EA042139B1 - PHARMACEUTICAL COMPOSITIONS AND METHODS FOR CANCER TREATMENT - Google Patents

Abstract

Presented are pharmaceutical compositions and kits comprising a tyrosine hydroxylase inhibitor; melanin, a melanin promoter, or a combination thereof; a p450 3A4 promoter, and a leucine aminopeptidase inhibitor. Also presented are methods for treating a malignant neoplasm in a subject, comprising administering to a subject in need thereof an effective amount of a tyrosine hydroxylase inhibitor, a melanin promoter, a p450 3A4 promoter, and a leucine aminopeptidase inhibitor. Also provided are methods for reducing cell proliferation in a subject, comprising administering to a subject in need thereof an effective amount of a tyrosine hydroxylase inhibitor, a melanin promoter, a p450 3A4 promoter, and a leucine aminopeptidase inhibitor.

Description

Technical field

The present invention generally relates to compositions, kits and methods for reducing cell proliferation, such as in the treatment of cancer.

State of the art

According to the U.S. database National Cancer Institute's Surveillance Epidemiology and End Results (SEER) In 2008, the most recent year for which incidence data are available, 1,1958,000 Americans have invasive cancer. Cancer is the second most common cause of death in the United States, behind only cardiovascular disease, and is responsible for one in four deaths. It has been estimated that approximately 1,600 Americans die of cancer every day. In addition to the medical, emotional and psychological costs associated with cancer, there are significant financial costs to both the individual and society. The National Institutes of Health estimated the total cost of cancer care in 2010 to be $263.8 billion. In addition, an additional $140.1 billion is estimated to be lost in productivity due to premature death.

To date, cancer treatment includes surgery, hormone therapy, radiation therapy, chemotherapy, immunotherapy, targeted therapy, and combinations thereof. Surgical removal of a malignant neoplasm has advanced considerably; however, there remains a high chance of recurrence of the disease. Hormone therapy using drugs such as aromatase inhibitors and analogues and inhibitors of luteinizing hormone releasing factor has been relatively effective in the treatment of prostate and breast cancer. Radiation and related methods of conformal proton beam therapy, stereotactic radiosurgery, stereotactic radiotherapy, intraoperative radiotherapy, chemical modifiers, and radiosensitizers are effective in killing cancer cells, but can also kill and alter surrounding normal tissue. Chemotherapy drugs such as aminopterin, cisplatin, methotrexate, doxorubicin, daunorubicin, and others, alone or in combination, are effective in killing cancer cells by often altering the DNA replication process. Biological response modifier (BRM) therapy, biological therapy, biotherapy, or immunotherapy alters the growth of cancer cells or affects the natural immune response and includes the administration of biological agents to the patient, such as interferons, interleukins, and other cytokines, and antibodies, such as rituximab and trastuzumab, and even cancer vaccines such as sipuleucel-T.

Recently, new targeted therapies have been developed to combat cancer. These targeted therapies differ from chemotherapy in that they have a greater effect on cancer cells, while chemotherapy works by killing both cancerous and normal cells. Targeted therapy works by interfering with the processes that control the growth, division, and spread of cancer cells and the signals that cause cancer cells to naturally die. One type of targeted therapy includes growth signal inhibitors such as trastuzumab, gefitinib, imatinib, centuximab, dasatinib, and nilotinib. Another type of targeted therapy includes angiogenesis inhibitors such as bevacizumab, which inhibit malignancies associated with increased vasculature and blood supply. The latter type of targeted therapy includes drugs that induce apoptosis, which can cause direct death of cancer cells.

While all of these treatments have been effective to varying degrees, they all have disadvantages and limitations. In addition to being expensive, many treatments are also often too imprecise or the cancer is able to adapt to them and become resistant.

Thus, there is a great need for additional methods of treating malignant neoplasms. In particular, there is a need to treat cancer that has become resistant to other forms of treatment.

The essence of the invention

The present invention provides compositions, combination therapy, kits, and methods for reducing excessive cell proliferation, including those associated with the treatment of cancer. In some embodiments, the invention relates to pharmaceutical compositions containing at least one tyrosine hydroxylase inhibitor; at least one of melanin, a melanin promoter, or a combination thereof; at least one p450 3A4 promoter; at least one leucine aminopeptidase inhibitor; and optionally at least one growth hormone inhibitor. In other embodiments, the invention relates to kits that include these components along with suitable packaging. Also provided are methods for reducing cell proliferation and/or methods for treating cancer, comprising administering to a subject in need thereof an effective amount of at least one tyrosine hydroxylase inhibitor; at least one of melanin, a melanin promoter, or a combination thereof; at least one p450 3A4 promoter; at least one leucine aminopeptidase inhibitor; and optionally at least one growth hormone inhibitor.

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Detailed description of illustrative embodiments of the invention

The present subject matter may be more easily understood by reference to the following detailed description, which forms part of the information set forth herein. It should be understood that this invention is not limited to the specific products, methods, conditions, or parameters described and/or shown herein, and that the terminology used herein is intended to describe specific embodiments by way of example only, and is not intended to limitations of the claimed invention.

Unless otherwise indicated herein, scientific and technical terms used in connection with this application have the meanings traditionally understood by those of ordinary skill in the art. In addition, unless the context otherwise requires, singular terms include the plural, and terms in the singular include the plural.

As used above and throughout the description, the following terms and abbreviations, unless otherwise indicated, are to be understood as having the following meanings.

As used herein, the singular forms include a reference to a plural, and a reference to a specific numerical value includes at least that specific value, unless the context clearly indicates otherwise. Thus, for example, a reference to a compound is a reference to one or more such compounds and their equivalents known to those skilled in the art, and so on. The term set, as used herein, means more than one. When a range of values is expressed, another embodiment includes one specific value and/or another specific value. Likewise, if values are expressed as approximations using the preceding term approximately, then it should be understood that a particular value constitutes another embodiment. All ranges are included and can be combined.

As used herein, the terms component, composition, composition of compounds, compound, drug, pharmacologically active agent, active agent, therapeutic, therapy, treatment, or drug are used interchangeably herein to refer to a compound or compounds or compound composition that, when administered to a subject ( human or animal) produce the desired pharmacological and/or physiological effect through local and/or systemic action.

As used herein, the terms treatment or therapy (as well as their various forms) include preventive (eg, prophylactic), therapeutic, or palliative care. As used herein, the term treatment includes alleviating or reducing at least one adverse or negative effect or symptom of a condition, disease, or disorder. This condition, disease or disorder may be a malignancy.

As used above and throughout the description, the term effective amount refers to an amount effective in doses and for the necessary periods of time to achieve the desired result in relation to the treatment of the corresponding disorder, condition or side effect. It is understood that the effective amount of the components of the present invention will vary from patient to patient, not only depending on the particular compound, the selected component or composition, the route of administration and the ability of the components to produce the desired result in the individual, but also on factors such as the stage of the disease or the severity condition to be alleviated, hormone levels, age, sex, weight of the individual, the condition of the patient and the severity of the pathological condition being treated, the concomitant medications or special foods that a particular patient should be taking, and other factors that will be clear to specialists in this field of technology, at an appropriate dosage at the discretion of the attending physician. Dosing regimens can be adjusted to provide an improved therapeutic response. An effective amount is also one in which any toxic or detrimental effects of the components are outweighed by the therapeutically beneficial effects.

