures 7A-7C; Figure 8A depicts experimental animals subcutaneously injected with L1210 cells with NOC15 or NC15 according to an embodiment of the present invention. Survival curve of the experimental animal after treatment; FIG. 8B depicts the experimental animal after intraperitoneal injection of L1210 cells treated with NOC15 or NC15 according to an embodiment of the present invention, Survival curve of the experimental animal; FIG. 9 depicts experimental animals in which L1210 cells were injected subcutaneously or intraperitoneally, respectively, in vivo (A) tumor, (B) liver and (C) spleen weight according to an embodiment of the present invention. Curves with time; FIG. 10A is a graph showing changes in the number of white blood cells in the experimental animals after subcutaneous injection of L1210 cells in the experimental animals treated with NOC15 or NC15 according to an embodiment of the present invention; FIG. 10B is a graph showing the basis In one embodiment of the present invention, the number of white blood cell cells in the experimental animal after the intraperitoneal injection of the L1210 cell is treated with NOC15 or NC15; FIG. 11A depicts the experimental animal treated with NOC15 according to an embodiment of the present invention. The body weight is plotted against time; and FIG. 11B is a graph showing the body weight of the experimental animals treated with NC15 according to an embodiment of the present invention; the various features and components in the drawing according to the conventional operation mode The drawings are not to scale, the manner In addition, similar elements/components are referred to by the same or similar element symbols throughout the different drawings.

The description of the embodiments of the present invention is intended to be illustrative and not restrictive. The features of various specific embodiments, as well as the method steps and sequences thereof, are constructed and manipulated in the embodiments. However, other specific embodiments may be utilized to achieve the same or equivalent function and sequence of steps.

Although numerical ranges and parameters are used to define a broad range of values for the present invention, the relevant values in the specific embodiments have been presented as precisely as possible. However, any numerical value inherently inevitably contains standard deviations due to individual test methods. As used herein, "about" generally means that the actual value is within plus or minus 10%, 5%, 1%, or 0.5% of a particular value or range. Alternatively, the term "about" means that the actual value falls within the acceptable standard error of the average, depending on the considerations of those of ordinary skill in the art to which the invention pertains. Except for the experimental examples, or unless otherwise explicitly stated, all ranges, quantities, values, and percentages used herein are understood (eg, to describe the amount of material used, the length of time, the temperature, the operating conditions, the quantity ratio, and the like. Are all modified by "about". Therefore, unless otherwise indicated to the contrary, the numerical parameters disclosed in the specification and the appended claims are intended to be At a minimum, these numerical parameters should be understood as the number of significant digits indicated and the values obtained by applying the general carry method.

The scientific and technical terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the invention pertains, unless otherwise defined herein. In addition, the singular noun used in the specification encompasses the plural of the noun, and the plural noun of the noun is also included in the plural noun.

In the following, the term "inhibits, inhibiting, suppresses, suppressing" means applying the formula (I) to a body. a compound such that at least 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 99% of the cancer cells are stagnant and no longer replicated, Therefore, the number of cancer cells can be reduced or the tumor volume can be reduced. Therefore, the term "inhibition" is also used herein to mean killing cancer cells or inducing apoptosis of cancer cells.

As used herein, the term "treating or treating" means a condition that partially or completely prevents, slows, reduces, and/or manages a secondary disease or condition associated with a tumor. The term "treatment" means the administration or administration of a compound of formula (I) of the present invention to an individual exhibiting a condition associated with a secondary disease or condition associated with a tumor, to partially or completely prevent, slow, alleviate, delay the onset, inhibit progression. The severity is reduced and/or the occurrence of one or more symptoms of a secondary disease or characteristic associated with the tumor is reduced. Symptoms of secondary diseases or characteristics associated with the tumor include, but are not limited to, unexplained weight loss, fever, fatigue, pain, and changes in body function. Treatment may be administered to an individual who exhibits early signs of the above mentioned signs, diseases or conditions in order to reduce the risk of developing a second disease or condition associated with the tumor. If the one or more signs or clinical markers are reduced, the treatment is said to be effective. Alternatively, the treatment is said to be effective if the progression of the one or more signs or clinical markers is reduced or stopped.

The term "an effective amount" means that the dosage can achieve the desired therapeutic effect for cancer treatment after an appropriate administration period. In other words, the effective amount can achieve the desired effect of inhibiting cancer cells.

"compound", "composition The words "composition", "agent" or "medicine or medicament" are used interchangeably herein, and are meant to be applied locally when applied to a body (human or animal). / or a compound or composition that induces a desired pharmaceutical and/or physiological response systemically.

The term "administered, administered" or "administration" as used herein refers to the direct administration of the compound or composition, or the administration of a prodrug, derivative, or analog of the active compound. Alternatively, a substantial amount of the active compound can be formed in the subject to be administered.

The words "subject" or "patient" are used interchangeably herein to mean an animal (including a human) that is treatable by the compounds and/or methods. "Individual" or "patient" is used herein to encompass both male and female genders unless otherwise specified. Thus "individual" or "patient" encompasses any mammal, preferably a human, which may benefit from treatment with the compound.

At least a portion of the disclosure is due to the accidental discovery of two lipophilic N-substituted cantharidin derivatives which are not cytotoxic in themselves and which delay the growth characteristics of cancer cells, particularly white blood cells. The above findings suggest that the two N-substituted desmethylcantharidins in the present disclosure are useful as lead compounds for the therapeutic agents of cancer, including leukemia.

One aspect of the present disclosure is directed to a compound having the structure of formula (I): Wherein R is farnesyl or farnesyloxy.

