Link:  Pharm/Biotech Resources

Title:  Combination therapies using vitamin B12 and interferon for treatment of viral, proliferative and inflammatory diseases

United States Patent:  6,908,611

Issued:  June 21, 2005

Inventors:  Cruz; Tony (Toronto, CA); Pastrak; Aleksandra (Toronto, CA)

Assignee:  Transition Therapeutics Inc. (Ontario, CA)

Appl. No.:  167765

Filed:  June 11, 2002

Pharmaceutical compositions for treating viral, proliferative and inflammatory diseases are disclosed comprising an amount of pharmaceutically acceptable vitamin B12 compound in combination with an interferon compound. Vitamin B12 compounds are administered separately, simultaneously or in combination with interferon compounds to provide an enhanced therapeutic effect for treating viral, proliferative and inflammatory diseases.

FIELD OF THE INVENTION

The present invention provides pharmaceutical compositions and methods of use for the treatment of viral, inflammatory or proliferative diseases with a vitamin B12 compound in combination with an interferon compound.

BACKGROUND OF THE INVENTION

Review of Viral, Inflammatory and Proliferative Diseases

Diseases and disorders that have significant inflammatory, viral and/or proliferative components are widespread and affect millions of people worldwide. Selected examples of inflammatory, viral and/or proliferative diseases include multiple sclerosis, diabetes, restenosis, cancer, hepatitis B, hepatitis C, HIV/AIDS and genital warts. These types of diseases share common disease processes and as a result often share or possess related/common therapies.

Viral Diseases

Viruses are potent infectious pathogenic agents that cause important functional alterations of the invaded cells, often resulting in cellular death. It is generally acknowledged that the cell injury in viral diseases includes not only direct damages inflicted by the proliferation of viruses but also various immunologic reactions elicited by infection with viruses. The consequences of a viral disease depend upon several viral and host factors such as the quantity of infecting viral particles, the speed of viral multiplication and spread, the impact on cell functions, the host's secondary responses to the cellular injury, and both the immunologic and the non-specific defenses of the host. In general, the effects of a viral disease include asymptomatic infections, both acute and chronic clinical diseases and induction of various types of cancers. One example of a well known viral disease is viral hepatitis infection which results in chronic or acute inflammation of the liver and can lead to hepatocellular carcinoma in some cases. To date, there are several types of hepatitis viruses that have been identified including hepatitis viruses A, B, C, D, E and G; such viruses are prevalent among the population. For instance, it has been estimated by Liver Foundation International that there are more than 520 million individuals worldwide that suffer from either hepatitis B or hepatitis C.

Inflammatory Diseases

Inflammatory diseases are a class of diseases and disorders that are characterized by the influx of certain cell types and mediators, the presence of which can lead to tissue damage and sometimes death. Inflammatory diseases trigger what is known as the inflammatory cascade, a complex process that involves the triggering of immunological response, the release of chemokines, cytokines and toxic agents by the activated cells, the up-regulation of cell surface adhesion molecules and trans-endothelial cell migration. Typically, inflammation occurs as a defensive response to invasion of the host by foreign material, bacteria or to mechanical trauma, toxins and neoplasia. Autoimmune responses by intrinsic stimulation also can induce inflammatory responses.

One example of inflammatory disease is multiple sclerosis ("MS"). MS is a multi-factorial inflammatory disease of the human central nervous system resulting in the slowing of electrical conduction along the nerve. It is estimated that close to a third of a million people in the United States have MS. MS is believed to result from inflammation and breakdown in the myelin surrounding the nerve fibers of the central nervous system. The disease is characterized by an increase in the infiltration of inflammatory cells, loss of oligodendrocytes, and increased gliosis (astrocyte hypertrophy and proliferation). (For review see Amit et al., 1999; Pouly et al., 1999; Steinman et al., 1993; Miller, 1994).

