Using A Trans-dermal Growth Hormone Releasing Hormone Peptide

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Using A Trans-dermal Growth Hormone Releasing Hormone Peptide Combination (TD-GHRH-A) For Prevention of Aging Fifth International Congress on Aging Skin May 17 – 20, 2001 Session Loews Coronado Bay Resort, San Diego, CA Rashid A. Buttar, DO, FAAPM, FACAM 1 ©V-SAB Medical Labs, Incorporated

Lecture Objectives: 1. ) Explain the hypothalamic-pituitary axis clearly 2. ) Understand why GHRH is a better treatment option than GH injections to increase GH levels 3. ) Present research data on a highly effective and only transdermal longevity medical therapeutic 4. ) Explain the IGF-1 myth and support with clinical data, published research & physiologic concepts 5. ) Present initial findings of multi-centered, double blind, placebo controlled, cross over study 6. ) Review protocols & compare to GH injection tx Rashid A. Buttar, DO, FAAPM, FACAM 2 ©V-SAB Medical Labs, Incorporated

Human Growth Hormone (h. GH) Has Many Beneficial Effects Such As: Ø Increase in lean body (muscle) mass Ø Decrease in body fat Ø Improved cognitive function Ø Increase in sexual vigor Ø Increase in energy Ø Increase in stamina Ø Improved endurance Ø Increased immune system response Ø Faster resolution of acute injuries Rashid A. Buttar, DO, FAAPM, FACAM 3 ©V-SAB Medical Labs, Incorporated

Dietary Carbohydrates Dietary Protein Hypothalamus GHRH Somatostatin Adrenal Output Exercise Sex Hormones Pituitary h. GH Liver Stress Multi Organ Influence Rashid A. Buttar, DO, FAAPM, FACAM 4 IGF-1 ©V-SAB Medical Labs, Incorporated

h. GH Injections vs GHRH Treatments ……long-term stimulation of pituitary cells with GHRH will shift the GHRH/somatostatin tone by exogenous [injection] therapy to increase GHRH responsivity and pituitary GH stores. It is predicted that this therapy will reverse the chronic inhibitory state induced by long-term somatostatin domination and create an environment now responsive to the endogenous GHRH tone…. and allow for the normal [physiological] pulsatile GH release to reappear. This would produce a greater therapeutic benefit and a better safety profile compared with once daily injections of a bolus of recombinant GH …. . [the need for] repeated stimulation of GHRH receptors is required in a patient friendly format…. efforts are ongoing…. . format Scott Chappel, Ph. D Serono Laboratories, Norwell, MA Clinical Endocrinology (1999) 50, 547 -556 Rashid A. Buttar, DO, FAAPM, FACAM 5 ©V-SAB Medical Labs, Incorporated

Only Two GHRH Products Available on the Market 1. ) Geref ® by Serono® (Depot Injection Delivery) 2. ) Trans-D Tropin® by Balance Dermaceuticals® (Transdermal Delivery) Rashid A. Buttar, DO, FAAPM, FACAM 6 ©V-SAB Medical Labs, Incorporated

"Delivery System of the Future" Doctors around the world are calling trans-dermal delivery the 'delivery system of the future…. high absorption rate of many supplements is achieved when delivered through the skin…the physical size of the supplement’s molecule plays an important role…when delivered trans-dermally, they directly enter the vascular system, initially bypassing the liver …. The result can mean as much as 95% of the supplements get to the cellular level where they are needed. . Conversely, studies show as little as 5% of supplements taken orally, make it to the cellular level…. because the GI and hepatic systems are degrading and eliminating the majority of the active substances. A 95% absorption v. s. 5% absorption is why we believe trans-dermals to be a far more effective method of drug/supplement delivery. *. * Source: Dept. of Pharmacology, Univ. of Dublin Rashid A. Buttar, DO, FAAPM, FACAM 7 ©V-SAB Medical Labs, Incorporated

Trans-D Tropin ® NDC # 65448 -2115 -1 A Transdermal Growth Hormone Releasing Hormone Analog Rashid A. Buttar, DO, FAAPM, FACAM 8 ©V-SAB Medical Labs, Incorporated

Hypothalamus GHRH Somatostatin Trans-D Tropin / Geraf Pituitary Recombinant h. GH Rashid A. Buttar, DO, FAAPM, FACAM V. S. 9 Endogenous h. GH ©V-SAB Medical Labs, Incorporated

Endogenous h. GH Levels (2 weeks apart) per h. GH Radio-Immunoassay (ng/ml) Rashid A. Buttar, DO, FAAPM, FACAM 10 ©V-SAB Medical Labs, Incorporated