The phrase pharmaceutically acceptable, as used herein, refers to those compounds, substances, compositions and/or dosage forms which, within the limits of sound medical judgment, are suitable for use in contact with human and animal tissues without undue toxicity, irritation, allergic reaction. or other problem or complication, commensurate with a reasonable benefit/risk ratio.

Within the scope of the present invention, the described compounds can be obtained in the form of pharmaceutically acceptable salts. The term pharmaceutically acceptable salts refers to derivatives of the described compounds in which the parent compound is modified to form salts with acids or bases. Examples of pharmaceutically acceptable salts include, but are not limited to, salts of inorganic or organic acids with base residues such as amines; alkali or organic salts of acid residues such as carboxylic acids; etc. Pharmaceutically acceptable salts include conventional non-toxic or quaternary ammonium salts of the parent compound, formed from, for example, non-toxic inorganic or organic acids. For example

- 2 042139 measures, such conventional non-toxic salts include salts derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like; and salts derived from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethanedisulfonic, oxalic, isethionic and the like. These physiologically acceptable salts are prepared by methods known in the art, for example by dissolving the free amine bases with an excess of acid in an aqueous alcohol, or by neutralizing the free carboxylic acid with an alkali metal base such as hydroxide or with an amine.

The compounds described herein can be obtained in alternative forms. For example, many amine-containing compounds can be used or prepared as acid addition salts. Often such salts improve the properties of the compound for isolation or use. For example, depending on the reagents, reaction conditions, and the like, the compounds described herein can be used or obtained, for example, in the form of their hydrochloride or tosylate salts. Isomorphic crystalline forms, all chiral and racemic forms, Noxide, hydrates, solvates and hydrates of acid salts are also within the scope of the present invention.

Some acidic or basic compounds of the present invention may exist as zwitterions. All forms of the compounds, including free acid, free base, and zwitterions, are intended to be within the scope of the present invention. It is well known in the art that compounds containing both amino and carboxy groups often exist in equilibrium with their zwitterionic forms. Thus, any of the compounds described herein, which contain, for example, both amino and carboxy groups, also include a reference to their respective zwitterions.

The term stereoisomers refers to compounds that have an identical chemical structure but differ in the arrangement of atoms or groups in space.

The term administration means either the direct administration of a compound or composition of the present invention, or the administration of a prodrug, derivative or analog that forms an equivalent amount of the active compound or substance in the body.

The terms subject, individual, and patient are used interchangeably herein and refer to an animal, such as a human, for which treatment, including prophylactic treatment, is contemplated with a pharmaceutical composition of the present invention. As used herein, the term subject refers to animals and non-human animals. The terms non-human animals and non-human mammals are used interchangeably herein and include all vertebrate animals, e.g., mammals such as non-human primates (especially higher primates), sheep, dogs, rodents (e.g., mice or rats), guinea pigs, goats, pigs, cats, rabbits, cows, horses, and non-mammals such as reptiles, amphibians, chickens, and turkeys.

The term inhibitor as used herein includes compounds that inhibit the expression or activity of a protein, polypeptide, or enzyme, and optionally means complete inhibition of expression and/or activity. More specifically, inhibition includes inhibiting the expression and/or activity of a protein, polypeptide, or enzyme to an extent and for a time sufficient to produce the desired effect.

The term promoter, as used herein, includes compounds that stimulate the expression or activity of a protein, polypeptide, or enzyme, and optionally means complete activation of expression and/or activity. More specifically, promotion includes stimulating the expression and/or activity of a protein, polypeptide, or enzyme to an extent and for a time sufficient to produce the desired effect.

In one embodiment, the present invention relates to a combination therapy that alters the protection of cancer cells from oxidative stress. One class of such therapy increases the availability of free radicals to cancer cells. A representative subclass of such therapy includes the administration of pharmaceutical compositions comprising tyrosine hydroxylase inhibitors, a melanin or melanin promoter, a p450 3A4 promoter, a leucine aminopeptidase inhibitor, and optionally a growth hormone inhibitor. Another subclass includes the administration of a pharmaceutical composition comprising melanin and a tyrosine hydroxylase inhibitor. Specific components of the pharmaceutical composition are described below.

While not wishing to be bound by any particular mechanism of action, the tyrosine hydroxylase inhibitors of the present invention function by accumulating in cancer cells and preventing them from forming a coating of lipids or hyaluronan. By preventing cancer cells from forming a coating of lipids or hyaluron, it is believed that cancer cells become more accessible to oxidative stress. Typical tyrosine hydroxylase inhibitors include tyrosine derivatives, which are usually rapidly absorbed by most cancerous and inflamed tissues.

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Representative tyrosine derivatives include one or more of the following: methyl-(2R)-2-amino3-(2-chloro-4-hydroxyphenyl)propanoate, D-tyrosine ethyl ester hydrochloride, methyl-(2R)-2-amino-3( 2,6-dichloro-3,4-dimethoxyphenyl)propanoate, HD-Tyr(TBU)allyl ether-HCl, methyl-(2R)-2-amino-3(3-chloro-4,5-dimethoxyphenyl)propanoate, methyl -(2R)-2-amino-3-(2-chloro-3-hydroxy-4-methoxyphenyl)propanoate, methyl-(2R)-2-amino-3-(4-[(2-chloro-6-fluorophenyl) )methoxy]phenyl)propanoate, methyl-(2R)-2-amino3-(2-chloro-3,4-dimethoxyphenyl)propanoate, methyl-(2R)-2-amino-3-(3-chloro-5-fluoro -4-hydroxyphenyl)propanoate, diethyl-2-(acetylamino)-2-(4-[(2-chloro-6-fluorobenzyl)oxy]benzylmalonate, methyl-(2R)-2-amino-3(3-chloro- 4-methoxyphenyl)propanoate, methyl-(2R)-2-amino-3-(3-chloro-4-hydroxy-5-methoxyphenyl)propanoate, methyl-(2R)-2-amino-3-(2,6- dichloro-3-hydroxy-4-methoxyphenyl)propanoate, methyl-(2R)-2-amino3-(3-chloro-4-hydroxyphenyl)propanoate, H-DL-Tyr-OMe-HCl, H-3,5-diiodine -Tyr-OMeHCl, HD-3,5-diiodineTyr-OMeHCl, HD-Tyr-OMe-HCl, methyl eff. hydrochloride D-tyrosine hydrochloride, D-tyrosine-OMeHCl, methyl-D-tyrosinate hydrochloride, HD-Tyr-OMe-HCl, D-tyrosine methyl ester-HCl, HD-Tyr-OMe-HCl, (2R)-2-amino- 3-(4-hydroxyphenyl)propionic acid, (2R)-2-amino-3-(4-hydroxyphenyl)methyl ester hydrochloride, methyl-(2R)-2-amino-3-(4-hydroxyphenyl)propanoate hydrochloride, hydrochloride methyl-(2R)-2-azanyl-3-(4-hydroxyphenyl)propanoate, 3-chloro-L-tyrosine, 3-nitro-L-tyrosine, 3-nitro-L-tyrosine ethyl ester hydrochloride, DL-m- tyrosine, DL-o-tyrosine, Boc-Tyr(3,5-I ₂ )-OSu, FmocTyr(3-NO ₂ )-OH, and α-methyl-DL-tyrosine (also known as DL-2-methyl-3 -(4-hydroxyphenyl)alanine).

