Patent Description:
Malignant gliomas, the most common form of central nervous system (CNS) cancers, are currently considered essentially incurable. Among the various malignant gliomas, anaplastic astrocytomas (Grade III) and glioblastoma mul- tiforme (GBM; Grade IV) have an especially poor prognosis due to their aggressive growth and resistance to currently available therapies. The present standard of care for malignant gliomas consists of surgery, ionizing radiation, and chemotherapy. Despite recent advances in medicine, the past <NUM> years have not seen any significant improvement in prognosis for malignant gliomas.

The poor response of tumors, including malignant gliomas, to various types of chemotherapeutic agents are often due to intrinsic drug resistance. Additionally, acquired resistance of initially well-responding tumors and unwanted side effects are other problems that frequently thwart long-term treatment using chemotherapeutic agents. Hence, various analogues of chemotherapeutic agents have been prepared in an effort to overcome these problems. The analogues include novel therapeutic agents which are hybrid molecules of at least two existing therapeutic agents. For example, cisplatin has been conjugated with cytotoxic codrugs, or conjugated with bioactive shuttle components such as porphyrins, bile acids, hormones, or modulators that expedite the transmembrane transport or the drug accumulation within the cell. (<NUM>-Aminomethylnicotinate) dichloridoplatinum (II) complexes esterified with terpene alcohols were tested on a panel of human tumor cell lines. The terpenyl moieties in these complexes appeared to fulfill a transmembrane shuttle function and increased the rate and extent of the uptake of these conjugates into various tumor cell lines.

Perillyl alcohol (POH), a naturally occurring monoterpene, has been suggested to be an effective agent against a variety of cancers, including CNS cancer, breast cancer, pancreatic cancer, lung cancer, melanomas and colon cancer. Hybrid molecules containing both perillyl alcohol and retinoids were prepared to increase apoptosis-inducing activity.

In order to improve performance over perillyl alcohol and its derivatives, there is a need to prepare isomers or analogs including isoperillyl alcohol conjugated with other therapeutic agents, and use this material in the treatment of cancers such as malignant gliomas, as well as other brain disorders such as Parkinson's and Alzheimer's disease.

These compounds may be administered alone or in combination with other treatment methods including radiation, standard chemotherapy, and surgery. The administration can also be through various routes including intranasal, oral, oral-tracheal for pulmonary delivery, and transdermal.

<CIT> discloses a composition comprising a fatty alcohol carbamate conjugated to a therapeutic agent such as the DNA alkylating agent temozolomide utilized for treating human cancers.

Any subject-matter falling outside the scope of the claims is provided for information purposes only.

Any references in the description to methods of treatment refer to the pharmaceutical compositions and medicaments of the present invention for use in a method for treatment of the human (or animal) body by therapy.

The method may further comprise the step of treating the mammal with radiation, and/or further comprise the step of delivering to the mammal a chemotherapeutic agent. The diseases treated may be cancer, including a tumor of the nervous system, such as a glioblastoma. The routes of administration include inhalation, intra- nasal, oral, intravenous, subcutaneous or intramuscular administration.

The present invention provides for a pharmaceutical composition comprising particular isoperillyl alcohol carbamates being iso-perillyl alcohols conjugated with a therapeutic agent, which is temozolomide (TMZ) The pharmaceutical compositions of the present invention may be administered before, during or after radiation. The pharmaceutical compositions may be administered before, during or after the administration of a chemotherapeutic agent.

A process for making an isoperillyl alcohol tarbamate comprises the step of reacting a first reactant of isoperillyl chloroformate with a second reactant, which is temozolomide (TMZ).

<FIG> shows the results of the MTT cytotoxicity assays demonstrating the efficacy of different types of POH and iso-POH in killing LN229 human glioma cells. <FIG> shows the results of the MTT cytotoxicity assays demonstrating the efficacy of different types of POH and iso- POH in killing U251 human glioma cells. <FIG> shows the results of the MTT cytotoxicity assays demonstrating the efficacy of different types of POH and iso- POH in killing A172 human glioma cells. <FIG> show the results of the MTT cytotoxicity assays demonstrating the efficacy of different types of POH and iso-POH in killing A172 human glioma cells (temozolomide-sensitive) (<FIG>) and A172 temozolomide-resistant cells (<FIG>). SGP-<NUM>-<NUM> is the POH purified to the GLP quality (with a GC relative area purity (area under the curve) of about <NUM>%). SGP-<NUM>-79P and SGP-<NUM>-65P are two different batches of Iso-POH. SGP-<NUM>-79P is more purified than SGP-<NUM>-65P and the details are as follows.

<FIG> show the results of the MTT cytotoxicity assays demonstrating the efficacy of different types of POH and iso-POH in killing U251 human glioma cells (temozolomide-sensitive) (<FIG>) and U251 (temozolomide-resistant) (<FIG>). <FIG> show the results of the MTT cytotoxicity assays demonstrating the efficacy of different types of POH and iso-POH in killing LN229 human glioma cells (temozolomide-sensitive) (<FIG>) and LN229 temozolomide-resistant cells (<FIG>). <FIG> show the results of the MTT cytotoxicity assays demonstrating the efficacy of different types of POH and iso-POH in killing U87 human glioma cells (temozolomide-sensitive) (<FIG>) and U87 temozolomide-resistant cells (<FIG>). <FIG> and <FIG> show the results of the MTT cytotoxicity assays demonstrating the efficacy of different types of POH and iso-POH in killing U251 human glioma cells (temozolomide-sensitive) (<FIG>) and U251 temozolomide- resistant cells (<FIG> and <FIG>). U251-TR1 and U251-TR1 refer to two temozolomide-resistant U251 cell lines. Sigma POH is the POH purchased from Sigma Chemicals. GLP POH is the POH purified to the GLP quality (with a GC relative area purity (area under the curve) of about <NUM>%). Iso-POH65 and Iso-POH79 are different batches of iso-POH (see details above for <FIG>). <FIG> shows the results of the MTT cytotoxicity assays demonstrating the efficacy of POH and iso-POH in killing USC04 glioblastoma cancer stem cell line. <FIG> show Western blot performed after an <NUM>-hour treatment of U251 glioma cells with Sigma POH (<NUM>, having a purity of about <NUM>% (AUC)) or ultrapure POH ("GLP-POH", <NUM>, having a purity of about <NUM>% (AUC)) in both U251 TMZ-sensitive and TMZ-resistant (U251-TR1, U251-TR2) cells demonstrating increased expression of glucose-regulatory protein <NUM> (GRP-<NUM>) and the apoptosis marker CHOP, showing increased endoplasmic reticulum (ER) stress after treatment (<FIG>). Under the same conditions, iso-POH (iso-POH65, iso-POH79) also increased ER stress (<FIG>). <FIG> shows Western blot performed after a <NUM>-hour treatment of U251 glioma cells with <NUM> Sigma POH, GLP POH or iso-POH (isoPOH65, isoPOH79) demonstrating that all treatments decreased Kras expression.