What Level of h. GH Increase Is Necessary In Order To Achieve Therapeutic Benefit? Endocrinology and Physiology Texts indicate that an absolute level of h. GH above 5 ng/ml is needed before efficacy can be attained. This “efficacy” referred to, is a DIAGNOSTIC response (for purposes of diagnosis), not a THERAPEUTIC response. • Diagnostic – To elicit a response far beyond the normal physiological range by taxing and overloading the system. • Therapeutic – To elicit a subtle response well within the normal physiological range to achieve therapeutic effects. Rashid A. Buttar, DO, FAAPM, FACAM 11 ©V-SAB Medical Labs, Incorporated

Endogenous h. GH Means & % Change 53 Specimens via h. GH Radio-Immunoassay (ng/ml) Rashid A. Buttar, DO, FAAPM, FACAM 12 ©V-SAB Medical Labs, Incorporated

Statistical Analysis of Data Baseline standard deviation 0. 464112 Baseline mean 0. 295918367 90 minute mean 2. 213636364 90 minute standard deviation 2. 673173 P < 0. 001 Rashid A. Buttar, DO, FAAPM, FACAM 13 ©V-SAB Medical Labs, Incorporated

Specific Changes Noted With Use of Trans-D Tropin®: Ø Change in facial/body contour Ø Decreased serum glucose levels Ø Increase in energy, stamina, endurance Ø Healing of old injuries with an increase in ROM Ø Increase in strength within 24 to 36 hours Ø Improved sleep patterns with increase in REM sleep Ø Improved mental focus and concentration Ø Decrease in depression with improved coping ability Ø Increased force of ejaculation and quality of erection Ø Improvement in LFT’s & other blood chemistries Ø Marked improvement in CHF patients and some MS patients Rashid A. Buttar, DO, FAAPM, FACAM 14 ©V-SAB Medical Labs, Incorporated

Response in Serum Glucose Levels • Trans-D Tropin® appears to have a distinct “euglycemic” effect on serum glucose. • Glucose level modulation was evidenced by glucose levels below 75 mg/dl to ~100 mg/dl levels and above 150 mg/dl to ~110 mg/dl levels. • Pt’s with IDDM had 50 to 70 mg/dl in glucose, 90 minutes after Trans-D Tropin® usage. Rashid A. Buttar, DO, FAAPM, FACAM 15 ©V-SAB Medical Labs, Incorporated

Case Report - 2 Patients with IDDM • 2 brittle insulin dependent diabetic patients showed the following response : Baseline Pre Trans-D Tx Glucose - 247 mg/dl - 190 mg/dl 90 Min Post Trans-D Tx Glucose - 160 mg/dl - 116 mg/dl Rashid A. Buttar, DO, FAAPM, FACAM 16 Both diabetic patients did NOT show response in h. GH levels during the first blood draw (first day of Trans-D Tropin® use) or the second blood draw (after 2 weeks of Trans-D Tropin® usage). However, at the 5 week blood draw, there was an average of over 400 % increase in levels of ENDOGENOUS h. GH in both IDDM patients. ©V-SAB Medical Labs, Incorporated

Response in Serum IGF-1 Levels • Trans-D Tropin® causes a measurable DECREASE in IGF-1 levels (ng/ml) on a consistent basis. • Efficacious h. GH therapy DOES NOT increase IGF-1. • An inverse correlation of IGF-1 and h. GH efficacy has clearly been established in our previous studies, in published literature, current research and in clinical observation. Rashid A. Buttar, DO, FAAPM, FACAM 17 ©V-SAB Medical Labs, Incorporated

Response in Serum IGF-1 Levels Rashid A. Buttar, DO, FAAPM, FACAM 18 ©V-SAB Medical Labs, Incorporated

Evidence of Unreliable Relationship between h. GH and IGF-1 levels 1. Jorgensen JO, Pedersen SB, Borglum J, Frystyk J, Ho KK, Christiansen JS, et al: Serum concentrations of insulin-like growth factors (IGFs), IGF binding proteins 1 and 3 and growth hormone binding protein in obese women and the effects of growth hormone administration: a double-blind, placebo- controlled study. European Journal of Endocrinology 1995 July; 133(1): 65 -70. 2. Chapman IM, Hartman ML, Pieper KS, Skiles EH, Pezzoli SS, Hintz RL, et al: Recovery of growth hormone release from suppression by exogenous insulin-like growth factor I (IGF-I): evidence for a suppressive action of free rather than bound IGF-I. Journal of Clinical Endocrinology and Metabolism 1998 August; 83(8): 2836 -42. Rashid A. Buttar, DO, FAAPM, FACAM 19 ©V-SAB Medical Labs, Incorporated

