New compositions are disclosed that comprise the phytochemical Diindolylmethane (DIM), as well as its precursor, Indole-3-carbinol (I3C), and cogener, 2-(Indol-3-ylmethyl)-3,3'diindolylmethane (LTR-1), acceptable carriers and/or excipients. These compositions are administered to prevent or reduce symptoms associated with mastalgia and endometriosis.

Description of the Invention

SUMMARY OF THE INVENTION

The present invention relates to compositions and methods for prevention or reduction of symptoms associated with mastalgia and endometriosis by administering phytochemicals, e.g., dietary indoles. In a preferred embodiment, the pain associated with endometriosis and mastalgia is prevented or reduced. In another embodiment, the presence of a marker associated with endometriosis is reduced through phytochemical treatment. Among the phytochemicals useful in the compositions and methods of the invention are dietary indole, Diindolylmethane (DIM), as well as its precursor, Indole-3-carbinol (I3C), and cogener, 2-(Indol-3-ylmethyl)-3,3'-diindolylmethane (LTR-1).

Also according to the present invention, a pharmaceutical composition is provided, which comprises a phytochemical, preferably I3C, DIM and/or LTR-1, and, optionally, pharmaceutically acceptable carriers.

4. DETAILED DESCRIPTION OF THE INVENTION

The present invention is based upon the observation that administration of phytochemicals, in particular, I3C, DIM, and LT-1, has improved symptoms of mastalgia and endometriosis.

The facilitated delivery of DIM and related indoles as dietary supplements may be accomplished according to formulations and methods described in U.S. patent application Ser. No. 09/053,180. The effectiveness of supplemental DIM is further supported by co-administration of phytochemicals (e.g., DIM, LTR-1) with grapefruit concentrate, which additionally facilitates the absorption of phytochemicals (e.g., DIM, LTR-1).

4.1. Methods of Treating Mastalgia

The invention provides compositions and methods for prevention or reduction of symptoms associated with mastalgia. In a preferred embodiment, breast pain of subjects suffering from mastalgia is prevented, reduced and/or eliminated through the administration of phytochemicals, e.g., dietary indoles, in a pharmaceutically acceptable fashion. In preferred embodiments, the phytochemicals are DIM, I3C and LTR-1. In particular embodiments, I3C, DIM, or LTR-1, alone or in combination with each other or other dietary supplements, are administered orally in, for example, the form of encapsulated dietary supplements.

The I3C is preferably administered at a dose of 200-500 mg per day. In alternative embodiments, I3C is administered at doses of 200-300 mg per day, 300-400 mg per day and 400-500 mg per day.

DIM is administered providing 150-500 mg per day of DIM. In preferred embodiments, the dose of DIM, I3C or LTR-1 is 150-200 mg per day, 200-300 mg per day, 300-400 mg per day, and 400-500 mg per day.

In a preferred embodiment, DIM is administered in an absorption enhancing formulation, as described in U.S. patent application Ser. No. 09/053,180, providing 60-500 mg per day of DIM suspended as microparticles in a starch carrier matrix. In preferred embodiments, the dose of processed DIM is 60-100 mg per day, 100-200 mg per day, 200-300 mg per day, 300-400 mg per day, and 400-500 mg per day.

The LTR-1 is preferably administered in an absorption enhancing formulation providing 60-500 mg per day of LTR-1 suspended as microparticles in a starch carrier matrix as previously described, however, the present invention contemplates the administration of any preparation of LTR-1. In a preferred embodiment, the dose of LTR-1 is 150-200 mg per day. In preferred embodiments, the dose of processed LTR-1 is 60-100 mg per day, 200-300 mg per day, 300-400 mg per day, and 400-500 mg per day.

Doses of the phytochemicals of the invention can also be calculated based upon the body weight of the subject to be treated. Doses of phytochemicals between 0.5 and 3 mg per kg of body weight per day are preferred. In another preferred embodiment, the phytochemicals are administered at a dose of between 0.5 and 2.0 mg per kg per day, preferably 1.5 mg per kg per day.

Alternatively, the co-administration of grapefruit, grapefruit concentrate, grapefruit juice, or grapefruit juice concentrate, or other grapefruit-derived composition along with a dietary indole (e.g., I3C, DIM or LTR-1) can be used to increase absorption of the phytochemicals and promote even more efficient relief from the symptoms of mastalgia.

