HK1096382A - Jasmonate derivative compounds, pharmaceuticals compositions and methods of use thereof - Google Patents

Abstract

The present invention provides novel jasmonate derivative compounds, methods for their preparation, pharmaceutical compositions including such compounds, and methods of using these compounds and compositions, especially as chemotherapeutic agents for treatment of cancers, especially mammalian cancers.

Description

Jasmonate derivative compounds, pharmaceutical compositions and methods of use thereof

Technical Field

The present invention relates to the field of jasmonate derivative compounds, methods of preparing the same, pharmaceutical compositions comprising these compounds, and methods of using these compounds and compositions, particularly as chemotherapeutic agents for treating cancer, especially in mammals.

Background

Jasmonates are a family of plant stress hormones, derived from linolenic acid by the stearic acid pathway and found in trace amounts in many edible plants. Stress hormones, such as the jasmonate family, have evolved in plants and are released in response to conditions such as excessive ultraviolet radiation, osmotic stimuli, heat shock, and pathogen attack to initiate a cascade of responses that are appropriate as a result. Examples of members of the jasmonate family are jasmonic acids that are crucial for intracellular signal transduction in response to injury, and methyl jasmonate that causes induction of protease inhibitors to allow them to accumulate at low concentrations in response to traumatic and pathogenic attacks. Jasmonates have been patented for many uses in plant growth and crop improvement, but their use as pharmaceuticals has not been previously known. The use of jasmonates as a treatment for cancer in mammals is disclosed in U.S. Pat. No.6,469,061, which is incorporated herein by reference in its entirety. In U.S. Pat. No.6,469,061, jasmonates have been shown to be directly cytotoxic to different types of human cancer cells derived from breast, prostate, skin and blood cancers. Although jasmonates cause Molt-4 cell death in human leukemias, they do not damage normal lymphocytes.

The data collected later also showed that jasmonates did not damage healthy red blood cells (see WO 02/080890). In U.S. Pat. No.6,469,061, in particular, a jasmonate compound methyl jasmonate was shown to be effective in preventing the development of murine lymphomas. See also fingurut, o. and e.flescher.2002. "plant stress hormones inhibit proliferation and induce apoptosis in human cancer cells", leukemia 16: 608-.

The pharmacological activity of jasmonates makes it an attractive candidate for therapeutic drugs for cancer therapy. Since only a few jasmonate derivatives have been reported (see, for example, Ishii et al, Leukemia, 1-7(2004)) (Ishii et al, Leukemia, 1-7(2004)), there is a need in the art to develop jasmonate derivative compounds that are effective chemotherapeutic agents and have high specificity for cancer cells.

The present invention addresses this need while providing other benefits.

Summary of The Invention

The present invention refers to jasmonate derived compounds, especially those that are halogenated. Such compounds include "methyl dibromojasmonate" or "MJDB", and "methyl tetrabromo jasmonate" or "MJTB". These compounds are significantly more effective than the most effective jasmonate disclosed in U.S. patent No.6,469,061, methyl jasmonate. Jasmonate derivatives, such as MJDB and MJTB, exert selective cytotoxicity on cancerous lymphocytes from patients while leaving normal lymphocytes, as described below.

The invention also includes salts, hydrates, solvates, polymorphs, optical isomers, diastereomers, and any mixtures thereof, of the compounds of the invention, particularly MJDB and MJTB.

The compounds of the present invention have the following general formula:

general formula I

Wherein:

n is 0, 1 or 2;

R₁is OH, C₁To C₁₂Alkoxy radical, C₁To C₁₂Substituted alkoxy, aryloxy, O-glucosyl or imino;

R₂is OH, C₁To C₁₂Alkoxy radical, C₁To C₁₂Substituted alkoxy, O-glucosyl, oxo, alkyl or imino;

R₃、R₄、R₅、R₆、R₇a, B, C, D and E are each independently H, halogen, OH, C₁To C₁₂Alkoxy radical, C₁To C₁₂Substituted alkoxy, aryloxy, O-glucosyl, C₁To C₁₂Alkyl or C₁To C₁₂A substituted alkyl group;

wherein R is₁And R₂Or R₁And R₄May together form an optionally substituted lactone;

wherein C is₃:C₇、C₄:C₅And C₉:C₁₀The bonds between may independently be double or single bonds;

provided that R is₃、R₄、R₅、R₆、R₇At least one of A, B, C, D and E is a halogen; and provided that, if A is the only halogen in the compound, A is not fluorine;

or a derivative of said formula, wherein the derivative has at least one of the following:

is located at C₃Lower acyl side chain (free acid or ester or conjugate) of (a) at C₆Or a hydroxy (free hydroxy or ester) moiety of, or at C₇N-pentenyl or n-pentyl side chains;

including salts, hydrates, solvates, polymorphs, optical isomers, enantiomers, diastereomers, and mixtures thereof.

More particularly, preferred compounds of the invention (formula I) are those C₉And C₁₀The bond between (A) and (B) is a single bond. Other preferred compounds are R₂Is oxo. Still other preferred compounds are at least R₆And R₇One is bromine, iodine, fluorine or chlorine. Even more preferred is R₆And R₇All selected from the group consisting of bromine, iodine, fluorine and chlorine. However, more preferably R₆And R₇All bromine compounds.

A further preferred aspect of the invention is A, B, R₆And R₇Is a compound of bromine, iodine, fluorine or chlorine. A, B, R is even more preferred₆And R₇Each of which is bromine.

Other preferred compounds of the invention are R₁Is alkoxy. And on the other hand, R₃、R₄And R₅Each is H (hydrogen). Meanwhile, C, D and E are each H.

One of the most preferred compounds of the present invention is methyl dibromojasmonate (MJDB). According to formula I, MJDB is: n is 0; c₃:C₇、C₄:C₅And C₉:C₁₀The bond between (A) and (B) is a single bond; r₁Is methoxy; r₂Is oxo; r₃、R₄、R₅A, B, C, D and E are each H; r₆And R₇Each is bromine.

Another most preferred compound of the invention is methyl tetrabromo-jasmonate (MJTB). According to formula I, MJDB is: n is 0; c₃:C₇、C₄:C₅And C₉:C₁₀The bond between (A) and (B) is a single bond; r₁Is methoxy; r₂Is oxo; r₃、R₄、R₅C, D and E are each H; and A, B, R₆And R₇Each is bromine.

Other preferred compounds of the invention are those wherein n is 0; c₃:C₇、C₄:C₅And C₉:C₁₀The bond between (A) and (B) is a single bond; r₁Is methoxy; r₂Is oxo; r₃、R₄、R₅A, B, C, D and E are each H; and any of the following:

e)R₆and R₇Each is fluorine (designated compound "MJS 99");

b)R₆and R₇Each iodine (designated compound "MJS 85fl 4");

c)R₆and R₇Each is chloro (designated compound "MJS 81fl 3");

d)R₆and R₇One is bromo and the other is hydroxy (designated compound "NJ-63"); or

e)R₆And R₇One is iodine and the other is methoxy (designated compound "MJS 72f 5").

The present invention also refers to pharmaceutical compositions containing a pharmaceutically acceptable carrier and having as active ingredient a compound of the invention as described above. Preferred compositions have an active ingredient MJDB or MJTB. Preferably, in the pharmaceutical composition, the active ingredient is dissolved in any acceptable lipid carrier. Further, according to a preferred embodiment of the present invention, the composition further comprises at least one other chemotherapeutic agent.

The present invention also provides a method of reducing the growth of cancer cells comprising exposing the cancer cells to a therapeutically effective amount of a compound of the invention as described herein.

Further, the present invention provides a method of treating cancer comprising administering to a patient a pharmaceutical composition comprising as an active ingredient a therapeutically effective amount of a compound of the present invention as described herein. According to a preferred embodiment, the cancer is a cancer of a warm-blooded vertebrate, more preferably a mammal, most preferably a human.

The cancer includes: carcinomas (carcinoma), sarcomas, adenomas, hepatocellular carcinomas, hepatoblastoma, rhabdomyosarcoma, esophageal carcinoma, thyroid carcinoma, malignant gangliomas, fibrosarcomas, myxosarcomas, liposarcomas, chondrosarcomas, osteosarcomas, chordoma, angiosarcomas, endotheliosarcomas, lymphangiosarcomas, synoviomas (synovioamas), Ewing's tumors, leiomyosarcomas, rhabdoid intraepithelial sarcomas (rhabdotheliospora), colon carcinomas, pancreatic carcinomas, breast carcinomas, ovarian carcinomas, prostate carcinomas, squamous cell carcinomas, basal cell carcinomas, adenocarcinomas, renal cell carcinomas, hematomas, bile duct carcinomas, melanomas, choriocarcinomas, seminoma, embryonal carcinomas, Wilms' tumors, cervical carcinomas, testicular tumors, lung carcinomas, small and non-small cell lung carcinomas, bladder carcinomas, epithelial carcinomas, gliomas, astrocytomas, medulloblastomal cell carcinomas, craniopharyngioblastoma, carcinoma of the head of the eye, and non-small cell lung carcinomas of the head, Ependymoma (ependonoma), pinealoma, retinoblastoma, rectal cancer, thyroid cancer, head and neck cancer, brain tumor, peripheral nervous system cancer, central nervous system cancer, neuroblastoma, endometrial carcinoma, lymphoproliferative disease, hematopoietic malignancies including various types of leukocytosis (leukamia), and lymphomas, including: acute myeloid leukemia, acute myelogenous leukemia, acute lymphocytic leukemia, chronic myelogenous leukemia, chronic lymphocytic leukemia, mast cell leukemia, multiple myeloma, myeloid lymphoma (myeloid lymphoma), hodgkin's lymphoma, non-hodgkin's lymphoma, and metastases of all of the above cancers.

These and further features of the present invention will be better understood in conjunction with the following drawings, detailed description, and claims.

Drawings

The invention is herein described, by way of example only, with reference to the accompanying drawings, wherein:

figure 1 shows the cytotoxicity of MJDB against: a) human CLL lymphocytes (labeled "patient 1", "patient 2", and "patient 3"); and b) lymphocytes from healthy donors (labeled "normal").