Evidence of Unreliable Relationship between h. GH and IGF-1 levels 3. Juul A, Andersson AM, Pedersen SA, Jorgensen JO, Christiansen JS, Groome NP, et al: Effects of growth hormone replacement therapy on IGF-related parameters and on the pituitary-gonadal axis in GH-deficient males. A double- blind, placebocontrolled crossover study. Hormonal Research 1998; 49(6): 269 -78. 4. Yohay D, Lunenfeld E, Giat Y, Levy J, Sharoni Y, Potashnik G, et al: Do changes in growth hormone levels correlate with IGF-I levels in patients undergoing IVFET? Gynecological Endocrinology 1997 August; 11(4): 269 -74. 5. Aimaretti G, Corneli G, Razzore P, et al: Usefulness of IGF-1 assay for the diagnosis of GH deficiency in adults. Journal of Endocrinology Investigation 1998 September; 21(8): 506 -511. Rashid A. Buttar, DO, FAAPM, FACAM 20 ©V-SAB Medical Labs, Incorporated

Evidence of Unreliable Relationship between h. GH and IGF-1 levels 6. Murphy LJ, Seneviratne C, Moreira P, Reid RE, et al: Enhanced expression of IGF binding protein-I in the fasted rat: the effects of insulin and growth hormone administration. Endocrinology 1991 February; 128(2): 689 -96. 7. Norrelund H, Fisker S, Vahl N, Borglum J, Richelsen B, Christiansen, JS, et al: Evidence supporting a direct suppressive effect of GH on serum IGFBP-1 levels. Growth Hormone IGF Research (Denmark) 1999 February; 9(1): 52 -60. 8. Borges MH, Pinto AC, Di. Ninno FB, Camacho-Hubner C, Grossman A, Kater CE, et. al: IGF-I levels rise and GH responses to GHRH decrease during longterm prednisone treatment in man. Journal of Endocrinological Investigation 1999 January; 22(1): 12 -7. Rashid A. Buttar, DO, FAAPM, FACAM 21 ©V-SAB Medical Labs, Incorporated

Evidence of Unreliable Relationship between h. GH and IGF-1 levels 9. Mazzoccoli G, Giuliani A, Bianco G, De Cata A, Balzanelli M, Carella AM, et al: Decreased serum levels of insulin-like growth factor (IGF)-I in patients with lung cancer: temporal relationship with growth hormone (GH) levels. Anticancer Research (Italy) 1999 March-April; 19(2 B): 1397 -9. 10. Furlanetto RW: Insulin-like growth factor measurements in the evaluation of growth hormone secretion. Hormonal Research 1990; 33 Suppl 4: 25 -30. Rashid A. Buttar, DO, FAAPM, FACAM 22 ©V-SAB Medical Labs, Incorporated

Possible Explanation of IGF-1 Levels (Basic Physiology Questions) IGF-1 = Insulin Like Growth Factor, Type I Question # 1: Do sedentary people or athletes have lower glucose levels? Question # 2: Are people who exercise, biologically (physiologically) younger or older? Rashid A. Buttar, DO, FAAPM, FACAM 23 ©V-SAB Medical Labs, Incorporated

Exercise Insulin Sensitivity (or Insulin Levels) Exercise sensitizes cells to the effects of insulin. - Body needs less insulin to accomplish same effect. Efficient use of glucose in patients who exercise. Higher metabolism, lean body mass + more activity lead to levels of circulating glucose, in turn reducing insulin requirements = Insulin Levels. Rashid A. Buttar, DO, FAAPM, FACAM 24 ©V-SAB Medical Labs, Incorporated

Exercise Younger Physiology Exercise has always been considered a natural form of anti-aging therapy. • Exercise h. GH, Testosterone, and improves overall hormonal response in the entire system. • Exercise BP, heart rate, respiratory rate, and peripheral vascular resistance, making the system more efficient. • Exercise Endorphins, Lean Body Mass, Immunity, Range of Motion, Endurance, Stamina, Libido, etc. Rashid A. Buttar, DO, FAAPM, FACAM 25 ©V-SAB Medical Labs, Incorporated

Possible Explanation of IGF-1 Levels (Basic Physiology Continued) Answer # 1: Athletes have lower serum glucose levels 2 o to insulin sensitivity. Answer # 2 : Exercise Younger physiological age ( lean body mass, insulin sensitivity, h. GH, etc. ) Rashid A. Buttar, DO, FAAPM, FACAM What is IGF-1? Insulin Like Growth Factor Type 1 is one of many growth factors. Why "Insulin Like” GF-1? The poly-peptide sequence is very similar to Insulin. IGF-1 and Insulin also appear to share many of the same properties & characteristics. 26 ©V-SAB Medical Labs, Incorporated

