Humanetics Corporation 1550 Utica Avenue South

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7-Keto® for Weight Management An Innovative, All-Natural Response to Diet- and Age-Related

Metabolic Decline

John L. Zenk, MD Chief Medical and Scientific Officer, Humanetics Corporation

A Humanetics Corporation White Paper

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Abstract Aging is associated with decreased resting metabolic rates (RMR) and,

subsequently, weight gain. In humans, resting metabolic rate (RMR) represents

60% of total energy expenditure -- compared to 30% as non-resting energy

expenditure and 10% as heat. Thus, small increases in RMR may result in

considerable energy consumption over time.

As obesity is an epidemic born from energy imbalance, the ability to positively

impact RMR has noteworthy implications in the fight against obesity and calls

into question the assumption that aging necessarily coincides with inevitable

weight gain.

7-oxo DHEA (7-Keto), a naturally occurring substance in the body, is a

metabolite of the adrenal hormone dehydroepiandrosterone (DHEA) and, like

DHEA, the natural production of 7-Keto declines with age. Double-blind, placebo-

controlled studies demonstrate that supplementing with 7-Keto results in a)

measurable increases in RMR, and b) measurable weight loss compared with

diet and exercise alone.

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Problem Statement A significant problem in the management of body weight is a result of changes in

metabolism over time. A key contributing factor in this challenge is that resting

metabolic rate (RMR); which accounts for more than half of daily energy

expenditure decreases markedly with advancing age in sedentary populations

[13][see fig. 1].

Source: Shock, 1962

In the United States, 68% of adults are currently considered overweight and 34%

are classified as obese [1] -- and increased age is accompanied by an increased

prevalence of obesity in nearly every adult age/gender group [see fig. 2].

Source: CDC, Nat. Center for Health Stats, 2000

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It can be hypothesized that age-related weight gain is highly influenced by the

phenomenon of age-related decline in RMR.

In addition to limiting mobility and daily activities, obesity is a risk factor for a host

of chronic disorders including hypertension, hyperlipidemia, diabetes mellitus and

osteoarthritis; obesity has been associated with premature death from all causes

[1-6]. Consequently, obesity and physical inactivity is the second leading cause

of premature death in this country and is currently responsible for 400,000 deaths

annually [7].

Obesity is a disorder of energy balance, occurring when energy expenditure is no

longer in equilibrium with daily energy intake to ensure body weight homeostasis.

Accordingly, efforts to treat obesity must create a negative energy balance,

utilizing stored fat as an energy source. Although approximately 74% of

Americans are engaged in an active attempt to lose or maintain body weight at

any given time [2], the majority of these efforts will not be successful.

Background Information In most weight loss programs, major emphasis is placed on manipulating diet

and appetite; however, interest in pharmacologically increasing energy

expenditure is increasing as such agents represent a new tool for the treatment

of obesity [8]. Increasing energy expenditure may be accomplished by activating

the central or sympathetic nervous systems, thyroid hormones, or other

thermogenically futile cellular mechanisms [8,18].

Specifically, growing evidence supports the hypothesis that individuals with a

low-energy phenotype or advancing age may be predisposed to weight gain and

obesity, as a result of low energy output caused by a low RMR, lack of physical

activity, or both [8,9].

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Fat Free Mass and Resting Metabolic Rate

The fat-free mass (FFM) represents the most metabolically-active body tissue

and is the major determinant of RMR [10,11]; however, weight loss may be

associated with a reduction in metabolic rate due to a decrease of FFM, partially

explaining the difficulty in achieving and maintaining a lower body weight in some

individuals [10-12].

Aging is also associated with: a) a decrease in RMR and FFM, and b) overall

weight gain [13]. Fukagawa and colleagues found that when RMR is adjusted for

FFM it was still lower in older subjects than the mean RMR of younger subjects.

Therefire, the differences in FFM cannot fully account for the lower RMR in older

individuals, suggesting that aging is associated with an alteration in energy

metabolism [13].

Source: Perichon, 1996

Alterations in energy metabolism at the cellular level are difficult to measure in

humans; however, preclinical evidence indicates that there is indeed an age-

related decrease in the peroxisomal fatty acid oxidizing system in mice,

specifically acyl–CoA oxidase activity [14][see fig. 3]. This would suggest an age-

related decline in capacity to convert stored body fat to energy. These

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physiologic changes eventually result in age-related weight gain, which is

documented extensively by the CDC Vital and Health Statistics.