Proliferative Diseases

Cancer is the most well known proliferative disease. Specifically, cancer is a generic term representing a collection of diseases arising from mutations of key molecules that regulate cell proliferation, invasion, and metastasis. The ability of tumor cells to metastasize involves deregulation via overproduction or mutation of genes that allow cells to invade out of the tissue of origin, survive in a contact-independent manner, escape immune recognition, lodge at a distant site, then invade to a suitable place within the new tissue and grow there. The molecules that are commonly involved in tumor initiation, progression and metastasis include adhesion molecules, growth factor receptors, factors regulating the cytoskeleton, master switches regulating cell cycle, proliferation repressor genes, proteases and transcription factors. The ability of most tumors to kill is directly related to their capacity to invade and ultimately to metastasize.

Viral, Inflammatory and Proliferative Diseases Share Common Disease Processes

There are several classes of molecules and disease processes that are common to viral, inflammatory, and proliferative diseases. These include elevated expression of adhesion molecules, cytokines and matrix metalloproteinases, increased cell proliferation and migration, increased inflammatory cell activation and infiltration, increased angiogenesis, and increased tissue destruction and dysfunctional matrix remodeling. Consequently, compounds for the treatment of these diseases are aimed at altering the immune system, cell proliferation, cell adhesion and migration, cytokine levels or activities or viral replication.

Interferons

Interferons are multi-functional cytokines that are capable of producing pleitrophic effects on cells, such as inhibition of virus replication (anti-viral effects), inhibition of cell proliferation (anti-proliferative effects) and inhibition of inflammation (anti-inflammatory effects). Because of these cellular responses to interferons, interferon-alpha and interferon-beta have been found to be clinically useful in the treatment of viral, proliferative and inflammatory diseases such as multiple sclerosis, hepatitis B/C and a number of cancers. Interferon therapies may also have potential use for the treatment of other inflammatory diseases, viral diseases and proliferative diseases.

Need for Enhancing Compounds to Decrease Side Effects

Although interferon compounds are used or potentially can be used to treat inflammatory, viral and proliferative diseases, these compounds have many undesirable side effects, which are exacerbated at high doses. These include local injection reactions, flu-like syndrome and depression. Accordingly, some patients are unable to tolerate the doses needed to achieve a therapeutic effect.

In addition, although many anti-viral, anti-inflammatory or anti-proliferative compounds may have shown promise in vitro against inflammatory, viral, or proliferative diseases, very high doses may be required in vivo; such doses often being toxic or causing severe side effects. Therefore, the limitation of these therapies may be in using sufficient levels of the compound to achieve maximal efficacy in the absence of side effects.

Thus, compounds that can be combined with anti-viral, anti-proliferative or anti-inflammatory compounds to increase effectiveness of treatments for inflammatory, viral or proliferative diseases are necessary. In addition, compounds that can reduce the doses and/or reduce frequency of administration to reduce side effects and to maintain or improve efficacy are necessary.

For example, a compound that can improve efficacy of interferon-beta in the treatment of MS would potentially reduce the amount and/or frequency of interferon-beta administration, reducing side effects induced by interferon-beta and possibly reducing the occurrence of neutralizing antibodies to interferon-beta in MS patients.

Neutralizing antibodies to interferon-beta occur in about one third of MS patients treated with interferon-beta and are positively correlated with a loss of therapeutic efficacy of interferon-beta Deisenhammer et al., 1999).

In addition, said compounds could not only enhance the efficacy in current or potential anti-viral, anti-proliferative or anti-inflammatory therapies but could also broaden use of these therapies into many other inflammatory, proliferative and viral diseases.

There have been reports of other therapeutic agents being tested in combination with interferon for treatment of inflammatory diseases such as multiple sclerosis. However, these combination therapies have resulted in limited efficacy or no reduction in side effects. For example, combination therapy comprising copaxone and interferon-alpha did not improve clinical scores in EAE-treated mouse (Brod et al., 2000). While novantrone, an anti-proliferative drug, is currently being tested in clinical trials for combination therapy with interferon-beta, novantrone is known to have a significant side effect profile including serious cardiac toxicity. Novantrone is restricted in its use because the risk of heart disease increases with the cumulative dose. According to the Food and Drug Administration, patients with MS should ordinarily not receive more than 8 to 12 doses of novantrone, administered over two to three years.