In one embodiment, the compound of formula (I) is N-farneoxy-7-oxabicyclo[2,2,1]heptane-2,3-dimethylimine (Compound 9 or NOC 15) . In another embodiment, the compound of formula (I) is N-farnesyl-7-oxabicyclo[2,2,1]heptane-2,3-dimethylimine (Compound 18 or NC15) .

NOC 15 and NC 15 can be produced by the synthesis method shown in Example 1 of the present disclosure, respectively.

According to one embodiment, a NO-substituted demethylcantharidin derivative, including NOC15, is produced in the manner described in Scheme 1. In particular, using furan (3) and maleic anhydride (4) as starting materials, 5,6-anhydrodemethylcantharidin is produced by Diels-Alder reaction. 5). Next, the compound (5) is hydrogenated to obtain demethylcantharidin (2), which is further reacted with hydroxylamine hydrogen chloride in the presence of sodium methoxide in anhydrous methanol at room temperature to produce N-hydroxydemethylated quinone. Prime (6). Next, N-hydroxydesmethylcantharidin (6) is reacted with farnesyl bromide to form N-farnesyloxy-7-oxobicyclo[2,2,1 in the presence of potassium carbonate. Heptane-2,3-dimethylimine (Compound 9 or NOC 15).

Process 1

The reagents and conditions in Scheme 1 are as follows: (a) diethyl ether, room temperature, 48 hours; (b) 3 atmospheres of hydrogen, 10% Pd/C, THF, room temperature, 8 hours; (c) NaHCO ₃ , NH ₂ OH. HCl, MeOH, room temperature, 20 hours; (d) K ₂ CO ₃ , hexanes bromide, acetone, reflux for 8-10 hours.

According to another embodiment, the N-substituted demethylcantharidin derivative, including NC15, is made according to the procedure described in Scheme 2. In particular, furan (3) and maleic anhydride (4) are reacted in toluene to form dihydroxydesmethylcantharidin (14). Next, the compound (14) is hydrogenated to obtain the compound (15), which is further reacted with farnesyl bromide in the presence of potassium carbonate at room temperature to form N-farnesyl-7-oxygen. Ring [2,2,1]heptane-2,3-dimethylimine (Compound 18 or NC15).

Process 2

The reagents and conditions in Scheme 2 are as follows: (a) toluene, 80 ° C, 6 hours; (b) 3 atmospheres of hydrogen, 10% Pd / C, THF, room temperature, 8-48 hours; (c) K ₂ CO ₃ , farnesene bromide, acetone, reflux for 8-10 hours.

Such as leukemia, liver cancer, bladder cancer, rectal cancer, kidney cancer, ovarian cancer, cervical cancer, endometrial cancer, prostate cancer, pancreatic cancer, lung cancer, breast cancer, melanoma and squamous cell cancer (squamous cell cancer, growth of cancer cells SCC) or the like, may be between 1-20 μ M concentration of compound to inhibit the formula (the I), for example 1,2,3,4,5,6,7,8,9,10,11 , 12, 13, 14, 15, 16, 17, 18, 19 or 20 μM . According to one embodiment, 50% of the in vitro growth of liver cancer cells can be NOC15 8.2 μ M (Compound 9) inhibited; however, this concentration but does not harm normal liver cells. In another example, 50% of in vitro growth of liver cancer cells can be NOC15 10.7 μ M (Compound 9) inhibited; In yet another example, NOC15 8.3 μ M (Compound 9) inhibit 50% of the leukemic cells Growth in vitro. According to another embodiment, 50% of leukemia cells in vitro (L1210) growth may be NC15 1.6 μ M (Compound 18) inhibited. In another example, 50% of leukemia cells in vitro (L1210) growth may be NC15 2.6 μ M (Compound 18) inhibited

Accordingly, the present invention also provides a pharmaceutical or pharmaceutical composition useful for treating cancer. The cancer includes, but is not limited to, leukemia, liver cancer, bladder cancer, rectal cancer, kidney cancer, ovarian cancer, cervical cancer, endometrial cancer, prostate cancer, pancreatic cancer, lung cancer, breast cancer, melanoma, and scale Squamous cell cancer (SCC). In a preferred embodiment, the cancer is leukemia; in another preferred embodiment, the cancer is liver cancer.

The pharmaceutical compositions of the present invention comprise an effective amount of a compound of formula (I) and a pharmaceutically acceptable adjuvant. Generally, the compound of formula (I) of the present invention will comprise from about 0.1% to about 99% by weight based on the total weight of the pharmaceutical composition, based on the total weight of the pharmaceutical composition. In certain embodiments, the amount of the compound of formula (I) of the present invention is at least about 1% by weight based on the total weight of the pharmaceutical composition. In a particular embodiment, the amount of the compound of formula (I) of the present invention is at least about 5% by weight based on the total weight of the pharmaceutical composition. In other embodiments, the amount of the compound of formula (I) of the present invention is at least about 10% by weight based on the total weight of the pharmaceutical composition. In a further embodiment, the amount of the compound of formula (I) of the present invention is at least about 25% by weight based on the total weight of the pharmaceutical composition.

The pharmaceutical compositions of the present invention can be made according to methods commonly used in pharmacy, such as the Remington Pharmacopoeia (17th Ed., Alfonoso R. Gennaro, ed., Mack Publishing Company, Easton, Pa (1985). Pharmaceutically acceptable adjuvants refer to pharmaceutically acceptable adjuvants. Those that are compatible with the other ingredients in the formulation and are acceptable to biological systems.

