AU773179B2 - Homeopathic preparations of purified growth hormone - Google Patents

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BARBARA A BREWITT Peter Maxwell Associates Level 6 60 Pitt Street SYDNEY NSW 2000 HOMEOPATHIC PREPARATIONS OF PURIFIED GROWTH HORMONE 09/251,820 17 February 1999 USA DETAILS OF ASSOCIATED APPLICATION NO: The following statement is a full description of this invention including the best method of performing it known to us:- HOMEOPATMIC PREPARATIONS OF PURTIEDGROWTH

HOMONE

Field of the Invention This invention relates to homeopathic prcparations comprising purified growth hormone and, optionally, purified growth factors and other constituents, as well as mcthods and systems for delivery of such prcparations and treatment of disorders and conditions by administering such preparations.

Background of the Invention Hormones and polypeptide growth factors are important regulatory substances that are involved in the regulation of cell growth and differentiation, as well as in the control of specific metabolic processes. Hormones are defined as chemical messengers that arc synthesized in the endocrine glands of multicellular organisms and secreted into 15 cxtracellular body fluids. Hormones are transported to hormone-responsive cells, where they bind to a hormone receptor, and the hormone-receptor complex regulates and modulates differentiated functions. Polypeptide growth factors, including cytokines, are produced and secreted by cells from a variety of tissues, and are generally involved in paracrine and autocrine responses. Growth factors are involved in cell survival as well as protection from cell death and play a crucial role in the control mechanisms governing the development and maintenance of organs and tissues. In addition to their growth promoting and differentiation inducing effects, growth factors are also involved in important physiological processes such as inflammation, immune reactions, and tissue repair.

25 Specific hemopoietic growth factors have been used to treat diseases such as AIDS and cancer. Hemopoietic growth factors are logical therapeutic immunomodulators to use for treatment of chronic viral infections and other diseases for several reasons.

Endogenous growth factors such as graulocyte-macrophage colony stimulating factor S(GM-CSF) and macrophage colony stimulating factor (M-CSF) stimulate proliferation of hemopoictic progenitor cells. Lymphocytes, macrophages and natural killer cells that normally produce these factors are quantitatively and qualitatively defective after infection by HIV, HH6V or EBV.

I 11 Clinical studies On AIDS patients Using the growth fcosG-S n -S at, pharmacological concentrations (ugfkg/day) have producedr Gmxed reslts ForS example, injections or intravenous administration of GM-CSF at concentrations of 0.5-0.8 ug/kglday transiently increased leukocyte, neutrophil, eosinophii, and monocyte counts i AIDS paticnts with no significant rise in platelet counts or change in reticulocyte and lymphocyte counts (Miles, S. 1992 AIDS Res. Run. Retroviruses -8:1073-1080)- Subcutaneous injections of 0.25-4.0 ug/kg/day improved leukocyte counts with no improvement in hemoglobin or platelet counts. However, the side effects included increascd [MV replication, increased levels of P24 antigen, chills, nausea, rnyalgia and flu-like symptoms (Poll. G. et ad. 1991 J. Exp. Med. 1,73:589-5s97; Scadden, D.T. 1990 J-etnatopoietic Gr-owth Factors in Trans. Med., Wilcy-Liss Inc., New York, pp. 163-176), GM-CSF also occasionally caused Ithromnbocytopenia. Granulocyte colony stimulating factor (G-CSF) has been effective in correcting neutroperia with some minor increases in lymphocyte counts. Additionally, hemoglobin and reticulocytes increased in numbers in patients given G-'.Sr alone or in combination with elyulropoietin. H-owever, resumption of treatment with A.ZT after use of these growth factors led to severe anemia.

Pharmracological doses of growth factors often have harsh side effects.

Following puberty, there is an. exponential decline in. growth hormone (Rudman, 1985, II. A. Ger. Soc., 33:800-807). By thirty years of age, the normal physiological 2o concentration found in the blood stream is 20 rig/nil (Corpas, Harman, Pineyro,

M.,

Robertson, Blackman, 1992, J. Clin. Endocrinol. Metab., 75:530-535). This is reduced to 10 ng/mil by age 60, and continues to decline 2-4 ng/ml each decade (Irranmianesh. ILisarraide, Veidhuis, ii., 1991, 1 Gun. Endocrinol. Metal,., 73:1081 -1088). Additional studies have shown that growth hormone secretion peaks at 25 approximately 31 years of age and then continues to decline by 14 to 50% per decade, .dependent on gender, activity level and diet, or with the onset of chronice disease (no, K., Veidhuis, Endocrinol. Metal,., 1994 1 (Suppl A):61-63). While the definition of GH deficiency is not absolute, symptomis associated with age-related declines in hGH are often used to define GH deficiency. The American Association of Clinical Endocrinology and the American College of Endocrinology suggest that growth hormone deficiency is characteristically dcfined as a cluster of self perceived symptoms which include fatigue, decreased lean body mass, decreased muscle mass, abdominal obesity, reduced cardiac performance, impaired sense of well being, increased social isolation,

I

anxiety, emotional lability, reduced vitality and energy, cold extremities, poor sleep, and decreased physical strength, exercise capacity and physical performance.