Accordingly, there is a need to develop therapies for viral, inflammatory or proliferative diseases that result in sufficient levels of anti-viral, anti-inflammatory and anti-proliferative compounds to achieve maximal efficacy with minimal side effects. There is a need to develop anti-viral, anti-inflammatory and anti-proliferative combination therapies for the treatment of viral, inflammatory or proliferative diseases wherein the combined elements have an enhancing therapeutic effect, while minimizing side effects.

Vitamin B12

Vitamin B12 is a cobalt-containing B complex vitamin that has various effects on biological processes in vivo. Vitamin B12 has a well-elucidated family of analogues and conjugates. Vitamin B12 compounds have been known to be involved in metabolic processes; and deficiency of vitamin B12 has been known to provoke pernicious anemia and neurological disorders. It is a co-factor essential in the metabolic pathway leading to synthesis of DNA, cell division, as well as cellular metabolism. Biochemical evidence suggests that vitamin B12 compounds may up-regulate gene transcription and thereby protein synthesis (Watanabe et al, 1994). It has been also suggested that vitamin B12 compounds play an important role in immune system regulation (Tamura et al, 1999, Sakane et al, 1982).

Specifically, vitamin B12 compounds have been suggested to possess some or limited anti-viral (Weinberg et al, 1995, 1998, Lott et al, 2001, Poydock, 1979, Tsao et al., 1990), anti-proliferative (Nishizawa et al, 1997, Shimizu, 1987, Poydock et al., 1979, 1985) and anti-inflammatory activities (U.S. Pat. No. 5,508,271, U.S. Pat. No. 5,964,224, U.S. Pat. No. 5,716,941) on their own. Vitamin B12 has also been tried in combination with other therapeutic agents (EP Patent # 0835660, U.S. Pat. No. 6,096,737) for the treatment of specific inflammatory diseases or proliferative diseases. However, to date, there has been no scientific evidence demonstrating an enhanced therapeutic effect of vitamin B12 in combination with interferon compounds for treatment of viral, inflammatory or proliferative disease.

It is therefore an object of the present invention to provide a combination therapy using vitamin B12 compounds with interferon compounds for the enhanced treatment of viral, inflammatory or proliferative diseases.

SUMMARY OF THE INVENTION

The invention comprises a pharmaceutical composition for the treatment of a disease selected from the group consisting of viral diseases; proliferative diseases; inflammatory diseases; proliferative and inflammatory diseases; proliferative and viral diseases; viral and inflammatory diseases; and proliferative, viral and inflammatory diseases; comprising: (1) at least one vitamin B12 compound; and (2) at least one interferon compound.

Another embodiment of this invention comprises a pharmaceutical composition for the treatment of proliferative diseases, such as cancer comprising at least one vitamin B12 compound and at least one interferon compound.

Yet another embodiment of this invention comprises a pharmaceutical composition for the treatment of viral diseases, such as hepatitis B, hepatitis C, herpes, or vesticular stomatitis comprising at least one vitamin B12 compound and at least one interferon compound.

Yet another embodiment of this invention comprises a pharmaceutical composition for the treatment of inflammatory diseases comprising at least one vitamin B12 compound and at least one interferon compound.

Yet another embodiment of this invention comprises a pharmaceutical composition for the treatment of astrocytoma and glioma comprising at least one vitamin B12 compound and at least one interferon compound.

Another embodiment of this invention comprises a pharmaceutical composition for the treatment of multiple sclerosis comprising at least one vitamin B12 compound and at least one interferon compound, such as interferon-alpha or interferon-beta.

Another aspect of this invention is a pharmaceutical composition for the treatment of hepatitis B comprising vitamin B12 and interferon-alpha or interferon-beta.

Another aspect of this invention is a pharmaceutical composition for the treatment of hepatitis C comprising vitamin B12 and interferon-alpha or interferon-beta.

Another aspect of this invention is a pharmaceutical composition according to any of the aspects outlined above wherein the vitamin B12 compound is conjugated to the second compound.