The invention relates to a pharmaceutical composition comprising an isoperillyl alcohol carbamate, wherein the isoperillyl alcohol carbamate is an isoperrilyl alcohol covalently bound via a carbamate linking group to temozolomide (TMZ), wherein the isoperillyl alcohol is selected from the group consisting of (<NUM>-isopropyl cyclohexa-<NUM>,<NUM>-dienyl)methanol, (<NUM>-isopropyl cyclohexa-<NUM>,<NUM>- dienyl)methanol, (<NUM>-isopropylphenyl)methanol and (<NUM>-isopropenylphenyl)methanol. Routes of administration include inhalation, intranasal, oral transdermal, intravenous, subcutaneous and intramuscular injection.

The isoperillyl alcohols POH) which are conjugated according to the present invention are (<NUM>-isopropylidene cyclohex-<NUM>-enyl)methanol. Isoperillyl alcohols of the present invention include, but are not limited to, (<NUM>-isopropyl cyclohexa-<NUM>,<NUM>-dienyl)methanol, (<NUM>-isopropyl cyclohexa-<NUM>,<NUM>-dienyl)methanol, (<NUM>-isopropylphenyl)methanol and (<NUM>-iso- propenylphenyl)methanol.

An exemplary isoperillyl alcohol, (<NUM>-isopropylidene cyclohex-<NUM>-enyl)methanol, is shown below:
<CHM>.

The compounds of the present invention may be used for the treatment of nervous system cancers, such a malignant glioma (e.g., astrocytoma, anaplastic astrocytoma, glioblastoma multiforme), retinoblastoma, pilocytic as- trocytomas (grade I), meningiomas, metastatic brain tumors, neuroblastoma, pituitary adenomas, skull base meningi- omas, and skull base cancer. The present invention also provides methods of treating CNS (central nervous system) disorders, including, without limitation, primary degenerative neurological disorders such as Alzheimer's, Parkinson's, psychological disorders, psychosis and depression.

In the claimed composition, the isoperillyl alcohol carbamate is present in amounts ranging from about <NUM> % (w/w) to about <NUM>% (w/w), from about <NUM>% (w/w) to about <NUM>% (w/w), from about <NUM>% (w/w) to about <NUM>% (w/w), from about <NUM>% (w/w) to about <NUM>% (w/w), or from about <NUM> % (w/w) to about <NUM>% (w/w). The present compositions can be administered alone, or may be co-administered together with radiation or another agent (e.g., a chemotherapeutic agent), to treat a disease such as cancer. Treatments may be sequential, with the isoperillyl alcohol carbamate being administered before or after the administration of other agents. For example, said isoperillyl alcohol carbamate may be used to sensitize a cancer patient to radiation or chemotherapy. Alternatively, agents may be administered concurrently. The route of administration may vary, and can include, inhalation, intranasal, oral, transdermal, intravenous, subcutaneous or intra- muscular injection.

Carbamate refers to a class of chemical compounds sharing the functional group
<CHM>
based on a carbonyl group flanked by an oxygen and a nitrogen. R<NUM>, R<NUM> and R<NUM> can be a group such as alkyl, aryl, etc., which can be substituted. The R groups on the nitrogen and the oxygen may form a ring. R<NUM>-OH may be a monoterpene. The R<NUM>-N-R<NUM> moiety may be a therapeutic agent.

Carbamates may be synthesized by reacting isocyanate and alcohol, or by reacting chloroformate with amine. Carbamates may be synthesized by reactions making use of phosgene or phosgene equivalents. For example, car- bamates may be synthesized by reacting phosgene gas, diphosgene or a solid phosgene precursor such as triphosgene with two amines or an amine and an alcohol. Carbamates (also known as urethanes. ) can also be made from reaction of a urea intermediate with an alcohol. Dimethyl carbonate and diphenyl carbonate are also used for making carbamates. Alternatively, carbamates may be synthesized through the reaction of alcohol and/or amine precursors with an ester-substituted diaryl carbonate, such as bismethylsalicylcarbonate (BMSC).

Carbamates may be synthesized by the following approach:
<CHM>
Suitable reaction solvents include, but are not limited to, tetrahydrofuran, dichloromethane, dichloroethane, acetone, and diisopropyl ether. The reaction may be performed at a temperature ranging from about -<NUM> to about <NUM>. or from about -<NUM> to about <NUM>. The molar ratio of isoperillyl chloroformate to the substrate R - NH<NUM> may range from about <NUM>:<NUM> to about <NUM>:<NUM>, from about <NUM>:<NUM> to about <NUM>:<NUM>, from about <NUM>:<NUM> to about <NUM>:<NUM>, or from about <NUM>:<NUM> to about <NUM>:<NUM>. Suitable bases include, but are not limited to, organic bases, such as triethylamine, potassium carbonate, N,N'-diisopropylethyl- amine, butyl lithium, and potassium-t-butoxide.

Alternatively, carbamates may be synthesized by the following approach:
<CHM>.

Suitable reaction solvents include, but are not limited to, dichloromethane, dichloroethane, toluene, diisopropyl ether, and tetrahydrofuran. The reaction may be performed at a temperature ranging from about <NUM> to about <NUM>, or from about <NUM> to about <NUM>, or about <NUM>. The molar ratio of isoperillyl alcohol to the substrate R-N=C=O may range from about <NUM>:<NUM> to about <NUM>:<NUM>, from about <NUM>:<NUM> to about <NUM>:<NUM>, from about <NUM>: <NUM> to about <NUM>:<NUM>, or from about <NUM>:<NUM> to about <NUM>:<NUM>.

In certain embodiments, an iso-POH carbamate is synthesized by a process comprising the step of reacting a first reactant of isoperillyl chloroformate with temozolomide (TMZ) The reaction may be carried out in the presence of tetrahydrofuran and a base such as n-butyl lithium.

Isoperillyl chloroformate may be made by reacting iso-POH with phosgene. For example, iso-POH conjugated with temozolomide through a carbamate bond may be synthesized by reacting temozolomide with oxalyl chloride followed by reaction with isoperillyl alcohol. The reaction may be carried out in the presence of <NUM>,<NUM>-dichloroethane.

The purity of a compound comprised in the compositions of the invention may be assayed by gas chromatography (GC) or high pressure liquid chromatography (HPLC). Other techniques for assaying the purity of the compounds of the present invention and for determining the presence of impurities include, but are not limited to, nuclear magnetic resonance (NMR) spectroscopy, mass spectrometry (MS), GC-MS, infrared spectroscopy (IR), and thin layer chromatography (TLC). Chiral purity can be assessed by chiral GC or measurement of optical rotation.