The pharmaceutical compositions of the present invention can be formulated into solid dosage forms including, but not limited to, tablets, capsules and sachets (or sachets). Each tablet may contain various adjuvants (eg, microcrystalline cellulose, sodium citrate, calcium carbonate, dicalcium phosphate, and glycine); and various disintegrating agents (such as starch, alginic acid, and specific citrate). ) with particle binders (polyvinylpyrrolidone, sucrose, gelatin and acacia). In addition, a lubricant such as magnesium stearate, sodium lauryl sulfate, and talc may be contained in the tablet. Preferred materials associated therewith include sugars in lactose or milk as well as high molecular weight polyethylene glycols. The above solid dosage forms may also optionally include a coating or shell, such as a casing coating, and a coating to improve the rate of release of any of the pharmaceutically active ingredients. Examples of such coatings are well known to those skilled in the art. In one example, the pharmaceutical composition is formulated as a tablet. In another example, the pharmaceutical composition is a granule formulated to be filled in a soft or hard gelatin capsule or encapsulated in a biodegradable kit. When used in the form of oral suspensions and / or special effects (elixirs), the active ingredients may be combined with various sweeteners or flavors, colorants or dyes, and emulsifiers and/or suspensions may be added if desired. And diluents such as water, alcohol, propylene glycol, glycerin.

In certain embodiments, the pharmaceutical compositions of the invention are formulated as liquid dosage forms suitable for oral administration. Such liquid formulations may further include a buffer to maintain the pH. This liquid formulation can also be filled in a soft capsule. The liquid dosage form can be a solution, suspension, emulsion, microemulsion, precipitate or can carry the compound of the formula (I) of the present invention, its pharmaceutically acceptable derivatives, isomers, metabolites, salts Or any liquid medium of the solvate. This liquid can be designed to improve the formula (I) of the present invention The solubility of the compound or a pharmaceutically acceptable salt thereof to form a drug-containing emulsion or dispersion. When such a dosage form is employed, it is formed by mixing the active compound with at least one pharmaceutically acceptable adjuvant, including, but not limited to, the above adjuvants.

If administered parenterally, the compound of the invention may be formulated as a liquid pharmaceutical composition which may be sterile solution or suspension which can be administered by intravenous, intramuscular, subcutaneous or intraperitoneal injection. liquid. Diluents which can be used in the manufacture of such sterile injectable solutions or suspensions include, but are not limited to, 1,3-butanediol, mannitol, water, Ringer's solution, isotonic sodium chloride solution. Fatty acids (e.g., oleic acid) and their glyceride derivatives, or natural pharmaceutically acceptable oils (e.g., olive oil or rapeseed oil) can also be used to make solutions or suspensions for injectable use. Alcohols or carboxymethylcellulose or similar dispersing agents for dilution may also be included in such oily solutions or suspensions. Other commonly used surfactants (e.g., Tweens or Spans series) or emulsifiers, or agents commonly used in the pharmaceutical arts to enhance bioavailability can be used.

In certain other embodiments, the above-described pharmaceutical or pharmaceutical compositions of the present invention may also be formulated into a variety of dosage forms suitable for mucosal applications, such as buccal and/or sublingual drugs. A dosage unit to deliver a drug through the oral mucosa. A wide variety of biodegradable and pharmaceutically acceptable polymeric adjuvants can be used which provide the pharmaceutical compositions with acceptable adsorption and desired drug release patterns, as well as buccal and/or sublingual drugs. The active ingredient or other ingredients to be administered in the dosage unit are compatible. Generally speaking, the above high scores The sub-adjuvant comprises a hydrophilic polymer that adheres to the wet surface of the oral mucosa. Examples of polymeric adjuvants include, but are not limited to, acrylic acid polymers and copolymers; hydrolyzed polyvinyl alcohol; polyethylene oxides; polyacrylates; Vinyl polymers and copolymers; polyvinylpyrrolidone; dextran; guar gum; pectins; starch; and cellulosic polymers.

In addition, the pharmaceutical compositions of the present invention may also be formulated in a form suitable for topical application, which may be made using various dermatologically acceptable inert adjuvants known in the art. Dosage forms suitable for transdermal, topical, and mucosal administration include, but are not limited to, solutions, creams, lotions, ointments, gels, sprays, aerosols, dermal patches, and the like, which are known to those of ordinary skill in the art. . Typical inert adjuvants can be water, alcohol, polyvinylpyrrolidone, polyethylene glycol, mineral oil, stearic alcohol, and gel materials. All of the above dosage forms and adjuvants are well known in the pharmaceutical arts, and the choice of dosage form is not critical to the pharmaceutical compositions of the present invention.

Accordingly, a third object of the present disclosure is to provide a method of treating an individual suffering from or likely to have cancer. The method comprises administering to the individual a pharmaceutical composition of the invention described above (which comprises an effective amount of a compound of formula (I)) for inhibiting the growth or metastasis of cancer cells in the subject. Cancers suitable for treatment by the methods of the invention include, but are not limited to, leukemia, liver cancer, bladder cancer, rectal cancer, kidney cancer, ovarian cancer, cervical cancer, endometrial cancer, prostate cancer, pancreatic cancer, lung cancer, breast cancer, melanin Cell tumors and squamous cell carcinoma. In one example, the individual has leukemia; in another example, the individual has liver cancer.

In certain embodiments, the pharmaceutical compositions of the present invention comprising an effective amount of a compound of formula (I) can be administered to a subject via oral, intravenous or intramuscular injection at an effective level of about 1-100 mg / kg body weight. An effective amount to be administered to the individual per day is about 10, 20, 30, 40, 50, 60, 70, 80, 90 or 100 mg/kg body weight; preferably about 30-70 mg/kg body weight, for example about 30, 40, 50, 60, 70 mg / kg body weight; more preferably about 50 mg / kg body weight. These doses can be administered in a single administration or divided into multiple administrations within one day.