As RMR represents 60% of total energy expenditure -- compared to 30% as non-

resting energy expenditure and 10% as heat [12] -- small increases in RMR may

result in considerable energy consumption over time. Thus, even minimal

increases in daily energy expenditure of 2-3% may have clinical relevance in

preventing the decline in RMR associated with weight loss and aging, aiding in

weight loss and decreasing the risk of regaining the lost weight [8].

7-Keto – A Natural Approach to Combating Age Related Weight Gain 7-Keto is a naturally occurring endogenous metabolite of

dehydroepiandrosterone (DHEA) and its natural production declines with age

similar to DHEA [16][see fig. 4].

Source: Marenich, 1979

7-Keto is structurally different from DHEA and has unique characteristics that

render 7-Keto functionally distinct from DHEA [see fig. 5]. Unlike DHEA, 7-Keto

does not convert to testosterone or estrogens in the body, [20, 21] 7-Keto is not a

steroid hormone precursor, nor does it have androgenic or anabolic potential

[19].

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Fig. 5: 7-Keto

7-Keto demonstrates documented thermogenic activity. This is accomplished

through the activation of three thermogenic enzymes: Glycerol-3-Phosphate

Dehydrogenase, Malic Enzyme and Fatty Acyl CoA Oxidase [18, 21, 37, 38].

In keeping with the biological definition of thermogenesis, all three of these

enzyme activations drive energy-producing substrates in a direction of less

efficient ATP production relative to heat production. The enzymes also promote

the utilization of fat stores for energy and heat production. This is the basis for 7-

Keto’s ability to enhance thermogenesis and, through that mechanism,

accelerate the utilization of fat stores for energy.

Preclinical studies in rats have shown that 7-Keto is more potent than DHEA for

inducing these thermogenic enzymes [17]. Additionally, 7-Keto increases the rate

of mitochondrial substrate oxidation, liver catalase activity and fatty acyl-CoA

oxidase activity [18] without activating the androgen receptor [19].

The age-related decline in key compounds like 7-Keto is thought to play a role in

the decrease in metabolic enzyme activity with advancing age.

Based on these findings of 7-Keto-induced thermogenic activity, clinical research

was performed to further document these changes in humans. A placebo-

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controlled study showed an increase in RMR in overweight individuals taking 7-

Keto [24].

Additionally, two placebo-controlled studies demonstrated 200% greater weight

loss in subjects using 7-Keto (100 mg twice daily) than those using placebo over

8 weeks when both were used in conjunction with a calorie-restricted diet and

exercise program [22,23].

Clinical Studies

RMR Study The results of this study published in 2007 revealed that administration of 7-Keto

to overweight adults in conjunction with a calorie restricted diet did effectively

reverse the decline in resting metabolic rate (RMR) normally associated with

dieting. This study was randomized, double-blind and placebo controlled with a

crossover design to decrease inter-subject variability.

Source: Zenk, 2007

As expected, the individuals enrolled in this study demonstrated a substantial

decline in RMR (-3.9%) when subjected to a calorie-restricted diet; however, the

twice-daily administration of 7-Keto for 7 days resulted in a substantial increase

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in RMR of 1.4% above baseline levels when added to the same calorie-restricted

diet [see fig. 6].

Compared to placebo treatment periods, the mean increase in RMR following 7-

Keto treatment was 96 kcal/day (5.4%). No significant differences between

treatment periods with respect to the quantities of carbohydrate, protein, fat or

total calories consumed were found. Therefore, the significant increase in RMR

associated with the administration of 7-Keto in this study appear to be due to

pharmacologically-induced increase in resting energy expenditure in these

individuals. 7-Keto achieved this thermogenic effect without cardiovascular or

central nervous system side effects [24].

Weight Loss Study #1 In this double-blind, placebo controlled protocol, 30 adults with a mean body

mass index of 31.9 ± 6.2 kg/m2 were randomly divided into 2 groups of 15: Group

1 received 7-Keto 100 mg twice daily and Group 2 received placebo for 8 weeks.

All subjects participated in an exercise training program 3 times per week.

Exercise session consisted of 60 minutes of cross training under the supervision

of an exercise physiologist. In addition, each subject was instructed to follow a

diet of ~ 1800 kcal/day by a registered dietitian.

Subjects received biweekly dietary counseling to encourage compliance. Study

participants underwent serum multiple-assay chemistry testing, as well as body

composition, blood pressure, and dietary analysis at baseline, week 4 and week

8 [22].