Said compounds may be purified by methods such as crystallization.

The invention also provides for the claimed compositions for use in treating a disease, such as cancer or other nervous system disorders. The compounds of the present invention may be administered alone, or in combination with radiation, surgery or chemotherapeutic agents. Said compositions may also be co-administered with antiviral agents, anti-inflammatory agents or antibiotics. The agents may be administered concurrently or sequentially. The compounds of the present invention can be administered before, during or after the administration of the other active agent(s).

The compounds and methods of the present invention may used to inhibit the Ras protein. The Ras family is a protein family of small GTPases that are involved in cellular signal transduction. Activation of Ras signaling causes cell growth, differentiation and survival. Mtations in ras genes can permanently activate it and cause inappropriate transmission inside the cell even in the absence of extracellular signals. Because these signals result in cell growth and division, dysregulated Ras signaling can ultimately lead to oncogenesis and cancer. Activating mutations in Ras are found in <NUM>-<NUM>% of all human tumors and up to <NUM>% in specific tumor types. Goodsell DS (<NUM>). Ras family members include, but are not limited to, HRAS; KRAS; NRAS: DIRAS1; DIRAS2: DIRAS3; ERAS; GEM; MRAS; NKIRAS1: NKIRAS2: NRAS: RALA: RALB: RAP1A; RAP IB: RAP2A: RAP2B: RAP2C: RASD1: RASD2: RASL10A: RASL10B; RASL11A, RASL11B; RASL12; REM1; REM2; RERG; RERGL; RRAD; RRAS: and RRAS.

The claimed compositions may be used in combination with radiatior therapy, in one embodiment treating tumor cells, such as malignant glioma cells, with radiation, where the cells are treated with an effective amount of the claimed composition and then exposed to radiation. Treatment by the compounds of the present invention may be before, during and/or after radiation. For example, the compounds of the present invention may be administered continuously beginning one week prior to the initiation of radiotherapy and continued for two weeks after the completion of radiotherapy. <CIT> and <CIT>.

In one embodiment, the present invention provides for a method of treating tumor cells, such as malignant glioma cells, with chemotherapy, where the cells are treated with an effective amount of an isomer or analog of monot- erpene or sesquiterpene (or a derivative of the isomer or analog of monoterpene or sesquiterpene), such as isoperillyl alcohol, and then exposed to chemotherapy. Treatment by the compounds of the present invention may be before, during and/or after chemotherapy.

The compounds of the present invention may be used for the treatment of nervous system cancers, such as a malignant glioma (e.g., astrocytoma, anaplastic astrocytoma, glioblastoma multiforme), retinoblastoma, pilocytic as- trocytomas (grade I), meningiomas, metastatic brain tumors, neuroblastoma, pituitary adenomas, skull base meningi- omas, and skull base cancer. As used herein, the term "nervous system tumors" refers to a condition in which a subject has a malignant proliferation of nervous system cells.

Cancers that can be treated by the present compounds include, but are not limited to, lung cancer, ear, nose and throat cancer, leukemia, colon cancer, melanoma, pancreatic cancer, mammary cancer, prostate cancer, breast cancer, hematopoietic cancer, ovarian cancer, basal cell carcinoma, biliary tract cancer; bladder cancer, bone cancer; breast cancer; cervical cancer; choriocarcinoma; colon and rectum cancer; connective tissue cancer; cancer of the digestive system; endometrial cancer, esophageal cancer; eye cancer; cancer of the head and neck; gastric cancer; intra-epithelial neoplasm; kidney cancer; larynx cancer; leukemia including acute myeloid leukemia, acute lymphoid leukemia, chronic myeloid leukemia, chronic lymphoid leukemia; liver cancer; lymphoma including Hodgkin's and Non- Hodgkin's lymphoma; myeloma; fibroma, neuroblastoma; oral cavity cancer (e.g., lip, tongue, mouth, and pharynx); ovarian cancer; pancreatic cancer; prostate cancer; retinoblastoma; rhabdomyosarcoma; rectal cancer; renal cancer; cancer of the respiratory system; sarcoma; skin cancer; stomach cancer; testicular cancer; thyroid cancer; uterine cancer; cancer of the urinary system, as well as other carcinomas and sarcomas.

The present invention also provides methods of treating CNS disorders, including, without limitation, primary degenerative neurological disorders such as Alzheimer's, Parkinson's, psychological disorders, psychosis and depres- sion. Autism may also be treated by the present compositions and methods. Treatment may consist of the use of a compound of the present invention alone or in combination with current medications used in the treatment of Parkinson's, Alzheimer's, or psychological disorders.

The present invention also provides a method of improving immunomodulatory therapy responses comprising the steps of exposing cells to an effective amount of a compound of the present invention, such as isoperillyl alcohol, before or during immunomodulatory treatment. Preferred immunomodulatory agents are cytokines, such interleukins, lymphokines, monokines, interfereons and chemokines.

The present composition may be administered by any method known in the art, including, without limitation, intranasal, oral, transdermal, ocular, intraperitoneal, inhalation, intravenous, ICV, intracisternal injection or infusion, subcutaneous, implant, vaginal, sublingual, urethral (e.g., urethral suppository), subcutaneous, intramuscular, intrave- nous, rectal, sub-lingual, mucosal, ophthalmic, spinal, intrathecal, intra-articular, intra-arterial, sub-arachinoid, bronchial and lymphatic administration. Topical formulation may be in the form of gel. ointment, cream, aerosol, etc; intranasal formulation can be delivered as a spray or in a drop; transdermal formulation may be administered via a transdermal patch or iontorphoresis; inhalation formulation can be delivered using a nebulizer or similar device. Compositions can also take the form of tablets, pills, capsules, semisolids, powders, sustained release formulations, solutions, suspensions, elixirs, aerosols, or any other appropriate compositions.

To prepare such pharmaceutical compositions, one or more of compound of the present invention may be mixed with a pharmaceutical acceptable carrier, adjuvant and/or excipient, according to conventional pharmaceutical compounding techniques. Pharmaceutically acceptable carriers that can be used in the present compositions encompass any of the standard pharmaceutical carriers, such as a phosphate buffered saline solution, water, and emulsions, such as an oil/water or water/oil emulsion, and various types of wetting agents. The compositions can additionally contain solid pharmaceutical excipients such as starch, cellulose, talc, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, magnesium stearate, sodium stearate, glycerol monostearate, sodium chloride, dried skim milk and the like. Liquid and semisolid excipients may be selected from glycerol, propylene glycol, water, ethanol and various oils, including those of petroleum, animal, vegetable or synthetic origin, e.g., peanut oil, soybean oil, mineral oil, sesame oil, etc. Liquid carriers, particularly for injectable solutions, include water, saline, aqueous dextrose, and glycols. For examples of carriers, stabilizers and adjuvants, see <NPL>). The compositions also can include stabilizers and preservatives.