Growth hormone has been isolated and purified from mammalian sources and has been produced recombinantly. Administration of pharmacological dosages of growth hormone are best known for the treatment of growth hormone deficiency disorder in children. Administration of higher than physiological concentrations of growth hormone may produce serious side effects, including tissue turgor, neuropathy, back pain, increase in liver enzymes aspartate aminotransferase (SGOT) and alanine aminotransferase

SPGT,

increased sweating, headache, skin and joint problems, hypertension, and edema. It would thus be desirable to identify compositions or means of administration that, when administercd, produce the benefits of growth hormone without producing the serious side effects.

Amndt formulated one of the earliest laws of pharmacology representing the homeopathic effect, the Amdt-Schultz law in 1888. The Arndt-Schultz law states: every 15 stimulus on a living cell elicits an activity, which is inversely proportional to the intensity of the stimulus (Martius F. Das Ardt-Schultz Grundgesetz, Muench Med. Wschr., 1923, 70(31):1005-1006). This law was later restated by Hueppe as: for every substance, small doses stimulate, moderate doses inhibit, large doses kill. Allopathic medicine, with its emphasis on moderate drug doses, works to inhibit undesired physical symptoms and to kill undesired pathogens. Homeopathic medicine begins with small doses and moves towards higher and higher dilutions to stimulate the body's own natural electromagnetic forces.

One of the basic tenets of homeopathic medicine is that a cure for a disease can be evoked by using a high dilution medicine that resembles but is different from the cause of 25 the disease. Homeopathy is widely accepted as a useful therapeutic throughout Europe, the British Commonwealth countries and India, and has been demonstrated to have characteristic and reproducible effects. A critical review of more than 100 controlled and/or clinical studies of homeopathy and meta-analysis of 89 blinded, randomized, placebo-controlled clinical trials dctermined that patients received 2.45 times greater positive healing benefits from homeopathy beyond the placebo effect (Kleijnen, J. et al.

1991 Brit. Med. J. 302:316-323; Linde, K, Clausius, Ramirez, Melchart, Eitel, Hedges, Jonas, 1997, Lancet, 350:834-843; Reilly, et al, 1994, Lancet, 344:1601-1608).

I Many homeopathic medicines are used at concentrations ofmicrograms (10.6 M) and nanorams (10-1 however, in other homeopathic preparations, the dilutions exceed Avogadro's number (6.023 x When homeopathic compounds are diluted 1:10, with repeated succussions (similar to vortexing) and repetitively diluted by this procedure at least 24 times, a potency is achieved (10- 2 1) that is so highly dilute that the probability of a single molecule of the original substance remaining in the volume used is .less than x 10 Homeopathic practitioners believe that the potency of a compound increases with increasing dilutions. In traditional homeopathic practice, the standard homeopatlic dosage is 10-15 drops of a 10- molar, or 6C, solution administered two to three times per day. A 10-o molar or 30C may be given one to three time per day. A molar or 200C may be given only one time per month or year. A 6C dilution approximates I pg/ml. Highly dilute homeopathic medicines have been effective in treating some conditions, including viral infections, in vivo. Homeopathic dilutions of I x 10- to 1 x 10-'0' of typhoidinum, hydrophobinum, tuberculinum, nux vomica and S 15 malandrinum 100% inhibited pock-like lesions caused by a chicken embryo DNA virus on the chorio-allantoic membrane compared to controls (Singh, L.M. and Gupta, G. 1985 Brit. Homeopathy 24:168-174). Other homeopathic medicines, the same medicines at different homeopathic concentrations, or control phosphate buffered solution (PBS), had lesser or no effect.

It is clear that the mechanism of action of homeopathics is of a non-molecular origin. Yet homeopathics exert potent biological activities that are clinically effective. It has been postulated that highly dilute compounds transfer biological activity to cells by electromagnetic fields (Benvenistc, J. 1993 Frontier Perspectives 3:13-15). Del Giudice et al. have hypothesized that interactions between the electric dipoles of water and the 25 radiation fields of a charged molecule generate a permanent polarization of water which o becomes coherent and has the ability to transmit specific information to cell receptors, somewhat like a laser (Del Giudice, Preparata, Vitiello, G. 1988, Phys. Rev. Lctt.

61:1085-1088).