The present invention includes the use of at least one of melanin, a melanin promoter, or a combination thereof. Thus, melanin, one or more melanin promoters can be used, and both melanin and one or more melanin promoters can be used (either in separate dosage forms or in the same dosage form). The melanin promoters of the present invention are chemical compounds that increase the production and/or activity of melanin. An increase in melanin levels is thought to reduce inflammation (eg, by suppressing TNF) and eliminate the organ-specific lymphatic system. Melanin is a photocatalyst and therefore can promote chemical reactions that generate free radicals, which in turn can become available to cancer cells. Melanin promoters are represented by methoxsalen and melanotan II.

In some cases, the tyrosine hydroxylase inhibitor is mixed with melanin in the same dosage form. This association of melanin with the tyrosine hydroxylase inhibitor is believed to facilitate the uptake of melanin into cancer cells, since tyrosine hydroxylase inhibitors are more readily absorbed by such cells. In some embodiments, melanin is dissolved in a solubilizing agent and then mixed with a tyrosine hydroxylase inhibitor by methods known in the art. The solubilizing agent can be removed by standard methods such as evaporation, drying, etc. The solubilizing agent may be a non-toxic solubilizing agent such as hydrogen peroxide or other solubilizing agents commonly known in the art. The melanin and/or pharmaceutical composition may be further processed to optimize the effect of the pharmaceutical composition on cancer cells. In another embodiment, the pharmaceutical composition may include additional active agents and/or pharmaceutical excipients.

The pharmaceutical compositions of the invention also include the p450 3A4 promoter. Cytochrome p450 3A4 (which can be abbreviated as p450 3A4) is a member of the cytochrome p450 superfamily of enzymes and is a mixed function oxidase that is involved in the metabolism of xenobiotics in the body. It has the widest spectrum of substrates of all cytochromes. The function of the p450 3A4 promoter in the pharmaceutical compositions of the invention is to increase the expression and/or activity of p450 3A4. It is believed that the increased expression and/or activity of p450 3A4 reduces the level of cortisone and estrogen in the patient. In addition, increased expression and/or activity of p450 3A4 also slightly lowers blood pH, which is thought to help maintain or increase melanin activity. Exemplary p450 3A4 promoters are 5,5-diphenylhydantoin (sold as, for example, dilantin), valproic acid, and carbamazepine, which are believed to cause expression of the p450 3A4 enzyme.

The present pharmaceutical compositions further include leucine aminopeptidase inhibitors (alternatively known as leucyl aminopeptidase inhibitors).

Leucine aminopeptidases are enzymes that preferentially catalyze the hydrolysis of leucine residues at the N-terminus of peptides and/or proteins. It is assumed that inhibition of the expression and/or activity of leucine aminopeptidases promotes tumor reabsorption by increasing the transport of cholesterol to the liver. It is generally believed that aminopeptidase inhibitors, including aminopeptidase inhibitors, deplete sensitive tumor cells of specific amino acids, preventing protein recycling, thus creating an antiproliferative effect. Representative leucine aminopeptidase inhibitors are N-[(2S,3R)-3-amino-2-hydroxy-4-phenylbutyryl]-L-leucine and rapamycin.

The composition of instant pharmaceutical compositions also optionally includes inhibition

- 4 042139 tor of growth hormone. Growth hormone (such as, for example, pancreatic growth hormone) causes cells to replicate. Inhibition of growth hormone expression and/or activity is thought to preclude normal cells from rapidly replicating while allowing cancer cells to continue to rapidly replicate and turn on the tyrosine derivative. Typical growth hormone inhibitors are octreotide, somatostatin and seglitide.

Pharmaceutical compositions of the invention may further include D-leucine. D-leucine is a stereoisomer of naturally occurring L-leucine, the form of leucine incorporated into polypeptides and proteins. D-leucine cannot be incorporated into polypeptides and/or proteins. Along with the leucine aminopeptidase inhibitor, D-leucine is thought to create a physiological environment that mimics leucine deficiency. Thus, the presence of D-leucine allows the use of lower doses of the leucine aminopeptidase inhibitor in the pharmaceutical composition.

Also provided herein are combination therapy kits that alter the protection of cancer cells from oxidative stress. A contemplated suitable embodiment is a kit that includes a combination therapy that increases the availability of free radicals to cancer cells. Exemplary kits include a tyrosine hydroxylase inhibitor, a melanin and/or melanin promoter, a p450 3A4 promoter, a leucine aminopeptidase inhibitor, and optionally a growth hormone inhibitor of the type described above, along with packaging for them. The kit may include one or more individual containers, dividers or compartments and optionally informational material such as instructions for use. For example, each inhibitor or promoter (or various combinations thereof) may be contained in a bottle, vial, or syringe, and informational material may be contained in a plastic case or bag, or on a label. In some embodiments, a kit includes a collection (eg, packaging) of individual containers, each containing one or more unit dosage forms of a compound described herein. For example, a kit may include a collection of syringes, ampoules, foil pouches, or blister packs, each containing one single dose of a compound described herein, or any of various combinations thereof. The kit containers may be airtight, watertight (eg, impervious to moisture fluctuations or evaporation), and/or lighttight. The kit optionally includes a device suitable for administering the composition, such as a syringe, inhaler, pipette, tongs, measuring spoon, dropper (eg, eye dropper), swab (eg, cotton swab or Q-tip), or any similar delivery device.

Also provided are methods for treating cancer in a subject, as well as methods for reducing excessive cell proliferation. Such methods may include administering an effective amount of a combination therapy that modifies the protection of cancer cells from oxidative stress. Representative methods of treating cancer include administering an effective amount of a combination therapy that increases the availability of free radicals to cancer cells. Suitable embodiments are methods that include administering an effective amount of the aforementioned tyrosine hydroxylase inhibitor, melanin and/or melanin promoter, p450 3A4 promoter, leucine aminopeptidase inhibitor, and optionally a growth hormone inhibitor. Other suitable methods include administering an effective amount of melanin and a tyrosine hydroxylase inhibitor.