As used herein, the term "therapeutically effective amount" is an amount sufficient to treat a specified disorder or disease or alternatively to obtain a pharmacological response treating a disorder or disease. Methods of determining the most effective means and dosage of administration can vary with the composition used for therapy, the purpose of the therapy, the target cell being treated and the subject being treated. Treatment dosages generally may be titrated to optimize safety and efficacy. Single or multiple administrations can be carried out with the dose level and pattern being selected by the treating physician. Suitable dosage formulations and methods of administering the agents can be readily determined by those of skill in the art. For example, the composition are administered at about <NUM>/kg to about <NUM>/kg, about <NUM>/kg to about <NUM>/kg, or about <NUM>/kg to about <NUM>/kg. When the compounds described herein are co-administered with another agent or therapy, the effective amount may be less than when the agent is used alone.

Transdermal formulations may be prepared by incorporating the active agent in a thixotropic or gelatinous carrier such as a cellulosic medium, e.g., methyl cellulose or hydroxyethyl cellulose, with the resulting formulation then being packed in a transdermal device adapted to be secured in dermal contact with the skin of a wearer. If the composition is in the form of a gel, the composition may be rubbed onto a membrane of the patient, for example, the skin, preferably intact, clean, and dry skin, of the shoulder or upper arm and or the upper torso, and maintained thereon for a period of time sufficient for delivery of the present compound to the blood serum of the patient. The composition of the present invention in gel form may be contained in a tube, a sachet, or a metered pump. Such a tube or sachet may contain one unit dose, or more than one unit dose, of the composition. A metered pump may be capable of dispensing one metered dose of the composition.

This invention also provides the compositions as described above for intranasal administration. As such, the compositions can further comprise a permeation enhancer. The present compound may be administered intranasally in a liquid form such as a solution, an emulsion, a suspension, drops, or in a solid form such as a powder, gel, or ointment.

Devices to deliver intranasal medications are well known in the art. Nasal drug delivery can be carried out using devices including, but not limited to, intranasal inhalers, intranasal spray devices, atomizers, nasal spray bottles, unit dose containers, pumps, droppers, squeeze bottles, nebulizers, metered dose inhalers (MDI), pressurized dose inhalers, insufflators, and bidirectional devices. The nasal delivery device can be metered to administer an accurate effective dosage amount to the nasal cavity. The nasal delivery device can be for single unit delivery or multiple unit delivery. In a specific example, the ViaNase Electronic Atomizer from Kurve Technology (Bethell, Washington) can be used in this invention (http://www. The compounds of the present invention may also be delivered through a tube, a catheter, a syringe, a packtail, a pledget, a nasal tampon or by submucosal infusion. <CIT>, <CIT>, <CIT>and <CIT>.

The present compound can be formulated as aerosols using standard procedures. The compound may be formulated with or without solvents, and formulated with or without carriers. The formulation may be a solution, or may be an aqueous emulsion with one or more surfactants. For example, an aerosol spray may be generated from pressurized container with a suitable propellant such as, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, hydrocarbons, compressed air, nitrogen, carbon dioxide, or other suitable gas. The dosage unit can be determined by providing a valve to deliver a metered amount. Pump spray dispensers can dispense a metered dose or a dose having a specific particle or droplet size. As used herein, the term "aerosol" refers to a suspension of fine solid particles or liquid solution droplets in a gas. Specifically, aerosol includes a gas-borne suspension of droplets of a monoterpene (or sesquiterpene), as may be produced in any suitable device, such as an MDI, a nebulizer, or a mist sprayer. Aerosol also includes a dry powder composition of the composition of the instant invention suspended in air or other carrier gas.

The present compound may be delivered to the nasal cavity as a powder in a form such as microspheres delivered by a nasal insufflator. The present compound may be absorbed to a solid surface, for example, a carrier. The powder or microspheres may be administered in a dry, air-dispensable form. The powder or microspheres may be stored in a container of the insufflator. Alternatively the powder or microspheres may be filled into a capsule, such as a gelatin capsule, or other single dose unit adapted for nasal administration.

The pharmaceutical composition can be delivered to the nasal cavity by direct placement of the composition in the nasal cavity, for example, in the form of a gel, an ointment, a nasal emulsion, a lotion, a cream, a nasal tampon, a dropper, or a bioadhesive strip. In certain embodiments, it can be desirable to prolong the residence time of the pharmaceutical composition in the nasal cavity, for example, to enhance absorption. Thus, the pharmaceutical compo- sition can optionally be formulated with a bioadhesive polymer, a gum (e.g., xanthan gum), chitosan (e.g., highly purified cationic polysaccharide), pectin (or any carbohydrate that thickens like a gel or emulsifies when applied to nasal mucosa), a microsphere (e.g., starch, albumin, dextran, cyclodextrin), gelatin, a liposome, carbamer, polyvinyl alcohol, alginate, acacia, chitosans and/or cellulose (e.g., methyl or propyl; hydroxyl or carboxy; carboxymethyl or hydroxylpropyl).

The composition containing the present compound can be administered by oral inhalation into the respiratory tract, i.e., the lungs.

Typical delivery systems for inhalable agents include nebulizer inhalers, dry powder inhalers (DPI), and metered- dose inhalers (MDI).

Nebulizer devices produce a stream of high velocity air that causes a therapeutic agent in the form of liquid to spray as a mist. The therapeutic agent is formulated in a liquid form such as a solution or a suspension of particles of suitable size. In one embodiment, the particles are micronized. The term "micronized" is defined as having about <NUM>% or more of the particles with a diameter of less than about <NUM>. Suitable nebulizer devices are provided commercially, for example, by PARI GmbH (Stamberg, Germany). Other nebulizer devices include Respimat (Boehringer Ingelheim) and those disclosed in, for example, <CIT> and <CIT>, and <CIT>. The present com- pound can be formulated for use in a nebulizer device as an aqueous solution or as a liquid suspension.