In an alternative embodiment, the dietary indole (e.g., DIM or LTR-1) is administered in the form of a liposomal sublingual spray applied directly to the oral mucosa. This liposomal suspension provides phytochemical loaded liposomes to administer the phytochemicals and create a sustained delivery system. Dietary indole (e.g., DIM or LTR-1) containing liposomes are sequestered in the oral mucosa, allowing absorption which avoids "first pass" hepatic metabolism. The liposomal spray uses standard liposomal preparation and structural lipid ingredients (Ranade, V. V., "Drug delivery systems. 1. Site-specific drug delivery using liposomes as carriers," J. Clin. Pharmacol. 29(8):685-94, 1989). In a preferred embodiment, the liposomal spray is administered at a dose of 5-30 mg of dietary indole (e.g., DIM or LTR-1) daily delivered in 2-12 sprays of a typical liposomal preparation.

Alternatively the phytochemicals (e.g., DIM or LTR-1) may be administered in the form of a transdermal cream applied directly to the skin. This cream utilizes various absorption enhancing emollients and consists of phytochemical (e.g., DIM or LTR-1) in a concentration of 1-3% by weight. It is designed as a moisturizing cosmetic that is formulated to allow application directly to painful breasts in women not wishing to take phytochemicals orally. Formulations are also made with the neurohormone, melatonin, to provide a nighttime cosmetic mosturizer offering the benefits of melatonin in combination with the phytochemical (e.g., DIM or LTR-1). This allows application of cruciferous indoles and melatonin directly to underlying breast tissue with the added benefit of sleep regulating action from melatonin. In a particular embodiment, application of from 5-10 cc of the transdermal preparation daily is used to administer from 5-30 mg of DIM or other dietary indole per day, and optionally, 3-10 mg of melatonin per day.

Alternatively, the phytochemical (e.g., DIM or LTR-1) may be administered in the form of a vaginal cream or suppository containing microcrystalline phytochemical (e.g., DIM or LTR-1) in a combined dose of 20-100 mg. This allows application of cruciferous indoles directly to vaginal and cervical mucosa for the benefit of cervical dysplasia.

The phytochemicals of the invention may be administered in any appropriate amount in any suitable galenic formulation and following any regime of administration.

The actual administered amount of phytochemical may be decided by a supervising physician and may depend on multiple factors, such as, the age, condition, file history, etc., of the patient in question.

The subject, or patient, to be treated using the methods of the invention is an animal, e.g., a mammal, and is preferably human, and can be a fetus, child, or adult. In a preferred embodiment, the subject is a human female.

4.2. Methods of Treating Endometriosis

The invention provides compositions and methods for reduction or prevention of symptoms associated with endometriosis. In a preferred embodiment, the pain of subjects suffering from endometriosis is prevented, reduced and/or eliminated through the administration of phytochemicals in a pharmaceutically acceptable fashion. In another preferred embodiment, the levels of an endometriosis marker (e.g., Ca-125 antigen, a serum marker of endometriosis) in subjects suffering from endometriosis is lowered through the administration of phytochemicals in a pharmaceutically acceptable fashion. In preferred embodiments, the phytochemicals are DIM, I3C and LTR-1. In particular embodiments, I3C, DIM, or LTR-1, alone or in combination with each other or other dietary supplements, are administered orally in, for example, the form of encapsulated dietary supplements.

The 13C is preferably administered at a dose of 200-500 mg per day. In alternative embodiments, I3C is administered at doses of 200-300 mg per day, 300-400 mg per day and 400-500 mg per day.

DIM is administered providing 30-500 mg per day of DIM. In preferred embodiments, the dose of DIM, I3C or LTR-1 is 30-100 mg per day, 100-200 mg per day, 200-300 mg per day, 300-400 mg per day, and 400-500 mg per day.

In a preferred embodiment, DIM is administered in an absorption enhancing formulation, as described in U.S. patent application Ser. No. 09/053,180, providing 30-500 mg per day of DIM suspended as microparticles in a starch carrier matrix. In preferred embodiments, the dose of processed DIM is 30-100 mg per day, 100-200 mg per day, 200-300 mg per day, 300-400 mg per day, and 400-500 mg per day.

The LTR-1 is preferably administered in an absorption enhancing formulation providing 30-400 mg per day of LTR-1 suspended as microparticles in a starch carrier matrix as previously described, however, the present invention contemplates the administration of any preparation of LTR-1. In a preferred embodiment, the dose of LTR-1 is 100-200 mg per day. In preferred embodiments, the dose of processed LTR-1 is 30-100 mg per day, 200-300 mg per day, 300-400 mg per day, and 400-500 mg per day.