Figure 2 shows MJDB and Methyl Jasmonate (MJ) acting on four human cancer cell lines: comparison of toxicity levels in lymphoblastic leukemia (Molt-4), lung cancer (3LL), melanoma (B16) and colon cancer (HCT 116).

FIG. 3 shows the cytotoxicity of MJDB against cells expressing wild-type or mutant p 53.

FIG. 4 shows a comparison of the effect of MJDB and MJ on ATP levels in Molt-4 leukemia cells.

Figure 5 shows MJTB versus different cell lines showing multidrug resistance: molt4 (leukemia), D122 (lung cancer), B16 (melanoma), and B16MDR (melanoma).

Figure 6 compares the cytotoxicity of MJTB and MJ. Cytotoxicity was calculated as a percentage (%) of untreated medium as a control, mean ± standard error (mean ± SE). n is 3.^***Indicating an effect comparing MJ and MJTB, P < 0.001.

Figure 7 compares the effect of MJTB on normal lymphocytes with leukemia cells. Comparing the effect of MJTB on normal lymphocytes and leukemia cells,^*the expression P is less than 0.05,^**the expression P is less than 0.01,^***represents P < 0.001. Cytotoxicity was calculated as a percentage (%) of untreated medium as a control, mean ± standard error (mean ± SE). n is 3.

FIG. 8 shows cytotoxicity of other MJ derivatives of the present invention against leukemia cells.

Detailed Description

The present invention provides novel jasmonate derivatives, compositions comprising these compounds, and methods of using these compositions to treat cancer.

The compounds of the present invention have the following general formula:

general formula I

Wherein:

n is 0, 1, or 2;

R₁is OH, C₁To C₁₂Alkoxy radical, C₁To C₁₂Substituted alkoxy, aryloxy, O-glucosyl or imino;

R₂is OH, C₁To C₁₂Alkoxy radical, C₁To C₁₂Substituted alkoxy, O-glucosyl, oxo, alkyl or imino;

R₃、R₄、R₅、R₆、R₇a, B, C, D and E are each independently H, halogen, OH, C₁To C₁₂Alkoxy radical, C₁To C₁₂Substituted alkoxy, aryloxy, O-glucosyl, C₁To C₁₂Alkyl or C₁To C₁₂A substituted alkyl group;

wherein R is₁And R₂Or R₁And R₄May together form an optionally substituted lactone;

wherein C is₃:C₇、C₄:C₅And C₉:C₁₀The bonds between may independently be double or single bonds;

provided that R is₃、R₄、R₅、R₆、R₇At least one of A, B, C, D and E is a halogen; and provided that, if A is the only halogen in the compound, A is not fluorine;

or a derivative of said formula, wherein the derivative has at least one of the following:

is located at C₃Lower acyl side chain (free acid or ester or co-ester)Yoke) located at C₆Or a hydroxy (free hydroxy or ester) moiety of, or at C₇N-pentenyl or n-pentyl side chains;

including salts, hydrates, solvates, polymorphs, optical isomers, enantiomers, diastereomers, and mixtures thereof.

More particularly, preferred compounds of the invention (formula I) are those C₉And C₁₀The bond between (A) and (B) is a single bond. Other preferred compounds are R₂Is oxo. Still other preferred compounds are at least R₆And R₇One is bromine, iodine, fluorine or chlorine. Even more preferred is R₆And R₇All selected from the group consisting of bromine, iodine, fluorine and chlorine. However, more preferably R₆And R₇All bromine compounds.

A further preferred aspect of the invention is A, B, R₆And R₇Is a compound of bromine, iodine, fluorine or chlorine. More preferably A, B, R₆And R₇Each of which is bromine.

Other preferred compounds of the invention are R₁Is alkoxy. And on the other hand, R₃、R₄And R₅Each is H (hydrogen). Meanwhile, C, D and E are each H.

One of the most preferred compounds of the present invention is methyl dibromojasmonate (MJDB). According to formula I, MJDB is: n is 0; c₃:C₇、C₄:C₅And C₉:C₁₀The bond between (A) and (B) is a single bond; r₁Is methoxy; r₂Is oxo; r₃、R₄、R₅A, B, C, D and E are each H; and R₆And R₇Each is bromine.

Another most preferred compound of the invention is methyl tetrabromo-jasmonate (MJTB). According to formula I, MJDB is: n is 0; c₃:C₇、C₄:C₅And C₉:C₁₀The bond between (A) and (B) is a single bond; r₁Is a firstAn oxy group; r₂Is oxo; r₃、R₄、R₅C, D and E are each H; and A, B, R₆And R₇Each is bromine.

Other preferred compounds of the invention are those wherein n is 0; c₃:C₇、C₄:C₅And C₉:C₁₀The bond between (A) and (B) is a single bond; r₁Is methoxy; r₂Is oxo; r₃、R₄、R₅A, B, C, D and E are each H; and any of the following:

a)R₆and R₇Each is fluorine (designated compound "MJS 99");

b)R₆and R₇Each iodine (designated compound "MJS 85fl 4");

c)R₆and R₇Each is chloro (designated compound "MJS 81fl 3");

d)R₆and R₇One is bromo and the other is hydroxy (designated compound "NJ-63"); or

e)R₆And R₇One is iodine and the other is methoxy (designated compound "MJS 72f 5").

The present invention also refers to pharmaceutical compositions containing a pharmaceutically acceptable carrier and having as active ingredient a compound of the invention as described above. Preferred compositions have an active ingredient MJDB or MJTB. Preferably, in the pharmaceutical composition, the active ingredient is dissolved in any acceptable lipid carrier. Further in accordance with a preferred embodiment of the present invention, the composition further comprises at least one other chemotherapeutic agent.

The present invention also provides a method of reducing the growth of cancerous cells in a mammal comprising applying to the cancerous cells a therapeutically effective amount of a compound of the present invention as described herein.

Further, the present invention provides a method of treating cancer in a warm-blooded vertebrate, especially a mammal, comprising administering to the patient a pharmaceutical composition comprising as an active ingredient a therapeutically effective amount of a compound of the invention as described herein. The term "mammal" includes non-human mammals and humans.

It will be understood that when the terms "treating or inhibiting a cancerous cell proliferative disease or disorder", "treating or inhibiting a non-solid cancer", "treating or inhibiting a tumor" and "treating or inhibiting a tumor" appear in the specification and claims, they are meant to include tumor formation, primary tumors, tumor progression or tumor metastasis.

The term "reduced growth" in the context of the present invention with respect to cancerous tumor cells refers to a reduction in at least one of: cell number compared to control (which may be necrotic, apoptotic, or any other type of cell death or combination thereof due to cell death); decrease in cell growth rate, i.e.: the total number of cells may increase but at a lower level or rate than the control; a decrease in the invasiveness of the cells (as determined by, e.g., soft agar analysis) compared to the control, even though their total number is unchanged; progression from more differentiated cell types to less variant cell types; a reduced rate of neoplasia; or slow the progression of cancer cells from one stage to the next.

Reduction of growth of cancerous tumor cells can be used to treat cancer by administering to a subject in need of such treatment a therapeutically effective amount of a compound of the present invention as described herein.

In a preferred embodiment, the process of the invention comprises the use of a compound of formula I, wherein at least R₆And R₇One is Br. In a further preferred embodiment, the compound is MJDB or MJTB.

As mentioned above, the present invention also discloses the use of a composition of formula I for the manufacture of a medicament for the treatment of cancer in a mammal.

The term "treatment of cancer" in the context of the present invention includes at least one of the following: a decrease in the rate of growth of the carcinoma (i.e., the carcinoma, while still growing, grows at a slower rate); growth arrest in cancerous growths (cancerous growth), i.e.: arrest of tumor growth, and, in preferred instances, tumor reduction or size reduction. The term also includes reducing the number of metastases (metastasis), reducing the number of newly formed metastases, slowing the progression of the carcinoma from one stage to another, and reducing angiogenesis induced by the carcinoma. In the most preferred example, the tumor is completely cleared. The term also includes extending the survival time of a patient undergoing treatment. The term also includes prophylaxis for prophylactic conditions (therapeutic issues) or those individuals susceptible to infection of tumors. Administration of the compounds of the present invention will reduce the likelihood of infection of the individual with the disease. In the best case, the subject is not infected with the disease.

The term "carcinoma" in the context of the present invention includes all types of tumors, whether in the form of solid or non-solid tumors from various origins, and includes both malignant or premalignant conditions and their metastases. This term particularly refers to: carcinomas (carcinoma), sarcomas, adenomas, hepatocellular carcinomas, hepatoblastoma, rhabdomyosarcoma, esophageal carcinoma, thyroid carcinoma, malignant gangliomas, fibrosarcomas, myxosarcomas, liposarcomas, chondrosarcomas, osteosarcomas, chordoma, angiosarcomas, endotheliosarcomas, lymphangiosarcomas, synoviomas, Ewing's tumors, leiomyosarcomas, rhabdoid intraepithelial sarcomas (rhabdotheliosacoma), colon carcinomas, pancreatic carcinomas, breast carcinomas, ovarian carcinomas, prostate carcinomas, squamous cell carcinomas, basal cell carcinomas, adenocarcinomas, renal cell carcinomas, hematomas, bile duct carcinomas, melanomas, choriocarcinomas, seminoma, embryonal carcinomas, Wilms' tumors, cervical carcinomas, testicular tumors, lung carcinomas, small and non-small cell carcinomas, bladder carcinomas, epithelial carcinomas, gliomas, astrocytomas, medulloblastoma, craniopharyngiomas, ependymomas (epynomas), epynomas, ependymomas, ependymoma, ep, Pinealoma, retinoblastoma, rectal cancer, thyroid cancer, head and neck cancer, brain tumor, peripheral nervous system cancer, central nervous system cancer, neuroblastoma, endometrial cancer, lymphoproliferative disease, hematopoietic malignancies including various types of leukocytosis (leukamia), and lymphomas, including: acute myeloid leukemia, acute myelogenous leukemia, acute lymphocytic leukemia, chronic myelogenous leukemia, chronic lymphocytic leukemia, mast cell leukemia, multiple myeloma, myeloid lymphoma (myeloid lymphoma), hodgkin's lymphoma, non-hodgkin's lymphoma, and metastases of all of the above cancers.