Certain honnones have been prepared and used homeopathically. Adrenalinum, or ephinephrine, a sympathomimetic hormone produced by the medulla of the adrenal glands, thyroidinum, a preparation from the thyroid gland, and adrenocorticotrophin, or cortocotropin, a polypeptide hormone that increases the rate of secretion of the adrenal corticosteroids, are included in the official Homeopathic Monographs from the General Pharmacy of the Homeopathic Pharmacoepia of the United States. Insulin, an active molecule found in the pancreas which affects sugar metabolism, is listed in Boericke's Materia Medica, and is noted for its applicability for skin conditions. Parathyroid hormone, an extract from the parathyroid gland; thyreotrophic hormone, an extract from the anterior lobe of the pituitary gland; Corticotrophin, also extracted from the anterior lobe of the pituitary gland; cortisone and corticoids, which are steroid hormones; and folliculinum, a hormone secreted by the ovaries, are listed in the Materia Medica of New Homleopatic Remedies by Julian. The clinical symptomatology for parathyroid hormone includcs general weakness, depression, asthenia, hypotonia, fatigue, pallor and emaciation. The clinical symptomatology for thyreotrophic hormone include various conditions of the mind, digestive system, circulatory system, respiratory system, sense organs, and urinary and genital organs. The clinical symptomatology for corticotrophin include various psychological and nervous conditions. The symptomatology of cortisone and corticoids includes vanous psychological, nervous, endocrine and digestive system 15 conditions. The clinical symptomatology for folliculinum includes various conditions of the mind, digestive system and circulatory system.

Summary of the Invention Growth hormone, or somatotropin, is a well-characterized single chain alphahelical polypeptide containing a discrete receptor binding domain. Human growth hormone is the most abundant hormone secreted by the anterior pituitary gland and has significant anabolic and anti-catabolic effects on the body. Cells of the immune system, such as macrophages and lymphocytes, also produce and secrete growth hormone.

Growth hormone stimulates the liver to produce somatomcdins which, in turn, promote 25 bone, muscle, cartilage, kidney, liver and skin growth. Two cleaved forms of human growth hormone have been isolated. One has prolactin-like activity and the other has growth promoting activity greater than that of the uncleaved molecule.

The present invention comprehends homeopathic preparations comprising purified growth hormone that may be administered orally, nasally, topically or by injection, and are effective in one or more of the following: reducing blood pressure and improving cardiovascular function; increasing serum IGF-l levels; treating growth deficiency disorders; increasing lean body mass; increasing muscle mass and physical strength; improving pulmonary function; improving vascular and intracellular nutrient support; improving endurance; increasing mental clarity; improving skin tone and increasing skin thickness; revitalizing liver, spleen, and brain functions; increasing libido and sex hormones; improving lipoprotein balance and fatty acid levels; increasing energy levels, oxygen uptake, nitrogen retention, physical mobility and exercise performance; reducing Sjoint, back and knee pain; reducing joint swelling; eliminating cellulite; improving cholesterol profile; promoting hair growth and color change; reducing bleeding of the gums; reducing nasal and sinus congestion; improving dermal cellularity and collagenicity; increasing cartilage strength; increasing the size and function of the thymus and spleen; enhancing immune system function and lymphocyte count; enhancing white 0 blood cell and natural killer cell counts; and reducing fat, in particular, hip size upper arm size and waist size. Homeopathic preparations of the present invention furthermore may further be effective to: improve short term memory; reduce the manifestations of anger, anxiety, depression, social isolation, mood swings and sleeping disorders; improve vision; remove wrinkles; accelerate wound healing; facilitate breast enlargement; and generally is contribute to a feeling of well-being. Additionally, homeopathic preparations of the present invention may be used to treat headaches, AIDS wasting syndrome, Turner syndrome, osteoporosis, Parkinson's, Alzheimer's disease and dementia.

Homeopathic preparations of the present invention are non-toxic and do not produce undesirable side effects. They can be formulated and provided to a large patient S 20 population at a reasonable cost by means of delivery systems that are convenient and safe.

Homeopathic preparations of the present invention are preferably administered via oral or topical delivery systems, or using eye drops, nasal or throat sprays, skin creams, transdermal delivery, or other routes of administration that do not involve injection and that do not require sterile equipment or the participation of health care professionals.

2 Alternatively, the present invention may also be administered by means of intracutaneous, Sintramuscular, intravenous, or subcutaneous injection.

The homeopathic preparations of the present invention preferably comprise one or more potencies of purified growth hormone equivalent to a 10- or less molar concentration, generally equivalent to a molar concentration of between about 106 molar and about 10-"00.0 molar. Many of the homeopathic preparations thus contain few or no molecules of growth hormone. Homeopathic preparations of the present invention are defined as comprising purified growth hormone when the preparation originates from a preparation comprising a measurable quantity or activity of purified growth hormone, Preferably, recombinant human growth hormone having a bioactivity of about 2.6 Units/mg based on World Health Organization (WHO) reference standard. The term "purified growth hormone,"' as used in' this applicatio inldsfl egto oeua growth hormone, as well as fragments or epitopes derived from or present in growth hormone.