In certain embodiments, the method further comprises administering to the patient another drug known to improve the therapeutic effect, simultaneously with, before or after administration of the compound of formula (I) of the invention. The drug which can be administered together with the compound of the formula (I) of the present invention may be a chemotherapeutic drug such as an alkylating agent, a tocopherylase inhibitor, a cytotoxic agent, an anti-microtubule agent and the like.

In general, the pharmaceutical compositions of the invention (or compounds of formula (I)) can be administered by any suitable route, for example, orally (e.g., orally, in capsules, sachets, suspensions or elixirs), or parenterally. (parenterally) and the like are applied by appropriate methods. Parenterally administered means include systemic administration such as intramuscular injection, intravenous injection, subcutaneous injection or intraperitoneal injection. Alternatively, it may be applied by means of a film, such as topical skin application or inhalation (such as intrabronchial, intranasal, intraoral or nasal drops); or intrarectal administration. Administration may be carried out alone or in combination with conventional pharmaceutically acceptable adjuvants. in In a preferred embodiment, a pharmaceutical composition of the invention (comprising a compound of formula (I)) can be administered to an individual via oral means (e.g., through food).

In certain embodiments, an individual treated with a pharmaceutical composition of the invention has a lifespan of at least 10-40% longer than the control group, and both the tumor volume and the number of white blood cells are greater than the control group (ie, no Individuals treated with the compounds of the invention are small.

It will be understood that the dosage of the compound of the invention will vary from individual to individual, not only because of the particular compound or composition employed, the route of administration, the compound (either alone or in combination with one or more drugs) Other factors may also be affected by other factors, such as the state or severity of the disease to be treated; the age, sex or weight of the patient, the health of the patient; the severity of the pathological condition to be treated, and the patient at the same time Other medical or special dietary content to be performed; and other factors conceivable by those of ordinary skill in the art; and the medical personnel responsible for care may ultimately determine the appropriate dosage based on these factors. The dosage and form of administration can be adjusted to provide a preferred therapeutic response. A therapeutically effective amount also refers to a toxic or detrimental effect of a compound or composition that is less than the therapeutic benefit it brings. In preferred instances, the compounds or compositions of the invention, when administered, should be administered in an appropriate dosage for a period of time to reduce the number and/or severity of symptoms.

The following examples are presented to illustrate certain aspects of the present disclosure and to assist those skilled in the art to understand and practice the present disclosure. However, the scope of the disclosure is not limited to these embodiments.

Example

Materials and Methods

Cell culture and animals

Cell lines used in the present invention include human hepatoma cell line HepG2, human bladder cancer cell line BFTC905, human rectal cancer cell lines HT29 and SW480, and human leukocyte cell lines HL60 and Jurkat T; and mouse lymphocytic leukemia cell line L1210. Each cell lines were cultured and maintained in a DU will be Igor's modified medium (Dulbecco 's modified Eagle media ( DMEM)) or RPMI 1640 medium, added with 10% heat-inactivated fetal calf serum, 1% penicillin / streptavidin And cultured in 5% CO ₂ /95% air at 37 °C.

DBA/2 mice (approximately 7 weeks old, 18-22 grams) were purchased from Lesco Biotech and were maintained in an environment free of pathogens and were free to consume water and feed. L1210 cells are implanted into the animal by subcutaneous or intraperitoneal injection to establish a homologous transplant tumor. The management and use of all experimental animals are in accordance with the rules set by the Animal Management Committee of Taipei Veterans General Hospital.

MTT analysis

MTT assay is a colorimetric assay that quantifies the enzyme (ie, reductase) in a living cell with a yellow tetrazolium compound - 3-(4,5-dimethylthiazole)-2,5-diphenyl bromide. Reduction of 3-[4,5-dimethylthiazol]-2,5-diphenyltetrazolium bromide (MTT) into a (formazane) activity. This reduction can only be carried out when the cells are alive; therefore, MTT assays are often used to assess the ability of cells to survive and proliferate. Briefly, the cells and test compounds at various concentrations (e.g., of Formula (1) compound) 48 hours; after addition of MTT dye (500 μ g / ml), and so that the reduction reaction for 1 hour; followed by addition of 500 μ L Isopropyl alcohol to terminate the reduction reaction. The absorbance of the sample at a wavelength of 540 nm was measured using a spectrophotometer. When the cell survival rate of the untreated drug is 100%, the drug concentration which can inhibit 50% of cell growth is the IC ₅₀ value. All values are expressed as the mean ± standard deviation of 3 independent experiments.

Jurkat T cell pattern activated by crotonol-12-tetradecanoate-13-acetate (PMA) plus ionomycin

Cell viability experiments were performed using 96-well plates. 100 μl of cell suspension (NHL or Jurkat T cells, 5×10 ³ cells/well, serum-free medium) was inoculated into the wells, and the 96-well plates were cultured in an incubator for 24 hours (pre-culture for 22 hours, then Incubation with PMA 5 ng/ml plus ionomycin 250 ng/ml for 2 hours). Different concentrations of demethylcantharidin (0, 2, 4, 15, 30 and 60 μM) or NOC 15 (0, 0.25, 0.5, 1, 2 and 4 μM) were then added to the medium in the wells. After 24 hours of drug action, the cell viability of NHL and Jurkat T cells was measured by Cell Counting Kit-8 (CCK-8, Sigma), and methylcantharidin and NOC15 were extracted to NHL cells and Jurkat T cells. The half-inhibitory concentration (IC ₅₀ ).

Hearst 33528 staining (Hoechst 33528 staining)

The Hurst 33528 staining was carried out according to a known method (Huang et al., Cytotechnology 2009, 59, 201-208). Briefly, 40 μ M to demethyl-to cantharidin, NOC15 or NC15 cells were treated for 48 hours and then rinsed with PBS and then stained in Hoechst 33528 (final concentration 10 μ g / mL), and then fluorescence View the slide under the microscope. Cells that exhibit small nuclei or cells that exhibit high fluorescence intensity or nucleus fragments are considered to belong to apoptotic cells.