More preferably, the carcinoma is selected from the group consisting of prostate, breast, skin, colon, lung, pancreatic, lymphoma, leukocytosis (leukamia), head and neck, kidney, ovary, bone, liver or thyroid cancer. More preferably, the cancer is selected from the group consisting of leukocytosis (leukamia), lung cancer, melanoma and colon cancer, including lymphocytic leukemia. Additionally, in a preferred embodiment of the method, the compound is administered in a dose selected from the group consisting of 1 μ g to 1000mg/kg body weight.

In other embodiments of the use for the preparation of a medicament, the medicament comprises, in addition to the compound of formula I, at least one effective chemotherapeutic agent. In certain embodiments, the novel compounds may be used in combination with conventional effective chemotherapeutic agents that have non-negligible side effects. The combination of the compounds of the present invention with conventional drugs allows the dosage of conventional drugs to be reduced and thus the side effects experienced by the patient to be significantly reduced, yet still achieve adequate chemotherapeutic efficacy.

Also disclosed is a method of treating a patient having a cancer in need of treatment thereof, comprising administering to the mammal a therapeutically effective amount of a pharmaceutical composition comprising a compound of the present invention as described above as an active ingredient.

The present invention also provides a pharmaceutical composition for treating cancer in a mammal comprising as an active ingredient a therapeutically effective amount of a compound of the present invention as described above.

The invention also provides the use of a compound of the invention for the preparation of a medicament for reducing the growth of cancerous cells, as described herein.

The invention further discloses a method for preparing MJDB

Or MJTB

Comprises the following steps:

i. CCl at methyl jasmonate₄Adding bromine into the solution; then the

Evaporate CCl₄。

Other jasmonate derivatives of the invention can be prepared as described in the examples below. As further illustration, an alkyl moiety may be added to R₂And (c) a compound of formula (I): by using Grignard reagents at low temperatures.

Alternatively, the 9, 10-epoxide can be obtained from methyl jasmonate using a peroxyacid. The epoxide can be reacted with a compound of the general formula RMgX to give the 9-and 10-positions (R, respectively)₆And R₇) Compounds that are hydroxyl and alkyl. Substituted alkyl groups may be added in a similar manner.

In addition, under acidic or basic conditions, epoxides are reacted with compounds of the general formula ROH or ArOH, where R is alkyl and Ar is aryl, optionally at the 9-and 10-positions (R, respectively)₆And R₇) To give hydroxyl and alkoxy or acyloxy groups. Furthermore, the 5, 6-enamine to produce methyl jasmonate will be at C₅The position (B position) gives an alkyl group. Substituted alkyl groups may be added in a similar manner.

The present invention also provides a pharmaceutical composition for treating cancer in a mammal comprising as an active ingredient a therapeutically effective amount of a compound of the present invention, as described herein, and a pharmaceutically acceptable carrier.

The term "pharmaceutically acceptable" means approved by a regulatory agency of the federal or a state government or listed in the U.S. pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.

The term "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the compound is administered. Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like, polyethylene glycol, glycerol, propylene glycol or other synthetic solvents. Water is the preferred carrier when the pharmaceutical composition is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions may also be employed as liquid carriers, particularly as injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene glycol, water, ethanol and the like.

The composition may also contain minor amounts of wetting or emulsifying agents, or pH buffering agents such as acetates, citrates or phosphates, if desired. Antibacterial agents such as benzyl alcohol or methyl parabens are also contemplated; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; and tonicity (tonicity) adjusting agents such as sodium chloride or glucose.

These compositions may take the form of solutions, suspensions, emulsions, tablets, pills, capsules, powders, sustained release formulations and the like. The composition can be formulated into suppository with conventional binder and carrier such as triglyceride, microcrystalline cellulose, gum tragacanth (gum tragacanth) or gelatin. Oral dosage forms (Oralformulation) may include standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate, and the like. Examples of suitable Pharmaceutical carriers are described in reishi general Pharmaceutical Sciences, e.w. martin ("Remington's Pharmaceutical Sciences"). Such pharmaceutical compositions will contain a therapeutically effective amount of a compound of the invention, preferably in a substantially purified form, together with a suitable amount of carrier so as to provide suitable administration to the patient.

In some cases, the carrier may be selected according to the desired dosage form. In some cases, the carrier may also facilitate the delivery and penetration of the active ingredient to the target tissue, while improving the stability of the drug, reducing clearance, imparting sustained release properties to the drug, reducing undesirable side effects, and the like. The carrier may also be a substance that stabilizes the dosage form (e.g., a preservative), imparts an edible taste to the dosage form, and the like.

The carrier may be those commonly used and limited only by chemical-physical considerations, such as solubility and non-reactivity with the compounds of the invention, and the route of administration. The choice of carrier will be determined by the particular method of administration used to administer the pharmaceutical composition. Accordingly, the carrier may include additives, colorants, diluents, buffering agents, disintegrating agents, wetting agents, preservatives, flavoring agents, and pharmacologically compatible carriers. Alternatively, the carrier may be an adjuvant, which is defined as a substance that affects the action of the active ingredient in a predictable manner.

Methods of introducing pharmaceutical compositions containing compounds of the present invention include, but are not limited to: topical, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, ocular, and oral routes. The compounds may be administered by any convenient route, for example by infusion or bolus injection, by absorption through the mucosal epithelium (e.g., oral mucosa, rectal and intestinal mucosa, etc.), and may be administered with other therapeutically active agents. Administration is preferably topical, but may also be systemic. In addition, it may be desirable to introduce the pharmaceutical compositions of the present invention into the central nervous system by any suitable route of administration, including intraventricular and intrathecal injection; intraventricular injections can be administered by one of: an intraventricular catheter connected to a reservoir (reservoir). Pulmonary administration may also be employed, for example, by: using an inhaler or nebulizer, and a dosage form with an aerosolizing agent.

It would be desirable to administer the pharmaceutical compositions of the present invention locally in the area in need of treatment; this may be achieved by, but is not limited to, for example: local infusion during surgery, local application, e.g. in combination with wound dressings after surgery, by injection, by means of a catheter, by means of a suppository, or by means of an implant, which is a porous, non-porous, or gelatinous material. According to some preferred embodiments, the injection may be directly into the tumor site or tumor tissue or tissue prior to tumorigenesis, such as: the administration is carried out using a syringe.

Pharmaceutical compositions suitable for oral administration may consist of: (a) liquid solutions in which an effective amount of active substance is dissolved in a solvent such as: water, saline, natural fruit juice, alcohols, syrup, and other diluents; (b) solid dosage forms such as capsules (e.g., of the conventional hard-or soft-shelled gelatin type containing, for example, surfactants, lubricants, and inert fillers), tablets, lozenges (wherein the active ingredient is flavored with, for example, sucrose and acacia or tragacanth, or the active ingredient is in an inert base such as gelatin or glycerin), and troches, each containing a predetermined amount of the active ingredient as a solid or granules; (c) a powder formulation; (d) a suspension in a suitable liquid; (e) a suitable emulsion; (f) a liposome formulation; and other dosage forms.

In yet another embodiment, the composition is for topical administration, such as an ointment, gel, drop or cream. The compounds of the present invention may be prepared and applied in physiologically acceptable diluents, with or without pharmaceutical carriers, for topical application to body surfaces, such as: creams, gels, drops, ointments and the like. The invention is useful for the topical or transdermal treatment of cancer, such as melanoma. Adjuvants or gel matrix forms for topical use also include, for example: sodium carboxymethylcellulose, polyacrylates, polyoxyethylene polyoxypropylene block polymers, polyethylene glycol and wood wax alcohols.

For topical application directed to the body, the pharmaceutical composition may take the form of a tablet or capsule, which may contain any of the following ingredients, or compounds with similar properties: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose; a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterotes; or a glidant such as colloidal silicon dioxide. When the unit dosage form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier such as a fatty oil. In addition, the unit dosage form may contain many other materials which modify the physical form of the unit dosage, such as: sugar, shellac, or other enteral absorbents.

The compounds of the present invention may be released through a controlled release system. In one embodiment, infusion pumps may be used to administer the compounds of the invention, such as those used to release chemotherapeutic agents to specific organs or tumors (see Buchwald et al, 1980, Surgery 88: 507; Saudek et al, 1989, N.Engl. J. Med. ("New England medical journal) 321: 574). In preferred forms, administration of the compounds of the invention is combined with a biodegradable, biocompatible polymeric implant that releases the compound at a selected site over a controlled period of time. Examples of preferred polymeric materials include polyanhydrides, polyorthoesters, polyglycolic acid, polylactic acid, polyvinyl ethyl acetate, and copolymers and blends thereof (see Medical applications of controlled release, Langer and Wise, 1974, CRCPres, Boca Raton, Fla.). In yet another embodiment, the controlled release system may be placed close to the therapeutic target, requiring only a fraction of the systemic dose.

In some cases, the active compound may be administered as an aerosol by inhalation. These aerosol formulations, such as dichlorodifluoromethane, propane, nitrogen, and the like, may be placed in pressurized acceptable propellants. They may also be formulated as pharmaceutical formulations in non-pressurized dosage forms, such as in a nebulizer or atomizer.

In addition, the pharmaceutical compositions may, in some cases, be formulated for parenteral administration (subcutaneous, intravenous, intraarterial, or intramuscular injection) and may include aqueous and non-aqueous isotonic sterile injection solutions which may contain antioxidants, buffers, bacteriostats, and solutes which render the formulation isotonic with the blood of the intended recipient, as well as aqueous and non-aqueous sterile suspensions which include suspending agents, solubilizers, thickening agents, stabilizers, and preservatives. Fats and oils such as petroleum, animal, vegetable, or synthetic fats and oils and fatty acid salts such as fatty alkali metal, ammonium, and triethanolamine salts, and suitable detergents may also be used for parenteral administration. The above dosage forms can also be used for direct intratumoral injection. In addition, the composition may contain one or more nonionic surfactants in order to minimize or eliminate irritation at the injection site. Suitable surfactants include polyethylene sorbitol fatty acid esters, such as sorbitol monooleate and the high molecular weight adducts of ethylene oxide with one hydrophobic group (base) formed by the condensation of propylene oxide with propylene glycol.

