Title: Therapeutic uses of
United States Patent: 7,264,813
Issued: September 4, 2007
Inventors: Shaish; Aviv
(Tel Hashomer, IL), Harats; Dror (Ramat Ran, IL)
Assignee: Nikken Sohonsha
Corporation (Hashima, JP)
Tel Hashomer Medical Research Infrastructure and Services Ltd. (Tel
Appl. No.: 10/668,601
Filed: September 24, 2003
Master of Science in Law
A method for treating a disease selected
from diabetes mellitus and atherosclerosis, and a method for reducing
triglycerides and/or increasing HDL cholesterol levels in the plasma of a
subject. The method comprises administrating. to a subject an effective
amount of crude Dunaliella powder, optionally together with an activator
of nuclear receptors.
Description of the Invention
FIELD OF THE
This invention relates to the use of
crude Dunaliella bardawil powder in the treatment of a number of
conditions including high triglyceride and low high-density lipoprotein (HDL)
cholesterol levels, atherosclerosis and diabetes.
The following is a list of references
believed to be pertinent as a background to the present invention: 1. Ben-Amotz,
A. and Shaish, A., Carotene Biosynthesis in Dunaliella: Physiology,
Biochemistry and Biotechnology, Ed. Avron, M. and Ben-Amotz., 9:206-16,
1992. 2. U.S. Plant Pat. No. 4,511, issued Mar. 18, 1980. 3. U.S. Pat. No.
4,199,895, issued Apr. 29, 1980. 4. Ben-Amotz, A., Mokady, S., Edelstein,
S. and Avron, M., Bio-availability of a natural isomer mixture as compared
with synthetic all-trans beta-carotene in, rats and chicks, J. Nutrition,
119:1013-1019, 1989. 5. Day, C., Thiazolidinediones: a new class of
antidiabetic drugs Diabetic Med. 16:179-192, 1999. 6. Mukherjee, R.,
Davies, P. J. A. et al., Sensitization of diabetic and obese mice to
insulin by retinoid X receptor agonists. Nature, 386:407-410, 1997.
BACKGROUND OF THE
Two strains of Dunaliella, a unicellular,
biflagellate, wall-less green alga, are capable of producing very large
amounts of .beta.-carotene, Dunaliella salina Teod. and Dunaliella
bardawil.sup.(1). D. bardawil is a halotolerant alga whose .beta.-carotene
content comprises about 50% all-trans-.beta.-carotene with the remainder
composed mostly of 9-cis .beta.-carotene and a few other .beta.-carotene
isomers.sup.(2). A process has been described for cultivating D. bardawil
so as to obtain algae containing up to about 5% by weight of .beta.-carotene.sup.(3).
Later developments of the process increased the percentage to more than
8%. It has been shown that the natural isomer mixture of .beta.-carotene
which is accumulated in the alga Dunaliella bardawil is accumulated in
fatty tissues of rats and chicks to an extent which is about 10 fold
higher than that observed by feeding the synthetic all-trands .beta.-carotene.sup.(4).
Various carotenoid-enriched Dunaliella commercial products are available
such as Betatene.TM. (produced by the Henkel Corp., Germany) and
Nutrilite.TM. (Amway, Inc., U.S.A.). These products are oil extracts of
carotenoids from, Dunaliella.
Retinoids are essential for a living organism for vision, cellular growth,
differentiation, and to maintain the general health of the organism.
9-cis-retinoic-acid and all-trans-retinoic-acid are produced in the body
by the cleavage of .beta.-carotene (BC). The retinoid nuclear receptors,
also known as orphan receptors RAR and RXR, have distinct physiologic
properties by activating transcriptional. factors. All-trans retinoic-acid
binds to RAR but not to RXR, while 9-cis retinoic-acid binds to RXR, which
plays a key role in important biological processes.
The activation of nuclear receptors is essential to cell metabolism.,
especially with respect to lipids and glucose. Several nuclear receptors
have been implicated in cholesterol homeostasis. These receptors include:
the liver X receptors (LXRa/NR1H3 and LXRb/NR1H2) and farnesoid X receptor
(FXR/NR1H4) that are bound and activated by oxysterols and bile acids,
respectively. LXR and FXR form obligate heterodimters with RXR that is
activated by 9-cis retinoic acid and synthetic agonists (termed rexinoids).