Cell cycle analysis was acquired from the culture medium of cells incubated (compound of the present invention, whether or not through (i.e., after 10-60 μ M of demethylated cantharidin, NOC15 or NC15 for about 48 hours) pre-treatment), then, in The cells were treated with 75% ice-cold alcohol at 4 ° C for at least one night to immobilize the cells. Thereafter, rinsed with PBS and RNA enzyme (DNAase, 200μg / mL, 500 μ L) at 37 [deg.] C cells were treated for about 1.5 hours, after centrifugation the cell pellet was resuspended in an appropriate amount of propidium iodide (propidium iodide, PI (40 μg/ml), and then incubated at 20-25 ° C for about 20 minutes in the dark, and then cells of different cycles were measured by flow cytometry and counted.

The percentage of apoptotic cells was determined by flow cytometry with a fifth membrane-fixing protein kit (annexin-V/FITC kit/propidium iodine (PI)) with green fluorescent protein according to the manufacturer's protocol. To help detect apoptotic cells, the treated cells were first centrifuged at 1,000 g for 5 minutes, then resuspended and washed with PBS, followed by a fifth membrane-adhesin/propidium iodide (annexin-V/PI). ) to perform dyeing.

Produce homologous tumors and in vivo therapy

Start of the experiment on day 1, intraperitoneally or subcutaneously injecting the L1210 cells (1x10 ⁶ cells per injection) into the experimental animals. After 7 days, the test group animals (i.e., NOC15 or NC15) were administered by intraperitoneal injection for 7 consecutive days. The body weight of the experimental animals was measured and recorded daily during the test.

After the test was completed, the experimental animals were sacrificed, and the tumor, liver, and spleen were taken out and weighed, and the number of white blood cells was measured.

Example 1 Synthesis of a Compound of the Invention

1.1 Synthesis of N-farnesyloxy-7-oxabicyclo[2,2,1]heptane-2,3-dimethylimine (Compound 9 or NOC15)

The NOC15 compound was synthesized in the manner described in Scheme 1 below.

Wherein (a) diethyl ether, room temperature, 48 hours; (b) 3 atmospheres of hydrogen, 10% Pd/C, THF, room temperature, 8 hours; (c) NaHCO ₃ , NH ₂ OH. HCl, MeOH, room temperature, 20 hours; (d) K ₂ CO ₃ , hexanes bromide, acetone, reflux for 8-10 hours.

7-oxobicyclo[2,2,1]heptene-2,3-dicarboxylic anhydride (compound 5)

Furan (40 ml, 550 mmol) and maleic anhydride (10 g, 102 mmol) were mixed in diethyl ether and reacted at room temperature for about 48 hours, then the precipitate was collected and dried. The colorless solid of Compound 5 (yield: 93.3%) having a melting point of about 121-122 ° C and its structure was identified by ¹ H-NMR and ¹³ C-NMR. The spectral data are as follows: ¹ H NMR (CDCl ₃ ) δ 6.58 (s, 2H, H-5, 6), 5.47 (s, 2H, H-1, 4), 3.19 (s, 2H, H-2, 3); ¹³ C NMR (CDCl ₃ ) δ 170.1 (C =O), 137.2 (C-5, C-6), 82.4 (C-1, C-4), 48.9 (C-2, C-3).

^{LC-MS (ESI -, m} / z) C 8 H 6 O 4: theoretical value of 166.03, the actual measurement value of 164.90 [MH] ^-.

7-oxobicyclo[2,2,1]heptane-2,3-dicarboxylic anhydride (compound 2)

10% Pd/C (470 mg) was added to a solution of the above compound 5 (4.7 g, 28.3 mmol) in THF (200 ml), and the mixture was stirred at room temperature under atmospheric pressure of 3 atm. hour. Celite 545® reaction mixture was filtered, and dried in vacuo to give compound 2 may be of a colorless solid (4.3 g, yield: 91.4%) having a melting point of about 116-118 deg.] C, and in ¹ H-NMR and ¹³ C-NMR identified Its structure and spectral data are as follows: ¹ H NMR (CDCl ₃ ) δ 5.05 (dd, J = 2.3, 2.2 Hz, 2H, H-1, 4), 3.18 (s, 2H, H-2, 3), 1.91 1.89 (m, 2H, H-5, 6), 1.65-1.63 (m, 2H, H-5, 6); ¹³ C NMR (CDCl ₃ ) δ 171.1 (C=O), 80.2 (C-1, C -4), 50.6 (C-2, C-3), 28.1 (C-5, C-6); LC-MS (ESI ^- , m/z) C ₈ H ₈ O ₄ : theoretical value 168.04, actual The measured value is 166.86 [MH] ^- .

N-hydroxy-7-oxobicyclo[2,2,1]-5-heptane-2,3-dimethylimine (Compound 2)

To a solution of the aforementioned compound 2 (5.04 g, 30 mmol) in anhydrous methanol (200 ml), sodium methoxide (1.62 g, 30 mM) and hydroxylamine hydrogen chloride (2.08 g, 30 mM) . The reaction mixture was stirred at room temperature for about 20 hours, the reaction mixture was filtered, dried and concentrated in vacuo to CHCl ₃ Recrystallization available compound 6 as colorless crystals (yield: 63%). Its melting point is about 168-169 ° C, and the isolated compound is analyzed and identified by ¹ H-NMR and ¹³ C-NMR. The spectral data are as follows: ¹ H NMR (D ₂ O) δ 4.93 (s, 2H, H-1, 4), 3.21 (s, 2H, H-2, 3), 1.89-1.86 (m, 2H, H-5) , 6), 1.78-1.74 (m, 2H, H-5, 6); ¹³ C NMR (D ₂ O) δ 177.0, 78.8, 46.9, 28.0; LC-MS (ESI ^- , m/z) C ₈ H ₉ NO ₄ : The theoretical value is 183.05, and the actual measured value is 206.00 [MH] ^- .