Parenteral dosage forms can be presented in unit-dose or multi-dose sealed containers, such as ampules and vials, and can be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier for the injection solution, e.g., water, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules, and tablets of the type previously described and known in the art.

The amount of a compound of the invention that will be effective in treating a particular disorder or condition, including cancer, will depend on the nature of the disorder or condition and can be determined by standard clinical techniques. In addition, in vitro assays may optionally be used to help determine the desired dosage range. The precise dose to be employed in the formulation will also depend on the route of administration, and the severity of the disorder or condition, and should be determined at the discretion of the attendant physician and in the individual patient's circumstances. Preferred dosages will be in the range of 0.01-1000mg/kg body weight, more preferably, 0.1mg/kg to 100mg/kg and still more preferably 1mg/kg to 10 mg/kg. Effective doses can be inferred from dose-response curves obtained from bioassays or systems (systems) in vitro or in animal model trials.

A "therapeutic" treatment is a treatment administered to a patient exhibiting signs of pathology with the aim of diminishing or eliminating those signs. A "therapeutically effective amount" of a compound of the present invention is an amount of the compound sufficient to provide a beneficial effect to the patient to whom the compound is administered.

The patient in need of treatment may suffer from a disease such as cancer or has been determined to be very susceptible to such diseases. Thus, the methods according to the invention encompass both therapeutic and prophylactic uses.

The compounds of the invention may also be tested in vivo for the desired therapeutic or prophylactic activity and to determine the effective therapeutic dose. For example: these compounds may be tested in a suitable animal model system prior to testing in humans, including, but not limited to: mice, chickens, cows, monkeys, rabbits, and the like. In vivo testing prior to administration to humans, any animal model system known in the art may be used.

When the above compounds include one or more chiral centers, the stereochemistry of these chiral centers may independently be in the R or S configuration, or a mixture of both. The chiral center may be further designated as R or S or R, S or D, D, L, L or D, L, D, L.

The term "C₁To C₁₂Alkyl "denotes such groups as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, tert-pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl and the like. Preferred is "C₁To C₁₂Alkyl "radicals are methyl, ethyl, isobutyl, sec-butyl and isopropylAnd (4) a base. Similarly, the term "C₁To C₁₂Alkenyl "means a group of 1 to 12 carbons that links two other moieties in a compound.

The term "C₁To C₁₂Substituted alkyl "denotes a group substituted by one or more, while preferably one or two substituents are selected from halogen, hydroxy, protected hydroxy, oxo, protected oxo, C₃To C₇Cycloalkyl, phenyl, substituted phenyl, naphthyl, amino, protected amino, (mono) amino, protected (mono) amino, (di) amino, guanidino, protected guanidino, heterocycle, substituted heterocycle, imidazolyl, indolyl, pyrrolidinyl, C₁To C₁₂Alkoxy radical, C₁To C₁₂Acyl radical, C₁To C₁₂Acyloxy, nitro, carboxyl, protected carboxyl, carbamoyl, carboxamide, protected carboxamide, N- (C)₁To C₁₂Alkyl) carboxamide group, protected N- (C)₁To C₁₂Alkyl) carboxamide group, N-di (C)₁To C₁₂Alkyl) carboxamide group, cyano group, methanesulfonamide group, mercapto group, C₁To C₁₀Alkylthio or C of₁To C₁₀An alkylsulfonyl group of (a). The substituted alkyl group may be substituted one or more times, preferably one or two times, with the same or different substituents. Preferred substituents are halogen.

The term "protected oxo" means that one carbon atom is bonded to two alkoxy groups, thereby forming an acyclic or cyclic acetal or ketal moiety. The term "C" as used herein₁To C₁₂Alkoxy "denotes, for example, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy and the like. The most preferred alkoxy group is methoxy. The term "C₁To C₁₂By substituted alkoxy "is meant that the alkyl portion of the alkoxy group may be substituted with C₁To C₁₂Substituted alkyl groups of (a) are substituted in the same manner. Preferred substituents are halogen. Similarly, the term "C" as used herein₁To C₁₂Phenylalkoxy "Is denoted by "C₁To C₁₂The "alkoxy group of (a)" is bonded to one phenyl group.

The term "C" herein₁To C₁₂Acyloxy "denotes, for example, formyloxy, acetyloxy, propionyloxy, butyryloxy, valeryloxy, pivaloyloxy, hexanoyloxy, heptanoyloxy, octanoyloxy, nonanoyloxy, decanoyloxy, undecanoyloxy, dodecanoyloxy and the like.

Similarly, the term "C" is used herein₁To C₁₂Acyl "includes groups such as formyl, acetyl, propionyl, butyryl, pentanoyl, pivaloyl, hexanoyl, heptanoyl, octanoyl, nonanoyl, decanoyl, undecanoyl, dodecanoyl, benzoyl and the like. Preferred acyl groups are acetyl and benzoyl.

The term "C₁To C₁₂Substituted acyl "means that the acyl group is substituted with one or more, and preferably one or two, of the substituents identified above for alkyl. The term "C" for a substituent₃To C₇Cycloalkyl "includes cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl rings. Similarly, the substituent may be "C₃To C₇Cycloalkyl radicals ", which may also be" C₅To C₇Cycloalkyl "including cyclopentyl, cyclohexyl or cycloheptyl rings.

The term "C" for a substituent₃To C₇Substituted cycloalkyl "or" C₅To C₇Substituted cycloalkyl "means that the aforementioned cycloalkyl ring is substituted with one or two of the substituents identified above for alkyl. The term "cycloalkylene" refers to a cycloalkyl group as described above bonded at two positions, together with two separate additional groups. Similarly, the term "substituted cycloalkene" refers to a cycloalkene, which is bonded at two positions, connects two separate groups together and has at least one additional substituent.

Similarly, the terms"substituted C₅To C₇Cycloalkenylene "means a cycloalkylene further substituted by halogen, hydroxy, protected hydroxy, C₁To C₁₀Alkylthio of, C₁To C₁₀Alkylsulfono of (A), C₁To C₁₀Alkanesulfonyl group of (A), C₁To C₁₀Substituted alkylthio, C₁To C₁₀Substituted alkylsulfono, C₁To C₁₀Substituted alkanesulfonyl, C₁To C₁₂Alkyl radical, C₁To C₁₂Alkoxy radical, C₁To C₁₂Substituted alkyl, C₁To C₁₂Alkoxy, oxo, protected oxo, (mono) amino, (di) amino, trifluoromethyl, carboxy, protected carboxy, phenyl, substituted phenyl, thiophenyl, phenylsulfinyl, phenylsulfonyl, amino, or protected amino groups.

The term "heterocycle" or "ring of different ring type" denotes an optionally substituted five-to eight-membered ring having 1 to 4 heteroatoms, such as oxygen, sulfur and/or nitrogen, especially nitrogen, either alone or in combination with sulfur or oxygen atoms, forming a ring. These five to eight membered rings may be saturated, fully unsaturated or partially unsaturated, with fully saturated rings being preferred. Preferred heterocycles include morpholino, piperidinyl, piperazinyl, 2-amino-imidazolyl, tetrahydrofuranyl, pyrrolo, tetrahydrothiophenyl, hexylmethyleneimino, and heptylmethyleneimino.

The term "substituted heterocycle" or "substituted different cyclic ring" means that the above-mentioned heterocycle is substituted, for example, with one or more, preferably one or two of the above-identified substituents for alkyl. The term "heteroaryl" refers to an aromatic heterocyclic derivative which is a five or six membered ring system having 1 to 4 heteroatoms, such as oxygen, sulfur and/or nitrogen, especially nitrogen, either alone or in combination with sulfur or oxygen atoms to form a ring. Examples of heteroaryl groups include pyridyl, pyrimidinyl, and pyrazinyl, pyridazinyl, pyrrolo, furo, oxazolo, isoxazolo, phthalimidyl, thiazolyl, and the like.

The term "substituted heteroaryl" means that the heteroaryl is substituted with, for example: one or more, preferably one or two, identical or different substituents as defined above for the alkyl radical. The term "substituted phenyl" designates a phenyl group substituted with one or more, preferably one or two, moieties selected from the group of substituents identified above for alkyl.

The term "phenoxy" means that a phenyl group is bonded to an oxygen atom, wherein the remainder of the molecule is bound through the oxygen atom. The term "substituted phenoxy" designates a phenoxy group substituted with one or more, preferably one or two moieties selected from the group of substituents identified above for alkyl.

The term "aryl" refers to an aromatic group having at least one carbocyclic or heterocyclic aromatic group, which may be unsubstituted or substituted with one or more groups selected from halogen, haloalkyl, hydroxy, alkoxycarbonyl, amido, alkylamido, dialkylamido, nitro, amino, alkylamino, dialkylamino, carboxy or thio or alkylthio. Non-limiting examples of aromatic rings are phenyl, naphthyl, pyranyl, pyrrolyl, pyrazinyl, pyrimidinyl, pyrazolyl, pyridyl, furanyl, thienyl, thiazolyl, imidazolyl, isoxazolyl, and the like. The term "aryloxy" refers to an "aryl" group bonded to an oxygen atom, wherein the remainder of the molecule is bonded through an oxygen atom.

The terms "halo" and "halogen" refer to a fluorine, chlorine, bromine or iodine atom. There may be one or more of the same or different halogens.

The term "(monosubstituted) amino" means that an amino group has one substituent selected from the group consisting of phenyl, substituted phenyl, C₁To C₁₂Alkyl radical, C₁To C₁₂Substituted alkyl, C₁To C₁₂Acyl radical, C₁To C₁₂Substituted acyl, C₂To C₁₂Alkenyl radical, C₂To C₁₂Substituted alkenyl, C₂To C₁₂Alkynyl, C₂To C₁₂Substituted alkynyl, C₇To C₁₈Phenylalkyl, C₇To C₁₈Substituted phenylalkyl, heterocycle, substituted heterocycle, C₁To C₁₂Heterocycloalkyl and C₁To C₁₂A substituent of the group consisting of substituted heterocycloalkyl groups. The (monosubstituted) amino group may additionally have an amino protecting group, as encompassed by the term "protected (monosubstituted) amino".