It was recently found that the activation of RXR by its ligands can affect
two central processes of cholesterol metabolism: I. Cholesterol absorption
in the intestine(activation of RXR/FXR heterodimer repressed cholesterol
7.alpha.-hydroxylase (CYP7A1) expression, resulting in decreased bile acid
synthesis and cholesterol absorption). II. Reverse cholesterol transport
from peripheral tissues (activation of RXR/LXR heterodimer inhibited
cholesterol absorption and induced reverse cholesterol, transport in
The following nuclear receptors are known to form heterodimers with RXR
and hence, can be potentially activated by 9-cis retinoic acid
administration: thyroid hormone (TRa/b); vitamin, D (VDR); fatty acid/eicosanoids
(PPAR.alpha./.beta..gamma.); oxysterols (LXRa/b); bile acids (FXR);
pregnanes/bile acids/xenobiotics (PXR); androstanes/xenobiotics (CAR).
Fibrates (clofibrate, fenofibrate, bezafibrate, ciprofibrate, beclofibrate
and gemfibrozil) are currently recommended for the treatment of patients
with hypertriglyceridemia (high plasma TG). The treatment of
hypertriglycerdemia results in decreased plasma levels of
triglyceride-rich lipoproteins. HDL cholesterol levels are usually
increased when the baseline levels are low. The increase in HDL
cholesterol is usually concomitant with, increased levels of the HDL
apolipoprotein A-I and A-II. In addition, fibrates reduce the atherogenic
apoC-III-containing particles, effect post-prandial lipemia and some
fibrate lower plasma fibrinogen and CRP levels.
As a consequence of their effect on lipid metabolism, fibrates have been
shown to affect specific lipoprotein disorders such as in combined
hyperlipidemia, primary hypertriglyceredemia, type III
dysbetalipoprotenemia, and non-insulin dependent diabetes mellitus (NIDDM).
Fibrates are considered to be well tolerated, with an excellent safety
profile. A low toxicity has been, reported in almost every organ system.
Fenofibrate treatment revealed low frequency of side effects. Long-term
administration revealed no effect on peroxisome proliferation in human
liver and no evidence for carcinogenesis. Clinically relevant interaction
of fibrates with other anti-hyperlipedemic drugs include rhabdomyolysis
and decreased bioavailibility when combined with some bile acid
sequestrants. Potentiation of the anticoagulant effect of coumarin may
Fibrates mediate at least part of their effects by peroxisome proliferator-activated
receptor .alpha. (PPAR-.alpha.). Upon activation with fibrates, PPARs
heterodimerize with another nuclear receptor, the 9-cis retinoic acid
receptor (RXR). The dimer binds to specific response elements, termed
peroxisome proliferator response elements (PPREs) and regulates gene
Low plasma levels of high-density lipoprotein (HDL) and high triglyceride
(TG) plasma levels are risk factors for atherosclerosis. Low levels of
plasma HDL cholesterol, apolipoprotein AI (apoA-T) and high levels of
triglycerides (TG) are associated with increased risk for atherosclerosis,
the major cause of morbidity and mortality in Western societies. A recent
study showed that the rate of coronary events is reduced by 22 percent by
raising HDL cholesterol levels and lowering the plasma TG levels in
patients treated with the fibrate gemfibrozil.
Several modes of action were proposed for the fibrate beneficial effects
on atherosclerosis: Induction of lipoprotein lipolysis by reducing apoC-III
levels and/or by increasing lipoprotein lipase activity. Induction of
hepatic fatty acid (FA) uptake by the induction of FA-transporter protein
and acyl-CoA synthetase. Decrease apoB and VLDL production. Reduction of
hepatic TG production by induction of peroxisome (in rodents only) or
mitochondrial .beta.-oxidation pathway and inhibition of FA synthesis.
Most important, fibrates increase the production of apoA-I and apoA-II in
the liver, which probably contribute to the process of reverse cholesterol
Type 2 diabetes mellitus is a serious health problem. It arises when,
resistance to the glucose lowering effects of insulin combines with
impaired insulin secretion to raise the levels of glucose in the blood
beyond the normal range. Thiazolidinediones are a new class of
antidiabetic agents that improve insulin sensitivity and reduce plasma
glucose and blood pressure in subjects with type 2 diabetes.sup.(5). The
drugs bind and activate PPAR.gamma. that binds to DNA as heterodimers with
a common partner, retinoid X receptor (RXR) to regulate transcription. RXR
agonists have been shown to function as insulin sensitizers, markedly
decreasing serum glucose, triglycerides and insulin in obese mice.sup.(6).
WO 93/24454 describes a carotenoid composition derived from Dunaliella
algae in which the .beta.-carotene content is predominantly 9-cis
.beta.-carotene. There is no mention of any medical applications.
