rexresearch.com
Christopher Rinsch,
et al.
Pomegranate Anti-Aging ( Urolithin-A )
Youtube.com : Pomegranates
reveal its powerful anti-aging secret
Ecole Polytechnique Fédérale de Lausanne
: Pomegranate Reveals its Powerful Anti-aging
Secret
US8933217 : COMPOUNDS, COMPOSITIONS AND METHODS
FOR PROTECTING BRAIN HEALTH IN NEURODEGENERATIVE DISORDERS
US2012164243 : Compositions and methods for
improving mitochondrial function and treating
neurodegenerative diseases and cognitive disorders
US2016000753 : Enhancing Autophagy or
Increasing Longevity by Administration of Urolithins or
Precursors Thereof
US2015183758 : Process-Scale Synthesis of
Urolithins
WO2015097231 : PRODRUGS OF UROLITIHNS AND USES
THEREOF
Pomegranate Patents : Extraction &
Cultivation
Pomegranate Cultivation
https://www.youtube.com/watch?v=Lf1vCyfaosE
Pomegranates reveal its powerful
anti-aging secret
École polytechnique fédérale de Lausanne (EPFL)
Scientists have managed to prove pomegranates anti-aging
potential: intestinal bacteria transform a molecule contained in
the fruit with spectacular results. Although tests in humans are
still underway, scientists have already published the initial
promising results from animal studies in the journal Nature
Medicine.
Nature Medicine, July 2016
'Urolithin A induces mitophagy and prolongs lifespan in C. elegans
and increases muscle function in rodents'
Dongryeol Ryu, Laurent Mouchiroud, Pénélope A Andreux, Elena
Katsyuba, Norman Moullan, Amandine A Nicolet-dit-Félix, Evan G
Williams, Pooja Jha, Giuseppe Lo Sasso, Damien Huzard, Patrick
Aebischer, Carmen Sandi, Chris Rinsch & Johan Auwerx
http://www.biosciencetechnology.com/news/2016/07/pomegranate-reveals-its-powerful-anti-aging-secret
http://actu.epfl.ch/news/pomegranate-finally-reveals-its-powerful-anti-agin/
Press kit (photos, video, article, TV B-roll with 3D
animation): http://bit.ly/pomegranate2016
Pomegranate Reveals its Powerful Anti-aging Secret
by
Ecole Polytechnique Fédérale de Lausanne
Are pomegranates really the superfood we've been led to believe
will counteract the aging process? Up to now, scientific proof has
been fairly weak. And some controversial marketing tactics have
led to skepticism as well. A team of scientists from EPFL and the
company Amazentis wanted to explore the issue by taking a closer
look at the secrets of this plump pink fruit. They discovered that
a molecule in pomegranates, transformed by microbes in the gut,
enables muscle cells to protect themselves against one of the
major causes of aging. In nematodes and rodents, the effect is
nothing short of amazing. Human clinical trials are currently
underway, but these initial findings have already been published
in the journal Nature Medicine.
As we age, our cells increasingly struggle to recycle their
powerhouses. Called mitochondria, these inner compartments are no
longer able to carry out their vital function, thus accumulate in
the cell. This degradation affects the health of many tissues,
including muscles, which gradually weaken over the years. A
buildup of dysfunctional mitochondria is also suspected of playing
a role in other diseases of aging, such as Parkinson's disease.
One molecule plays David against the Goliath of aging
The scientists identified a molecule that, all by itself, managed
to re-establish the cell's ability to recycle the components of
the defective mitochondria: urolithin A. "It's the only known
molecule that can relaunch the mitochondrial clean-up process,
otherwise known as mitophagy," says Patrick Aebischer, co-author
on the study. "It's a completely natural substance, and its effect
is powerful and measurable."
The team started out by testing their hypothesis on the usual
suspect: the nematode C. elegans. It's a favorite test subject
among aging experts, because after just 8-10 days it's already
considered elderly. The lifespan of worms exposed to urolithin A
increased by more than 45% compared with the control group.
These initial encouraging results led the team to test the
molecule on animals that have more in common with humans. In the
rodent studies, like with C. elegans, a significant reduction in
the number of mitochondria was observed, indicating that a robust
cellular recycling process was taking place. Older mice, around
two years of age, showed 42% better endurance while running than
equally old mice in the control group.
Human testing underway
Before heading out to stock up on pomegranates, however, it's
worth noting that the fruit doesn't itself contain the miracle
molecule, but rather its precursor. That molecule is converted
into urolithin A by the microbes that inhabit the intestine.
Because of this, the amount of urolithin A produced can vary
widely, depending on the species of animal and the flora present
in the gut microbiome. Some individuals don't produce any at all.
If you're one of the unlucky ones, it's possible that pomegranate
juice won't do you any good.
For those without the right microbes in their guts, however, the
scientists are already working on a solution. The study's
co-authors founded a start-up company, Amazentis, which has
developed a method to deliver finely calibrated doses of urolithin
A. The company is currently conducting first clinical trials
testing the molecule in humans in European hospitals.