General Physiological Principals Thus, athletes have lower insulin levels and are biologically younger than non-exercisers. Example: A 79 year-old who exercises is biologically younger than a 79 year-old who rocks in a rocking chair all day. So, Exercise = Anti-Aging Rashid A. Buttar, DO, FAAPM, FACAM 27 Exercise/Anti-Aging causes: in Lean Body Mass in Hormonal levels in h. GH levels in Insulin Sensitivity in Physiological Age in Insulin levels And since Insulin is very similar to IGF-1, then …. . IGF-1 should in Athletes ©V-SAB Medical Labs, Incorporated

IGF-1 in Athletes v. s. Sedentary Patients (3 Week, 38 Patient, Outcome Based Study) 21 Sedentary ♂ & ♀ patients (with no previous history of regular exercise) 17 Athletic ♂ & ♀ patients (regularly exercising for a minimum of 2 years) Age Range Mean Age Median Age IGF-1 Range IGF-1 Mean IGF-1 Median Age Range 25 to 42 yr Mean Age 34. 1 yr Median Age 33. 5 yr IGF-1 Range 88 to 196 IGF-1 Mean 149. 4 ng/ml IGF-1 Median 142. 0 ng/ml 30 to 84 yr 55. 3 yr 57. 0 yr 61 to 304 153. 0 ng/ml 182. 5 ng/ml Rashid A. Buttar, DO, FAAPM, FACAM 28 ©V-SAB Medical Labs, Incorporated

IGF-1 in Athletes v. s. Sedentary Patients (3 Week, 38 Patient, Outcome Based Study) 21 Sedentary ♂ & ♀ Older Group Age Range Mean Age 30 to 84 yr 55. 3 yr Higher IGF-1 Range IGF-1 Mean Age Range Mean Age 25 to 42 yr 34. 1 yr Lower IGF-1 61 to 304 153. 0 ng/ml Rashid A. Buttar, DO, FAAPM, FACAM 17 Athletic ♂ & ♀ Younger Group 29 IGF-1 Range IGF-1 Mean 88 to 196 149. 4 ng/ml ©V-SAB Medical Labs, Incorporated

Growing Consensus that IGF-1 Levels are NOT Related to h. GH Levels Janssen YJ, Helmerhorst F, Frolich M, Roelfsema F, et al: A switch from oral (2 mg/day) to trans-dermal (50 micro/day) 17 beta-estradiol therapy increases serum IGF-I levels in recombinant h. GH substituted women with GH deficiency. Journal of Clinical Endocrinology and Metabolism. 2000 January; 85(1): 464 -6. . …found direct relationship between serum levels of estradiol and IGF-1 levels, independent of h. GH levels. Inuki T, Takanashi K, Takebayashi K, Fuiwara Y, Tayama K, Takemura Y, et al: Thyroid hormone modulates insulin-like growth factor-1(IGF-1) and IGF-binding protein-3, without mediation by growth hormone, in patients with autoimmune thyroid disease. Metabolism Research 1999 October; 31 (10): 576 -9. ……. . found that thyroid hormone modulates IGF-1 and IGF-BP 3, without mediation by h. GH. Rashid A. Buttar, DO, FAAPM, FACAM 30 ©V-SAB Medical Labs, Incorporated

Growing Consensus that IGF-1 Levels are NOT Related to h. GH Levels Furlanetto RW: Insulin-like growth factor measurements in the evaluation of growth hormone secretion. Hormonal Research 1990; 33 Suppl 4: 25 -30. …. . found that IGF-I levels are age dependent and subject to regulation by other hormones and nutritional variables; these features complicate the interpretation of IGF-I levels in individual patients and greatly limit the usefulness of these measurements, especially in establishing GH deficiency or [assessing GH replacement efficacy] …. . . Circulating IGF-II levels are not GH dependent and, therefore, their measurement is of little clinical utility in assessing GH secretion. Department of Pediatrics, University of Rochester Medical Center, N. Y. Rashid A. Buttar, DO, FAAPM, FACAM 31 ©V-SAB Medical Labs, Incorporated