Suitable methods include the simultaneous or at least concurrent administration of at least two of the tyrosine hydroxylase inhibitors, melanin or melanin promoter, the p450 3A4 promoter and the leucine aminopeptidase inhibitor, at least three of them or each of them (in each case, optionally with growth hormone inhibitor). It is believed that it is desirable that an effective concentration of these groups be present in the subject's blood stream at the same time, and any drug regimen that achieves this is within the scope of the present invention. The desired number of inhibitors and promoters may be provided in a single dosage form or any number of desired dosage forms, including separate dosage forms. Typical dosage forms include tablets, capsules, caplets, sterile aqueous or organic solutions, reconstituted powders, elixirs, liquids, colloidal or other types of suspensions, emulsions, beads, grains, granules, microparticles, nanoparticles, and combinations thereof. The amount of composition administered will, of course, depend on the subject being treated, the weight of the subject, the severity of the condition being treated, the route of administration, and the judgment of the prescribing physician.

The introduction of melanin, promoters and/or inhibitors can be carried out in various ways, including oral, nasal, subcutaneous, intravenous, intramuscular, transdermal, vaginal, rectal, or any combination thereof. Transdermal administration can be carried out using, for example, oleic acid, 1-methyl-2-pyrrolidone or dodecylnonoxyethylene glycol monoester.

Melanin, promoters and/or inhibitors can be administered over a cycle of five to seven days of melanin, promoters and/or inhibitors and one to two days without melanin, promoters and/or inhibitors. Melanin, promoters and/or inhibitors can be administered for

- 5 042139 at least six of the indicated cycles. It may be desirable to administer these components approximately two hours between meals to facilitate absorption.

The subject to whom the instant compositions are administered may be a mammal, preferably a human.

In one exemplary method, 60 mg of the tyrosine derivative is administered orally and 0.25 ml of a 2 mg/ml suspension of the tyrosine derivative is administered subcutaneously; 10 mg of methoxsalen is administered orally and 0.25 ml of a 1 mg/ml suspension of methoxsalen is administered subcutaneously; 30 mg of 5,5-diphenylhydantoin is administered orally, and 20 mg of N-[(2S,3R)-3-amino-2-hydroxy-4-phenylbutyryl]-L-leucine is administered orally.

In some embodiments, the combination therapy includes:

(i) a dosage form containing melanin (50 μg) and α-methyl-DL-tyrosine (75 mg);

(ii) a dosage form containing 5,5-diphenylhydantoin (15 mg) and α-methyl-DL-tyrosine (75 mg);

(iii) a dosage form containing 3-amino-2-hydroxy-4-phenylbutyryl]-b-leucine (50 μg) and α-methyl-DL-tyrosine (75 mg);

(iv) a dosage form containing 3-amino-2-hydroxy-4-phenylbutyryl]-L-leucine (5 µg), melanotan II (10 µg) and 5,5-diphenylhydantoin (2 mg) and (v) a dosage form containing α-methyl-DL-tyrosine (5 mg) in bacteriostatic water with NaCl.

In other embodiments, the combination therapy includes:

(i) a dosage form containing melanin (50 μg) and α-methyl-DL-tyrosine (75 mg);

(ii) a dosage form containing 5,5-diphenylhydantoin (15 mg) and α-methyl-DL-tyrosine (75 mg);

(iii) a dosage form containing rapamycin (0.2 mg) and α-methyl-DL-tyrosine (75 mg);

(iv) a dosage form containing rapamycin (0.15 μg), melanotan II (10 μg) and 5.5 diphenylhydantoin (2 mg); and (v) a dosage form containing α-methyl-DL-tyrosine (5 mg) in bacteriostatic water with NaCl.

Doses that are twice this and even four times this are considered safe and effective.

Representative methods include those wherein the cancer is non-small cell lung cancer. In some embodiments, the non-small cell lung cancer is stage IV non-small cell lung cancer. In still other embodiments, the cancer is ovarian cancer, breast cancer, cervical cancer, pancreatic cancer, gastric cancer, brain cancer, liver cancer, testicular cancer, leukemia, lymphoma, appendix cancer, biliary tract cancer, cholangiocarcinoma , colon cancer, colorectal cancer, germ cell tumor, glioma, Hodgkin's lymphoma, lung cancer, neuroblastoma, prostate cancer, kidney cancer, sarcoma, thyroid cancer, tongue cancer, tonsil squamous cell carcinoma, or urothelial cancer.

In some embodiments, one or more tyrosine hydroxylase inhibitors; melanin promoter; p450 3A4 promoter and leucine aminopeptidase inhibitor is a nucleic acid, protein, antibody or antigen-binding fragment of an antibody.

The presented methods may include not only the described step of administering, but also the step of assessing the progression of the level of cancer in the specified subject and/or the degree of cell proliferation. The evaluation step may be performed before or after the administration step.

Suitable embodiments may include a pharmaceutical composition comprising a tyrosine hydroxylase inhibitor, a melanin and/or melanin promoter, a p450 3A4 promoter, and a leucine aminopeptidase inhibitor. The pharmaceutical composition may further comprise a growth hormone inhibitor. The growth hormone may be pancreatic growth hormone. The growth hormone inhibitor can be octreotide or somatostatin. The tyrosine hydroxylase inhibitor may be a tyrosine derivative. The tyrosine derivative may be one or more of the following: methyl-(2R)-2-amino-3-(2-chloro-4-hydroxyphenyl)propanoate, D-tyrosine ethyl ester hydrochloride, methyl-(2R)-2-amino -3-(2,6-dichloro-3,4-dimethoxyphenyl)propanoate, H-D-Tyr(TBU)-allyl ether-HCl, methyl-(2R)-2-amino-3-(3-chloro-4,5 -dimethoxyphenyl)propanoate, methyl-(2R)-2-amino-3-(2-chloro-3-hydroxy-4-methoxyphenyl)propanoate, methyl-(2R)-2-amino-3-(4-[(2 -chloro-6-fluorophenyl)methoxy]phenyl)propanoate, methyl-(2R)-2-amino-3-(2-chloro-3,4-dimethoxyphenyl)propanoate, methyl-(2R)-2-amino-3- (3-chloro-5fluoro-4-hydroxyphenyl)propanoate, diethyl 2-(acetylamino)-2-(4-[(2-chloro-6-fluorobenzyl)oxy]benzylmalonate, methyl-(2R)-2-amino- 3-(3-chloro-4-methoxyphenyl)propanoate, methyl-(2R)-2-amino-3-(3-chloro-4hydroxy-5-methoxyphenyl)propanoate, methyl-(2R)-2-amino-3- (2,6-dichloro-3-hydroxy-4-methoxyphenyl)propanoate, methyl-(2R)-2-amino-3-(3-chloro-4-hydroxyphenyl)propanoate, H-DL-Tyr-OMe-HCl, H-3,5diiodine-Tyr-OMe-HCl, H-D-3,5-diiodine-Tyr-OMe-HCl, H-D-Tyr-OMe-HCl, hydrochloride methyl ester D