Another aspect of this invention is a method of treating a viral, proliferative or inflammatory disease, including MS, hepatitis B and hepatitis C, comprising the step of administering to a patient any of the pharmaceutical compositions outlined above.

An other aspect of this invention is a method of treating a viral, proliferative, or inflammatory disease, including MS, hepatitis B and hepatitis C, comprising the steps of administering the vitamin B12 compound at a frequency selected from the group consisting of: more than once daily, daily, more than once weekly, weekly, more than once monthly and monthly, and administering the second compound at a frequency selected from the group consisting of: more than once daily, daily, more than once weekly, weekly, more than once monthly, and monthly.

An other aspect of this invention is a method of treating a viral, proliferative, or inflammatory disease, including MS, hepatitis B and hepatitis C, comprising the steps of administering to a patient, either together or separately, one or more vitamin B12 compound, and one or more interferon compound.

Encompassed within this invention are any of the above treatments wherein the dose of vitamin B12 compound is between 10 and 2500 mg, and/or the dose of interferon compound is similar or less than the interferon therapeutic dose range used.

Another aspect of this invention is the use of any of the compounds described above to treat a viral, proliferative, or inflammatory disease.

DETAILED DESCRIPTION OF THE INVENTION

The current invention discloses the effect of vitamin B12 compounds alone and in combination with interferon compounds for treatment of viral, inflammatory or proliferative diseases. Vitamin B12 compounds alone show efficacy and clearly show an enhancing or synergistic effect in combination with interferon compounds for treating viral, inflammatory, and proliferative diseases.

Pharmaceutical Composition

The present invention provides pharmaceutical compositions for enhancing anti-viral, anti-proliferative and anti-inflammatory effects and pharmaceutical compositions for treatment of viral, proliferative and inflammatory diseases. The pharmaceutical composition comprises a first compound that is a vitamin B12 compound and a second compound that is an interferon compound.

Excipients

The pharmaceutical compositions of the invention preferably contain a pharmaceutically acceptable carrier or excipient suitable for rendering the compound or mixture administrable orally, intranasally, or parenterally, intravenously, intradermally, intramuscularly or subcutaneously, rectally, via inhalation or via buccal administration, or transdermally. The active ingredients may be admixed or compounded with any conventional, pharmaceutically acceptable carrier or excipient. It will be understood by those skilled in the art that any mode of administration, vehicle or carrier conventionally employed and which is inert with respect to the active agents may be utilized for preparing and administering the pharmaceutical compositions of the present invention. Illustrative of such methods, vehicles and carriers are those described, for example, in Remington's Pharmaceutical Sciences, 4th ed. (1970). Those skilled in the art, having been exposed to the principles of the invention, will experience no difficulty in determining suitable and appropriate vehicles, excipients and carriers or in compounding the active ingredients therewith to form the pharmaceutical compositions of the invention.

The compositions of the invention may also be conjugated to transport molecules, monoclonal antibodies or transport modalities such as vesicles and micelles that preferentially target recipient cells.

Administration

The compounds of the present invention in the described dosages are administered orally, intranasally, by inhalation, intraperitoneally, subcutaneously, intramuscularly, transdermally, sublingually or intravenously. For oral administration the pharmaceutical composition can be prepared, for example, in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, chewing gum or the like prepared by procedures known to those skilled in the art. The amount of active compound in such therapeutically useful compositions or preparations is such that a suitable dosage will be obtained.

The therapeutically effective amount of compound to be included in the pharmaceutical composition of the invention depends, in each case, upon several factors, e.g., the type, size and condition of the patient to be treated, the intended mode of administration, the capacity of the patient to incorporate the intended dosage form, the severity of the disease progression, etc.

The dosages used for each interferon compound are similar to those dosages known to those skilled in the art and used in pre-clinical and clinical studies and in commercial use. The concentrations may be lower than the currently used dosages as the combination of these compounds with vitamin B12 increases efficacy of these compounds. Indeed, vitamin B12 may be combined with interferon compounds with the objective to reduce the dosages of the interferon compounds, in order to achieve both effective treatment and to lessen the negative effects of the interferon compounds.