Doses of the phytochemicals of the invention can also be calculated based upon the body weight of the subject to be treated. Doses of phytochemicals between 1 and 3 mg per kg of body weight per day are preferred. In another preferred embodiment, the phytochemicals are administered at a dose of between 1.5 and 2.5 mg per kg per day, preferably 2.0 mg per kg per day.

Alternatively, the co-administration of grapefruit, grapefruit concentrate, grapefruit juice, or grapefruit juice concentrate, or other grapefruit-derived composition along with I3C, DIM or LTR-1 can be used to increase absorption of the phytochemicals and promote even more efficient relief from the symptoms of endometriosis, including the reduction of markers associated with endometriosis.

In an alternative embodiment, the phytochemical (e.g., DIM or LTR-1) is administered in the form of a liposomal sublingual spray applied directly to the oral mucosa. This liposomal suspension provides phytochemical loaded liposomes to administer the phytochemicals and create a sustained delivery system. DIM and LTR-1 containing liposomes are sequestered in the oral mucosa, allowing absorption which avoids "first pass" hepatic metabolism. The liposomal spray uses standard liposomal preparation and structural lipid ingredients. (Ranade, V. V., "Drug delivery systems. 1. Site-specific drug delivery using liposomes as carriers, J. Clin. Pharmacol. 29(8):685-94, 1989; Crommelin, D. J. A. and Schreir, H., "Liposomes", Colloidal Drug Delivery Systems, Kreuter, J. editor, Marcel Dekker, N.Y., 1994, p. 85). In a preferred embodiment, the liposomal spray is administered at a dose of 5-30 mg of phytochemical daily delivered in 2-12 sprays of a typical liposomal preparation.

Alternatively the phytochemicals (e.g., DIM or LTR-1) may be administered in the form of a transdermal cream applied directly to the skin. This cream utilizes various absorption-enhancing emollients and consists of DIM or LTR-1 in a concentration of 1-3% by weight. It is designed as a moisturizing cosmetic which is formulated to allow application directly to the skin of women not wishing to take phytochemicals orally. Formulations are also made with the neurohormone, melatonin, to provide a nighttime cosmetic moisturizer offering the benefits of melatonin in combination with the phytochemical (e.g., DIM or LTR-1). This provides, the added benefit of sleep regulating action from melatonin. In a particular embodiment, application of from 5-10 cc of the transdermal preparation daily is used to administer from 5-30 mg of phytochemical (e.g., DIM or LTR-1) per day, and optionally, 3-10 mg of melatonin per day.

Alternatively the phytochemical (e.g., DIM or LTR-1) may be administered in the form of a vaginal cream or suppository containing microcrystalline DIM or LTR-1 in a combined dose of 20-100 mg. This allows application of cruciferous indoles directly to vaginal and cervical mucosa for the benefit of cervical dysplasia.

The phytochemicals of the invention may be administered in any appropriate amount in any suitable galenic formulation and following any regime of administration.

The actual administered amount of phytochemical may be decided by a supervising physician and may depend on multiple factors, such as, the age, condition, file history, etc., of the patient in question.

The subject, or patient, to be treated using the methods of the invention is an animal, e.g., a mammal, and is preferably human, and can be a fetus, child, or adult. In a preferred embodiment, the subject is a human female.

4.3. Pharmaceutical Compositions

The pharmaceutical compositions according to the present invention preferably comprise one or more pharmaceutically acceptable carriers and the active constituents. The carrier(s) must be "acceptable" in the sense of being compatible with the other ingredients of the composition and not deleterious to the recipient thereof.

It will be appreciated that the amounts of phytochemical required for said treatment will vary according to the route of administration, the disorder to be treated, the condition, age, and file history of the subject, the galenic formulation of the pharmaceutical composition, etc.

Preferably, the phytochemical used in the invention has been processed to enhance bioavailability, as is described in U.S. patent application Ser. No. 09/053,180. So processed DIM or LTR-1 is referred to as "processed DIM" and "processed LTR-1", respectively. However, any suitable preparation of phytochemical can be used in the methods and compositions of the invention.