Of the 30 subjects who entered the study, 23 completed the 8-week protocol.

Seven subjects dropped out for personal reasons unrelated to the study. Group 1

lost a significant amount of body weight compared with Group 2 (-2.88 kg vs –

0.97 kg; p = 0.01) over the 8 weeks [see fig. 7]. Group 1 also achieved a

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significant reduction in body fat compared with Group 2 (-1.8% vs. –0.28%; p <

0.01).

Fig. 7: Weight Loss Study #1

Source: Kalman, 2000

There were no significant changes in levels of thyroid-stimulating hormone (TSH)

or thyroxine (T4) in either group. In addition, no significant changes were

observed in vital signs, blood sugar, testosterone and estradiol levels, liver and

renal function, or overall caloric intake during the study. No subjective adverse

effects were reported throughout the study. The results of the study suggest that

7-Keto combined with moderate exercise and a reduced-calorie diet significantly

reduces body weight and body fat compared with exercise and a reduced-calorie

diet alone [22].

Weight Loss Study #2 This study assessed the effects of a formula containing 7-Keto, (200 mg/day) on

weight loss, body composition, and RMR in overweight patients following a

weight-reduction diet and exercise regimen. It is worth noting that the formula

used for this clinical trial contained no elements -- other than 7-Keto -- with

proven weight loss effects.

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In this prospective, randomized, double-blind, placebo-controlled trial, healthy,

overweight adults were given the formula containing 7-Keto or a placebo and

followed a calorie restricted diet (~1800 kcal/d) and an exercise program for 8

weeks. Body weight, body composition (by bioelectric impedance), and RMR (by

indirect calorimetry) were measured at baseline, week 4 and week 8. A thyroid

panel was done at baseline and week 8 [23].

Of 35 healthy, overweight adults enrolled, 33 completed the study (12 men, 21

women; age, 40-69 years; body mass index [BMI], 27.0-42.7 kg/m2). Patients

taking 7-Keto lost significantly more weight after 8 weeks than those taking

placebo, 2.15 ± 2.38 kg and 0.72 ± 2.12 kg, respectively) (p=0.038)[see fig. 8].

The change in BMI in the 7-Keto-treated group was significant compared with the

change in the placebo group decrease, 0.71 ± 0.79 kg/m2 and 0.01 ± 1.05 kg/m2,

respectively) (p=0.036).

Fig. 8: Weight Loss Study #2

Source: Zenk, 2002

There were no other statistically significant differences in any of the other

measured variables. 7-Keto was well tolerated, and there were no significant

adverse events [23].

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Discussion

In the United States, the obesity epidemic is widely acknowledged to be just that:

an epidemic. With prevalence increasing along with age [1] -- and the myriad of

associated health-related complications -- there is a clear need for safe, effective

solutions. As obesity is an issue of energy imbalance, the challenge is thus to

address that imbalance and move toward equilibrium. As RMR represents well

over half of total energy expenditure [12], and RMR is known to decline with age,

it is a key player in this effort.

The three studies detailed in this paper offer compelling evidence to support the

following conclusions:

a) Increased RMR is associated with weight loss among overweight and obese

adults; and

b) 7-Keto has been shown to positively impact RMR and ultimately contribute to

three times more weight loss when compared with diet and exercise efforts

alone.

As much of this discussion revolves around age-related weight gain, it is

important to acknowledge that any recommended solution must be feasible for

adults of all ages. As a dietary supplement, 7-Keto offers a non-invasive option

for which advanced age and limited mobility are not preclusions.

While a thorough review of the extensive toxicological evaluations of 7-Keto is

outside the scope of this paper, it is worth noting that such analyses have taken

place with no adverse reactions reported in any of the studies [35, 36].1

In addition, a complete pharmacokinetic analysis for 7-Keto has been completed.

This pharmacokinetic analysis describes exactly how the body absorbs,

metabolizes, distributes and excretes 7-Keto. It reveals that 7-Keto is rapidly 1 To date, consumers have taken nearly one billion doses of 7-Keto. No serious adverse events have ever been reported.

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absorbed and converted to its sulfate derivative, it reaches peak plasma

concentrations in 2.2 hours and has a half-life of 2.17 hours and there is no

accumulation with repeated dosing [20].

7-Keto is considered a dietary ingredient under the framework of the Dietary

Supplement Health and Education Act of 1994 (DSHEA). Dietary ingredients are

regulated by FDA and in accordance with its lawful sale under DSHEA, 7-Keto

has been the subject of two successfully filed New Dietary Ingredient (NDI)

notifications with the FDA. It has been sold in the United States in dietary

supplements since 1998.