Darwin at your service: parallel evolution makes good dinner
partners According to study co-author Johan Auwerx, it would be
surprising if urolithin A weren't effective in humans. "Species
that are evolutionarily quite distant, such as C elegans and the
rat, react to the same substance in the same way. That's a good
indication that we're touching here on an essential mechanism in
living organisms."
Urolithin A's function is the product of tens of millions of years
of parallel evolution between plants, bacteria and animals.
According to Chris Rinsch, co-author and CEO of Amazentis, this
evolutionary process explains the molecule's effectiveness:
"Precursors to urolithin A are found not only in pomegranates, but
also in smaller amounts in many nuts and berries. Yet for it to be
produced in our intestines, the bacteria must be able to break
down what we're eating. When, via digestion, a substance is
produced that is of benefit to us, natural selection favors both
the bacteria involved and their host. Our objective is to follow
strict clinical validations, so that everyone can benefit from the
result of these millions of years of evolution."
The EPFL scientists' approach provides a whole new palette of
opportunities to fight the muscular degeneration that takes place
as we age, and possibly also to counteract other effects of aging.
By helping the body to renew itself, urolithin A could well
succeed where so many pharmaceutical products, most of which have
tried to increase muscle mass, have failed. Auwerx, who has also
published a recent discovery about the anti-aging effects of
another molecule in the journal Science, emphasizes the
game-changing importance of these studies. "The nutritional
approach opens up territory that traditional pharma has never
explored. It's a true shift in the scientific paradigm."
US8933217
COMPOUNDS, COMPOSITIONS AND METHODS FOR PROTECTING BRAIN
HEALTH IN NEURODEGENERATIVE DISORDERS
Aspects of the invention relate to compounds, extracts and
compositions thereof, and methods of using of the same, to treat
neurodegenerative disorders and/or improve brain health. In
certain embodiments, said compounds are pomegranate flavonoids.
US2012164243
Compositions and methods for improving mitochondrial
function and treating neurodegenerative diseases and
cognitive disorders
Provided are compositions comprising compounds or precursors to
compounds which may be used for a variety of therapeutic
applications including, for example, treating and/or preventing a
disease or disorder related to reduced or inadequate mitochondrial
activity, including aging or stress, diabetes, obesity, and
neurodegenerative diseases. The compounds relate generally to
urolithins and precursors thereof, including but not limited to
ellagitannins and urolithin A. In certain embodiments the
compositions are presented in or as food products or nutritional
supplements. These same compounds and compositions can also be
used advantageously in generally healthy individuals to increase
or maintain metabolic rate, decrease percent body fat, increase or
maintain muscle mass, manage body weight, improve or maintain
mental performance (including memory), improve or maintain muscle
performance, improve or maintain mood, and manage stress.
BACKGROUND OF THE INVENTION
[0002] Ellagitannins are monomeric, oligomeric, and polymeric
polyphenols that are abundant in some fruits, berries and nuts,
such as pomegranates, raspberries, strawberries, black
raspberries, walnuts and almonds. The fruits and berries are
widely consumed fresh and as beverages, such as juice, and these
have been reported to promote health.
[0003] In commercial fruit juice processing methods,
ellagitannins, which are particularly abundant in some fruit
peels, are extracted in large quantities into the juice.
Ellagitannins belong to the chemical class of hydrolyzable
tannins, which release ellagic acid upon hydrolysis. In vitro
studies have suggested that ellagitannins, at concentrations in
the range of 10-100 micromolar (μM), have potential anti-oxidant,
anti-atherogenic, anti-thrombotic, anti-inflammatory, and
anti-angiogenic effects. Fruits may have different ellagitannins
that are predominant, for example, in fruit juice prepared from
pomegranate, the predominant ellagitannin is punicalagin [2,3
hexahydroxydiphenoyl-4,6-gallagylglucose], which occurs as a
mixture of isomers. The reported potent anti-oxidant properties of
pomegranate juice have been attributed to the high content of
punicalagin isomers, which can reach levels >2 g/L of juice.
Ellagitannins have also been identified as the active
anti-atherogenic compounds in pomegranate juice. It has also been
suggested that pomegranate ellagitannins and pomegranate fruit
extracts inhibit the proliferation of human cancer cells and
modulate inflammatory sub-cellular signaling pathways and
apoptosis. See, for example, Seeram et al. (2005) J Nutr Biochem.
16:360-7; Adams et al. (2006) J Agric Food Chem. 54:980-85; Afaq
et al. (2005) Photochem Photobiol. 81:38-45; Afaq et al. (2005)
Int J Cancer. 113:423-33. Pomegranate fruit extract has also been
reported to reduce prostate tumor growth and prostate serum
antigen (PSA) levels in athymic nude mice implanted with CWR22Rv1
prostate cells. Malik et al. (2005) Proc Natl Acad Sci.
102:14813-8.
[0004] Unfortunately, for the most part ellagitannins are poorly
absorbed by the human gut. However, a number of metabolites
derived from ellagitannins are absorbed by the human gut,
including certain metabolites ultimately formed in the gut by
commensal microorganisms (i.e., intestinal microflora).