The term "(disubstituted) amino" means that one amino group has two substituents selected from the group consisting of phenyl, substituted phenyl, C₁To C₁₂Alkyl radical, C₁To C₁₂Substituted alkyl, C₁To C₁₂Acyl radical, C₂To C₁₂Alkenyl radical, C₂To C₁₂Alkynyl, C₇To C₁₈Phenylalkyl, C₇To C₁₈Substituted phenylalkyl, C₁To C₁₂Heterocycloalkyl and C₁To C₁₂A substituent of the group consisting of substituted heterocycloalkyl groups. The two substituents may be the same or different.

The term "amino protecting group" as used herein refers to a substituent of an amino group that is typically used to block or protect an amino functionality when reacting other functional groups of a molecule. The term "protected (monosubstituted) amino" means a protecting group having an amino group on the nitrogen atom of the monosubstituted amino group. In addition, the term "protected carboxamide group" means that there is an amino protecting group on the nitrogen of the carboxamide group. Similarly, the term "protected N- (C)₁To C₁₂Alkyl) carboxamide "means that there is an amino protecting group on the nitrogen of the carboxamide group.

The type of amino protecting group used is not critical, so long as the derivatized amino group is stable to the reaction conditions thereafter and can be removed at the appropriate site without cleaving the remainder of the compound. Preferred amino protecting groups are Boc, Cbz and Fmoc. Further examples of amino protecting groups encompassed by the above terms are well known in organic synthesis and peptide technology and are described, for example, in: T.W.Greene and P.G.M.Wuts, "Protective Groups in Organic Synthesis," ("Protective Groups in Organic Synthesis") 2nded., "John Wiley and Sons, New York, NY, 1991, Chapter 7, M.Bodanzsky," Prnciples of Peptide Synthesis, "(" principles of Peptide Synthesis) 1st and 2nd revisised ed., Springer-Verlag, New York, NY, 1984and 1993, and Stewart and Young, "Solid Phase Peptide Synthesis," ("2 nd Chemical ed.," Pierce Co., Rockford, IL, 1984, all of which are incorporated herein by reference. The relative term "protected amino" is defined as an amino group substituted with the amino protecting group discussed above.

The term "protected guanidino" as used herein refers to an "amino protecting group" on the nitrogen atom of one or both guanidino groups. Examples of "protected guanidino" are given by t.w.greene and p.g.m.wuts; m. bodanzsky; and Stewart and Young, previously described.

The term "thio" refers to-SH or, if between two other groups: -S-. The term "C₁To C₁₀The "thioalkenyl group" refers to an alkenyl chain of 1 to 10 carbons having a sulfur at any position (point) along the chain. The term "C₁To C₁₀Substituted thioalkenyl "means a C₁To C₁₀Substituted at one or more alkenyl positions (as described above, in the same manner as "substituted alkenyl").

The term "sulfonyl" refers to-S (O)₂-. The term "C₁To C₁₀Sulfonylalkenyl "refers to an alkenyl chain of 1 to 10 carbons having a sulfonyl group at any position (point) along the chain. The term "C₁To C₁₀Substituted sulfonylalkenyl "refers to a C₁To C₁₀The sulfonylalkenyl group of (a) is substituted at one or more alkenyl positions (as described above, in the same manner as "substituted alkenyl").

The term "sulfinyl" refers to-S (O) -. The term "C₁To C₁₀Sulfinylalkenyl "refers to an alkenyl chain of 1 to 10 carbons having a sulfinyl group at any position (point) along the chain. The term "C₁To C₁₀Substituted sulfinenyl "refers to a C₁To C₁₀The sulfinyl alkenyl group of (a) is substituted at one or more alkenyl positions (as described above, in the same manner as "substituted alkenyl").

The term "oxo" refers to-O-. The term "C₁To C₁₀Oxoalkenyl group "," C₁To C₁₀Dioxoenyl "and" C₁To C₁₀Trioxenyl "refers to an alkenyl chain of 1 to 10 carbons, each having one, two, or three-O-atoms at any position along the chain, provided that no two oxygen atoms are adjacent, and provided that any two oxygen atoms are separated by at least two carbons. The term "C₁To C₁₀Substituted oxoalkenyl group "," C₁To C₁₀Substituted dioxoenyl "and" C₁To C₁₀Substituted trioxenyl "refers to" C "substituted at one or more alkenyl positions (as described above, in the same manner as" substituted alkenyl ") respectively₁To C₁₀Oxoalkenyl group "," C₁To C₁₀Dioxoenyl "and" C₁To C₁₀A trioxenyl group.

As used herein, the term "carboxyl protecting group" refers to an ester derivative of a carboxylic acid group that is commonly used to block or protect the carboxylic acid group when other functional groups on the compound are reacted. Examples of such Groups are found in E.Haslam, "Protective Groups in Organic Chemistry," ("protecting Groups in Organic Chemistry") J.G.W.McOmie, Ed., Plenum Press, New York, NY, 1973, Chapter 5, and T.W.Greene and P.G.M.Wuts, "Protective Groups in Organic Synthesis," ("protecting Groups in Organic Synthesis") 2nd ed., John and sons, New York, NY, 1991, Chapter 5, all of which are incorporated herein by reference. The relative term "protected carboxy" means that the carboxy group is replaced with a carboxy protecting group of one of the above.

The term "hydroxy protecting group" refers to a group that is readily cleavable to form a bond with a hydroxy group. The type of hydroxy protecting group is not critical so long as the derivatized hydroxy group is stable to the reaction conditions thereafter and can be removed at the appropriate site without cleaving the remainder of the molecule. Examples of hydroxy protecting Groups are described in C.B.Reese and E.Haslam, "Protective Groups in Organic Chemistry," ("protecting Groups in Organic Chemistry") J.G.W.McOmie, Ed., Plenum Press, New York, NY, 1973, Chapters 3 and 4, and T.W.Greene and P.G.M.Wuts, "Protective Groups in Organic Synthesis," ("protecting Groups in Organic Synthesis") 2nd ed., John Wiley and Sons, New York, NY, 1991, Chapters2 and 3, respectively. The relative terms are "protected hydroxy" and "protected hydroxymethyl," which means that either the hydroxy or hydroxymethyl group is substituted with a hydroxy protecting group of one of the above.

The term "C₁To C₁₀Alkylthio "refers to a sulfide group, such as: methylthio, ethylthio, n-propylthio, isopropylthio, n-butylthio, t-butylthio and the like.

The term "C₁To C₁₀Alkylsulfinyl "denotes a sulfoxide group, such as: methyl sulfoxide, ethyl sulfoxide, n-propyl sulfoxide, isopropyl sulfoxide, n-butyl sulfoxide, sec-butyl sulfoxide, and the like. The term "C₁To C₁₀Alkylsulfonyl "includes groups such as: methylsulfonyl, ethylsulfonyl, n-propylsulfonyl, isopropylsulfonyl, n-butylsulfonyl, t-butylsulfonyl, and the like. It is to be understood that the above-mentioned thio, sulfoxide or sulfonyl group can be anywhere on the alkyl chain (e.g., 2-methylthioethyl).

The term "C₁To C₁₀Substituted alkylthio group "," C₁To C₁₀Substituted alkylsulfidenyl 'and' C₁To C₁₀Substituted alkylsulfonyl "denotes C of these radicals₁To C₁₀The alkyl moiety may be substituted as described above for "substituted alkyl".

The terms "thiophenyl", "phenylsulfonyl" and "phenylsulfonyl" designate a mercapto group, a sulfoxide group, or a sulfone group, respectively, containing a phenyl group. The terms "substituted phenylthio", "substituted phenylsulfonyl" and "substituted phenylsulfonyl" mean that the phenyl groups of these groups may be substituted as described above in connection with "substituted phenyl".

The term "C₁To C₁₂Alkylaminocarbonyl "means a C₁To C₁₂Is bonded to the nitrogen of one of the aminocarbonyl groups. C₁To C₁₂Examples of alkylaminocarbonyl groups include methylaminocarbonyl, ethylaminocarbonyl, propylaminocarbonyl and butylaminocarbonyl. The term "C₁To C₁₂Substituted alkylaminocarbonyl "denotes a substituted alkyl group bonded to the nitrogen atom of an aminocarbonyl group, which alkyl group may be as defined above and C₁To C₁₂Substituted alkyl groups are substituted as for substituted alkyl groups.

The term "C₁To C₁₂Alkoxycarbonyl "means a" C₁To C₁₂The "alkoxy" group of (a) is bound to a carbonyl group. The term "C₁To C₁₂Substituted alkoxycarbonyl "denotes a bond of a substituted alkoxy group to a carbonyl group, the alkoxy group being as defined above for" C₁To C₁₂Substituted alkyl "is substituted as relevant.

The term "phenylaminocarbonyl" means that one phenyl group is bonded to the nitrogen atom of one aminocarbonyl group. The term "substituted phenylaminocarbonyl" denotes a substituted phenyl group bonded to the nitrogen atom of an aminocarbonyl group, which phenyl group may be substituted as described above in connection with the substituted phenyl group.

The term "C₁To C₁₂Alkylthiocarbonyl "means a C₁To C₁₂Is bonded to an aminothiocarbonyl group, wherein the alkyl group has the same meaning as defined above.

The term "C₁To C₁₂Substituted alkylaminothiocarbonyl "denotes a bond of a substituted alkyl group to an aminothiocarbonyl group, wherein alkyl may be as defined aboveAbove and C₁To C₁₂Substituted alkyl groups are substituted as for substituted alkyl groups.

The term "phenylaminothiocarbonyl" means that a phenyl group is bonded to an aminothiocarbonyl group, wherein the phenyl group has the same meaning as defined above.

The term "substituted phenylaminothiocarbonyl" denotes a bond between a substituted phenyl group and an aminothiocarbonyl group, wherein the phenyl group may be substituted as described above in connection with the substituted phenyl group.

The term "substituted C₁To C₁₂Alkenyl "means a C₁To C₁₂The alkyl group of (1) which is bonded at two positions, is commonly bonded to two other independent groups, and has one substituent in addition.