The following is a list of ingredients useful for formulating processed DIM or LTR-1: 1. About 10 to about 40 percent by weight of LTR-1 or DIM. 2. About 10 to about 40 percent by weight of the following, alone or in combination: vitamin E succinate polyethylene glycol 1000; vitamin E succinate Polyethylene glycols with polyethylene glycol. (with a molecular weight range of 400-2000); other polyethylene glycol esters such as those formed by fatty acids such as oleic acid or stearic acid; polyvinylpyrrolidones; polyvinylpolypyrrolidones; Poloxamer 188, Tweens; or Spans. 3. About 5 to about 20 percent by weight of the following, alone or in combination: phosphatidyl choline (derived from soy lecithin and supplied as Phospholipon 50 G from Rhone Poulenc Rorer); dioleoyl phosphatidylcholine; phoshatidylglycerol; dioleoylphosphatidylglycerol; dimyristoylphosphatidylcholine; dipalmitoylphosphatidylcholine; phosphatidylethalolamines; phosphatidylserines; or sphingomyelins; or other sources of phospholipids as those from purified Milk Fat Globule Membrane; glycerolesters; poly glycerol esters; or ethoxylated castor oil. 4. About 15 to about 30 percent by weight of the following, alone or in combination: hexanol; ethanol; butanol; heptanol; 2-methyl-1-pentanol; various ketone solvents that would be acceptable in foods such as methyl ethyl ketone, acetone and others; propylene glycol; and certain ester solvents such as ethyl acetate. 5. About 20 to about 40 percent by weight of the following, alone or in combination: modified starch such as Capsul.TM. Starch from National Starch, Inc.; methylcellulose; hydroxypropyl methylcellulose; hydroxyethylcellulose; hydroxypropylethylcellulose; pectin; gum arabic; gelatin; or other polymeric matrix-forming preparation known to those skilled in the art, soluble in water and, suitable for spray drying. 6. About 0.5 to about 35 percent by weight of the following, alone or in combination: aerosil 200; or any other flow enhancing excipient from silica, or related salt, known to those skilled in the art.

The following is a detailed method of formulating processed DIM: 1. 6.75 kg of TPGS is heated just beyond its melting point with constant stirring in a heated vessel ("First vessel"). 2. 9.38 kg of hexanol and 9.83 kg of jet milled DIM is added to the first vessel and the mixture stirred to a uniform suspension at 70.degree. C. 1.4 kg of phosphatidyl choline is then added. 3. In a second larger vessel, 185 L of water and 10.7 kg of starch are thoroughly mixed with a Cowles blade. This mixture is neutralized to pH 7 with a small amount of sodium carbonate and then heated to 75.degree. C. and stirred for 1 hour. 4. The contents of the first vessel is added to the starch mixture in the second larger vessel and thoroughly mixed with a homogenizing rotor/stator type mixer at moderate speed for 15 minutes. 5. The mixture from step 4 is spray dried with a small amount (approximately 0.5%) of hydrophilic silica to provide a free flowing powder of finely dispersed microparticles containing the co-precipitated TPGS, phosphatidyl choline and DIM in an amorphous, non-crystalline structure. 6. The flowable powder product is collected and stored in evacuated foil sacks, after de-aerating and flushing with nitrogen. 7. Analysis of presence of unchanged dietary ingredient, reveals about 30 to about 33 percent by weight of DIM.

The procedure of making processed DIM may be summarized as follows: (a) heating one or more solubilizing emulsifiers selected from the group consisting of vitamin E succinate polyethylene glycol 1000, polyvinylpyrrolidone, polyoxyethylene stearate, sodium cholate, deoxycholate and taurocholate; (b) adding to the product of step (a) a solvent and a surfactant phospholipid selected from the group consisting of phosphatidyl choline, dioleoyl phosphatidyl choline, phosphatidylglycerol, dioleoylphosphatidylglycerol, dimyristoylphosphatidylcholine, dipalitoylphosphatidylcholine, phosphatidylethanolamine, phosphatidylserine and sphingomyelin to produce a solution; (c) dissolving in the solution of step (b) LTR-1 and/or DIM; (d) adding to the solution of step (c) a solution containing an encapsulator; (e) mixing the solution produced in step (d) to produce a microdispersion with a particle size of 5 microns or less; and (f) spray drying the resulting mixture to leave a solid hydrophobic phytochemical composition.

In general, a suitable (therapeutically effective) amount of I3C is 300-500 mg per day. DIM is preferably administered in an absorption enhancing formulation, as described in U.S. patent application Ser. No. 09/053,180, at 50-200 mg per day as a suspension of microparticles in a starch carrier matrix. The LTR-1 is preferably administered in an absorption enhancing formulation at 50-200 mg per day as a suspension of microparticles in a starch carrier matrix. The actually administered amounts of phytochemical may be decided by a supervising physician. The phytochemicals of the invention may be administered alone or in combination with one another, and/or with other dietary supplements. The combinations of phytochemicals and supplements can be in the same composition for administering in combination concurrently, or in different compositions for administering concurrently but separately, or sequentially.