N-farnesyloxy-7-oxabicyclo[2,2,1]heptane-2,3-dimethylimine (Compound 9 or NOC15)

To a solution of the aforementioned compound 6 (1 mmol) in dry acetone, bromofurane (1 mmol) and K ₂ CO ₃ (3 mmol) were added, and the mixture was refluxed for 8 to 10 hours. The reaction mixture was filtered, concentrated in vacuo purified by silica gel column (ethyl acetate / hexane as a lotion), a Compound 9 as a colorless liquid (yield: 28%). The isolated compound was subjected to structural analysis and identification by ¹ H-NMR and ¹³ C-NMR. The spectral data are as follows: ¹ H NMR (D ₂ O) δ 5.42 (td, J = 0.9, 7.7 Hz, 1H, H-2'), 5.11-5.07 (m, 2H, H-6', 10'), 4.85 (q, J = 2.4 Hz, 2H, H-1, 4), 4.62 (d, J = 7.7 Hz, 2H, H-1 '), 2.83 (s, 2H, H-2, 3), 2.09-2.04 (m, 6H, H-5, 6, H-4', 8'), 1.99-1.96 (m, 2H, H-5'), 1.89-1.86 (m, 2H, H-6'), 1.72 ( d, J-0.7Hz, 3H, CH ₃ ), 1.68 (s, 3H, CH ₃ ), 1.63-1.62 (m, 2H, H-5, 6), 1.60 (s, 3H, CH ₃ ), 1.59 ( s, 3H, CH ₃ ); ¹³ C-NMR (CDCl ₃ ): δ 171.6, 147.3, 135.3, 131.3, 124.3, 123.5, 116.1, 78.5, 73.0, 47.4, 39.65, 39.63, 28.7, 26.7, 26.1, 25.7, ^{17.7,16.6,16.0; LC-MS (ESI -} , m / z) C 23 H 33 NO 4: theoretical value of 387.24, the actual measurement value of 410.21 [m + Na] ^-.

1.2 Synthesis of N-farnesyl-7-oxabicyclo[2,2,1]heptane-2,3-dimethylimine (Compound 18 or NC15)

The NC15 compound was synthesized as described in Scheme 2 below.

Wherein: (a) toluene, 80 ° C, 6 hours; (b) 3 atmospheres of hydrogen, 10% Pd / C, THF, room temperature, 8-48 hours; (c) K ₂ CO ₃ , bromide bromide , acetone, reflux for 8-10 hours.

7-oxobicyclo[2,2,1]-5-heptene-2,3-dimethylguanamine (compound 14)

A solution of maleimide (Compound 13 ) (1.5 g, 15 mmol) in toluene (30 mL) was heated to 80 ° C then added furan (5.4 mL, 75 mmol) and at 80 ° C After stirring for 6 hours, the mixture was cooled to room temperature, and then the precipitate was collected, and then purified on a silica gel column (ethyl acetate / n-hexane as a washing liquid) to obtain a colorless crystal of compound 14 (2.1 g, yield: 87%) Its melting point is about 162-163 ° C, and its structure is identified by ¹ H-NMR and ¹³ C-NMR. The spectral data are as follows: ¹ H-NMIR (CDCl ₃ ): δ 8.13 (s, 1H, NH), 6.50 ( s, 2H, H-5, 6), 5.29 (s, 2H, H-1.4), 2.87 (s, 2H, H-2, 3); ¹³ C-NMR (CDCl ₃ ): δ 176.2, 136.8, 81.2 , 48.9; LC-MS (ESI ^- , m / z) C ₈ H ₇ NO _{3 The} theoretical value is 165.04, the actual measured value is 164.02 [MH] ^- .

7-oxobicyclo[2,2,1]heptane-2,3-dimethylguanamine (compound 15)

10% Pd/C (50 mg) was added to a solution of the above compound 14 (0.5 g, 2.8 mmol) in THF (20 ml), and the mixture was stirred for about 4 hours at room temperature in the presence of hydrogen. . Celite545® reaction mixture was filtered, and dried in vacuo to afford compound may be 15 (i.e., NC15) of a colorless solid (424 mg, yield: 84%) having a melting point of about 188-190 deg.] C, and at ¹³ and ¹ H-NMR The structure was identified by C-NMR, and the spectral data were as follows: ¹ H-NMR (CDCl ₃ ): δ 8.60 (s, 1H, NH), 4.91 (dd, J = 2.4, 3.3 Hz, 2H, H-1, 4), 2.92 (s, 2H, H-2, 3), 1.89-1.85 (m, 2H, H-5, 6), 1.61-1.57 (m, 2H, H-5, 6); ¹³ C-NMR (CDCl ₃ ): δ 177.2, 79.1, 51.3, 28.5; LC-MS (ESI ^- , m/z), C ₈ H ₉ NO _{3 The} theoretical value is 167.06, the actual measured value is 166.03 [MH] ^- .