Purified growth hormone for use in horneopathic preparations of the present invention may be isolated from natural sources, or it may be produced using recombinant techniques or other polypeptide synthesis technology. Growth hormone isolated from marmmaliant sources, or produced recornbinantly to havre substantially the same structure and activity as human growth hormone, is preferred. The purity of the growth hormone used in homeopathic preparations of flie present invention is preferably at least about and more preferably at least 95%. H~uman growth hormone is preferred, although growth hormone polypeptides that are different from but have a high degree of similarity to human growth hormone molecules are suitable. Purified recombinant human growth hormone (AiGii) is commercially available from several sources, including NovoNordjsk, Eli1 Lilly, Pharmnacia/rUpjoli Ares Serono and Genentech- Purified recombinant human growth hormone may be produced in bacteria or in multicellular organisms, such as .*see:plants. In addition to overall purity, the biological specific activity of recombinant human growth hormone is preferably approximately 2.6 Units/mg based on World Health Organization (WHO) reference standard. (Clackson, T. and Wells, 1995 Science, Vol. 267, pp. 383-386).

Detailed Derition Thc homeopathic preparations the present invention comprise one or more 25 homeopathic potencies of puriflcd growth hormone in a h~omeopathically acceptable a.....diluent. Preparations of growth hormone according to the present invention may contain multiple potencies of purified growth hormone and/or other purified constituents, such as purified growth factors, etc., in addition to the growth hormone. Homeopathic preparations comprising one or more of the following potencics of purified growth hormone are preferred: 3C, 6X; 7X; 4C; 8X; 9X; 5C; l OX; IIX); 6C; 7C;I 9C; 12C; 100C; 200C, and IM. Especially preferred homneopathic preparations of thc present invention comprise multiple potencies of pui-fied growth hormone, selected from the homeopathic potencies described above. Preferred combinations of potencies of purified growth hormone include 6X 1 2C potencies, as well as a combination of 6C I OoC -f 200C potencies, 8X 12C potencies, 6C IM potencies, 7C 9C potencies, 6X potencies, 9X 5C potencies, and 30-X 6C 30C IM potencies. Various potencies of growth hormone may additionally be combined with other purified constituents, such as growth factors, phoSpholipids, vitamins, minerals, amino acids, and/or traditional horneopathics.

In particular, the combination of a homecopathic preparation of one or more potencies of purified growth hormone with one or more potencies of purified growth factors is of great interest. Suitable growth factors include: gyrnulocyte mracrophagecoloniy stimulating factor (GM-CSF), granulocyte-colony stimulating factor

(G-CSF),

macrophage-colony stimulating factor tumnor necrosis factor (TNF-a), insulinlike growth factor (IGF), including insulin-lie growth factor-i (IGF,) transforming growth factors, including transforming growth factor-B (TGF-3) and transforming growth factor-ot (TGF-cx), nerve growth factor (NGF), epidermal growth factors (EGF), stem cell 1.5 factors (SCF), platelet-derived growth factors (PDGF), fibroblast growth factors

(FGF),

interleukin- I, initer!eukin-2, keratinocqte growth factor, ciliary neurotrophic growth ****factor, insulin, Schwanm cell-derived growth factor, vaccinia virus growth factor, S.....bombyxin, neu differentiation factor, v-Sis, glial growth factor/acetylchouine receptorinducing activity, cytokines; and other proteins belonging to their structural superfamilies.

20 More specifically, the following potcncies of purified growth factor(s) are preferred for combination with one or more home opathic potencies of purified growth hormone: IGF,: 6X or 6C or 30C or 200C or IM or 30C IM or 6X 12C 3CC; *eve GMCSF: 200C or 200C IM; MCSF: 1M; TNF-a: 6C or 200C or 200C IM; TGF-cx: 6C or 30C or 200C or IM or 200C 1IM; TGF-6 1 6C or 200C or IM or 30C IM or 6C +30C IM; EGE: IM or 30C I M or 200C IM; PDGF,,B: 30C or IM or PDGF,,,: 30C or IM or 30C IM; NGF:. 1M or 6C or 6C 1M; acidic FGF: IM or IlM; basic FOE:I lMor 200C± +IM.

In addition to the homeopathic combinations and formulations described above, a combination of one or more homeopathic potencies of growth hormone with one or more homneopathic potencies of IGF 1 PDGF,,L, TGF-B and GMCSF is especially preferred for use in treating IKIW infection and HIV and AIDS symptoms. More particularly, the following combinations of homeopathic potencies of growth hormone and the specified growth factors are preferred: GH: 6X 12c or 8X 12C or 6C 10C 200C; IGF 1M; PDGF,,: 30C M; TGF-8: 30C IM; and GMCSF: 200C.