IGF-1 Δ’s Seen in h. GH Injection Tx Current Theory: IGF-1 results from hepatic conversion of h. GH. The more h. GH injected into the body, the higher the IGF-1 levels accumulated. As the limited number of IGF receptor sites become saturated from the h. GH levels being injected (and being converted to IGF-1), IGF-1 serum levels start rising. Thus, assumption is h. GH = IGF-1= Desired Result. Problem with Theory: IGF-1 is similar to Insulin & insulin levels = Bad (cancer & heart dz. ) Our Postulate – Solution to Problem IGF-1 receptor sites should never be saturated to the point where excess serum IGF-1 ’s. Trans-D very effectively ’s number of IGF receptor sites but does not allows IGF-1 levels to exceed physiological limits. Rashid A. Buttar, DO, FAAPM, FACAM 32 ©V-SAB Medical Labs, Incorporated

Possible Reasons for IGF-1 Levels University of Washington, School of Medicine, Seattle, WA …. . “GH exerts its effects by binding to its own receptor sites as well as by stimulating the synthesis of IGF-1. The liver is the primary contributor to levels of IGF-1 in the systemic circulation. But IGF-1 is generated in many GH target tissues [and as a result], local effects may be more important than those of circulating IGF-1 of hepatic origin. ” Merriam GR, Kletke M, Barsness S, et al: GHRH in Normal Aging: An Update. Todays Therapeutic Trends Rashid A. Buttar, DO, FAAPM, FACAM 33 ©V-SAB Medical Labs, Incorporated

Normal Growth in Absence of IGF-1 Two separate studies show normal growth in mice in which hepatic IGF-1 synthesis was eliminated. Yaker S, et al: Normal growth and development in the absence of hepatic IGF-1. Proc. Natl. Acad. Sci. USA 96: 7324 -7329, 1999. Ohlsson C, Sjogren K, Jansson JO and Isaksson OG: The relative importance of endocrine versus autocrine/paracrine IGF-1 in the regulation of body growth. Pediatr. Nephrol. 14: 541 -543, 2000. Rashid A. Buttar, DO, FAAPM, FACAM 34 ©V-SAB Medical Labs, Incorporated

Postulate for IGF-1 Levels Possible reasons for the consistent drop in IGF-1 levels seen while using Trans-D Tropin® 1. Increase IGF-1 utilization leading to a decrease in free IGF-1 levels = circulating serum IGF-1 2. IGF-1 receptor site up regulation leading to an in IGF-1 binding = circulating serum IGF-1 IGF-BP 3 instead may be better for assessing efficacy of h. GH treatments. Further research to validate is necessary. Rashid A. Buttar, DO, FAAPM, FACAM 35 ©V-SAB Medical Labs, Incorporated

Possible correlation clinically observed between stress and IGF-1 levels: Significant mental/emotional stress (due to life style, vocational, situational factors) Higher level of physical stress (due to training/caloric restrictions as in athletes) Severe pain, chronic depression, chronic dz’s (due to immuno suppresion, adrenal exhaustion, late stages of cancer, etc. ) Rashid A. Buttar, DO, FAAPM, FACAM 36 ©V-SAB Medical Labs, Incorporated

Trans-D Tropin®, IGF-1 and Cancer • Kiaris, Schalty, Varga, et al, from Tulane University demonstrated GHRH inhibitors (opposite effect of Trans-D Tropin®) to suppress growth of various cancers, including small cell lung carcinoma. • Authors state clearly however, that NO studies have yet been able to show that GHRH stimulates the proliferation of cancer cells. • Mechanism cited by these authors for suppression of cancer cells was by reducing the levels of IGF’s, “which are known cancercausing agents secreted by the liver and by tumors themselves. ” • Mechanism of cancer suppression 2 o to ing IGF levels per authors. Rashid A. Buttar, DO, FAAPM, FACAM 37 ©V-SAB Medical Labs, Incorporated

IGF-1 and Cancer IGF-1’s are known to stimulate cancer cell proliferation Cohen, Pinchas, et al. Insulin-like growth factors (IGFs), IGF receptors, and IGFbinding proteins in primary cultures of prostate epithelial cells. Journal of Clinical Endocrinology and Metabolism, Vol. 73, No. 2, 1991, pp. 401 -07 Rosenfeld, R. G. , et al. Insulin-like growth factor binding proteins in neoplasia (meeting abstract). Hormones and Growth Factors in Development and Neoplasia, Fogarty International Conference, June 26 -28, 1995, Bethesda, MD, 1995, p. 24 Lippman, Marc E. The development of biological therapies for breast cancer. Science, Vol. 259, January 29, 1993, pp. 631 -32 Papa, Vincenzo, et al. Insulin-like growth factor-I receptors are overexpressed and predict a low risk in human breast cancer. Cancer Research, Vol. 53, 1993, pp. 3736 -40 Rashid A. Buttar, DO, FAAPM, FACAM 38 ©V-SAB Medical Labs, Incorporated