- 6 042139 tyrosine, D-tyrosine-OMe-HCl, methyl ^-tyrosinate hydrochloride, HD-Tyr-OMe-HCl, D-tyrosine methyl ether-HCl, HD-Tyr-OMe-HCl, (2R)-2-amino -3-(4-hydroxyphenyl)propionic acid, (2R)-2-amino-3-(4-hydroxyphenyl)methyl ester hydrochloride, methyl-(2R)-2-amino-3-(4hydroxyphenyl)propanoate hydrochloride, methyl hydrochloride -(2R)-2-azanyl-3-(4-hydroxyphenyl)propanoate, 3-chloroL-tyrosine, 3-nitro-L-tyrosine, 3-nitro-L-tyrosine ethyl ester hydrochloride, DL-m-tyrosine, DL -otyrosine, Boc-Tyr(3,5-I2)-OSu, Fmoc-Tyr(3-NO ₂ )-OH and α-methyl-DL-tyrosine. The melanin promoter can be methoxsalen or melanotan II. The p450 3A4 promoter may be 5,5-diphenylhydantoin. The p450 3A4 promoter can be valproic acid or carbamazepine. The leucine aminopeptidase inhibitor may be N-[(2S,3R)-3-amino-2-hydroxy-4-phenylbutyryl]-L-leucine or rapamycin. Pharmaceutical compositions of the invention may further include D-leucine.

Also provided herein are kits containing a tyrosine hydroxylase inhibitor, melanin and/or melanin promoter, a p450 3A4 promoter and a leucine aminopeptidase inhibitor, together with packaging for them. The kit may further comprise a growth hormone inhibitor. The growth hormone may be pancreatic growth hormone. The growth hormone inhibitor can be octreotide or somatostatin. The tyrosine hydroxylase inhibitor may be a tyrosine derivative. The tyrosine derivative may be one or more of the following: methyl-(2R)-2-amino-3-(2-chloro-4hydroxyphenyl)propanoate, D-tyrosine ethyl ester hydrochloride, methyl-(2R)-2-amino-3 -(2,6dichloro-3,4-dimethoxyphenyl)propanoate, HD-Tyr(TBU)-allyl ether-HCl, methyl-(2R)-2-amino-3-(3chloro-4,5-dimethoxyphenyl)propanoate, methyl -(2R)-2-amino-3-(2-chloro-3-hydroxy-4-methoxyphenyl)propanoate, methyl-(2R)-2-amino-3-(4-[(2-chloro-6-fluorophenyl) )methoxy]phenyl)propanoate, methyl-(2R)-2-amino3-(2-chloro-3,4-dimethoxyphenyl)propanoate, methyl-(2R)-2-amino-3-(3-chloro-5-fluoro -4-hydroxyphenyl)propanoate, diethyl-2-(acetylamino)-2-(4-[(2-chloro-6-fluorobenzyl)oxy]benzylmalonate, methyl-(2R)-2-amino-3(3-chloro- 4-methoxyphenyl)propanoate, methyl-(2R)-2-amino-3-(3-chloro-4-hydroxy-5-methoxyphenyl)propanoate, methyl-(2R)-2-amino-3-(2,6- dichloro-3-hydroxy-4-methoxyphenyl)propanoate, methyl-(2R)-2-amino3-(3-chloro-4-hydroxyphenyl)propanoate, H-DL-Tyr-OMe-HCl, H-3,5-diiodine -Tyr-OMe-HCl, HD-3,5-diiodineTyr-OMe-HCl, HD-Tyr-OMe-HCl, methyl hydrochloride D-tyrosine hydrochloride, D-tyrosine-OMeΉCl, methyl-D-tyrosinate hydrochloride, HD-Tyr-OMe-HCl, D-tyrosine methyl ester-HCl, HD-Tyr-OMe-HCl, (2R)-2-amino -3-(4-hydroxyphenyl)propionic acid, (2R)-2-amino-3-(4hydroxyphenyl)methyl ester hydrochloride, methyl-(2R)-2-amino-3-(4-hydroxyphenyl)propanoate hydrochloride, methyl hydrochloride -(2R)-2-azanyl-3-(4-hydroxyphenyl)propanoate, 3-chloro-L-tyrosine, 3-nitro-L-tyrosine, 3-nitro-L-tyrosine ethyl ester hydrochloride, DL-m-tyrosine, DL -o-tyrosine, Boc-Tyr(3,5I ₂ )-OSu, Fmoc-Tyr(3-NO ₂ )-OH and α-methyl-DL-tyrosine. The melanin promoter can be methoxsalen or melanotan II. The p450 3A4 promoter can be 5,5-diphenylhydantoin, valproic acid, or carbamazepine. The leucine aminopeptidase inhibitor may be N-[(2S,3R)-3-amino-2-hydroxy-4phenylbutyryl]-L-leucine or rapamycin. The kits of the invention may additionally contain D-leucine.

Also provided are methods of treating cancer in a subject, comprising administering to a subject in need thereof an effective amount of a tyrosine hydroxylase inhibitor, melanin and/or melanin promoter, p450 3A4 promoter, and a leucine aminopeptidase inhibitor. In a suitable embodiment, the method of treating cancer may further comprise a growth hormone inhibitor. In some embodiments, at least two components (ie melanin, promoters and/or inhibitors) are administered simultaneously. In other embodiments, at least three components are administered simultaneously. Each of the components can be entered simultaneously. In suitable embodiments, the components are administered orally, subcutaneously, intravenously, transdermally, vaginally, rectally, or any combination thereof. Transdermal administration can be done with oleic acid, 1-methyl-2-pyrrolidone, or dodecylnonoxyethylene glycol monoester. In other embodiments, the components are administered over a cycle of five to seven days of component administration and one to two days of no component administration. The components may be administered for at least six of these cycles. The tyrosine hydroxylase inhibitor may be a tyrosine derivative. The tyrosine derivative may be one or more of the following: methyl-(2R)-2-amino-3-(2-chloro-4-hydroxyphenyl)propanoate, D-tyrosine ethyl ester hydrochloride, methyl-(2R)-2-amino -3-(2,6-dichloro-3,4-dimethoxyphenyl)propanoate, H-D-Tyr(TBU)allyl ether-HCl, methyl-(2R)-2-amino-3-(3-chloro-4,5- dimethoxyphenyl)propanoate, methyl-(2R)-2-amino-3-(2-chloro-3-hydroxy-4-methoxyphenyl)propanoate, methyl-(2R)-2-amino-3-(4-[(2- chloro-6fluorophenyl)methoxy]phenyl)propanoate, methyl-(2R)-2-amino-3-(2-chloro-3,4-dimethoxyphenyl)propanoate, methyl-(2R)-2-amino-3-(3- chloro-5-fluoro-4-hydroxyphenyl)propanoate, diethyl 2-(acetylamino)-2-(4-[(2chloro-6-fluorobenzyl)oxy]benzylmalonate, methyl-(2R)-2-amino-3-( 3-chloro-4-methoxyphenyl)propanoate, methyl-(2R)-2-amino-3-(3-chloro-4-hydroxy-5-methoxyphenyl)propanoate, methyl-(2R)-2-amino-3-( 2,6-dichloro-3hydroxy-4-methoxyphenyl)propanoate, methyl-(2R)-2-amino-3-(3-chloro-4-hydroxyphenyl)propanoate, H-DLTyr-OMe-HCl, H-3.5 -diiodine-Tyr-OMe-HCl, H-D-3,5-diiodine-Tyr-OMe-HCl, H-D-Tyr-OMe-HCl, hydrochloride m D-tyrosine ethyl ester, D-tyrosine-OMe-HCl, methyl-D-tyrosinate hydrochloride, H-D-Tyr