Homeopathic preparations comprising purified growth hormone and, optionally, one or more purified growth factors of the present invention may also be combined with one or more phospholipids, with L-alpha-Glycerylphosphoryl choline being an especially preferred phospholipid. Examples of traditional homeopathics that may be used in combination with homeopathic preparations of growth hormone include insulin arsenicum, pulsatilla, aconite, hypericum and metabolic sarcodes. The same potencies, or combinations of potencies, of homeopathic preparations of purified growth hormone and additional constituents are used regardless of whether the administration is in liquid, solid, spray, topical, transdermal or injectable form.

Homeopathic preparations comprising a purified growth hormone are preferably administered orally, in liquid or solid form. Homeopathic preparations are traditionally provided in liquid form, or in solid fonn as pellets, chewable tablets, capsules, gel caps S is and the like. Techniques for formulating and manufacturing homeopathic preparations in liquid and solid form are well known. Oral administration is convenient and effective.

Alternative delivery systems, such as eye drops, nasal gels or sprays, throat sprays, topical preparations, transdermal delivery systems and injectables (intracutaneous, intramuscular, intravenous, or subcutaneous) may also provide convenient and effective delivery of the homeopathic preparations comprising growth hormone.

0o0 Oral delivery of polypeptides and proteins is generally ineffective, since the polypeptides and proteins are generally broken down and rendered inactive in the blood stream before they reach their desired target or exert their desired effect. Traditionally, polypeptides and proteins are thought to be effective only when delivered via injection, 25 intranasally or intravenously, and undesirably high dosages are administered because a Slarge proportion of the delivered dosage is destroyed prior to exerting its effect or reaching its target. Although the mechanism of action of the homeopathic preparations of the present invention is not well defined, the clinical effects of such preparations, delivered orally using liquid or chewable tablet formulations, have been demonstrated in numerous patients and have been demonstrated in a double-blind, placebo controlled study, as described below.

Various homcopathically acceptable diluents or substrates may be used, depending on the desired delivery system. Appropriate diluents for the following delivery systems are well known: oral administration in liquid or solid form; eye drops; nasal sprays; throat sprays; injectables; topical preparations; and transdermal preparations. One or more potencies of purified growth hormone and/or one or more additional constituents, such as a purified growth factor, phospholipid(s), vitamins, minerals, amino acids, or traditional homeopathic preparations, may be combined in a preparation. The preferred homeopathic diluents for oral administration are a solution of purified water, glycerin, citric acid and a preservative such as sodium benzoate, or a solution of purified water, glycerin, potassium sorbate, and/or a form ofproteinated-copper in a cationic state, and a prescrvative such as sodium benzoate. Other diluents for oral delivery, including various alcohol-containing solutions, are known in the art and may be employed in the present invention to increase solubility and stability of purified growth hormone.

The homeopathic preparations of the present invention are preferably administered orally, but may also be prepared in topical formulations for application to the skin: administered transdermally; by intracutaneous, intramuscular, intravenous, or 15 subcutaneous injection; or administered in the form of eye drops or nasal and throat sprays or gels. Lotions for topical and transdermal application and buffered salt solutions for eye applications are well known and widely used in the cosmetic industry, and are readily adaptable to the preparations of the present invention. Additionally, carrier 0. solutions for intranasal administration of substances are well known in the art and widely I 20 used in drug delivery systems.

The following example describes specific clinical studies and are not intended to limit the invention in any way.

oo ****Example 1 25 Studies were conducted on healthy adult subjects to ascertain the effect of various potencies of recombinant human growth hormone Specifically, three studies evaluating a total of 162 healthy individuals ages 18-72 were conducted in the format of double-blind, placebo controlled studies (DBPCS). Individuals who reported any chronic conditions, such as AIDS, cancer, or other diseases were excluded from the studies.

Individuals reporting any use of steroids, prednisone, or injectable growth hormone were also excluded.

The first study, referred to as the Seattle Study, was a thirty day DBPCS that included 15 subjects ranging in age from 18 to 57 years who exercised regularly three to U. five times per week. Subjects were randomly divided into two groups: one group received a combination 6C 100C 200C homeopathic hGH formulation (human growth hormone from Sigma and the other group received placebo.

The second study, referred to as the Santa Fe Proving Study, included 46 individuals aged 19 to 59 years old who enrolled for a classical Homeopathic proving, without knowing the test substance. The Proving Study was designed in accordance with the principles of classical homeopathy where all subjects were given a placebo in bottle A" and instructed to take onc chewable tablet three times per day for seven days: one in the morning between 7:00 and 8:30 am; one in the afternoon between 3:00 and 4:00 pm; 0o and one in the evening between 5:00 and 8:30 pm. The subjects were instructed to chew the tablet carefully and not swallow it. After seven days of treatment, there was a wash out period for an additional seven days where no substance was given. After this wash out period, subjects were given bottle which contained a single potency of either 6X or 6C or placebo.