N-farnesyl-7-oxabicyclo[2,2,1]heptane-2,3-dimethylimine (Compound 18 or NC15)

In a solution of the aforementioned compound 15 (1 mmol) in dry acetone (25 ml), br.nibrane (1 mmol) and K ₂ CO ₃ (3 mmol) were added and the mixture was refluxed. 10 hours. The reaction mixture was filtered, and then purified, mjjjjjjj The isolated compound was subjected to structural analysis and identification by ¹ H-NMR and ¹³ C-NMR. The spectral data are as follows: ¹ H-NMR (CDCl ₃ ): δ 5.10-5.02 (m, 3H, H-2', 6', 10'), 4.84 (dd, J = 2.2, 2.3 Hz, 2H, H-1 , 4), 4.03 (d, J = 7.0 Hz, 2H, H-1 '), 2.82 (s, 2H, H-2, 3), 2.06-2.01 (m, 4H, H-4', 8') , 1.97.1.22 (m, 4H, H-5', 9'), 1.84-1.81 (m, 2H, H-5, 6), 1.75 (s, 3H, CH ₃ ), 1.67 (s, 3H, CH ₃ ), 1.60 (s, 3H, CH ₃ ), 1.59 (s, 3H, CH ₃ ), 1.58-1.55 (m, 2H, H-5, 6); ¹³ C-NMR (CDCl ₃ ): δ 176.8, 140.9, 135.3, 131.2, 124.3, 123.7, 117.2, 78.9, 50.0, 39.7, 39.5, 37.0, 28.6, 26.8, 26.3, 25.7, 17.7, 16.4, 16.0; LC-MS (ESl ^- , m/z), C ₂₃ The theoretical value of H ₃₃ NO ₃ is 371.25, and the actual measured value is 394.2 [MH] ^- .

Example 2 Analysis of anti-proliferative activity of the compounds of the present invention

The cell survival assay was used to analyze the effects of the compound of the formula (I) of the present invention on the growth of various cancer cells including liver cancer HepG2 cells, bladder cancer BFTC905 cells, rectal cancer HT29 cells, SW480 cells and leukemia HL60 cells, and the compounds 2 and Other conventional anticancer drugs (including 5-FU, cisplatin and doxorubicin) were used as a control group. The results are shown in Table 1.

Table 1 Effect of the compound of formula (I) of the present invention on the growth of various cancer cells

As shown in Table 1, both NOC15 and NC15 compounds were toxic to the five cells tested, and their cytotoxicity was at least as low as that of the control compounds (eg, compound 2, 5-FU, cisplatin, and doxorubicin). Up and down, or better. In addition, 5-FU and cisplatin were also observed to inhibit the growth of leukemia cell line HL60, but were inhibited by both NOC15 and NC15 compounds.

Example 3 NOC15 and NC15 inhibit human liver cancer cell proliferation

3.1 Cell survival test

In this embodiment, the effects of NOC15 and NC15 compounds inhibiting the growth of human hepatoma cells HepG2 were tested according to the cell survival assay described in Materials and Methods. The results are shown in Table 2.

Table 2 NOC15 and NC inhibit liver cancer cell growth

As can be seen from the data in Table 2, the activity of NOC15 and NC15 is at least 3-4 times that of Compound 2, that is, only about 1/3 or 1/4 of Compound 2 is required, and NOC15 or NC15 can be the same as Compound 2. Degree of cytostatic effect.

3.2 Cell nuclear type changes

To further explore the causes of apoptosis in NOC15 and NC15, HepG2 cells were treated with Compound 2, NOC15 and NC15 for about 48 hours, respectively, and then stained with Hearst 33528 and examined for cell morphology by fluorescence microscopy.

Fig. 1 is a photograph of the nucleus after treatment with Compound 2, NOC15 and NC15. Compared with the control group, the cells treated with Compound 2, NOC15 or NC15 were observed to be significantly atrophied, and chromatin deposition and DNA fragmentation occurred.

3.3 Cell cycle and apoptosis analysis

Compound of 10-60 μ M 2, NOC15 or NC15 treated HepG2 cells for about 48 hours, followed by flow cytometry analysis of cell cycle and apoptosis of cells, and PI staining to the cells of the G0 / G1 period. The results are shown in Figures 2 and 3, respectively.

As shown in Figure 2, only a few cells in the control group (without compound treatment) developed apoptosis, as did the cell population treated with low concentration (10 μM) of Compound 2. As the concentration of Compound 2 increased from 20 μM to 40 μM, the apoptotic cell ratio increased from 1.6% to 16.7%. Similarly, when cells were treated with 10-60 μM of Compound 18, the apoptotic cell ratio increased from 3.7% to 19.8%. In addition, compounds 9 and 18 (i.e., NOC 15 and NC 15) can induce cells to enter the G2/M cycle.

The data in Figure 3 shows that the cell death caused by NOC15 and NC15 is related to the dose used, and the percentage of apoptotic cells increases with the concentration of NOC15 and NC15 used. As for Compound 2, the cytotoxicity of the Constitution was observed only when the concentration was as high as 60 μM.

Example 4 NOC15 inhibits proliferation of human leukemia cells

4.1 Cell survival test

In this embodiment, the Jurkat T cell pattern activated by crotyl alcohol-12-tetradecanoate-13-acetate (PMA) plus ionomycin is used to evaluate NOC15 on human leukemia cells (Jurkat T cells). ) and the effects of normal lymphocytes (NHL).

As shown in Figure 4A, Compound 2 is almost non-toxic to normal lymphocytes (NHL) but kills human leukemia cells (Jurkat T cells). In addition, if Jurkat T cells are previously treated with crotyl alcohol-12-tetradecanoate-13-acetate (PMA) plus ionomycin (known crotyl-12-tetradecanoate-13-B The acid ester (PMA) protects cells from apoptosis and slightly increases cell viability. Similar results can also be seen in the NOC15 treated cell population (Fig. 4B). The above results indicate that both compound 2 and NOC15 are also Tumor cell growth is inhibited via a mitogen-activated protein kinase (MAPK) pathway.