DPI devices typically administer a therapeutic agent in the form of a free flowing powder that can be dispersed in a patient's air-stream during inspiration. DPI devices which use an external energy source may also be used in the present invention. In order to achieve a free flowing powder, the present compound can be formulated with a suitable excipient (e.g., lactose). A dry powder formulation can be made, for example, by combining dry lactose having a particle size between about <NUM> and <NUM> with micronized particles of the present compound and dry blending. Alternatively, the compound can be formulated without excipients. The formulation is loaded into a dry powder dispenser, or into inhalation cartridges or capsules for use with a dry powder delivery device. Examples of DPI devices provided commer- cially include Diskhaler (GlaxoSmithKline, Research Triangle Park, N. ) (see, e.g., <CIT>); Diskus (GlaxoSmithKline) (see, e.g., <CIT>; Turbuhaler (AstraZeneca, Wilmington, Del. ) (see, e.g., <CIT>); and Rotahaler (GlaxoSmithKline) (see, e.g., <CIT>). Further examples of suitable DPI devices are described in <CIT>, <CIT>, and <CIT> and references therein.

MDI devices typically discharge a measured amount of the stored composition using compressed propellant gas. Formulations for MDI administration include a solution or suspension of an active ingredient in a liquefied propellant.

Examples of propellants include hydrofluoroalklanes (HFA), such as <NUM>,<NUM>,<NUM>,<NUM>-tetrafluoroethane (HFA 134a) and <NUM>,<NUM>,<NUM>,<NUM>,<NUM>,<NUM>,<NUM>-heptafluoro-n-propane (HFA <NUM>), and chlorofluorocarbons, such as CCl<NUM>F. Additional components of HFA formulations for MDI administration include co-solvents, such as ethanol, pentane, water; and surfactants, such as sorbitan trioleate, oleic acid, lecithin, and glycerin. (See, for example, <CIT>, <CIT>, and <CIT>). The formulation is loaded into an aerosol canister, which forms a portion of an MDI device. Examples of MDI devices developed specifically for use with HFA propellants are provided in <CIT> and <CIT>. For examples of processes of preparing suitable formulations and devices suitable for inhalation dosing see <CIT>,<CIT>, <CIT>, and <CIT>, and <CIT>, <CIT>, <CIT> and <CIT>.

The present compound may be encapsulated in liposomes or microcapsules for delivery via inhalation. A liposome is a vesicle composed of a lipid bilayer membrane and an aqueous interior. The lipid membrane may be made of phospholipids, examples of which include phosphatidylcholine such as lecithin and lysolecithin; acidic phospholipids such as phosphatidylserine and phosphatidylglycerol; and sphingophospholipids such as phosphatidylethanolamine and sphingomyelin. Alternatively, cholesterol may be added. A microcapsule is a particle coated with a coating material. For example, the coating material may consist of a mixture of a film-forming polymer, a hydrophobic plasticizer, a surface activating agent or/and a lubricant nitrogen-containing polymer. <CIT> and <CIT>.

The present compound may also be used alone or in combination with other chemotherapeutic agents via topical application for the treatment of localized cancers such as breast cancer or melanomas. The present compound may also be used in combination with narcotics or analgesics for transdermal delivery of pain medication.

This invention also provides the compositions as described above for ocular administration. As such, the com- positions can further comprise a permeation enhancer. For ocular administration, the compositions described herein can be formulated as a solution, emulsion, suspension, etc. A variety of vehicles suitable for administering compounds to the eye are known in the art. Specific non-limiting examples are described in <CIT>; <CIT>; <CIT>; <CIT>; <CIT>; <CIT>; <CIT>, <CIT>; <CIT>; <CIT>; and<CIT>.

The present compound can be given alone or in combination with other drugs for the treatment of the above diseases for a short or prolonged period of time. The present compositions can be administered to a mammal, preferably a human. Mammals include, but are not limited to, murines, rats. rabbit, simians, bovines, ovine, porcine, canines, feline, farm animals, sport animals, pets, equine, and primates.

The invention also provides a method for inhibiting the growth of a cell in vitro, ex vivo or in vivo, where a cell, such as a cancer cell, is contacted with an effective amount of the present compound as described herein.

Pathological cells or tissue such as hyperproliferative cells or tissue may be treated by contacting the cells or tissue with an effective amount of a composition of this invention. The cells, such as cancer cells, can be primary cancer cells or can be cultured cells available from tissue banks such as the American Type Culture Collection (ATCC). The pathological cells can be cells of a systemic cancer, gliomas, meningiomas, pituitary adenomas, or a CNS metastasis from a systemic cancer, lung cancer, prostate cancer, breast cancer, hematopoietic cancer or ovarian cancer. The cells can be from a vertebrate, preferably a mammal, more preferably a human.

In vitro efficacy of the present composition can be determined using methods well known in the art. For example, the cytoxicity of the present compound may be studied by MTT [<NUM>-(<NUM>,<NUM>-dimethylthiazol-<NUM>-yl)-<NUM>,<NUM>-diphenyl tetrazolium bromide] cytotoxicity assay. MTT assay is based on the principle of uptake of MTT, a tetrazolium salt, by metabolically active cells where it is metabolized into a blue colored formazon product, which can be read spectrometrically. The cytoxicity of the present compound may be studied by colony formation assay. Functional assays for inhibition of VEGF secretion and IL~<NUM> secretion may be performed via ELISA. Cell cycle block by the present compound may be studied by standard propidium iodide (PI) staining and flow cytometry. Invasion inhibition may be studied by Boyden chambers. In this assay a layer of reconstituted basement membrane, Matrigel, is coated onto chemotaxis filters and acts as a barrier to the migration of cells in the Boyden chambers. Only cells with invasive capacity can cross the Matrigel barrier. Other assays include, but are not limited to, cell viability assays, apoptosis assays, and morphological assays.

The following are examples of the present invention and are not to be construed as limiting.

The reaction scheme is the following:
<CHM>.

Isopropyltriphenylphosphonium iodide (<NUM>, <NUM> mmol) was added to NaH (<NUM>%, in mineral oil, <NUM>, <NUM> mmol) in dry dimethyl sulfoxide (<NUM>) at room temperature under a nitrogen atmosphere. The reaction mixture was slowly heated to <NUM> over a period of <NUM> and maintained at <NUM> until the reaction mass became a red color (approximately <NUM>). A solution of <NUM>,<NUM>-cyclohexanedione monoethylene ketal (<NUM>, <NUM>, <NUM> mmol) in dry dimethyl sulfoxide was added over a period of <NUM> while keeping the temperature below <NUM> and the reaction was maintained at <NUM> for <NUM>. The reaction mixture was cooled to room temperature, quenched with cold water (<NUM>), and extracted with ethyl acetate (<NUM>). The combined organic layer was washed with water (<NUM>), followed by brine (<NUM>%, <NUM>) and dried over sodium sulfate. The filtered organic layer was concentrated to give a solid which was triturated with hexanes (<NUM>) and the precipitated triphenylphosphine oxide was filtered off. The hexane layer was concentrated to give an oil which was purified by column chromatography. [Column dimensions: dia: <NUM>, height: <NUM>, silica: <NUM> mesh, eluted hexanes (<NUM>) followed by hexane: ethyl acetate (<NUM>:<NUM>, <NUM>)] The hexane: ethyl acetate fractions were combined and concentrated under vacuum to give an oil. Weight: <NUM>. Weight yield: <NUM>%. <NUM>H-NMR (<NUM>, CDCl<NUM>): δ <NUM>-<NUM> (t, <NUM>), <NUM> (s, <NUM>), <NUM> (m, <NUM>), <NUM> (s, <NUM>). MS (APCI method): No molecular ion peak was observed.