15 The third study, referred to as the Boulder Study, enrolled 101 individuals, ages 29 to 72 years old, who did not exercise regularly. This was a 42-day double-blind, Sp. lacebo controlled study with crossover after 21 days to the opposite test substance, i.e., treatment regimens are switched. For example, a group initially receives placebo and after 21 days "crosses over" to receiving treatment with a homeopathic preparation. Table 20 I describes the different groups and the treatment regimen. Specifically, test subjects received one of two formulations of homeopathic GH, a low potency 6X 12C (higher S. concentrations of hGH), or a higher potency, 6C 100C 200C (lower concentration of hGH), or placebo. Both formulations and the placebo were administered by chewable tablets. After three weeks, all subjects were crossed over to either placebo or one of the 25 two treatments, as described above. Furthermore, group 5 was administered a 6C I00C 200C potency combination of IHrhGH throughout the 42 day trial. Additionally, a control group consisting of three subjects who exercised regularly, ages 33, 35 and 62, were enrolled to determine whether exercise alone would increase IGF-1 levels and lean body mass (this group did not receive treatment or placebo). Each participant received instruction on the proper dosage (three times daily) and method of administration (chewable tablets) before being issued the 21-day supply of the substance. Group assignments were not revealed to the subjects or experimenters until the study was completed. Most subjects in the Seattle Study and all subjects in the Boulder Study had 11 S some knowledge of the perceived benefits of over-the-counter growth hormone related products. No one in the Santa Fe Proving Study knew the substance being tested.

Baseline assessments of body composition and specific laboratory measurements were made prior to the study and every 10.days until completion of the study. Final measurements were taken at the end of 42 days, after administration of the drug or placebo. Subjects completed an evaluation of self-perceived quality of life measurements before, every 10 days during, and upon completion of treatment.

In the Seattle Study, commercially available growth hormone (human, Sigma Chemicals) was serially diluted and hand succussed to obtain a final tablet of6C 100C 200C, or the active ingredient and hand succussion was withheld and a placebo was prepared. In the Santa Fe Study, single potencies of 6X (10- M) and 6C (10 M) homeopathic recombinant human growth hormone (HrhGH) were made, plus a placebo.

In the Boulder Study, two different combination formulations of HrhGH were used for 15 oral delivery in tablet form, the 6C 100C 200C, referred as the "high potency combination" and the 6X +12C, referred to as the "low potency combination." In addition, a placebo was prepared for the Santa Fe and Boulder studies.

Body compositions of participants, including lean body mass (LBM) and fat mass, were determined using bioelectric impedance analysis (BIA, Bioanalogics, Beaverton, OR). Blood pressure was monitored every ten days, as was body shape using tape measurements of the upper arm, upper chest, hips and waist. Total body weight (without shoes) and height measurements were also taken. To serve as a reference, ideal body weight was taken from standardized health charts.

In the Seattle Study, blood samples were taken to determine serum IGF-1 levels 25 before treatment, 14 days later and at the end of the end of the 30 day protocol. Subjects voluntarily arrived at the laboratory at a consistent time of day that was most convenient for them. At the end of the trial, subjects on placebo failed to arrive for their final blood draw. Tn the Boulder Study, potential diurnal fluctuations were controlled for by consistently taking blood samples between 5-7 pm. In addition to IGF-1, liver enzymes gamma-glutamyl-transpeptidase (GGPT), alanine aninotransferase (SGPT), and aspartate aninotransferase (SGOT) were monitored in order to detect potential toxicity associated with administering hGH.

1.1 Effects of Homeoathic Growth Hormone on IGF-1 levels.

IGF-1 has been cited most frequently as a reliable measure of hGH physiological activity. Figure -16 represents a scatter plot of response in subjects after 20 days of treatment or placebo. The groups receiving either the "high" or "low potency S combination" of HrhGH demonstrated a 12 to 78% increase in serum IGF-I in 28% of those subjects. In contrast, only 14% of the individuals receiving placebo increased in serum IGF-1 by 12 to 62%. A common 12% increase in serum IGF-1 levels occurred in both treatment and placebo groups.

Age-related changes in serum IGF-I levels within each test group were observed.

Baseline serum IGF-1 levels were generally within the normal range. In the Boulder Study, 56% of the individuals had above-average serum IGF-1 levels, however, 34% had levels below average, suggesting a potential age-related hGH deficiency in those individuals. In the groups receiving homeopathic treatments, 45-59% of individuals had serum IGF-1 baseline levels above the normal reference range compared to 67% of the 1l individuals in the placebo group. The baseline scrum IGF-1 values in the Boulder cohort ranged from 70 to 410 ng/ml. After 20 days of placebo, the range of serum IGF-1 was 52 to 382 ng/ml, demonstrating a slightly downward trend. Following 21 days of high potency combination treatment,. serum IGF-1 values ranged from 103 to 494 ng/ml, demonstrating an overall upward trend.