IGF-1 and Cancer Chan, June M. , et al. Plasma insulin-like growth factor I and prostate cancer risk: a prospective study. Science, Vol. 279, January 23, 1998, pp. 563 -66 Stoll, B. A. Breast cancer: further metabolic-endocrine risk markers? British Journal of Cancer, Vol. 76, No. 12, 1997, pp. 1652 -54 Le. Roith, Derek, et al. The role of the insulin-like growth factor-I receptor in cancer. Annals New York Academy of Sciences, Vol. 766, September 7, 1995, pp. 40208 Mantzoros, C. S. , et al. Insulin-like growth factor 1 in relation to prostate cancer and benign prostatic hyperplasia. British Journal of Cancer, Vol. 76, No. 9, 1997, pp. 1115 -18 Cascinu, S. , et al. Inhibition of tumor cell kinetics and serum insulin growth factor I levels by octreotide in colorectal cancer patients. Gastroenterology, Vol. 113, September 1997, pp. 767 -72 Rashid A. Buttar, DO, FAAPM, FACAM 39 ©V-SAB Medical Labs, Incorporated

Trans-D Tropin®, IGF-1 and Cancer • If cancer suppression is 2 o to IGF levels as per authors, then use of synthetic, recombinant GH injection therapy (which reportedly ’s IGF levels) may be carcinogenic and potentially dangerous. • Studies have shown possible correlations between GH injection therapy and cancer proliferation. • Trans-D Tropin® (GHRH analog) ’s IGF levels as demonstrated in 3 separate studies. Thus, conclusion is Trans-D Tropin® may be beneficial in cancer patients. Pending a trial with Cancer patients. • Clinical evidence supports conclusion. Trans-D Tropin® usage is helpful in anorexia, insomnia, pain, fatigue, malaise, ambulation. Rashid A. Buttar, DO, FAAPM, FACAM 40 ©V-SAB Medical Labs, Incorporated

IGF-1 and Cancer Yu, Herbert and Rohan, Thomas. Role of Insulin-Like Growth Factor Family in Cancer Development and Progression. Journal of the National Cancer Institute, Vol. 92, No. 18, September 20, 2000, pp. 1472 - 1489 …………. . “Laboratory studies have shown that IFGs exert strong mitogenic and antiapoptotic actions on various cancer cells……………. IGF’s also act synergistically with other mitogenic growth factors and steroids and antagonize the effect of antiproliferative molecules on cancer growth…. ……. . The role of IGFs in cancer is supported by epidemiologic studies, which have found that high levels of circulating IGF-1 and low levels of IGFBP-3 are associated with increased risk of [many] cancers …. …. IGFs are related to increased cell proliferation, suppression of apoptosis and increased cancer risk. ” Rashid A. Buttar, DO, FAAPM, FACAM 41 ©V-SAB Medical Labs, Incorporated

Multi Centered, Double Blind, Placebo Controlled, Cross Over Study Preliminary Results of Multi-Centered, Double Blind, Placebo Controlled, Cross-Over Study Evaluating Endogenous h. GH Levels with Serial h. GH Radio-Immunoassay Levels After Trans-Dermal GH Releasing Hormone Analog (Trans-D Tropin ) Administration Rashid Buttar, DO, James Biddle, MD, Rajiv Chandra, MD, Terry Grossman, MD, Clarence Norris, MD, James Smith, DO, Annette Stoesser, MD, Dean Viktora, Ph. D Rashid A. Buttar, DO, FAAPM, FACAM 42 ©V-SAB Medical Labs, Incorporated

Multi Centered, Double Blind, Placebo Controlled, Cross Over Study • Requirements were very stringent: – To insure accuracy of study due to transitory nature of h. GH. – To insure quality of study for establishing future protocols. • Patient Selection Criteria: Over 30 years old, non-gravid. • Placebo indistinguishable from Trans-D Tropin® (same carrier, consistency, smell, color, appearance, and bottle). • If study patient did not present as scheduled for blood draw, the patient was eliminated from the study. • Out of 46 centers, 25 separate study sites were selected. – 14 sites dropped out of study due to various reasons. – 4 sites eliminated due to not following study protocol. Rashid A. Buttar, DO, FAAPM, FACAM 43 ©V-SAB Medical Labs, Incorporated

Multi Centered, Double Blind, Placebo Controlled, Cross Over Study Blood Analysis Schedule: Blood Draw Intervals • Control Group (on placebo) • All study patients had Blood specimen analyzed at: blood drawn at Baseline. - 0 & 8 weeks • Trans-D Tropin®/ placebo (2 nd & 5 th week blood administered immediately specimens discarded) after baseline blood draw. • Experimental Group • All study patients had repeat blood draws at 30, (on Trans-D Tropin®) 60 and 90 minutes after Blood specimen analyzed at: treatment administration. - 0, 2, 5 & 8 weeks Rashid A. Buttar, DO, FAAPM, FACAM 44 ©V-SAB Medical Labs, Incorporated