- 7 042139

OMe-HCl, D-tyrosine methyl ester-HCl, HD-Tyr-OMe-HCl, (2R)-2-amino-3-(4-hydroxyphenyl)propionic acid, (2R)-2-amino-3-( hydrochloride 4-hydroxyphenyl)methyl ester hydrochloride, methyl-(2R)-2-amino-3-(4-hydroxyphenyl)propanoate, methyl-(2R)-2-azanyl-3-(4hydroxyphenyl)propanoate hydrochloride, 3-chloro^- tyrosine, 3-nitro-b-tyrosine, 3-nitro-L-tyrosine ethyl ester hydrochloride, DL-m-tyrosine, DL-o-tyrosine, Boc-Tyr(3,5-I ₂ )-OSu, Fmoc-Tyr(3 -NO ₂ )-OH and α-methylDL-tyrosine. In a suitable embodiment of the method, 60 mg of the tyrosine derivative is administered orally and 0.25 ml of a 2 mg/ml suspension of the tyrosine derivative is administered subcutaneously. The melanin promoter may be methoxsalen. In another suitable method, 10 mg of methoxsalene is administered orally and 0.25 ml of a 1 mg/ml suspension of methoxsalen is administered subcutaneously. The melanin promoter can also be melanotan II. The p450 3A4 promoter may be 5,5-diphenylhydantoin. In another suitable route, 30 mg of 5,5-diphenylhydantoin is administered orally. The p450 3A4 promoter can be valproic acid or carbamazepine. The leucine aminopeptidase inhibitor may be N-[(2S,3R)-3-amino-2hydroxy-4-phenylbutyryl]-L-leucine. In another suitable method, 20 mg of N-[(2S,3R)-3-amino-2hydroxy-4-phenylbutyryl]-L-leucine is administered orally. The leucine aminopeptidase inhibitor may also be rapamycin. The growth hormone may be pancreatic growth hormone. The growth hormone inhibitor can be octreotide or somatostatin. The method may further comprise administering an effective amount of D-leucine. The subject may be a mammal, and the mammal may be a human. Representative methods include those wherein the cancer is non-small cell lung cancer. In some embodiments, the non-small cell lung cancer is stage IV non-small cell lung cancer. In still other embodiments, the cancer is ovarian cancer, breast cancer, cervical cancer, pancreatic cancer, gastric cancer, brain cancer, liver cancer, testicular cancer, leukemia, lymphoma, appendix cancer, biliary tract cancer, cholangiocarcinoma , colon cancer, colorectal cancer, germ cell tumor, glioma, Hodgkin's lymphoma, lung cancer, neuroblastoma, prostate cancer, kidney cancer, sarcoma, thyroid cancer, tongue cancer, tonsil squamous cell carcinoma, or urothelial cancer. In other suitable embodiments, the tyrosine hydroxylase inhibitor, melanin promoter, p450 3A4 promoter, and leucine aminopeptidase inhibitor are one or more of nucleic acids, proteins, antibodies, or antigen-binding antibody fragments. Another suitable embodiment further includes assessing the progression of the level of malignancy in said subject. The evaluation step may be performed before said administration step, or the evaluation step may be performed after said administration step.

Also provided are methods for reducing cell proliferation in a subject, comprising administering to a subject in need thereof an effective amount of a tyrosine hydroxylase inhibitor; melanin and/or melanin promoter; p450 3A4 promoter; and a leucine aminopeptidase inhibitor. In a suitable embodiment, the method of treating cancer may further comprise a growth hormone inhibitor. In some embodiments, at least two components (ie melanin, promoters and/or inhibitors) are administered simultaneously. In other embodiments, at least three components are administered simultaneously. Each of the components can be entered simultaneously. In suitable embodiments, the components are administered orally, subcutaneously, intravenously, transdermally, vaginally, rectally, or any combination thereof. Transdermal administration can be done with oleic acid, 1-methyl-2-pyrrolidone, or dodecylnonoxyethylene glycol monoester. In other embodiments, the components are administered over a cycle of five to seven days of component administration and one to two days of no component administration. The components may be administered for at least six of these cycles. The tyrosine hydroxylase inhibitor may be a tyrosine derivative. The tyrosine derivative may be one or more of the following: methyl-(2R)-2-amino-3-(2-chloro-4-hydroxyphenyl)propanoate, D-tyrosine ethyl ester hydrochloride, methyl-(2R)-2-amino -3-(2,6-dichloro-3,4-dimethoxyphenyl)propanoate, H-D-Tyr(TBU)-allyl ether-HCl, methyl-(2R)-2-amino-3-(3-chloro-4,5 -dimethoxyphenyl)propanoate, methyl-(2R)-2-amino-3-(2-chloro-3-hydroxy-4-methoxyphenyl)propanoate, methyl-(2R)-2-amino-3-(4-[(2 -chloro-6fluorophenyl)methoxy]phenyl)propanoate, methyl-(2R)-2-amino-3-(2-chloro-3,4-dimethoxyphenyl)propanoate, methyl-(2R)-2-amino-3-(3 -chloro-5-fluoro-4-hydroxyphenyl)propanoate, diethyl-2-(acetylamino)-2-(4-[(2chloro-6-fluorobenzyl)oxy]benzylmalonate, methyl-(2R)-2-amino-3- (3-chloro-4-methoxyphenyl)propanoate, methyl-(2R)-2-amino-3-(3-chloro-4-hydroxy-5-methoxyphenyl)propanoate, methyl-(2R)-2-amino-3- (2,6-dichloro-3hydroxy-4-methoxyphenyl)propanoate, methyl-(2R)-2-amino-3-(3-chloro-4-hydroxyphenyl)propanoate, H-DLTyr-OMe-HCl, H-3, 5-diiodine-Tyr-OMe-HCl, H-D-3,5-diiodine-Tyr-OMe-HCl, H-D-Tyr-OMe-HCl, hydrochloride D-tyrosine methyl ester, D-tyrosine-OMe-HCl, methyl-D-tyrosinate hydrochloride, H-D-TyrOMe-HCl, D-tyrosine methyl ester-HCl, H-D-Tyr-OMe-HCl, (2R)-2-amino -3-(4-hydroxyphenyl)propionic acid, (2R)-2-amino-3-(4-hydroxyphenyl)methyl ester hydrochloride, methyl-(2R)-2-amino-3-(4-hydroxyphenyl)propanoate hydrochloride, methyl-(2R)-2-azanyl-3-(4hydroxyphenyl)propanoate hydrochloride, 3-chloro-L-tyrosine, 3-nitro-L-tyrosine, ethyl ester hydrochloride 3