p-Toluenesulfonic acid (<NUM>, <NUM><NUM> mmol) was added to a solution of ketal (<NUM>, <NUM>, <NUM> mmol) in acetone (<NUM>) and water (<NUM>). The reaction mixture was heated to reflux and maintained at reflux for <NUM>. The mixture was cooled to room temperature, treated with saturated sodium bicarbonate (<NUM>) and concentrated under vacuum. The resulting oily residue was extracted with ethyl acetate (<NUM>), washed with water (<NUM>), then brine (<NUM>), and dried over sodium sulfate. The filtered organic layer was concentrated under vacuum to give an oil. Weight:
<NUM>. Weight yield: <NUM>%. <NUM>H-NMR (<NUM>, CDCl<NUM>): δ <NUM> (s, <NUM>), <NUM> (t, <NUM>), <NUM> (t, <NUM>). MS (APCI method): No molecular ion peak was observed (Note: <NUM>H-NMR showed the presence of ~<NUM>% of ketal <NUM> but used without purification).

Potassium t-butoxide (<NUM>, <NUM> mmol) was added to a mixture of ketone (<NUM>, <NUM>, <NUM> mmol) and trimethylsul-foxonium iodide (<NUM>, <NUM> mmol) in dry dimethyl sulfoxide (<NUM>) under nitrogen atmosphere at room temperature. The mixture was stirred for <NUM> at room temperature. The reaction was quenched by the addition of cold water (<NUM>) and extracted with ethyl acetate (<NUM>). The combined organic layer was washed with water (<NUM>) followed by brine (<NUM>) and dried over sodium sulfate. The filtered organic layer was concentrated under vacuum to give an oil. Weight: <NUM>. Weight yield: <NUM>%. <NUM>H-NMR (<NUM>, CDCl<NUM>): δ <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> (t, <NUM>), <NUM> (s, <NUM>). MS (APCI method): No molecular ion peak was observed.

Aluminum isopropoxide (<NUM>, <NUM> mmol) was added to a mixture of epoxide (<NUM>. <NUM>, <NUM> mmol) in toluene (<NUM>) and the mixture was heated to reflux for <NUM>. The mixture was cooled to room temperature and quenched with saturated potassium sodium tartrate solution. The organic layer was separated, washed with water (<NUM>), followed by brine (<NUM>), and dried over sodium sulfate. The filtered organic layer was concentrated under vacuum to give crude isoperillyl alcohol (<NUM>) as an oil. Weight: <NUM>, Weight yield: <NUM>%, Purity: ~<NUM>-<NUM>% (by GC area percent, Actual yield ca; <NUM>%).

Triethylamine (<NUM>, <NUM> mmol) was added to a solution of crude isoperillyl alcohol (<NUM>, <NUM>, <NUM> mmol) in dichloromethane (<NUM>). After stirring for <NUM>. <NUM>,<NUM>-dinitrobenzoyl chloride (<NUM>, <NUM> mmol) was added over a period of <NUM>. The reaction mixture was stirred for <NUM> and quenched with water (<NUM>). The organic layer was separated, washed with water (<NUM>), and dried over sodium sulfate. The filtered organic layer was concentrated under vacuum to give a pale yellow solid (<NUM>), which was recrystallized from acetone to give pure ester <NUM> as a pale yellow solid. Mp: <NUM>-<NUM> (acetone). Weight: <NUM>, Yield: <NUM>% (from epoxide). <NUM>H-NMR (<NUM>, CDCl<NUM>): δ <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (t, <NUM>), <NUM> (t, <NUM>), <NUM> (br s, <NUM>), <NUM> (s, <NUM>); <NUM> (s, <NUM>), <NUM> (t, <NUM>), <NUM> (s, <NUM>). MS (APCI method): m/e: <NUM> (<NUM>%), <NUM> (<NUM>%), <NUM> (<NUM>%), <NUM> (<NUM>%).

Aqueous sodium hydroxide (<NUM>, <NUM> mmol, dissolved in <NUM><NUM> of water) was added to an ice cold solution of <NUM>, <NUM>-dinitrobenzoic acid <NUM>-isopropylidene-cyclohex-<NUM>-enylmethyl ester (<NUM>, <NUM>, <NUM> mmol) in methanol (<NUM>) over a period of <NUM>. The reaction mixture was allowed to warm to room temperature and then stirred for <NUM>. The methanol was concentrated under vacuum to a minimum stirring volume and the mixture was suspended in water (<NUM>). The resulting mixture was extracted with ethyl acetate (<NUM>). The organic layer was washed with water (<NUM>), then brine (<NUM>), and dried over sodium sulfate. The filtered organic layer was concentrated under vacuum to give pure isoperillyl alcohol as an oil. Weight: <NUM>, Yield: <NUM>%, Purity: <NUM>%. (by GC AUC). <NUM>H-NMR (<NUM>, CDCl<NUM>):.

<NUM> solution of n-Butyl lithium in hexanes (<NUM>, <NUM> mmol) was added to a solution of diisopropylamine (<NUM>, <NUM> mmol) in dry THF (<NUM>) at -<NUM> over a period of <NUM> hr. After stirring for <NUM> at -<NUM>, a solution of ketone (<NUM>, <NUM>, <NUM> mmol) in dry THF (<NUM>) was added over a period of <NUM> white maintaining the temperature below -<NUM>. The reaction mixture was stirred for <NUM> at -<NUM>. A solution of phenyltriflimide (<NUM>, <NUM> mmol) in THF (<NUM>) was added slowly while maintaining the temperature below -<NUM>. The reaction mixture was slowly warmed to <NUM>, maintained for <NUM> at <NUM> and then quenched with satd ammonium chloride solution. The separated organic layer was washed with water (<NUM>), brine (<NUM>) and dried over sodium sulfate. The filtered organic layer was concentrated under vacuum and the resulting residue was purified by column chromatography [Column dimensions:
dia: <NUM>, height: <NUM>, silica: <NUM> mesh, eluted with hexanes (<NUM>)] The similar fractions were combined and concentrated under vacuum which gave an oil. Weight: <NUM>. Weight yield: <NUM>%. <NUM>H-NMR (<NUM>, CDCl<NUM>): δ <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (m, <NUM>), <NUM> (m, <NUM>), <NUM> (m, <NUM>), <NUM> (m, <NUM>). MS (APCI method): No molecular ion peak was observed.