Therapeutic formulations include those suitable for parenteral (including intramuscular and intravenous), oral, rectal or intradermal administration, although oral administration is the preferred route. Thus, the pharmaceutical composition may be formulated as tablets, pills, syrups, capsules, suppositories, formulations for transdermal application, powders, especially lyophilized powders for reconstitution with a carrier for intravenous administration, etc.

The term "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the therapeutic is administered. The carriers in the pharmaceutical composition may comprise a binder, such as microcrystalline cellulose, polyvinylpyrrolidone (polyvidone or povidone), gum tragacanth, gelatin, starch, lactose or lactose monohydrate; a disintegrating agent, such as alginic acid, maize starch and the like; a lubricant or surfactant, such as magnesium stearate, or sodium lauryl sulphate; a glidant, such as colloidal silicon dioxide; a sweetening agent, such as sucrose or saccharin; and/or a flavoring agent, such as peppermint, methyl salicylate, or orange flavoring.

Therapeutic formulations suitable for oral administration, e.g., tablets and pills, may be obtained by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by mixing phytochemicals, and compressing this mixture in a suitable apparatus into tablets having a suitable size. Prior to the mixing, the phytochemical may be mixed with a binder, a lubricant, an inert diluent and/or a disintegrating agent.

In a preferred embodiment, phytochemical is mixed with a binder, such as microcrystalline cellulose, and a surfactant, such as sodium lauryl sulphate until a homogeneous mixture is obtained. Subsequently, another binder, such as polyvidone, is transferred to the mixture under stirring with a small amount of added water. This mixture is passed through granulating sieves and dried by desiccation before compression into tablets in a standard tableting apparatus.

A tablet may be coated or uncoated. An uncoated tablet may be scored. A coated tablet may be coated with sugar, shellac, film or other enteric coating agents.

Therapeutic formulations suitable for parenteral administration include sterile solutions or suspensions of the active constituents. An aqueous or oily carrier may be used. Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. Formulations for parenteral administration also include a lyophilized powder comprising phytochemical that is to be reconstituted by dissolving in a pharmaceutically acceptable carrier that dissolves said phytochemical.

When the pharmaceutical composition is a capsule, it may contain a liquid carrier, such as a fatty oil, e.g., cacao butter.

Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like. These compositions can take the form of solutions, suspensions, emulsion, tablets, pills, capsules, powders, sustained-release formulations and the like. The composition can be formulated as a suppository, with traditional binders and carriers such as triglycerides.

In yet another embodiment, the therapeutic compound can be delivered in a controlled release system. In one embodiment, a pump may be used (see Langer, supra; Sefton, CRC Crit. Ref. Biomed. Eng. 14:201 (1987); Buchwald et al., Surgery 88:507 (1980); Saudek et al., N. Engl. J. Med. 321:574 (1989)). In another embodiment, polymeric materials can be used (see Medical Applications of Controlled Release, Langer and Wise (eds.), CRC Pres., Boca Raton, Fla. (1974); Controlled Drug Bioavailability, Drug Product Design and Performance, Smolen and Ball (eds.), Wiley, N.Y. (1984); Ranger and Peppas, J. Macromol. Sci. Rev. Macromol. Chem. 23:61 (1983); see also Levy et al., Science 228:190 (1985); During et al., Ann. Neurol. 25:351 (1989); Howard et al., J. Neurosurg. 71:105 (1989)).

Other controlled release systems are discussed in the review by Langer (Science 249:1527-1533 (1990)).

In one embodiment of the pharmaceutical composition according to the invention, two or more active constituents are comprised as separate entities. The two entities may be administered simultaneously or sequentially.

The invention also provides a pharmaceutical pack or kit comprising one or more containers filled with one or more of the ingredients of the pharmaceutical compositions of the invention. Optionally associated with such container(s) can be a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals or biological products, which notice reflects approval by the agency of manufacture, use or sale for human administration.
 

Claim 1 of 19 Claims

1. A method of treating endometriosis in a subject having endometriosis comprising administering to the subject an amount of an indole selected from the group consisting of 3,3'-Diindolylmethane (DIM), 2-(Indol-3-ylmethyl)-3,3'-Diindolylmethane (LTr-1) and Indole-3-carbinol (I3C) effective to reduce one or more symptoms associated with endometriosis, wherein the indole is formulated as a tablet, pill, capsule, suppository, cream, parenteral suspension or liposomal spray, or is suspended as microparticles in a starch carrier matrix.

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