The dosages for interferon-alpha and interferon-beta are known in the art. For example, doses for interferon-beta typically range from 30 μg to 250 μg. The main differences between Avonex™, Betaseron™ and Rebif™ are the amount of interferon-beta given and the route and frequency of administration. Avonex™ is preferably administered in the amount of 30 μg by intramuscular injection once weekly; Betaseron™ is preferably administered in the amount of 250 μg by subcutaneous injection every other day; and Rebif™ is preferably administered in the amount of 44 μg by subcutaneous injection three times a week. Several clinical studies have used a dose of up to 9 MIU every other day or thrice weekly of recombinant interferon alpha-2a in treatment of multiple sclerosis (Durelli et al, 1996, Nyland et al, 1996). The dosage generally used for both non-pegylated and pegylated interferon-alpha-2a or 2b for clinical indications for hepatitis C range from 3MIU-10 MIU three times a week for non-pegylated form and ˜70 μg to 180 μg weekly (Komanduri and Cotler, 2002). Interferon dosage used in the treatment of proliferative diseases such as hairy cell leukemia range from 1-36 MIU, daily or twice weekly.

The preferred dosage of a vitamin B12 compound for the present invention is the maximum a patient requires to provide an optimal enhancing effect, such maximum being tempered by the absolute upper limit of vitamin B12 compound dosage being the maximum that a patient can tolerate and not develop any serious complications. Vitamin B12 compounds have been available for many years as an injectable treatment for pernicious anemia, with doses typically in the range of 1000 μcg. Vitamin B12 compounds also have a long history as a general oral health supplement with doses also in the "μcg" range. Hydroxocobalamin, a vitamin B12 compound, is available as a cyanide poisoning antidote, called Cyanokit® (U.S. Pat. No. 5,834,448). Cyanokit® is an acute one-time 5-gram dose of hydroxocobalamin administered for emergency purposes. Though vitamin B12 has been proposed for use for the therapeutic treatment for a few inflammatory diseases, no person has demonstrated that high doses of B12 in combination with an interferon compound would achieve an enhanced therapeutic effect. With our experimental studies, no toxicity has been found in animals, even at concentrations of 15 mg/kg of vitamin B12 in mice. In a previous acute toxicity study, no toxicities were observed in mice and rabbits at doses of vitamin B12 (hydroxocobalamin) up to 1 g/kg and 100 mg/kg body weight, respectively (Mizoule, 1966). The dosage of vitamin B12 compound for our invention are within the range of 10 mg to 2.5 g daily.

Those skilled in the art will be aware that the amounts of the various components of the compositions of the invention to be administered in accordance with the invention to a patient will depend upon those factors noted above.

Methods and Uses

The present invention provides methods to enhance or potentiate interferon compound-induced anti-viral, anti-proliferative and anti-inflammatory effects and methods of treating viral, proliferative or inflammatory diseases in patients by administering an amount of vitamin B12 compound in combination with an interferon compound. Vitamin B12 compounds can be administered simultaneously, separately or in combination with interferon compounds, under different dose and route regimens, to enhance the efficacy of interferon compounds in the treatment of viral, proliferative or inflammatory diseases in patients compared to when such compounds are administered alone.

The first method of treatment is the administration of a pharmaceutical composition including both a vitamin B12 compound and an interferon compound. An alternate method of treatment includes the step of the administration of a pharmaceutical composition including a vitamin B12 compound followed by the step of the administration of a second pharmaceutical composition including an interferon compound. Optionally, the administration of the vitamin B12 compound can follow the administration of the interferon compound. Optionally, the administration of the pharmaceutical compositions can occur separately or simultaneously.

Claim 1 of 17 Claims

1. A method of treating a viral disease comprising the step of administering to a patient a pharmaceutical composition comprising:

(i) hydroxocobalamin; and

(ii) at least one interferon compound selected from the group consisting of interferon alpha and interferon beta.

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