- 8 042139

HUTpo-L-tyrosine, DL-m-tyrosine, DL-o-tyrosine, Boc-Tyr(3,5-I2)-OSu, Fmoc-Tyr(3-NO2)-OH and α-methylDL-tyrosine. In a suitable embodiment of the method, 60 mg of the tyrosine derivative is administered orally and 0.25 ml of a 2 mg/ml suspension of the tyrosine derivative is administered subcutaneously. The melanin promoter may be methoxsalen. In another suitable method, 10 mg of methoxsalene is administered orally and 0.25 ml of a 1 mg/ml suspension of methoxsalen is administered subcutaneously. The melanin promoter can also be melanotan II. The p450 3A4 promoter may be 5,5-diphenylhydantoin. In another suitable route, 30 mg of 5,5-diphenylhydantoin is administered orally. The p450 3A4 promoter can be valproic acid or carbamazepine. The leucine aminopeptidase inhibitor may be N-[(2S,3R)-3-amino-2hydroxy-4-phenylbutyryl]-L-leucine. In another suitable method, 20 mg of N-[(2S,3R)-3-amino-2hydroxy-4-phenylbutyryl]-L-leucine is administered orally. The leucine aminopeptidase inhibitor may also be rapamycin. The growth hormone may be pancreatic growth hormone. The growth hormone inhibitor can be octreotide or somatostatin. The method may further comprise administering an effective amount of D-leucine. The subject may be a mammal, and the mammal may be a human. Representative methods include those wherein the cancer is non-small cell lung cancer. In some embodiments, the non-small cell lung cancer is stage IV non-small cell lung cancer. In still other embodiments, the cancer is ovarian cancer, breast cancer, cervical cancer, pancreatic cancer, gastric cancer, brain cancer, liver cancer, testicular cancer, leukemia, lymphoma, appendix cancer, biliary tract cancer, cholangiocarcinoma , colon cancer, colorectal cancer, germ cell tumor, glioma, Hodgkin's lymphoma, lung cancer, neuroblastoma, prostate cancer, kidney cancer, sarcoma, thyroid cancer, tongue cancer, tonsil squamous cell carcinoma, or urothelial cancer. In other suitable embodiments, the tyrosine hydroxylase inhibitor, melanin promoter, p450 3A4 promoter, and leucine aminopeptidase inhibitor are one or more nucleic acids, proteins, antibodies, or antigen-binding antibody fragments. Another suitable embodiment further includes assessing the progression of the level of malignancy in said subject. The evaluation step may be performed before said administration step, or the evaluation step may be performed after said administration step.

The following examples of specific embodiments of the present invention are offered for illustrative purposes only and are not intended to limit the scope of the present invention in any way.

Representative routes for administering pharmaceutical compositions and combination therapies are also provided. Various embodiments of the present invention further relate to methods for administering a pharmaceutical composition or combination therapy to a human patient for the treatment of cancer. The methods may include administering the pharmaceutical composition or combination therapy by conventional routes of administration (eg, oral, subcutaneous, parenteral, inhalation, topical, etc.). In some instances, the pharmaceutical composition or combination therapy may be administered orally and/or subcutaneously. In some cases, human patients may be given a pharmaceutical composition or combination therapy between meals.

In some embodiments of the present invention, the pharmaceutical composition or combination therapy can be administered to a human patient for 5 days per week over a period of 6 weeks, creating one cycle of 30 days of treatment. Depending on the result, after 6 weeks or one cycle of treatment, additional cycles of the pharmaceutical composition or combination therapy may be prescribed. The present invention also provides pharmaceutical compositions containing a tyrosine hydroxylase inhibitor; and melanin, a melanin promoter, or a combination thereof (preferably melanin);

pharmaceutical compositions containing a tyrosine hydroxylase inhibitor and a p450 3A4 promoter;

pharmaceutical compositions containing a tyrosine hydroxylase inhibitor and a leucine aminopeptidase inhibitor; and pharmaceutical compositions containing melanin, a melanin promoter, or a combination thereof (preferably melanotan II); p450 3A4 promoter; and leucine aminopeptidase inhibitor.

The tyrosine hydroxylase inhibitor in such compositions is preferably αmethyl-DL-tyrosine, the p450 3A4 promoter is preferably 5,5-diphenylhydantoin, and the leucine aminopeptidase inhibitor is preferably N-[(2S,3R)-3-amino-2-hydroxy4-phenylbutyryl ]-L-leucine. The invention also relates to kits containing each of these pharmaceutical compositions, along with a pharmaceutical composition containing a tyrosine hydroxylase inhibitor; as well as methods comprising administering each of the pharmaceutical compositions to a patient together with a pharmaceutical composition containing a tyrosine hydroxylase inhibitor, preferably such that they are administered to the patient over a 24 hour period.

- 9 042139

Example 1

A clinical study was performed to evaluate the efficacy, safety, acceptability and tolerability of combination therapy according to embodiments of the present invention as a treatment for metastatic cancer.

Combination therapy included the following:

(a) a capsule containing melanin (50 μg) and α-methyl-DL-tyrosine (75 mg) administered orally;

(b) a capsule containing 5,5-diphenylhydantoin (15 mg) and α-methyl-DL-tyrosine (75 mg) administered orally;

(c) a capsule containing 5,5-diphenylhydantoin (15 mg) and α-methyl-DL-tyrosine (75 mg) administered orally;

(d) a capsule containing rapamycin (0.2 mg) and α-methyl-DL-tyrosine (75 mg) administered orally;

(e) a suspension containing rapamycin (0.15 μg), melanotan II (10 μg) and 5,5-diphenylhydantoin (2 mg) administered subcutaneously; and (f) a suspension containing α-methyl-DL-tyrosine (5 mg) in NaCl bacteriostatic water administered subcutaneously.

Each patient was given combination therapy five days a week for six weeks. More than 200 patients were examined. Criteria included patients with metastatic cancer. Thirty patients meeting these criteria were accepted and participated in the study.

Treatment of metastatic breast cancer Fourteen patients in the study suffered from metastatic breast cancer. Patient information and results are shown in the table.

Table

Patient characteristics Average age 55 (40-70 years old) Women 14/14 Caucasoids 13/14 Before research Refusal of planned treatment before enrollment in studies All available treatments were used and found to be incurable Treatment results Improved ECOG rating by 1-3 points Improved EORTC rating by 1-5 points 4/14 10/14 11/14 10/14 (scale 1-7) Weight Weight gain Remained the same weight Weight loss 4/14 (1-5 pounds) 6/14 4/14 (1-2 pounds) Pain Pain reduction (scale 1 to 10) Admission to the study without pain and maintaining the same level Admission to research Pain medication Pain medication at the end of the cycle is no longer required 8/14 (1-9) 6/14 6/14 5/6 Signs of cancer Healthy on routine physical examination, systems review, and imaging Significant reduction in the number and/or size of the largest tumor Reducing the number and/or size of the largest tumor No cancer progression 3/14 5/14 2/14 4/14 Survival live 33-37 weeks 27-29 weeks 12-19 weeks 14/14 4/14 5/14 5/14 Current state Gone home Continuation of treatment 3/14 11/14

- 10 042139

One of the side effects of therapy was hyperpigmentation in all patients. In general, all patients tolerated combination therapy, there were no side effects.