4.2 Cell cycle analysis

Leukemia cells were treated with 1.4 μM NOC15 for about 24 or 48 hours, followed by analysis of cell cycle by flow cytometry and staining of G0/G1 cycle cells with PI. The results are shown in Figures 5A-5D, respectively.

It can be found that only a small number of control group cells are in the G0/G1 cycle (Fig. 5A), but if treated with NOC15 for 24 or 48 hours, the number of cells in the G0/G1 cycle will increase (Fig. 5B, 5C); After 48 hours of treatment of leukemia cells with NOC15, the number of cells remaining in the G2/M cycle was also increased. The data of Fig. 5A-5C is quantized and shown in Fig. 5D.

Example 5 NC15 inhibits proliferation of human leukemia cells

5.1 Cell survival test

The toxicity of NC15 on normal lymphocytes (NHL) and human leukemia cells is shown in Figure 6. Similar to the effects of Compound 2 and NOC15, NC15 is almost non-toxic to normal lymphocytes (NHL), but kills human leukemia cells (Jurkat T cells), and its toxicity is positively correlated with dose. After 24 hours of treatment with NC15, the viable cell ratio decreased as the concentration of NC15 increased (Fig. 6A), and cells were treated with NC15 for 48 hours, and the results were similar.

5.2 Cell cycle analysis

2.513 μ M to the NC15 treated leukemia cells of about 24 or 48 hours, followed by cell cycle analysis by flow cytometry and PI staining to the cells of G0 / G1 period. The results are shown in Figure 7, respectively.

Most of the control group cells (not receiving any drug treatment) were in the G0/G1 cycle, and the number of cells below the G0/G1 cycle (subG1 cycle) began to increase after the cells were treated with NOC15 for 24 hours (Fig. 7B). A significant amount was reached after 48 hours (Fig. 7C); it was also found that the number of cells remaining in the G0/G1 cycle was significantly reduced compared to the control group (Fig. 7D). The data of Fig. 7A-7C is quantized and shown in Fig. 7D.

5.3 NOC15 and / or NC15 can reduce tumors in animals implanted with human leukemia cells L1210

L1210 cells were seeded in experimental animals according to the procedures in the "Materials and Methods" section. In the in vivo treatment test, experimental animals were administered 18 mg/kg of NOC15 or NC15 via intraperitoneal injection for 7 consecutive days. Then, the tumor size, and the liver, spleen weight, and white blood cell count of the experimental animals were measured at the designated time.

The effect of NOC15 or NC15 on subcutaneous or intraperitoneal inoculation of L1210 cells is shown in Figure 8. It can be found that both NOC15 and NC15 can effectively prolong the life of the animal, and the life cycle of the animals receiving the subcutaneous injection of L1210 cells is extended by about 40%; for the animals inoculated with L1210 cells per abdomen, the life cycle is prolonged. About 14-18%. In addition, SC-tumor mice treated with NOC15 or NC15 showed a significant reduction in tumor weight (Fig. 9A), and their liver and spleen weights were significantly improved (compared to the control group) (9B) , 9C picture). In addition, the number of white blood cells decreased significantly on the 14th day after treatment compared to the control group (Fig. 10).

The biggest disadvantage of treating cancer with traditional chemotherapy drugs is that it will cause the patient to lose weight. Once the patient is unable to maintain normal weight, it will also be indirectly guided. The treatment effect is not good. In the present embodiment, the inventors have found that administering 18 mg/kg of NOC15 to a patient during a cancer treatment can gradually improve the patient's weight loss during the cancer treatment (Fig. 11A); in the same case, NC15 The animal's body weight was restored to the same extent as the control group animals (Fig. 11B). The data show that in addition to inhibiting tumor growth, NC15 does not have the disadvantage that conventional chemotherapeutic drugs cause weight loss in users. Therefore, NC15 has the potential to develop into a new generation of anticancer drugs (especially anti-leukemia).

In summary, the two novel lipophilic N-substituted demethylcantharidin derivatives synthesized by the present invention have the activity of being toxic to kill a plurality of cancer cells, and do not cause damage to normal cells, so they are developed into new ones. The potential of a generation of anticancer drugs (especially anti-leukemia).

It will be understood that the above-described embodiments and examples are merely illustrative, and that those skilled in the art can align various modifications. The description, examples, and materials set forth above are intended to be illustrative of the present invention and are illustrative of the invention. The present disclosure has been disclosed in the above embodiments, but it is not intended to limit the disclosure, and any person skilled in the art can make various changes and refinements without departing from the spirit and scope of the disclosure. The scope of protection of the disclosure is subject to the definition of the scope of the patent application.

Claims (7)

a compound of formula (I), Wherein R is farnesyl or farnesyloxy. A pharmaceutical composition for treating an individual having cancer comprising an effective amount of a compound of formula (I) as claimed in claim 1 and a pharmaceutically acceptable adjuvant therefor. The pharmaceutical composition according to claim 2, wherein the cancer is selected from the group consisting of leukemia, liver cancer, bladder cancer, rectal cancer, kidney cancer, ovarian cancer, cervical cancer, endometrial cancer, prostate Cancer, pancreatic cancer, lung cancer, breast cancer, melanoma, and squamous cell cancer (SCC). The pharmaceutical composition according to claim 3, wherein the cancer is leukemia. The pharmaceutical composition according to claim 3, wherein the cancer is liver cancer. The pharmaceutical composition according to claim 2, wherein the compound of the formula (I) is N-farnesyloxy-7-oxabicyclo[2,2,1]heptane-2,3-dimethylimine . The pharmaceutical composition according to claim 2, wherein the compound of the formula (I) is N-farnesyl-7-oxabicyclo[2,2,1]heptane-2,3-dimethylimine.