% Δ in Endogenous h. GH in 117 Patients Tested (Δ from Baseline to 90 Min. after Trans-D Tropin® Over 8 Wks) Rashid A. Buttar, DO, FAAPM, FACAM 45 ©V-SAB Medical Labs, Incorporated

Possible Reasons for Decrease in h. GH Levels During Week # 8 of Trans-D Tropin® Study • Somatostatin Issues Negative Inhibitory Feedback Loops become initiated…… Result is in Somatostatin (h. GH antagonist)…. . Result in decreased levels of endogenous h. GH. • Pituitary Reserve Issues Pituitary Gland holds h. GH in store, releasing it in a pulsatile manner. Due to effectiveness of Trans-D Tropin®, the pituitary reserves may become depleted and require time to replenish h. GH stores. Subjective improvements continue at 18 months post treatment initiation Rashid A. Buttar, DO, FAAPM, FACAM 46 ©V-SAB Medical Labs, Incorporated

Multi Centered, Double Blind, Placebo Controlled, Cross Over Study • 5 week increase in experimental group 1754 % but placebo was not drawn at 5 week interval. • Change in Endogenous h. GH measured at 8 weeks: Placebo group - 118. 13% in Endogenous h. GH (attributed to life style modification, exercise, diet, etc. ) Experimental group - 609. 04% in Endogenous h. GH (patients on Trans-D Tropin®) • All patients crossed over into experimental group at 8 weeks, subjectively followed for 8 more weeks Rashid A. Buttar, DO, FAAPM, FACAM 47 ©V-SAB Medical Labs, Incorporated

Trans-D Tropin® & h. GH Injections A study conducted by Roy Heerenveen, MD in the Dutch Antilles (Cura’cao, Aruba, Saba, Bonaire and St. Maarten), assessed concomitant h. GH injection use with Trans-D Tropin® : Pt’s treated with 2 iu h. GH injectable q day experienced same objective changes as pt’s treated with 0. 25 iu h. GH injectable + 6 gtts Trans-D Tropin® (0. 25 iu h. GH injectable q day) + (2 gtt Trans-D Tropin® tid) = 2 iu h. GH q day Conclusion based upon objective responses measured by IGF-1, IGF-BP 3, h. GH radio-immunoassay and SMAC 26 results. Synergism or does 1. 75 iu h. GH injection = 2 gtts Trans-D Tropin® tid? Official recommendation: Do not use h. GH injections with Trans-D Tropin® Rashid A. Buttar, DO, FAAPM, FACAM 51 ©V-SAB Medical Labs, Incorporated

Trans-D Tropin® & h. GH Injections Currently being studied at the Life Extension Institute in the Dutch Antilles and University of Munich in Germany Study conducted at these institutions is entitled The Role of Trans-D Tropin® As An Adjunct To h. GH Injections And should be available within the next 12 months. Rashid A. Buttar, DO, FAAPM, FACAM 52 ©V-SAB Medical Labs, Incorporated

Residual Effects / Responsivity Rene Martina MD, Ph. D and Arnold Segredo MD, Ph. D conducted a study assessing residual effects of Trans D-Tropin® Most people with idiopathic GH deficiency have an intact pituitary but one that does not release adequate GH. Trans D-Tropin® may be the answer in these cases. Trans D-Tropin® acts directly on the pituitary to stimulate synthesis and release of GH, definitively increasing pituitary responsivity as demonstrated. Rashid A. Buttar, DO, FAAPM, FACAM 53 Trans-D Tropin® administered to 12 patients at 9 am, noon, 3 pm and 6 pm. Residual effects were compared to 12 non-stimulated patients, monitored by hourly h. GH radio-immunoassays drawn from 8 pm to 8 am next morning. Rene Martina MD, Ph. D Arnold Segredo MD, Ph. D *Graphs reproduced by permission ©V-SAB Medical Labs, Incorporated

Residual Effects / Responsivity Non-Stimulated, Normal Physiology Stimulated with Trans-D Tropin® Reproduced by permission from Dr. Martina and Dr. Segredo Rashid A. Buttar, DO, FAAPM, FACAM 54 ©V-SAB Medical Labs, Incorporated