The age groups most responsive to the treatment after 21 days of testing were 57 years old. Also, in the Seattle Study, subjects over 32 years old increased serum IGF-1 levels 18% within the first 15 days, which then decreased to 7% by days. In contrast, subjects between 18 to 32 years showed no change in IGF-1 levels in the first 15 days, however this group had a 26% increase in serum IGF-1 levels S 25 after 30 days of treatment.

Reproducible rises in serum IGF-I were observed in all three studies. In the Boulder Study, the groups initially taking the low or high potency combination homeopathic formulas demonstrated a 4% and 10% increase in serum IGF-1 levels, respectively, after 21 days. The placebo group had no significant increase in serum IGF-1 at 21 days but did demonstrate a transient rise during the first 10 days. The control group not receiving treatment or placebo, but only exercising regularly, demonstrated a decrease of 28% in serum IGF-1 levels after 21 days and ended the study at 42 days with a decrease of 3% After the placebo group crossed over to treatment for 21 days, the group taking low potency combination HrhGH formulation increased in IGF-1 levels by 25% Serum IGF-1 levels rose 21% 13%) for the high potency combination after 21 days of treatment from the crossover point to the end of the study. There was a 16% increase in serum IGF-1 Icvels after 30 days of oral administration of the high potency combination formula compared to no increase with placebo after 15 days of use. In the Santa Fe Proving Study, there was an increase of 18% 10%) in serum IGF-1 serum levels from a single 6X potency of HrhGH after only seven days of oral administration.

1.2 Effects of Homeopathic Growth Hormone o Lean Body Mass and Tota Weight.

We observed changes in lean body mass over time for each treatment group compared to placebo (Boulder Study). During the first 21 days on treatment with the high potency combination formula, subjects gained 2.5 lbs. lbs.) of lean body mass.

15 These results closely approximate the total weight gained in those same groups. In contrast, the low potency combination stimulated no net gain in lean body mass (-1.6 lbs.)) or total weight gain after the same period. The placebo group also experienced no net gain in lean body mass, 0.05 lbs. Ibs.), yet gained 1.7 lbs. lbs.) total weight. Once the placebo group crossed over to the high potency combination formula, there was no significant lean body mass gain (2.1 lbs. 3.1 while total weight decreased approximately -0.6 lbs. 0.5 lbs.). Once the placebo group crossed over to the low potency combination formula, lean body mass increased 0.55 lbs. 1.1 Ibs.), while total body weight decreased 1.0 lbs. 0.9 lbs.). With regards to lean mass versus total mass, the high potency combination yielded lean mass/total weight ratios 25 ranging from 1.0 to 2.0, whereas the low potency combination yielded ratios ranging from to 1, and placebo yielded a ratio of zero. A positive ratio suggests greater gain in lean mass than total mass.

1.3 Effcts of Himeopathic Growth Hormone on Body Dm ensions.

Weight Changes Weight loss occurred during HhG treatment but not during placebo in the same subjects, P-0.03. Individuals on either HhGH treatment maintained -2.07 0.52 lb lower body weight/month versus their placebo period (P<0.002). Additionally, subjects of 6X 12C IIhGH tended to additionally lose -1.2 0.6 lbs/month versus subjects on 6F 100C 200C HhGh (P 0.05).

Body Shape We observed an upward trend in upper arm size (+0.29 0.09 inches) after HhGH compared to a downward trend on placebo (-0.21 0.11 inches) (P<0.0001). Trends in upper arm measurements had statistically divergent time-treatment differences between I-hGHI and placebo (P 0.01). Neither age nor gender affected outcome, only rhGH determined outcome.

We furthermore observed a decreasing trend in hip size on HhGH compared to an upward trend on placebo (P 0.02). At study end a time and treatment effect correlated to a loss of -2.09 0.50 inches/month versus placebo (P<0.001). Men on 6X 12C HhGH lost more hip inches than did women on the same formula In addition, baseline hip size was a highly significant parameter for responsiveness to 6X 12C HhGH (P<0.001). Chest measurement between treatment and placebo did not 15 statistically vary. However, both treatment groups statistically differed from each other.

Chest size of those on 6X 12C HhGH averaged +0.4 0.2 inches larger at study-end versus those on 6C 1 OOC 200C HhGH (P 0.02).

Waist measurements decreased continually by -0.9 0.3 inches over the 42-day period following treatment with 6C 1OOC 200C HhGH. Subjects on placebo decreased waist size minimally 0.3 inches). Waist size of subjects on 6X 12C HhGH decreased by 0.3 0.2 inches after 21 days and continued to lose inches once treatment stopped with a loss of-0.8 0.4 inches at study end. Three people who only exercised reduced waist size by -2.3 0.9 inches in 42 days.