The term "cyclic C₂To C₇Alkenyl group, substituted cyclic C₂To C₇Alkenyl group and cyclic C₂To C₇Heteroalkenyl "and" substituted cyclic C₂To C₇Heteroalkenyl "defines such a cyclic group bonded (" fused ") to a phenyl group to provide a bicyclic ring system. The cyclic group may be saturated or contain one or two double bonds. Furthermore, the cyclic group may have one or two methylene or methine groups substituted by one or two oxygen, nitrogen or sulfur atoms, that is to say a cyclic C₂To C₇A heteroalkenyl group.

The cycloalkenyl or heteroalkenyl group may be substituted once or twice by identical or different substituents, which may, if appropriate, be bonded to another moiety of a compound (e.g., alkenyl group) selected from the group consisting of: hydroxy, protected hydroxy, carboxy, protected carboxy, oxo, protected oxo, C₁To C₄Acyloxy, formyl, C₁To C₁₂Acyl radical, C₁To C₁₂Alkyl radical, C₁To C₇Alkoxy radical, C₁To C₁₀Alkylthio of, C₁To C₁₀Alkylsulfono of (A), C₁To C₁₀Alkanesulfonyl, halogen, ammoniaA protected amino group, a (mono) amino group, a protected (mono) amino group, a (di) amino group, a hydroxymethyl group or a protected hydroxymethyl group.

The cycloalkenyl or heteroalkenyl group fused to the phenyl group may contain two to ten membered rings, but preferably contains three to six membered rings. Examples of such saturated cyclic groups are the case when the bicyclic ring system obtained is a 2, 3-indanyl and a tetralin ring. When the cyclic groups are unsaturated, the examples are the case when the resulting bicyclic ring system is a naphthalene ring or an indole ring. When the phenyl group is fused to a pyrido, pyrano, pyrrolo, pyridyl, dihydropyrrolo or dihydropyridyl group, examples of fused cyclic groups contain one nitrogen atom and one or more double bonds, preferably one or two double bonds. Examples of fused cyclic groups when phenyl is fused to a furo, pyrano, dihydrofuro or dihydropyrano ring all contain one oxygen atom and one or two double bonds. Examples of fused cyclic groups when a benzene ring is fused with a thieno, thiopyrano, dihydrothieno or dihydrothiopyrano ring all contain one sulfur atom and one or two double bonds. When the phenyl ring is fused to a thiazolone, isothiazolone, dihydrothiazolone or dihydroisothiazolone ring, embodiments of the cyclic group contain two heteroatoms selected from sulfur and nitrogen and one or two double bonds. Examples of cyclic groups contain two heteroatoms selected from oxygen and nitrogen and one or two double bonds when a benzene ring is fused with an oxazolone, isoxazolone, dihydrooxazolone, or dihydroisoxazolone ring. Examples of cyclic groups contain two nitrogen heteroatoms and one or two double bonds when the phenyl ring is fused to a pyrazolone, imidazolone, dihydropyrazolone or dihydroimidazolone ring or a pyrazinyl group.

The term "carbamoyl" means a-NC (O) -group bonded at two positions to two other groups, respectively.

One or more of the compounds of the present invention may exist as salts. The term "salts" includes those formed with carboxylic acid anions and amine nitrogens (amine nitriles) as well as salts formed with organic and inorganic anions and cations as discussed below. In addition, the term includes salts formed by standard acid-base reactions with basic groups (such as amino groups) and organic or inorganic acids. Such acids include hydrochloric, hydrofluoric, trifluoroacetic, sulfuric, phosphoric, acetic, succinic, citric, lactic, maleic, fumaric, palmitic, cholic, pamoic, galactaric, D-glutamic, D-camphoric, glutaric, phthalic, tartaric, lauric, stearic, salicylic, methanesulfonic, benzenesulfonic, sorbic, picric, benzoic, cinnamic, and the like.

The term "organic or inorganic cation" refers to a counterion to the carboxylate anion of the carboxylate. Such counter ions are selected from the group consisting of alkali and alkaline earth metals, (e.g., lithium, sodium, potassium, barium, aluminum and calcium); ammonium and mono-, di-and trialkylamines such as triethylamine, cyclohexylamine; and organic cations such as dibenzylammonium, benzylammonium, 2-hydroxyethylammonium, bis (2-hydroxyethyl) ammonium, phenethylbenzylammonium, dibenzylethylenediammonium, and the like. See, for example: "Pharmaceutical Salts," ("pharmaceutically acceptable Salts") Berge et al, j.pharm.sci., 66: 1-19(1977), incorporated herein by reference. Other cations include the protonated forms of procaine, quinine and N-methylglucamine encompassed by the above terms, as well as protonated forms of basic amino acids such as glycine, ornithine, histidine, phenylglycine, lysine and arginine. In addition, any transient compound in zwitterionic form formed by one carboxyl group and one amino group is also suitable under this term. For example: when a position is substituted with a quaternary ammonium methyl group, a cation to the carboxylic acid anion will be present. For the carboxylic acid anion, the preferred cation is sodium.

The compounds of the present invention also exist as solvates and hydrates. Thus, these compounds may be used, for example: water of hydration, or one, more, or any fraction of the mother liquor solvent molecules. These solvates and hydrates of the compounds are included within the scope of the present invention.

One or more of the compounds of the invention, even when in a library (library), may be in the form of a biologically effective ester, such as: in the form of a non-toxic, readily metabolisable (metabiolicula-labile) ester. These ester forms cause increased blood levels and prolong the efficacy of the corresponding non-ester forms of the compounds. Useful ester groups include lower alkoxymethyl groups such as: methoxymethyl, ethoxymethyl, isopropoxymethyl and the like; - (C)₁To C₁₂) Alkoxyethyl groups, such as: methoxyethyl, ethoxyethyl, propoxyethyl, isopropoxyethyl and the like; 2-oxo-1, 3-dioxolan-4-ylmethyl groups, such as: 5-methyl-2-oxo-1, 3-dioxolan-4-ylmethyl, 5-phenyl-2-oxo-1, 3-dioxolan-4-ylmethyl, and the like; c₁To C₁₀Alkyl thiomethyl groups of (a), such as: dimethyl sulfide, ethyl thiomethyl, isopropyl thiomethyl, and the like; acyloxymethyl groups, such as: pivaloyloxymethyl, pivaloyloxyethyl, -acetoxymethyl, and the like; an ethoxycarbonyl-1-methyl group; -an acetoxyethyl group; 1- (C)₁To C₁₂Alkoxycarbonyloxy (alkyloxycarbonyloxy)) ethyl groups such as 1-ethoxycarbonyloxyethyl group; 1- (C)₁To C₁₂An alkylaminocarbonyloxy (alkylamino) ethyl group such as the 1-methylaminocarbonyloxyethyl group.

It should be understood that any position required by the present invention may achieve three consecutive "substitutions". For example: a "substituted alkyl" is substituted with a "substituted phenyl", which in turn is substituted with another "substituted alkyl", which in turn can be substituted with a group and can no longer be substituted. However, it is also to be understood that the invention contemplates more than three parallel substitutions if (subject matter) appropriate. For example: if appropriate, more than three hydrogens on one alkyl moiety may be substituted with any one or more different groups, including halogen and hydroxy.

Examples

Experimental procedure

MJDB Synthesis

Carbon tetrachloride (CCl) of methyl jasmonate at-20 deg.C₄) The solution was reacted with bromine until the yellow color remained for 5 minutes. The solvent was then distilled off and the yellow residue was chromatographed on a column of silica gel (VLC) washed with methanol (MeOH), eluting with hexane/5-10% ethyl acetate.

A1: 1 mixture of two possible racemic compounds is obtained.

Mass spectra (ms spectrum): m/z 384 (Br)₂) Rf 0.8 on silica gel eluted with hexane/ethyl acetate 1: 1.

C NMR (CDCl 3): (C-1 to C-13): 172.1/172.3; 37.1/37.3; 38.0/38.3; 29.5/29.6; 38.8/39.1; 218.4/219.1; 51.7 (both are); 27.0/27.2; 57.8/55.8; 60.7/60.2; 35.7/36.1; 12.4 (both are); 51.1/51.3 ppm.

H NMR (CDCl 3): 2.39-2.41(H-2 and 3); 1.94-2.14(H-4, H-7 and H-11); 2.74-2.75 (H-5); 1.61 (H-8); 4.62 and 4.89 (H-9); 4.14 (H-10); 1.12 (H-12); 3.75(OMe) ppm.

2) Chromatography analysis

Methyl jasmonate dibromo methyl jasmonate

Synthesis of MJTB

In CCl₄MJ was reacted with 10 equivalents of bromine overnight. The solvent and excess bromine were evaporated under vacuum. Recrystallization from isopropanol (iPrOH) gave one tetrabromo isomer and from ethanol (EtOH) gave the second isomer. The compounds were identified by NMR, MS, while one isomer was identified by X-ray diffraction analysis.

Synthesis of MJS72f5

To stirred (. + -.) -MJ (111mg, 0.49 mmo) at 0 deg.Cl) in methanol (5mL) iodine (I) was added dropwise₂) (580mg, 2.28mmol) in methanol (15 mL). After stirring the mixture at 0 ℃ for 0.5 h in the absence of light, it was warmed to room temperature and stirring was continued for 48 h. The solvent was then distilled off, and saturated sodium sulfite (Na) was added to the residue₂SO₃) Aqueous (10mL) and extracted with diethyl ether (2X 10 mL). The combined organic layers were washed with magnesium sulfate (MgSO)₄) Dried and concentrated in vacuo. The residue was purified by VLC (ethyl acetate/petroleum ether 1: 4) to give the product as a colorless oil (42mg, 22%).

Synthesis of MJS99f7

Fluorine in nitrogen (N)₂) The medium 3% concentration was passed through a cold (-75 ℃) air stream with vigorous stirring of 250mL trichlorofluoromethane (CFCl) with substrate (. + -.) -MJ (2.1gr, 9.37mmol) dissolved therein in a slow stream of air₃) 200mL of chloroform (CHCl)₃) And 50mL of ethanol. The reaction was allowed to proceed for 3 hours, then the mixture was poured into 500mL of water and saturated sodium bicarbonate (NaHCO) was used₃) Washed with aqueous solution and dichloromethane (CH)₂Cl₂) And (4) extracting. The combined organic layers were over MgSO₄Dried and concentrated in vacuo. The residue was purified by VLC (ethyl acetate/petroleum ether 1: 9) to give a diastereomeric mixture as a red oil (950mg, 39%).