U.S. Pat. No. 5,219,888 (Katocs) discloses a method to increase plasma HDL
levels for the treatment and prevention of coronary artery disease by
administrating a therapeutic amount of the retinoids all trans-retinoic
acid and 9-cis-retinoic acid.
U.S. Pat. No. 5,948,823 discloses use of a substantially crude Dunaliellia
algae preparation in protecting mammals against the detrimental effects of
medical and nuclear irradiation.
WO 97/10819 teaches a new treatment for non-insulin dependent diabetes
mellitus by administration of RXR agonists and optionally, peroxisome
proliferation activated receptor gamma agonists.
WO 99/50658 identifies compounds which modulate nuclear receptor activity,
used for treating e.g. cancer, cardiovascular disease, osteoporosis,
diabetes, postmenopausal disorders and inflammatory conditions.
WO 2001119770 reveals new retinoid X analogs, useful as retinoid X
receptor modulators for lowering blood glucose levels, for modulating
lipid metabolism and in treatment of e.g. diabetes, obesity,
cardiovascular diseases and breast cancer.
U.S. Pat. Nos. 5,972,881 and 6,028,052 (Heyman) disclose methods and
compositions for the treatment of non-insulin dependent diabetes using an
RXR agonist together with a PPAR.gamma. agonist.
WO 03/027090 provides novel retinoid compounds that have selectivity as
RXR agonists, and are effective in reducing blood glucose and maintaining
body weight, thus being useful for the treatment of diabetes (NIDDIM) and
Colagiuri S. and Best, J. Lipid-lowering therapy in people with type 2
diabetes. Current Opinion in Lipidology (2002) 13:617-623, teaches that
lipid powering with statins and fibrates is effective in improving
cardiovascular disease outcome in diabetes.
Levy, Y. et al. Dietary supplementation of a natural isomer mixture of
Beta-carotene inhibits oxidation of LDL derived from patients with
diabetes mellitus. Nutrition & Metabolism (2000) 44:54-60, describes
experiments in which Dunaliella bardawil-derived .beta.-carotene was
supplemented to the diet of diabetic patients. The dietary supplementation
normalized the enhanced LDL susceptibility to oxidation in these patients.
SUMMARY OF THE
It is an object of the invention to
provide a method for treating patients suffering from diabetes, low plasma
HDL and/or high plasma TG and/or atherosclerosis.
The present invention provides by one of its aspects, a method for
treating a disease selected from diabetes mellitus and atherosclerosis
comprising administrating to a subject an effective amount of crude
The present invention also provides by another aspect a method for
reducing triglycerides and/or increasing HDL cholesterol levels in the
plasma of a subject comprising administrating to the subject an effective
amount of crude Dunaliella powder.
In accordance with another aspect of the invention there is provided a
medicament for use in treating diabetes mellitus, low plasma HDL and/or
high plasma TG and/or atherosclerosis, comprising an effective amount of a
substantially crude Dunaliella algae preparation.
In accordance with a further aspect of the invention, there is provided
the use of an effective amount of a substantially crude Dunaliella algae
preparation in, the preparation of a pharmaceutical composition for use in
treating diabetes mellitus, low plasma HDL and/or high plasma TG and/or
The active ingredient in accordance with the invention is a substantially
crude Dunaliella algae preparation, typically dried Dunaliella algae. The
Dunaliella algae are preferably Dunaliella bardawil.
In a preferred embodiment, the crude Dunaliella powder is administered
together with one or more activators of nuclear receptors. The activators
of nuclear receptors are preferably peroxisome proliferator-activated
receptor .alpha. and .gamma. (PPAR.alpha. and PPAR.gamma.) agonists, such
as fibrates and thiazolidinediones. Non-limiting examples of fibrates are
clofibrate, fenofibrate, bezafibrate, ciprofibrate, beclofibrate and
gemfibrozil. Non-limiting examples of thiazolidinediones are troglitazone,
BRL 49653, pioglitazone, ciglitazone, WAY120,744, englitazone, AD 5075,
darglitazone and rosiglitazone.
The crude Dunaliella powder is preferably administered orally, for example
in an encapsulated form.
Claim 1 of 12 Claims
1. A method for reducing
insulin and/or glucose plasma levels in a subject afflicted with diabetes,
comprising administering to the subject an effective amount of dried
Dunaliella algae, thereby reducing plasma insulin and/or glucose plasma
levels in the subject.
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