1.4 Effects of Homeopathic Growth Hormone on Systolic Blood Pressure.

Systolic blood pressure was measured over the course of the studies. Initial treatment with high and low potency combinations for 21 days resulted in no significant change in systolic pressure. Prolonged treatment over a 42 day period with high potency combination resulted in an 8% decrease in systolic pressure. Following the placebo-treated group crossing over to the low potency combination formulation, the systolic pressure dropped by almost 10% after 21 days. The treatment group that crossed over to the high potency formulation for the same time period had no significant changes.

Within the treatment group on the low potency combination formulation there was a 4.5% decrease in systolic blood pressure.

Determination of Potential Toxicity of Homeopathic hGH.

No significant changes in liver enzyme levels for GGPT, SGOT and SGPT, as described above, were detected over the course of the studies. SGOT and SGPT enzyme levels remained within the normal range in all treatment and placebo groups. The average GGPT value for the treatment group that received placebo for the first three weeks, followed by a cross over to the 6C formulation for an additional three weeks, was somewhat elevated, although not to a significant extent. These results suggest the homeopathic formulations of the present invention are not toxic.

1.6 Effects of Homeopathic Growth Hormone on Self-Reporting Symptoms.

The primary self-reporting symptoms associated with age-related deficiency in 15 human growth hormone is fatigue. In the Boulder Study, 38% of enrollees reported this symptom preceding treatment protocols. Additional symptoms generally recognized as being associated with hGH deficiency were reported by 21-31% of individuals prior to treatment, including abdominal obesity, loss of strength (also detected as decreased lean 20 body mass), poor sleep, depression, and mood swings.

Although the present invention has been described in terms of specific embodiments, changes and modifications can be carried out without departing from the 0 scope of the invention which is intended to be limited only by the scope of the appended 25 claims.

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Claims (14)

1. A homeopathic preparation comprising at least one homeopathic potency of purified growth hormone and at least one component selected from the group consisting of: growth factors; insulin; vitamins; minerals; amino acids; and traditional homeopathics, wherein the concentration of the purified growth hormone is between about lx10 6 molar and about 1 x 10-100,000 molar, and wherein administration of the homeopathic preparation to a subject results in fewer undesirable side effects in the subject than administration of a pharmaceutical concentration of growth hormone.

2. A homeopathic preparation according to claim 1, wherein the at least one homeopathic potency of purified growth hormone has a concentration selected from the group consisting of: 3C; 6X; 7X; 4C; 8X; 9X; 5C; 10 OX; 11 X; 6C; 7C; 9C; 120; 30X; 30C; 1 00C; 200C; and 1M. oo••

3. A homeopathic preparation according to claim 1, comprising at least two homeopathic potencies of purified growth hormone.

4. A homeopathic preparation according to claim 1, wherein the purified growth hormone is purified, recombinant human growth hormone.

5. A homeopathic preparation according to claim 1, wherein the preparation is in a liquid form.

6. A homeopathic preparation according to claim 1, wherein the preparation is in a solid form. 03/03/04 18

7. A homeopathic preparation according to claim 1, wherein the preparation is in a topical application form.

8. A homeopathic preparation comprising at least two different homeopathic potencies of purified growth hormone, wherein the two different potencies are 6 X and 12 C, and wherein administration of the homeopathic preparation to a subject results in fewer undesirable side effects in the subject than administration of a pharmaceutical concentration of growth hormone.

9. A homeopathic preparation according to claim 8, wherein the preparation is in a liquid form.

A homeopathic preparation according to claim 8, wherein the preparation is in a solid form.

11. A homeopathic preparation according to claim 8, wherein the •preparation is in a topical application form.

12. A method for modulating the effects of aging of a treatment subject comprising administering a homeopathic preparation comprising at least one homeopathic potency of purified growth hormone, wherein the concentration of the purified growth hormone is between about 1x10 6 molar and about 1 x 10100000 molar, and wherein administration of the homeopathic preparation to the treatment subject results in fewer undesirable side effects in the subject than administration of a pharmaceutical concentration of growth hormone. 03/03/04

13. A method for modulating lean body mass of a treatment subject comprising administering a homeopathic preparation comprising at least one homeopathic potency of purified growth hormone, wherein the concentration of the purified growth hormone is between about lx1 06 molar and about 1 x 1 00000 molar, and wherein administration of the homeopathic preparation to the treatment subject results in fewer undesirable side effects in the subject than administration of a pharmaceutical concentration of growth hormone.

14. A method for modulating IGF-1 levels in serum of a treatment subject comprising administering a homeopathic preparation comprising at least one homeopathic potency of purified growth hormone, wherein the concentration of the purified growth hormone is between about lxi 06 molar and about 1 x 101 00 000 molar, and wherein administration of the homeopathic preparation to .the treatment subject results in fewer undesirable side effects in the subject than administration of a pharmaceutical concentration of growth hormone. :o e S° Dated this 3 day of March 2004 Biomed Comm Inmc. Patent Attorneys for the Applicant PETER MAXWELL ASSOCIATES 03/03/04