Over 200 cancer patients have been screened in clinical trials. Thirty (30) eligible subjects provided consent. The median age of the patients was 56 years, with a range of 30 to 70 years. Patients in the study received a treatment regimen that included a tyrosine hydroxylase inhibitor (i.e., α-methyl-DL-tyrosine), a melanin promoter (i.e., melanotan II), a p450 3A4 promoter (i.e., 5,5-diphenylhydantoin), and a leucine aminopeptidase inhibitor (i.e., rapamycin). These compounds were administered every five days a week for six weeks with one or two days off between weekly cycles.

After six weeks of treatment, 12 of 30 patients (40%) maintained the same Eastern Cooperative Oncology Group (ECOG) rating of 0-5 (see Oken, et al., Toxicity and Response Criteria Of The Eastern Cooperative Oncology Group, Am. J. Clin. Oncol., 5:649-655, 1982). Fourteen (14) out of 30 (46%) had a 1-3 point improvement in the ECOG rankings.

Fourteen (14) of 30 patients (46%) maintained the same rating of 1-7 on the European Organization for the Research and Treatment of Cancer Quality of Life Questionnaire Core (EORTC) scale (see, e.g., Bergman, et al ., The EORTC QLQ-LC13: a modular supplement to the EORTC Core Quality of Life Questionnaire (QLQ-C30) for use in lung cancer clinical trials, EORTC Study Group on Quality of Life, Eur. J. Cancer, 1994. 30A( 5): pp. 635-42). Sixteen (16) out of 30 (54%) had a 1-5 point improvement in the EORTC rankings.

Eleven (11) out of 30 patients gained 1 to 9 pounds, 17 out of 30 remained the same weight, and 2 out of 30 lost 1 to 2 pounds.

Thirteen (13) out of 30 (43%) had a reduction in pain levels. Seventeen (17) of 30 (57%) presented with minimal pain and maintained the same level. Nine (9) of 30 (30%) entered the study on pain medication and 8 of these nine (89%) no longer needed pain medication at the end of the cycle.

No disease was found in four (4) of 30 (13%) cases on routine physical examination, system review, and imaging. Eight (8) out of 30 (27%) had a significant reduction in the number of tumors and/or the size of the largest tumor. Eight (8) out of 30 (27%) experienced a decrease in the number of tumors and/or the size of the largest tumor. Ten (10) out of 30 (33%) showed no disease progression.

Twenty-nine (29) of 30 patients were alive with a median survival of 22 weeks. Thirteen (13) out of 30 (43%) were released and went home. Seventeen (17) of 30 (57%) continued treatment. All subjects developed hyperpigmentation.

In general, the above treatment was well tolerated by the subjects, with no treatment-related side effects, and 100% response to treatment was recorded.

Example 2

In one aspect, the present invention relates to methods for inducing melanin production in vivo with one or both of methoxsalen and melanotan and/or by administering melanin. Without wishing to be bound by any particular theory, melanin is believed to be useful due to its photocatalytic nature and its ability to convert various wavelengths of environmental or induced electromagnetic radiation into electrical energy, thus causing desired reactions or dislocations. In some patients, either due to genetic variability, weakness, the need for accelerated availability, or to maximize efficacy, it has been found that melanin is preferably combined mechanically or chemically with α-methyl-DL-tyrosine prior to administration.

It is believed that melanin as a photocatalyst has polarity at the points of its physical mass. It has been found that small melanin particles can produce less electrical energy than larger particles, and aggregates of melanin particles generally do not accumulate in polarity-specific formations. An effective way to obtain a high output of electrical energy from melanin is the formation of melanin in the form of large polarized particles.

It is implicitly believed that for cancer, nanoparticles, regardless of the number, are not as desirable as larger particles, and that larger particles have a greater ability to be perceived by cancer cell membranes.

According to some embodiments, melanin is combined with a-methyl-DL-tyrosine in at least three ways.

1) Melanin, either natural or synthetic, is mechanically mixed with a compressive force to coagulate melanin, which is insoluble in water and somewhat malleable, with a-methyl-DL-tyrosine. After initially combining these components, it is desirable to add additional α-methyl-DL-tyrosine until a substantial coverage of melanin is achieved.

2) Melanin can be solubilized in many ways, as described in US patent No. 5225435, the contents of which are incorporated herein by reference. one prefer

- 11 042139 The effective method includes mixing melanin with distilled water and hydrogen peroxide to achieve a melanin concentration of at least 5 wt.% and then placing the resulting composition in a microwave oven until the boiling point is reached. The dissolved melanin thus obtained is used for infusion or bulking of a-methyl-Bb-tyrosine. The composition is then dried and the dry powder is used.

3) a-Methyl-Bb-tyrosine is placed in distilled water with 5-benzyloxy-6-methoxyindole and sealed for up to a month. It is believed that the L-part of racemic a-methyl-BL-tyrosine is converted to (BOPA) melanin. The size of the melanin particles can be controlled by controlling the time of the growth period. The resulting powder is then purified and dried. The racemic ratio is not more than 50/50, but the use of chemically combined ingredients facilitates the penetration of melanin even with a reduced L-component and, apparently, provides a potentially sufficient benefit.

Claims (12)

CLAIM 1. A method of treating a malignant neoplasm in a patient, comprising administering to a patient in need thereof a pharmaceutical composition containing: a) a tyrosine hydroxylase inhibitor, which is a-methyl-BE-tyrosine; b) a melanin promoter which is methoxsalen; c) the p450 3A4 promoter, which is 5,5-diphenylhydantoin; And d) a leucine aminopeptidase inhibitor which is rapamycin; wherein said cancer is metastatic cholangiocarcinoma, metastatic lung cancer, metastatic kidney cancer, metastatic colon cancer, metastatic tonsil cancer, metastatic appendix cancer, metastatic sarcoma, metastatic thyroid cancer, and metastatic glioma. 2. The method of claim 1 wherein said pharmaceutical composition is administered to said patient over a 24 hour period. 3. The method of claim 1, wherein said pharmaceutical composition is administered to said patient five days a week for six weeks. 4. The method of claim 1 wherein the malignancy is metastatic cholangiocarcinoma. 5. The method of claim 1 wherein the malignancy is metastatic lung cancer. 6. The method of claim 1 wherein the malignant neoplasm is a metastatic kidney cancer. 7. The method of claim 1 wherein the cancer is metastatic colon cancer. 8. The method of claim 1 wherein the cancer is metastatic tonsil cancer. 9. The method of claim 1 wherein the malignant neoplasm is metastatic cancer of the appendix. 10. The method of claim 1 wherein the malignant neoplasm is a metastatic sarcoma. 11. The method of claim 1 wherein the malignancy is metastatic thyroid cancer. 12. The method of claim 1 wherein the malignancy is a metastatic glioma.