Astaxanthin and Human Health Medical Summary

(July, 2010)

(1) Introduction- Oxidative Stress

Oxidative stress in living cells arises both as a natural result of aerobic metabolism and as a result of environmental and pathologic conditions (Papas 1999). Reactive-oxygen species (ROS) are constantly being produced during normal metabolic processes (Reviewed in Davies 1995, Dore 2005), and tend to react with neighboring molecules such as proteins, DNA, RNA, carbohydrates, and lipids putting these molecules at risk. The results of such oxidative attack may include protein and lipid peroxidation and structural changes in DNA and RNA, which in turn may lead to damage, mutations, and even loss of function. Environmental (air pollution, tobacco smoke, toxic chemicals or ultraviolet (UV) light), as well as physiological (stress, inflammation, diabetes) conditions can enhance the production of ROS. Indeed, oxidative damage has been linked to aging, atherosclerosis, ischemia-reperfusion injury, macular degeneration of the eye, carcinogenesis, neurodegenerative diseases such as Alzheimers and Parkinsons diseases, bacterial and viral meningitis, and many other biological processes and diseases. The human body has evolved an array of endogenous antioxidant defenses against oxidative insult, however, these endogenous antioxidants do not completely protect against the sum of oxidative stresses challenging the body. Dietary antioxidants, particularly those that the body cannot synthesize, are crucial to the body's endogenous defenses against oxidative stress and prominent in protecting against disease and age-related phenomena (Ames 1993; Davies 1995, Halliwell 1996).

(2) The Carotenoids

The group of antioxidants mostly investigated for their potential health benefits, are the carotenoids (reviewed in Cooper 1999). Carotenoids are lipid soluble pigments produced in some plants, algae, fungi and bacterial species that account for their red, orange, or yellow hues. Carotenoids are antioxidants due to their ability to quench singlet oxygen, be oxidized, and isomerized (Edge 1997, Mortensen 1997). In addition, carotenoids absorb light, hence providing defense from photooxidative damage. Among the various carotenoids, Astaxanthin is the most potent natural antioxidant. This was shown by comparative studies in many in vitro as well as in vivo systems, by comparing the ability to neutralize singlet oxygen and to inhibit the oxidation of proteins and lipids (Kurashige 1990, Miki 1991, Shimidzu 1996, Oshima 1993, Barros 2001, Kim 2009, Manabe 2008, Choi 2008, Camera 2009, Wolf 2010).

(3) Astaxanthin chemistry

Astaxanthin (3,3-dihydroxy---carotene-4,4-dione ) is a carotenoid, commonly found in marine environments. The presence of the hydroxyl and keto endings on each ionone ring (Figure below), explains some unique features of Astaxanthin, such as its ability to be esterified, its high anti-oxidant potency and a more polar configuration than other carotenoids. Free Astaxanthin is extremely sensitive to oxidation, however its esterified form, as found in nature in the Haematococcus pluvialis (H. pluvialis) microalga, is stable and displays higher bioavailability and potency (Odberg 2003, Rufer 2008, Barbossa 1999).

Chemical structure of Astaxanthin

O

17 16

18' 19 20

H3 C
1 2 3
HO

CH3
7 6 5 4
O

CH3
9 8 10 11 12

CH3
13 14 15 15' 14' 13' 12' 11' 10' 9'

H3 C
8' 7'

4' 5' 6' 1' 3' 2'

OH

CH3
18

CH3
20'

CH3
19'

H3 C

16' 17'

CH3

CAS Number: 472-61-7. MW: 596.85

(4) Bioavailability
Astaxanthin is not soluble in water, therefore, its bioavailability is low and is determined by several factors, the most prominent one being its level of esterification. When it is esterified (bound to fat), or when taken with oil, Astaxanthin is readily absorbed in the digestive system and displays higher bioavailabilty (Odberg 2003, Rufer 2008, Barbossa 1999, Okada 2009). Other factors influencing its bioavailability include its level of oxidation, its chemical structure, the method (Mendes-Pinto 2001) and source (Bowen 1999) of industrial production, and additional dietary factors such as the presence of other carotenoids and smoking (Okada 2009).

(5) Pharmacokinetics - ADME: Absorption, Distribution, Metabolism and Elimination