Note-<NUM>: <NUM>H-NMR indicated the presence of aromatic peaks (~<NUM>%) between δ <NUM>-<NUM> which were attributed to the by-product trifluoro-N-phenylmethanesulfonamide.

Note-<NUM>: The compound <NUM> was also synthesized in low yield (<NUM>%) using triflic anhydride in the presence of <NUM>- di-tert-butyl-<NUM>-methylpyridine as a base.

To a solution of compound <NUM> (<NUM>, <NUM> mmol) in N'N-dimethylformamide (<NUM>) was added methanol (<NUM>), triethylamine (<NUM>, <NUM> mmol), <NUM>,<NUM>-bis(diphenylphosphino)propane (<NUM>, <NUM> mmol) and palladium acetate (<NUM>, <NUM> mmol). The reaction mixture was degassed and then stirred at room temperature under carbon monoxide (balloon pressure) for <NUM>. The reaction mixture was diluted with ethyl acetate (<NUM>) and washed with <NUM> N HCl (<NUM>), brine (<NUM>) and dried over sodium sulfate. The filtered organic layer was concentrated under vacuum and the resulting residue was purified by column chromatography. [Column dimensions: dia: <NUM>, height: <NUM>, silica: <NUM> mesh, eluted with hexanes (<NUM>) followed by ethyl acetate: hexanes (<NUM>%, <NUM>)] The similar fractions were combined and concentrated under vacuum which gave an oil. While TLC analysis showed only a single spot, <NUM>H-NMR and GC analysis indicated that the isolated material was a mixture of two primary components that co-eluted by TLC. Weight: <NUM>. Weight yield: <NUM>%. <NUM>H-NMR (<NUM>, CDCl<NUM>) indicated the presence of peaks corresponding to the methyl ester (<NUM>) along with an unknown impurity. GC analysis confirmed that it is mainly a mixture of two compounds with a ratio of <NUM>:<NUM>. MS (APCI method): m/e: <NUM> (M+, <NUM>%), <NUM> (M+<NUM>, <NUM> %). The other peaks (≤ <NUM>%) at M+: <NUM>,<NUM> & <NUM> were not characterized. The crude mixture was taken forward without purification.

Preparation of isoperillyl alcohol (<NUM>): Methyl ester (<NUM>, <NUM>, <NUM> mmol) in dry THF (<NUM>) was added to a cold solution of LAH (<NUM>, <NUM> mmol) in dry THF (<NUM>) over a period of <NUM>. The reaction mixture was slowly heated to reflux and maintained for <NUM>. The mixture was cooled to <NUM> and quenched with satd sodium sulfate (<NUM>). The precipitated lithium salts were filtered off and washed with hot ethyl acetate (<NUM>). The filtrate was dried over sodium sulfate. The filtered organic layer was concentrated under vacuum which gave an oil. Weight: <NUM>. Weight yield: <NUM>%. While TLC analysis showed only a single spot, <NUM>H-NMR and GC analysis indicated that the isolated material was a mixture of two primary components that co-eluted by TLC. <NUM>H-NMR (<NUM>, CDCl<NUM>) indicated the presence of peaks corresponding to the isoperillyl alcohol (<NUM>) along with an unknown impurity. MS (APCI method): m/e: <NUM> (M+<NUM>, <NUM>%), <NUM> (M+, <NUM> %), <NUM> (M-OH). The other peaks at M+: <NUM> (<NUM>%), <NUM>, (<NUM>%), <NUM> (<NUM>%), <NUM> (<NUM>%). <NUM> (<NUM>%), & <NUM> (<NUM>%) were not characterized. GC analysis confirmed the presence of isoperillyl alcohol (<NUM>%, (AUC)), compared with the iso-POH obtained from the epoxide route along with the unknown impurity (<NUM>%.

The MTT cytotoxicity assays were carried out after cells were treated with iso-POH (e.g., synthesized by the method in Example <NUM>) or other types of POH with different purity. <FIG> shows the results of the MTT cytotoxicity assays demonstrating the efficacy of different types of POH and iso-POH in killing LN229 human glioma cells. Sigma POH is the POH purchased from Sigma Chemicals having a purity of about <NUM>%. SGP-<NUM>-<NUM> was prepared from the WAKO POH by two-fold crystallization from di-isopropyl ether solvent, and has a GC relative area purity of about <NUM>% (area under the curve). KWH0744 is the crude POH purchased from Wako having a purity of about <NUM>%. SGP-<NUM>-<NUM> was prepared from the WAKO POH by single crystallization from di-isopropyl ether solvent, and has a GC relative area purity of about <NUM>% (area under the curve). <FIG> shows the results of the MTT cytotoxicity assays demonstrating the efficacy of different types of POH and iso-POH in killing U251 human glioma cells. <FIG> shows the results of the MTT cytotoxicity assays demonstrating the efficacy of different types of POH and iso-POH in killing A172 human glioma cells. The results suggest that iso-POH exhibited much better cytotoxicity than POH with different purity.

In vitro cytotoxicity of iso-POH in temozolomide-sensitive or temozolomide-resistant cells were also studied. Glioma cells were treated with Sigma POH, POH synthesized to GLP quality (SGP-<NUM>-<NUM>) having a purity of about <NUM>%, and iso-POH (SGP-<NUM>-79P, SGP-<NUM>-65P) for <NUM> hours and MTT assay performed. <FIG> demonstrates that A172 cells had the greatest cytotoxic response to iso-POH compared to GLP POH and Sigma POH (<FIG>). This response pattern was also seen in A172 temozolomide resistant cells (<FIG>). Similarly, <FIG> demonstrates that U251 cells had the greatest response to iso-POH (<FIG>), and that this response was also seen in U251 temozolomide resistant cells (<FIG>). The same response to iso-POH was seen in LN229 temozolomide sensitive (<FIG>) and temozolomide resistant cells (<FIG>). U87 cells, both temozolomide sensitive (<FIG>) and resistant (<FIG>), had the greatest response to iso-POH, albeit less than LN229 and U251 cells.