Ultimately, with the support of multiple double-blind, placebo-controlled studies

and an impeccable safety record, 7-Keto emerges as a formidable tool in the

fight against weight gain related to age and decreased metabolic rate.

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References 1. Flegal KM, Carroll MD, Ogden CL, Curtin LR. Prevalence and trends in obesity

among US adults, 1999-2008. JAMA. 2010; 303:235-241.

2. Mokdad AH, Bowman BA, Ford ES, Vinicor F, Koplan JP. The continuing

epidemics of obesity and diabetes in the United States. JAMA. 2001; 286:1195-

1200.

3. Calle EE, Thum MJ, Petrelli JM, Rodriguez C, Heath CW. Body-Mass Index and

mortality in a prospective cohort of US adults. N Engl J Med. 1999; 341:1097-

1105.

4. Markus RA, Mack WJ, Azen SP, Hodis HN. Influence of lifestyle modification on

atherosclerotic progression determined by ultrasonographic change in the

common carotid intimal-media thickness. Am J Clin Nutr. 1997; 65:1000-04.

5. Nisoli E, Carruba MO. Emerging aspects of pharmacotherapy for obesity and

metabolic syndrome. Pharmacol Res. 2004; 50:453-69.

6. Colditz GA. Economic costs of obesity. Am J Clin Nutr. 1992; 55:503S-507S.

7. Mokdad AH, Marks JS, Stroup DF, Gerberdine JL. Actual causes of death in the

United States, 2000. JAMA. 2004; 291:1238-45.

8. Astrup A. Thermogenic drugs as a strategy for treatment of obesity. Endocrine.

2000; 13:207-12.

9. Astrup A. Macronutrient balances and obesity: the role of diet and physical

activity. Public Health Nutrition. 1999; 2:341-7.

10. Ravussin E, Swinburn BA. Metabolic predictors of obesity: cross-sectional versus

longitudinal data. Int J Obes Relat Metab Disord. 1993;17(Suppl 3):S28-S31.

11. Cunningham JJ. Body composition as a determinant of energy expenditure: a

synthetic review and a proposed general prediction equation. Am J Clin Nutr.

1991; 54:963-9.

12. Leibel RL, Rosenbaum M, Hirsch J. Changes in energy expenditure resulting

from altered body weight. N Engl J Med. 1995; 332:621-8.

13. Fukagawa NK, Bandini LG, Young JB. Effect of Age on Body Composition and

Resting Metabolic Rate, Amer J Physio Endo. 1990; 259(2):E233-E238.

14. Perichon R, Bourre JM. Liver Peroxisomal Fatty Acid Oxidizing System During

Aging in Control and Clofibrate-Treated Mice. Biochem Mol Biol Int. 1995;

37(3):475-480.

15

15. Bray GA. The MONA LISA hypothesis. Most obesities known are low in

sympathetic activity. In, Oomura Y, Tarui S, Inoue S, Shimazu T, editors.

Progress in Obesity Research. London: John Libbey & Co.; 1990. pp.61-6.

16. Marenich LP. Secretion of Testosterone, Epitestosterone, Androstenedione, and

7-Keto Dehydroepiandrosterone in Healthy Men of Different Ages. Prob

Endokrinol. 1979;25:28-31.

17. Lardy H, Partridge B, Kneer N, Wei Y. Ergosteroids: induction of thermogenic

enzymes in liver of rats treated with steroids derived from

dehydroepiandrosterone. Proc Natl Acad Sci. 1995; 92:6617-9.

18. Bobyleva V, Bellei M, Kneer N, Lardy H. The effects of the ergosteroid 7-oxo-

dehydroepiandrosterone on mitochondrial membrane potential: possible

relationship to thermogenesis. Arch Biochem Biophys. 1997; 341:122-8.

19. Miyamoto H, Yeh S, Lardy H, Messing E, Chang C. Delta-5-androstenediol is a

natural hormone with androgenic activity in human prostate cancer cells. Proc

Natl Acad Sci. 1998; 95:11083-88.

20. Davidson M, Marwah A, Sawchuk RJ, Maki K, Marwah P, Weeks C, Lardy H.

Safety and pharmacokinetic study with escalating doses of 3-acetyl-7-oxo-

dehydroepiandrosterone in healthy male volunteers. Clin Invest Med. 2000;

23:300-10.