Synthesis of MJS85f4

To a stirred solution of (+ -) -MJ (246mg, 1.10mmol) in dry Tetrahydrofuran (THF) (6mL) was added I dropwise at 0 deg.C₂(1.17gr, 4.6mmol) in THF (15 mL). After the mixture was stirred at 0 ℃ for 3 hours in the absence of light, it was warmed to room temperature and stirred for another 48 hours. When complete, add saturated Na to the residue₂SO₃Aqueous (10mL) and extracted with diethyl ether (2X 10 mL). The combined organic layers were over MgSO₄Dried and concentrated in vacuo. The residue was purified by VLC (ethyl acetate/petroleum ether 1: 4) to give the iodinated product.

Synthesis of MJS81f3

To a stirred mixture of (+ -) -MJ (78mg, 0.348mmol) in CCl at-10 deg.C₄(5mL) was purged with a solution of concentrated hydrochloric acid (5mL) and potassium permanganate (KMnO)₄) (800mg) chlorine gas (Cl) prepared in situ₂). The mixture was stirred at-10 ℃ for 2 hours. The solvent is subsequently distilled off and the residue is purified by VLC (ethyl acetate/petroleum ether 1: 19) to give the chlorinated product.

Cytotoxicity assays for the examples

The reduction in the number of viable cells was measured by Cell titration 96-well nonradioactive Cell Proliferation Assay (Promega, Madison, Wis.). After completion of the given experiment, MTS (a tetrazolium compound) was added to each well of a 96-well plate at 333. mu.g/ml + phenazine methosulfate (at 25. mu.M) for 1 hour at 37 ℃. This is considered to take place in a colour reaction, the dehydrogenase reducing MTS in metabolically active cells. Because the cells are not washed prior to addition of MTS, there are no potentially loosely adherent or non-adherent cells that may be problematic. Soluble MTS formazane product was measured at 490nm wavelength using a CERES 900HDI ELISA reader (Bio-Tek Instruments, Inc, Highland Park, VT). The optical density is directly proportional to the number of living cells in the culture medium. Cytotoxicity (%) was calculated by the following method: [ (absorbance of control cell-absorbance of drug-treated cells)/absorbance of control cells ] × 100.

Example 1: MJDB is highly cytotoxic to leukemic cells and non-toxic to healthy lymphocytes

To test the toxicity of MJDB on human leukemia cells, peripheral blood lymphocytes were collected from chronic lymphocytic leukemia (CCL) patients. These cells appeared to contain virtually 100% cancer cells as determined by flow cytometric analysis of cell surface CD5 and CD19 markers. Peripheral blood lymphocytes from healthy donors were also collected. The cells were cultured at 1.5X 10⁴One/well in 96-well plates, with MJDB added for one day at several concentrations as shown in figure 1. The absorbance representing viable cells was measured by the Cell Titer96 Aqueous Non-Radioactive Cell proliferationAssay (Promega, Madison, WI); as described above, surviving cells produce a colored product in the assay. This assay is quantitative, as the amount of color is read using an ELISA reader. Cytotoxicity was calculated as a percentage of untreated media controls, mean ± standard deviation (mean ± SD); n is 3.

The cytotoxicity of MJDB was plotted against peripheral blood lymphocytes from CCL patients compared to that of healthy donors (indicated by diamonds). See fig. 1. MJDB is clear and significant (P < 0.05) is more cytotoxic to peripheral blood lymphocytes from chronic lymphocytic leukemia (CCL) patients than to peripheral blood lymphocytes from healthy donors. MJDB is highly and selectively cytotoxic to cancer cells from CCL patients, while its cytotoxicity is minimal for lymphocytes from healthy donors.

Example 2: MJDB is much more cytotoxic than the previously studied jasmonates, as shown by its inhibition of four different cancer cell lines in humans

The cytotoxicity of MJDB was compared to the previous study of jasmonates, namely Methyl Jasmonate (MJ), the most potent jasmonate disclosed in U.S. patent No.6,469,061. The cytotoxicity of these compounds was compared and as seen each was used in four human cancer cell lines derived from lymphoblastic leukemia, lung cancer, melanoma or colon cancer.

Molt-4 lymphoblastic leukemia cells (at 1.5X 10)⁴Perwell), 3LL Lung cancer cells (at 4X 10)³Well), B16 melanoma cells (at 4X 10)³Per well), or HCT116 colon cancer cells (at 4 × 10)³/well) was plated on a 96-well plate, Methyl Jasmonate (MJ) or MJDB was added at 0.5mM for 1 day. Absorbance representing viable cells was determined by the Cell Titer96 Aqueous Non-Radioactive Cell promotion Assay (supra). Cytotoxicity was calculated as a percentage of untreated media controls, mean ± standard deviation (mean ± SD); n is 3.

The percentage of cytotoxicity of methyl jasmonate (MJ, white column) or MJDB (packed column) is illustrated. Figure 2 clearly shows that the newly synthesized compound MJDB is greatly advantageous in its cytotoxic effect against different cancer cell lines compared to the previously studied methyl jasmonate. (statistical studies of these results showed P < 0.05).

Example 3: MJDB is effective against cellular expression of wild-type or mutant p53

The effect of MJDB on the cellular expression of the pro-apoptotic tumor suppressor, a variant of p53, is shown. Aberrant p53 expression occurs in approximately 50% of human cancers and contributes to drug resistance and subsequent failure of chemotherapy and radiation therapy in cancer patients. Thus, the ability to kill mutant p 53-expressing cells is of high clinical significance.

Two B lymphoma cloning systems derived from the same cell line (29M4.1) were used. These clones differed only in the expression of wild type or mutant p 53. More specifically, 29M4.1 cells were cultured at 2.5X 10⁴One/well in 96-well plates, with MJDB added at several concentrations for one day. Absorbance representing viable cells was determined by the Cell Titer96 Aqueous Non-Radioactive Cell Proliferation Assay (Promega, Madison, Wis.). Cytotoxicity was calculated as a percentage of untreated media controls, mean ± standard deviation (mean ± SD); n is 3. MJDB showed the same cytotoxicity against B lymphoma cells expressing wild-type or mutant p53, suggesting potential clinical use of MJDB against drug resistant tumors. See fig. 3.

Example 4: MJDB reduces ATP levels in cancer cells

An ATP assay was used to evaluate the effect of MJDB in reducing ATP levels in cancer cells. More specifically, Molt-4 cells (at 1X 10)⁴/well) were plated in 96-well opaque-walled plates with MJ added at 3mM or MJDB at 1mM for 10, 30 or 60 minutes at 37 ℃. Untreated cells grown in culture medium were used as controls. ATP levels through the CellTiter-Glo^TMLuminescennt Cell vitality Assay (Cell Titer-Glo)^TMAnalysis of luminescent cell viability); wherein the cells produce a luminescent signal equivalent to the concentration of ATP in the cells. ATP consumption was calculated as a percentage of untreated media control, mean ± standard deviation (mean ± SD); n is 3.

As seen in fig. 4, MJDB caused rapid and potent ATP depletion in Molt-4 leukemia cells to a greater extent than that caused by methyl jasmonate. It is important to note that MJDB is administered at a concentration 3-fold lower than methyl jasmonate. Thus, there is a correlation between the superior cytotoxic effects of MJDB (compared to methyl jasmonate) and its ability to reduce intracellular ATP levels.

Example 5: MJTB is cytotoxic to four different cancer cell lines in humans

Molt-4 (leukemia, at 2.5X 10)⁴mL), D122 (lung cancer, 5X 10)³mL), B16 (melanoma, at 2X 10)³mL), and B16MDR (melanoma showing multidrug resistance at 2 × 10³mL) cells were incubated in 96-well plates for one day in the presence of different concentrations of MJTB. As described above, the absorbance representing viable cells was determined by the Cell Titer96 Aqueous Non-Radioactive Cell promotion Assay (Promega, Madison, Wis.). Cytotoxicity was calculated as a percentage of untreated media controls, mean ± standard deviation (mean ± SD); n is 3. The IC50 levels of MJTB for these cell lines are shown in the table below:

	Molt-4	D122	B16	B16MDR
					IC50(mM)	0.008	0.05	0.08	0.08

as can be seen from the table and fig. 5, MJTB is effective against leukemia cells as well as cancer cells derived from various solid tumors. In addition, MJTB can kill cells that exhibit multidrug resistance, making it potentially useful in clinical situations where resistance is a major obstacle to successful chemotherapy.

Example 6: comparison of MJ and MJTB

Bioassay was carried out on a 1: 1 mixture of the above tetrabromo isomers. More specifically, Molt-4 leukemia cells (at 2.5X 10)⁴Ml) were cultured in 96-well plates for one day in the presence of MJ or MJTB at different concentrations. As described above, the absorbance representing viable cells was determined by the Cell Titer96 aqueous non-Radioactive Cell Proliferation Assay (Promega, Madison, Wis.). Cytotoxicity was calculated as a percentage of untreated media controls, mean ± standard deviation (mean ± SD); n is 3. IC50 levels for MJTB and MJ are shown in the following table:

	MJTB	MJ
			IC50(mM)	0.008	0.5

as can be seen from the table and fig. 5, MJTB has an IC50 (concentration showing 50% cytotoxicity) that is about 60 times less than MJ (the most potent jasmonate derivative to date). MJTB is therefore more efficient than MJ by more than an order of magnitude.

Example 7: MJTB is selective against cancer cells

Molt-4 (leukemia) cells (at 2.5X 10)⁴/ml), and normal peripheral blood lymphocytes (PBLs, (in 2X 10. sup. th.)⁵/ml) were cultured in 96-well plates for one day in the presence of MJTB at various concentrations. PBL were pre-incubated with 0.8. mu.g/ml phytohemagglutinin +5ng/ml TPA for 48 hours to induce cell cycle entry. These cells proliferate and thus are similar to cancer cells in this respect, making this comparison more effective. As described above, the absorbance representing viable cells was determined by the Cell Titer96 aqueous non-Radioactive Cell Proliferation Assay (Promega, Madison, Wis.). IC50 levels are shown in the table below:

	Molt-4	PBL
			IC50(mM)	0.008	0.25

as can be seen from the table and fig. 7, there is a satisfactory therapeutic window that allows MJTB to kill leukemic cells without exerting a realistic effect on normal cells. In fact, the IC50 of normal peripheral blood lymphocytes is an order of magnitude higher than that of leukemia cells. It should be noted that IC50 of PBL determined using higher MJTB concentrations is not present in fig. 7.