Residual Effects / Responsivity Trans D-Tropin® works to stimulate the pituitary to restore the natural physiologic pulsatile release of endogenous GH and is the only physiologic choice to mimic the body's natural pattern of GH release. * Rene Martina MD, Ph. D Arnold Segredo MD, Ph. D *Diagram reproduced by permission Rashid A. Buttar, DO, FAAPM, FACAM 55 ©V-SAB Medical Labs, Incorporated

Advantage of Trans-D Tropin® Besides endogenous GH, there is evidence that Trans-D Tropin® also receptor site activity: 1. ) 2. ) Either up-regulation of target tissue receptor sites or, Increase utilization of GH at local target tissue sites. Evidence found during 24 hour urinary GH collection: Urinary GH levels don’t measure total GH in body but rather, indicate level of GH exceeding renal thresh hold. (similar to glucose spilling over into urine of diabetic patients) Rashid A. Buttar, DO, FAAPM, FACAM 56 ©V-SAB Medical Labs, Incorporated

Evidence of Receptor Site Up-Regulation Baseline to Wk 1: All pts had a large drop in urinary GH representing an in activity of receptor sites / upregulation. Wk 1 to Wk 3: Despite receptor site activity ing, GH steadily ’s beyond baselines, showing in GH production/release Rashid A. Buttar, DO, FAAPM, FACAM 57 ©V-SAB Medical Labs, Incorporated

Testing Protocol for Trans-D Tropin® Safety goal is to rule out a hyper-functioning pituitary state or a salient pituitary tumor to prevent further stimulation. Obtain a serum/saliva IGF-1 or a prolactin level within 30 days. If level above 1 SD of normal, refer to endocrinologist to rule out pituitary adenoma and send for CT scan. STOP TX with TDT. Medical goal is to establish objective proof that the hormonal balance is positively impacted in a physiological manner. TX. If serum, measure h. GH radio-immuno assays prior to TX (baseline) and again at 30, 60 and 90 minutes post TX. If urine, measure h. GH 1 wk post TX and again at 3 wks post Consider CBC, Chem 21, Cortisol, C-reactive prot. , IL-6, NFk. B & hormonal profile Rashid A. Buttar, DO, FAAPM, FACAM 58 ©V-SAB Medical Labs, Incorporated

Use Protocol for Trans-D Tropin® Goal is to emulate natural physiology the way God designed: Maintain natural physiological pulsatile release of h. GH Improve pituitary function instead of creating atrophy Maintain natural physiological safety mechanisms (negative inhibitory feed back loops) 15 gtts – tid or 10 -11 gtts – qid , 5 days on, 2 days off (May want to cycle off every 6 to 8 weeks for 2 weeks) Higher frequency, lower dose seems to get best response. If patients do not see a response in 2 months, stop therapy. Rashid A. Buttar, DO, FAAPM, FACAM 59 ©V-SAB Medical Labs, Incorporated

Uniqueness of Trans-D Tropin® Ingredients – Amino Acids and Fatty Acids Uniqueness is NOT in constituents but rather, how those ingredients ARE COMBINED! Technology utilized to manufacture Trans-D Tropin® as well as delivery mechanism utilized to administer Trans-D Tropin® are what allow for the unique efficacy of this transdermal GHRH analog. Rashid A. Buttar, DO, FAAPM, FACAM 60 ©V-SAB Medical Labs, Incorporated

Uniqueness of Trans-D Tropin® Best results obtained in patients who have been “optimized”, ie, GI dysbiosis resolved, liver detoxified, heavy metals removed, viral loads reduced, systemic candidiasis tx, etc. You can only REBUILD a burning house……. AFTER you’ve PUT OUT the FIRE! Trans-D Tropin® efficacy in smokers, with estrogen replacement therapy, as well as with antidepressant and anxiolytic usage. Rashid A. Buttar, DO, FAAPM, FACAM 61 ©V-SAB Medical Labs, Incorporated

Anti-Aging Therapy Comparison Delivery Monthly $ Ratings Efficacy Availability Restrictions Side Effects Compliance h. GH Injection Tx Injections (SQ) $600 - $2000 50% to 70% 1 to 3 months Prescription Yes Painful / Injections Rashid A. Buttar, DO, FAAPM, FACAM 62 Trans-D Tropin® Transdermal (Drops) $175 87% to 94% 2 to 4 weeks Prescription Pituitary CA, Gravid None to date No pain / Convenient ©V-SAB Medical Labs, Incorporated

Trans-D ® Tropin The goal of therapy is NOT ONLY to EXTEND LIFE, But to IMPROVE THE QUALITY OF THAT EXTENDED LIFE. Rashid A. Buttar, DO, FAAPM, FACAM 65 ©V-SAB Medical Labs, Incorporated