Absorption of Astaxanthin starts with hydrolization of the ester moiety when the molecule crosses the apical membranes of the intestinal epithelial cells (Odeberg 2003, Coral-Hinostroza 2004). Following hydrolization, Astaxanthin digestion and intestinal absorption are closely associated with fatty acids uptake, transport and delivery, since it is a fat-soluble compound. Astaxanthin is incorporated into chylomicrons, and distributed between very low density lipoprotein (VLDL), low density lipoprotein (LDL) and high density lipoprotein (HDL) and transported to the tissues (Wang 1992). Following intake, plasma levels reach maximal values within a few hours (Okada 2010, Odeberg 2003, sterlie 2000, Coral-Hinostroza 2004) and both Tmax and Cmax are higher after meal or fat intake (Odeberg 2003, Okada 2010). Accumulation of Astaxanthin in plasma, following dose response, shows nonlinear kinetics (Rufer 2008, Coral-Hinostroza 2004). This may be explained by prolonged absorption through the Peyers patch route into the lymphatic system. Astaxanthin is distributed to all tissues, and is able to cross the blood-brain and other biological barriers (Aoi 2003). The limited studies in humans suggest that there is a specific accumulation of Astaxanthin in VLDL chylomicrons (36-64% of total Astaxanthin, after 6.7 h, Osterlie 2000, Coral-Hinstroza 2004). Four main free metabolites of Astaxanthin were found and their reduction products were also present as glucuronides (Kistler 2002). Astaxanthin does not affect CYP enzymes or metabolism of drugs. Elimination of Astaxanthin is relatively fast with elimination half life ranging between 15.9h (Odenberg 2003) to 52h (Coral-Hinostroza 2004). In addition, studies in rodents showed that the length of exposure to Astaxanthin is not related to plasma concentration (Stewart 2008), and that there is a rapid elimination or catabolism of Astaxanthin without long-term storage of astaxanthin in tissues (Petri 2007). Taken together, these results suggest that Astaxanthin does not accumulate in the body.

(6) Safety of Astaxanthin

Astaxanthin is naturally found in many animals and plants. In salmon flesh its concentration may rang from 3 to 40 mg/kg (Turujman 1997). The main Astaxanthin isomer identified in salmon was the 3S,3'S stereoisomer identical to that found in H. pluvialis. Astaxanthin is also found in crustacean species such as krill, shrimp, crab, lobster and crawfish. Thus, humans have been ingesting Astaxanthin for centuries. Astaxanthin was approved by the FDA as dietary supplement in aquaculture in 1987. Astaxanthin, which is produced by H. Pluvialis, is currently the only type of Astaxanthin that was approved by the FDA, in 2000, as a dietary supplement (Pashkew 2008). Astaxanthin has been marketed as a dietary supplement for approximately 10 years without any adverse effects. Furthermore, there is no evidence that consumption of Astaxanthin in foods or as a dietary supplement has any cumulative effect that would affect its safety. The published recommended daily intake of Astaxanthin varies among different regulatory authorities. It is 6 mg/day in Japan, 5 mg/day in the US and 4 mg/day in Europe. The safety of Astaxanthin either as an extract from natural sources, in its synthetic form, or as present in algal biomass has been documented in many pre-clinical and clinical studies. The evidences for its safety can be summarized as:
1) The long-term repeat dose exposure (1.5 and 3 months) of Astaxanthin did not result in its bioaccumulation or any adverse effects (Ono 1999, Nishikawa 1997). 2) Many animal studies support the safety of Astaxanthin. These include specially designed safety tests such as ADME studies, acute and subchronic (Takashi 2005) studies (LD50 > 2000 mg/kg body weight, NOEL = 50 mg /kg/day), repeated-dose toxicity and fertility study (Nishikawa 1997), embryotoxicity and teratology (Roche 1987), multi generation study (Roche 1987), micronucleus test (Scantox, 1998) and many efficacy-related experiments in which histological and biochemical follow-up was included. 3) In vitro studies did not report of any mutagenicity (AMES test) (Takahashi 2005). 4) In multiple human clinical studies (>25), the safety of Astaxanthin was confirmed at doses of up to 40 mg/day for 8 weeks or 4 mg/day for one year. Results from these trials provide a huge margin of safety above the currently recommended levels.

In conclusion: none of all the above mentioned studies revealed any adverse effects of Astaxanthin.

(7) Astaxanthin and human health Pre-clinical and clinical studies.

The physiological functions of Astaxanthin have been studied for more than 20 years. It was found to have beneficial effects supporting human health and well-being and by preventing pathologies. In the following pages the main studies, which have been conducted with Astaxanthin are summarized in a table. Studies are categorized according to the physiological system / organ / or pathology studied. Table rows that summarize clinical studies are highlighted:

Effect of Astaxanthin on various physiological systems

System
General anti-oxidation

Pre-clinical/clinical Effect followed

Vitamine E-deficient mice Rats Rats Diabetic rats

Main outcome
Reduction Reduction Inhibition Improved Reduction Reduction
levels of plasma 12and 15hydroxy fatty acids. Improvement in some markers

Reference
Nishigaki 1994, Kurashige 1990 Gross 2006 Hussein 2006 Nakano 2008

Lipid peroxidation Reduction Oxidation NO end products in plasma General oxidation Renal function DNA damage
Hepatocarcinogenesis

Cyclophosphamide-

Tripathi 2010 Krappi 2007

treated rats 20 healthy men

Lipid peroxidation Reduced

35 postmenopausal women Eye Rats Rats Mice Rats Diabetic patients 15 subjects with
nonadvanced AMD

Biochemical and physical markers Light damage to retina Uveitis CNV Selenite-induced cataract
Humoral superoxide scavenging activity