Example 8: cytotoxicity of other Compounds of the invention

Molt-4 (leukemia) cells (at 1.5 x 104/ml) were cultured for 1 day in the presence of different concentrations of MJ halo-derivative. Specifically, these derivatives are MJS72f5 (one of R6 and R7 is iodo and the other is methoxy), MJS99 (both R6 and R7 are fluoro), and NJ-63 (one of R6 and R7 is bromo and the other is hydroxy). As described above, the absorbance representing viable cells was determined by the Cell Titer96 Aqueous Non-Radioactive Cell Proliferation Assay (Promega, Madison, Wis.). As shown in fig. 8, all of these derivatives exhibited cytotoxicity greater than MJ. Another derivative is: MJS813, also has cytotoxic activity.

All references cited herein are incorporated by reference in their entirety. It should be understood that the detailed description and specific examples, while indicating preferred embodiments of the invention, are given by way of illustration only. And are not intended to limit the scope of the invention, as set forth in the following claims.

Claims (31)

1. A compound of formula I:

general formula I

Wherein:

n is 0, 1 or 2;

R₁is OH, C₁To C₁₂Alkoxy radical, C₁To C₁₂Substituted alkoxy, aryloxy, O-grapeA sugar or imino group;

R₂is OH, C₁To C₁₂Alkoxy radical, C₁To C₁₂Substituted alkoxy, O-glucosyl, oxo, alkyl or imino;

R₃、R₄、R₅、R₆、R₇a, B, C, D and E are each independently H, halogen, OH, C₁To C₁₂Alkoxy radical, C₁To C₁₂Substituted alkoxy, aryloxy, O-glucosyl, C₁To C₁₂Alkyl or C₁To C₁₂A substituted alkyl group;

wherein R is₁And R₂Or R₁And R₄May together form an optionally substituted lactone;

wherein C is₃:C₇、C₄:C₅And C₉:C₁₀Each of the bonds between may be a double bond or a single bond;

provided that R is₃、R₄、R₅、R₆、R₇At least one of A, B, C, D and E is a halogen; and provided that if a is the only halogen in the compound, then a is not fluorine;

or a derivative of the formula, wherein the derivative has at least one of the following:

is located at C₃Lower acyl side chain (free acid or ester or conjugate) of (a) at C₆Or a hydroxy (free hydroxy or ester) moiety of, or at C₇N-pentenyl or n-pentyl side chains;

including salts, hydrates, solvates, polymorphs, optical isomers, enantiomers, diastereomers, and mixtures thereof.

2. The compound of claim 1, wherein C₉And C₁₀The bond between (A) and (B) is a single bond.

3. The compound of claim 1, wherein R₂Is oxo.

4. The compound of claim 1, wherein R₆And R₇At least one is selected from the group consisting of bromine, iodine, fluorine and chlorine.

5. The compound of claim 1, wherein R₆And R₇Are all selected from bromine, iodine, fluorine and chlorine.

6. The compound of claim 1, wherein R₆And R₇Are all bromine.

7. The compound of claim 1, wherein A, B, R₆And R₇Bromine, iodine, fluorine and chlorine.

8. The compound of claim 1, wherein A, B, R₆And R₇Each is bromine.

9. The compound of claim 1, wherein R₁Is an alkoxy group.

10. The compound of claim 1, wherein R₃、R₄And R₅Each is H.

11. The compound of claim 1, wherein C, D and E are each H.

12. The compound of claim 1, wherein: n is 0; c₃:C₇、C₄:C₅And C₉:C₁₀The bond between (A) and (B) is a single bond; r₁Is methoxy; r₂Is oxo; r₃、R₄、R₅A, B, C, D and E are each H; and R₆And R₇Each is bromine.

13. The compound of claim 1, wherein: n is 0; c₃:C₇、C₄:C₅And C₉:C₁₀The bond between (A) and (B) is a single bond; r₁Is methoxy; r₂Is O connected to the carbon at position 6 via a double bond to form a carbonyl group; r₃、R₄、R₅C, D and E are each H; and A, B, R₆And R₇Each is bromine.

14. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a compound of claim 1 as an active ingredient.

15. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a compound of claim 12 as an active ingredient.

16. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a compound of claim 13 as an active ingredient.

17. The pharmaceutical composition of claim 14, wherein said active ingredient is dissolved in a pharmaceutically acceptable lipid carrier.

18. A method of reducing the growth of cancer cells comprising exposing the cancer cells to a therapeutically effective amount of the compound of claim 1.

19. The method of claim 18, wherein the carcinoma is a mammalian carcinoma.

20. The method of claim 19, wherein the mammal is a human.

21. The method of claim 18, wherein the compound is the compound of claim 4.

22. The method of claim 18, wherein the compound is the compound of claim 12.

23. The method of claim 18, wherein the compound is the compound of claim 13.

24. The method of claim 18, wherein the carcinoma is selected from the group consisting of: carcinomas (carcinoma), sarcomas, adenomas, hepatocellular carcinomas, hepatoblastoma, rhabdomyosarcoma, esophageal carcinoma, thyroid carcinoma, malignant gangliomas, fibrosarcomas, myxosarcomas, liposarcomas, chondrosarcomas, osteosarcomas, chordoma, angiosarcomas, endotheliosarcomas, lymphangiosarcomas, synoviomas (synovioamas), Ewing's tumors, leiomyosarcomas, rhabdoid intraepithelial sarcomas (rhabdotheliospora), colon carcinomas, pancreatic carcinomas, breast carcinomas, ovarian carcinomas, prostate carcinomas, squamous cell carcinomas, basal cell carcinomas, adenocarcinomas, renal cell carcinomas, hematomas, bile duct carcinomas, melanomas, choriocarcinomas, seminoma, embryonal carcinomas, Wilms' tumors, cervical carcinomas, testicular tumors, lung carcinomas, small and non-small cell lung carcinomas, bladder carcinomas, epithelial carcinomas, gliomas, astrocytomas, medulloblastomal cell carcinomas, craniopharyngioblastoma, carcinoma of the head of the eye, and non-small cell lung carcinomas of the head, Ependymoma (ependonoma), pinealoma, retinoblastoma, rectal cancer, thyroid cancer, head and neck cancer, brain tumor, peripheral nervous system cancer, central nervous system cancer, neuroblastoma, endometrial carcinoma, lymphoproliferative disease, hematopoietic malignancies including various types of leukocytosis (leukamia), and lymphomas including: acute myeloid leukemia, acute lymphocytic leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, mast cell leukemia, multiple myeloma, myeloid lymphoma (myeloid lymphoma), hodgkin's lymphoma, non-hodgkin's lymphoma.

25. The method of claim 18, wherein the carcinoma is selected from the group consisting of: prostate cancer, breast cancer, skin cancer, colon cancer, lung cancer, pancreatic cancer, lymphoma, leukocytosis (leukamia), head and neck cancer, kidney cancer, ovarian cancer, bone cancer, liver cancer and thyroid cancer.

26. The method of claim 18, wherein the cancer is selected from the group consisting of leukocytosis (leukamia), lung cancer, melanoma, and colon cancer.

27. A method of treating cancer comprising administering to a patient in need of treatment thereof a pharmaceutical composition comprising as an active ingredient a therapeutically effective amount of a compound according to any one of claims 1-13.

28. Use of a compound according to any one of claims 1 to 13 for the manufacture of a medicament for the treatment of cancer.

29. The use of claim 28, wherein the carcinoma is selected from the group consisting of: carcinomas (carcinoma), sarcomas, adenomas, hepatocellular carcinomas, hepatoblastoma, rhabdomyosarcoma, esophageal carcinoma, thyroid carcinoma, malignant gangliomas, fibrosarcomas, myxosarcomas, liposarcomas, chondrosarcomas, osteosarcomas, chordoma, angiosarcomas, endotheliosarcomas, lymphangiosarcomas, synoviomas (synovioamas), Ewing's tumors, leiomyosarcomas, rhabdoid intraepithelial sarcomas (rhabdotheliospora), colon carcinomas, pancreatic carcinomas, breast carcinomas, ovarian carcinomas, prostate carcinomas, squamous cell carcinomas, basal cell carcinomas, adenocarcinomas, renal cell carcinomas, hematomas, bile duct carcinomas, melanomas, choriocarcinomas, seminoma, embryonal carcinomas, Wilms' tumors, cervical carcinomas, testicular tumors, lung carcinomas, small and non-small cell lung carcinomas, bladder carcinomas, epithelial carcinomas, gliomas, astrocytomas, medulloblastomal cell carcinomas, craniopharyngioblastoma, carcinoma of the head of the eye, and non-small cell lung carcinomas of the head, Ependymoma (ependonoma), pinealoma, retinoblastoma, rectal cancer, thyroid cancer, head and neck cancer, brain tumor, peripheral nervous system cancer, central nervous system cancer, neuroblastoma, endometrial carcinoma, lymphoproliferative disease, hematopoietic malignancies including various types of leukocytosis (leukamia), and lymphomas, including: acute myeloid leukemia, acute myelogenous leukemia, acute lymphocytic leukemia, chronic myelogenous leukemia, chronic lymphocytic leukemia, mast cell leukemia, multiple myeloma, myeloid lymphoma (myeloid lymphoma), hodgkin's lymphoma, non-hodgkin's lymphoma, and metastases of all of the above cancers.

30. The use of claim 28, wherein the carcinoma is selected from the group consisting of: prostate cancer, breast cancer, skin cancer, colon cancer, lung cancer, pancreatic cancer, lymphoma, leukocytosis (leukamia), head and neck cancer, kidney cancer, ovarian cancer, bone cancer, liver cancer and thyroid cancer.

31. The use of claim 28, wherein the cancer is selected from the group consisting of leukocytosis (leukamia), lung cancer, melanoma, and colon cancer.