21. Lardy H, Kneer N, Wei Y, Partridge B, Marwah P. Ergosteroids II: biologically

active metabolites and synthetic derivatives of dehydroepiandrosterone. Steroids.

1998; 63:158-65.

22. Kalman DS, Colker CM, Swain MA, Torina GC, Shi Q. A randomized, double

blind, placebo controlled study of 3-acetyl-7-oxo-dehydroepiandrosterone in

healthy overweight adults. Curr Ther Res. 2000; 61:35-442.

23. Zenk JL, Helmer TR, Kassen LJ, Kuskowski MA. The effect of 7-Keto

Naturalean™ on weight loss: a randomized, double-blind, placebo-controlled

trial. Curr Ther Res. 2002; 63:263-72.

24. Zenk JL, Frestedt JL, Kuskowski MA. HUM5007, a Novel Combination of

Thermogenice Compounds, and 3-Acetyl-7-Oxo Dehydroepiandrosterone: Each

Increases the Resting Metabolic Rate of Overweight Adults. J Nutr Biochem

2007;18:629-634.

16

25. Davies KM, Heaney RP, Recker RR, Lappe JM, Barger-Lux MJ, Rafferty K,

Hinders S. Calcium intake and body weight. J Clin Endocrinol Metab. 2000;

85:4635-8.

26. Lin YC, Lyle RM, McCabe LD, McCabe GP, Weaver CM, Teegarden D. Dairy

calcium is related to changes in body composition during a two-year exercise

intervention in young women. J Am Coll Nutr. 2000; 19:754-60.

27. Zemel MB, Thompson W, Milstead A, Morris K, Campbell P. Calcium and dairy

acceleration of weight and fat loss during energy restriction in obese adults.

Obes Res. 2004; 12:582-90.

28. Zemel MB, Richards J, Mathis S, Milstead A, Gebhardt L, Silva E. Dairy

augmentation of total and central fat loss in obese subjects. Int J Obese Relat

Metab Disord. 2005; 29:391-7.

29. Marcus R. Agents affecting calcification and bone turnover - calcium, phosphate,

parathyroid hormone, vitamin D, calcitonin, and other compounds. In, Hardman

JG, Limbird LE, Molinoff PB, Ruddon RW, editors. Goodman & Gilman’s The

Pharmacological Basis of Therapeutics. New York: McGraw Hill; 1996. pp.1529-

36.

30. Anonymous. Osteoporosis prevention, diagnosis, and therapy. NIH Consensus

Development Panel on Osteoporosis Prevention, Diagnosis, and Therapy. JAMA.

2001; 285:785-95.

31. Mortensen L, Charles P. Bioavailability of calcium supplements and the effect of

vitamin D: comparisons between milk, calcium citrate, and calcium citrate plus

Vitamin D. Am J Clin Nutr. 1996; 63:354-57.

32. Bronner F. Mechanisms and functional aspects of intestinal calcium absorption. J

Exp Zoolog A Comp Exp Biol. 2003; 300:47-52.

33. Bouillon R, Van Cromphaut S, Carmeliet G. Intestinal calcium absorption:

molecular vitamin D mediated mechanisms. J Cell Biochem. 2003; 88:332-39.

34. Hoenderop JG, Nilius B, Bindels RJ. Calcium absorption across the epithelium.

Physiol Rev. 2005; 85:373-422.

35. Lardy H, Henwood SM, Weeks CE. An Acute Oral Gavage Study of 3-

acetoxyandrost-5-ene-7,17-dione (7-oxo-DHEA-acetate) in Rats. Biochem and

Biophys Res Comm. 1999;254:120-123.

17

36. Henwood SM, Weeks CE, Lardy H. An Escalating Dose Oral Gavage Study of 3-

acetoxyandrost-5-ene-7,17-dione (7-oxo-DHEA-acetate) in Rhesis Monkeys.

Biochem and Biophys Res Comm. 1999;254:124-126.

37. Bobyleva V, Kneer N, Bellei M, Battelli D, Lardy H. Concerning the Mechanism of

Increased Thermogenesis in Rats Treated with Dehydroepiandrosterone. J

Bioenerg Biomembr. 1993;25:313-321.

38. Lardy H, Partridge B, Kneer N, Wei Y. Ergosteroids: Induction of Thermogenic

Enzymes in Liver of Rats Treated with Steroids Derived from

Dehydroepiandrosterone. Proc Natl Acad Sci. 1995;92:6617-6619.