Iwabayashi 2009 Tso 1996 Ohgami 2003, Suzuki 2006 Izumi-Nagai 2008 Liao 2009 Hashimoto 2009 Parisi 2008 Sawaki 2002 Iwasaki 2006 Kenji 2005 Nakamura 2004
Nagaki 2002 Nagaki 2006

Reduction Reduction Reduction Inhibition Increased Improved Improved Improved

Improved

18 healthy men 10 healthy men 40 asthenopia patients 49 healthy men 87 visual display terminal (VDT) workers Hairless UV-irradiated mice

Skin

Healthy subjects

Central retina function Deep vision Eye function Accommodation power Uncorrected far visual acuity Accommodation amplitude Wrinkles formation Biochemical markers of skin photo damage Erythema

Improved improved Reduction Reduction

Arakane 2002 Savoure 1995

Reduction

Yamashita, 1995

46 healthy women Immune response Mice 14 healthy women

skin elasticity and moisture

Humoral responses to T-dependent antigens

Improved Improved Reduced Reduced Improved Reduced Reduced Reduced Reduced Improved Reduced No effect No effect Increased Improved Improved Reduced Reduced Reduced Reduced Reduced Reduced Improved Increased Improved Reduced Reduced Reduced Improved

Yamashita 2002, Yamashita 2007,Seki 2001 Jyonouchi 1994, Bennedsen 1999, Chew 1999

Inflammation

Rats
Mice injected with inflammation inducers

Gastric ulcer

H. pylori-infected mice

Chemically - induced gastric ulcers in rats 44 patients with functional dyspepsia

Oxidative stress markers Inflammation markers Immune response Inflammation of paw Inflammation markers in plasma Gastric mucus Bacterial load Immune response Gastric ulcer markers inflammatory markers Gastrointestinal discomfort HDL level Dyslipidemia Adiponectin levels Hypertension Stroke incidence
Artery wall thickness

Park 2010

Kurashige 1990 Lockwood 2006 Bennedsen 1999, Wang 2000

Kim 2005a, Kamath 2008, Kim 2005b, Yang 2009

Kupcinskas 2008, Andersen 2007

Cardiovascular

Normal Rats
Insulin resistant rats

system

Obese rats Hypertensive rats

Murillo 1992 Hussein 2007, Ikeuchi 2007

Yanai 2008

Rats Rabbits Dogs

Myocardial ischemiareperfusion damage

Dogs 61 mildly hyperlipidemic subjects

Muscle endurance

Exercising mice

Rethrombosis Dyslipidemia Adiponectin level Muscle lipid metabolism Oxidative damage Fatigue
Lactic acid accumulation after run Respiratory and sympathetic nervous system activities

Gross 2004 Lauver 2005 Gross 2005 Krtz 2004 Yoshida 2010

Aoi 2008 Aoi 2003

Ikeuchi 2006

16 non trained men 19 non trained men

Sawaki 2002 Nagata 2006

20 non trained men

Strength/explosivenes s

No effect Improved No effect Inhibited

Lignell 2001

Cancer

20 resistance trained men Tumor-transplanted mice Mice Chemically-induced urinary bladder cancer in mice Chemically-induced cancers of the oral cavity and colon Mice subjected to transient cerebral ischemia Rats subjected to transient cerebral ischemia Rats chronically fed with alcohol Healthy mice 10 healthy men (50-69 years old) 20 Sub-fertile men 30 Sub-fertile men

test Strength/endurance test Markers of skeletal muscle injury Tumor growth

Bloomer 2005 Sun 1998 Jyonouchi 2000, Chew 1999 Nishino 1998 Tanaka 1994

Spontaneous liver carcinogenesis Tumor growth

Reduced Reduced

Tumor growth

Reduced

Central nervous system

Learning and memory skills

Improved

Prabhu 2009, Nagendrapra-bhu1 2009 Hussein 2005a

Male fertility

Metabolic syndrome (MS)

Diabetic db/db mice Hypertensive rats Obese rats

Diabetic db/db mice

Locomotor activity Cerebral infarction Alcohol-induced neural marker Memory Response time and accuracy of several tasks Semen quality Pregnancy rate Sperm velocity Oxidation markers Pregnancy rate Plasma glucose level MS biochemical markers Body weight Adipose tissue weight MS markers Type 2 Diabetesinduced nephropathy

Improved Reduced Reduced Improved Improved

Shen 2009

Abadie-Guedes 2008

Zhang 2007 Satoh 2008

Improved Improved Reduced Improved Reduced Improved Reduced Reduced Improved Progression prevented

El-Garem 2002 Comhaire 2005

Uchiyama 2002 Hussein 2007 Ikeuchi 2007

Naito 2004

Additional to the studies mentioned above it is important to emphasize that Astaxanthin have been marketed and used for the last 15 years by many satisfied customers, consuming it as and in dietary supplements, cosmetics and various food products.

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