<FIG> shows the results of the MTT assays performed using TMZ sensitive U251 glioma cells over <NUM> hours using Sigma POH, GLP POH having a purity of about <NUM>%, iso-POH (Iso-POH65, Iso-POH79) all at <NUM> - <NUM> concentration. Iso-POH had greater cytotoxicity compared to Sigma POH and GLP POH (<FIG>). U251 temozolomide resistant cell line (U251-TR2) treated under same conditions demonstrated greater cytotoxicity with iso-POH compared to Sigma and GLP POH (<FIG>). Another U251 temozolomide resistant cell line (U231-TR1) treated under same conditions also demonstrated greater cytotoxicity with iso-POH (iso-POH65, iso-POH79) compared to GLP POH or Sigma POH (<FIG>).

<FIG> shows the results of the MTT assay performed using glioblastoma cancer stem cell line USC04 treated with both GLP-POH and iso-POH (1so-POH65) over <NUM> hrs. GLP POH and iso-POH demonstrated similar cytoxicity on the cells.

U251 TMZ-sensitive and TMZ-resistant (U251-TR1, U251-TR2) cells were treated with Sigma POH (<NUM>) or GLP POH (<NUM>) for <NUM> hours, then Western blot was performed. The results show that Sigma POH and GLP POH increased expression of glucose-regulatory protein <NUM> (GRP-<NUM>) and the apoptosis marker CHOP, suggesting increased endoplasmic reticulum (ER) stress after treatment (<FIG>). Under the same conditions, iso-POH (iso-POH65, iso-POH79) also increased ER stress (<FIG>).

U251 glioma cells were treated with <NUM> Sigma POH, GLP POH or iso-POH (isoPOH65, isoPOH79) for <NUM> hours, then Western blot was performed. The results (<FIG>) demonstrate that all treatments decreased Kras expression.

Oxalyl chloride (<NUM>, <NUM> mmol) will be added slowly to a mixture of Temozolamide (Source: OChem Incor- poration, Lot # 0711185A; <NUM>, <NUM> mmol) in <NUM>,<NUM>-dichloroethane (<NUM>) over a period of <NUM> while maintaining the temperature at <NUM> under N<NUM>. The reaction mixture will be allowed to warm to room temperature and then heated to reflux for <NUM>. The excess of oxalyl chloride and <NUM>,<NUM>-dichloroethane will be removed by concentration under vacuum The resulting residue will be redissolved in <NUM>,<NUM>-dichloroethane (<NUM>) and the reaction mixture cooled to <NUM> under N<NUM>. A solution of isoperillyl alcohol (<NUM>, <NUM> mmol) in <NUM>,<NUM>-dichloroethane (<NUM>) will be added over a period of <NUM>. The reaction mixture will be allowed to warm to room temperature and stirred for <NUM>. <NUM>,<NUM>-Dichloroethana will be concentrated under vacuum to give a residue which will be triturated with hexanes. The resulting pale yellow solid will be filtered and washed with hexanes.

The reaction scheme is as follows.

Phosgene (<NUM>% in toluene, <NUM>, <NUM> mmol) will be added to a mixture of isoperillyl alcohol (<NUM>, <NUM> mmol) and potassium carbonate (<NUM>, <NUM> mmol) in dry toluene (<NUM>) over a period of <NUM> while maintaining the temperature between <NUM>-<NUM>. The reaction mixture will be allowed to warm to room temperature and stirred for <NUM> under N<NUM>. The reaction mixture will be quenched with water (<NUM>) and the organic layer separated. The aqueous layer will be extracted with toluene (<NUM>) and the combined organic layer washed with water (<NUM> <NUM><NUM>), brine (<NUM>%, <NUM>), and dried over sodium sulfate (<NUM>). The filtered organic layer will be concentrated under vacuum to give isoperillyl chloroformate as an oil.

Butyl lithium (<NUM>, <NUM>, <NUM> mmol) will be added to a solution of rolipram (Source: GL synthesis. Lot # GLS-SH-<NUM>: <NUM>, <NUM> mmol) in dry THF (<NUM>) at -<NUM> over a period of <NUM> under N<NUM>. After the reaction mixture being stirred for <NUM> at -<NUM>, isoperillyl chloroformate (<NUM>, <NUM> mmol, dissolved in <NUM> THF) will be added over a period of <NUM> while maintaining the temperature at -<NUM>. The reaction mixture will be stirred for <NUM> and quenched with saturated ammonium chloride (<NUM>). The reaction mixture will be allowed to warm to room temperature and extracted with ethyl acetate (2x20 mL). The combined organic layer will be washed with water (<NUM>), brine (<NUM>%, <NUM>), and dried over sodium sulfate. The filtered organic layer will be concentrated to give an oil which will be purified by column chromatography [Column dimensions: dia: <NUM>, height: <NUM>. silica: <NUM>-<NUM> mesh] and eluted with a mixture of <NUM>% ethyl acetate/hexanes (<NUM>) followed by <NUM>% ethyl acetate/hexanes (<NUM>). The <NUM>% ethyl acetate /hexanes fractions will be combined and concentrated under vacuum to give a gummy solid.

Phosgene (<NUM>% in toluene, <NUM>, <NUM> mmol) will be added to a mixture of isoperillyl alcohol (<NUM>, <NUM> <NUM> mmol) and potassium carbonate (<NUM>, <NUM> mmol) in dry toluene (<NUM>) over a period of <NUM> while maintaining the temperature between <NUM>-<NUM>. The reaction mixture will be allowed to warm to room temperature and stirred for <NUM> under N<NUM>. The reaction mixture will be quenched with water (<NUM>) and the organic layer separated. The aqueous layer will be extracted with toluene (<NUM>) and the combined organic layer washed with water (<NUM> <NUM><NUM>), brine (<NUM>%, <NUM>), and dried over sodium sulfate (<NUM>). The filtered organic layer will be concentrated under vacuum to give isoperillyl chloroformate as an oil.

Isoperillyl chloroformate (<NUM>, <NUM> mmol) will be added slowly to a mixture of dimethyl celecoxib (<NUM>, <NUM> mmol) and potassium carbonate (<NUM>. <NUM>, <NUM> mmol) in dry acetone (<NUM>) over a period of <NUM> under N<NUM>. The reaction mixture will be heated to reflux and maintained for <NUM> The reaction mixture will be cooled and the acetone concentrated under vacuum The resulting residue will be suspended in water (<NUM>) and extracted with ethyl acetate (3x20 mL).

Claim 1:
A pharmaceutical composition comprising an isoperillyl alcohol carbamate, wherein the isoperillyl alcohol carbamate is an isoperillyl alcohol covalently bound via a carbamate linking group to temozolomide (TMZ), wherein the isoperillyl alcohol is selected from the group consisting of (<NUM>-isopropyl cyclohexa-<NUM>,<NUM>-dienyl)methanol, (<NUM>-isopropyl cyclohexa-<NUM>,<NUM>-dienyl)methanol, (<NUM>-isopropylphenyl)methanol and (<NUM>-isopropenylphenyl)methanol.