[0005] Ellagitannins release ellagic acid under physiological
conditions in vivo, and ellagic acid is then gradually metabolized
by the gut microflora in the intestine to produce urolithin D,
urolithin C, urolithin A (UA) and urolithin B (UB). Once the
metabolites are absorbed, they undergo glucuronidation and once in
the liver, they are further metabolized to produce glucuronides,
and/or sulfates, to give a combination of metabolites secreted in
the bile.
[0006] Urolithins are metabolites of ellagic acid, punicalagin
(PA), punicalin (PB), tellimagrandin (TL), and other ellagitannins
(Cerda, Espin et al. 2004; Cerda, Periago et al. 2005). Ellagic
acid (EA) is abundant in pomegranate juice (Gil, Tomas-Barberan et
al. 2000). The ellagitannin tellimagrandin (TL) has been
previously isolated and characterized before from pomegranate and
other plants (Tanaka, Nonaka et al. 1986; Tanaka, Nonaka et al.
1986; Satomi, Umemura et al. 1993). Structural formulas for UA,
PA, PB, EA, and TL are presented in FIG. 1.
[0007] Considerable efforts have been made to understand the
mechanism of metabolic disorders, neurodegeneration and cognitive
decline, so as to better design treatment modalities including
those based on natural products. One of the key observations has
been therole of declining mitochondrial energy production,
corresponding with increased oxidative stress and apoptosis, plays
a significant role in degenerative diseases and the process of
aging. A variety of degenerative diseases have now been shown to
be caused by mutations in mitochondrial genes encoded by the
mitochondrial DNA (mtDNA) or the nuclear DNA (nDNA). Importantly,
somatic mtDNA mutations accumulate with age in post-mitotic
tissues in association with the age-related decline in
mitochondrial function and are thought to be an important factor
in aging and senescence. Inherited diseases can result from mtDNA
base substitution and rearrangement mutations and can affect the
CNS, heart and skeletal muscle, and renal, endocrine and
hematological systems.
[0008] Mitochondria generate most of the cellular energy by
oxidative phosphorylation (OXPHOS), and they produce most of the
toxic reactive oxygen species (ROS) as a by-product. Genetic
defects that inhibit OXPHOS also cause the redirection of OXPHOS
electrons into ROS production, thus increasing oxidative stress. A
decline in mitochondrial energy production and an increase in
oxidative stress can impinge on the mitochondrial permeability
transition pore (mtPTP) to initiate programmed cell death
(apoptosis). The interaction of these three factors is believed to
play a major role in the pathophysiology of degenerative diseases
and the aging process, which affects all tissues of the body.
[0009] In the normal brain, optimal cognitive function mainly
relies on the activity and communication between neurons, highly
complex cells able to convey electric signals and elicit chemical
neurotransmission. Neuronal function depends on long and complex
cellular processes that can extend over centimeters or even meters
to connect neurons or target cells, and can make more than 100,000
synaptic contacts. As such, neurons are highly dependent on energy
supply and, therefore, are exposed to oxidative stress damage.
Cognitive function is dependent on a careful balance of
intracellular signaling that takes place within a complex network
of neurons. Optimal cognitive function can be impaired by numerous
factors such as aging, cellular stress, chronic stress, and
neurodegenerative disorders. Cognitive decline may be
characterized by a decrease in performance in thinking, learning,
memory, alertness, and/or impaired psychological skills, as well
as by depression and anxiety.
[0010] Mitochondrial function has also been shown to be important
in metabolic disorders. Diabetes and obesity have been correlated
with compromises in mitochondrial function. It has been suggested
that the coupling efficiency in mitochondria, or the proportion of
oxygen consumption necessary to make ATP, is related to levels of
obesity, with high coupling efficiency possibly resulting in
higher deposition of fat stores (Harper, Green et al. 2008). In
diabetes, recent work has suggested that mitochondrial dysfunction
is a cause of insulin insensitivity in myocytes and adipocytes, as
a result of insufficient energy supply or defects in the insulin
signaling pathway (Wang, Wang et al. 2010).
SUMMARY OF THE INVENTION
[0011] The invention relates to compositions comprising compounds
or precursors to compounds which may be used for a variety of
therapeutic applications including, for example, treating and/or
preventing disease or disorders related to reduced or inadequate
mitochondrial activity, including aging or stress, diabetes,
obesity, and neurodegenerative diseases. These same compounds and
compositions can also be used advantageously in generally healthy
individuals to increase or maintain metabolic rate, decrease
percent body fat, increase or maintain muscle mass, manage body
weight, improve or maintain mental performance (including memory),
improve or maintain muscle performance, improve or maintain mood,
and manage stress….
[0140] Remarkably, the inventors have discovered that certain
compounds derived from ellagitannins are useful in the treatment
and prevention of physiological and psychological manifestations
of stress, including oxidative stress. Without meaning to be tied
to any particular mechanism of action, it is believed that the
compounds exert beneficial effects on mitochondria, promoting and
restoring crucial mitochondrial functions and counteracting
stress-induced mitochondrial dysfunction. These same compounds
have been discovered, in accordance with the instant invention, to
be useful in the treatment and prevention of any of a variety of
conditions, diseases, and disorders related to mitochondrial
dysfunction including, without limitation, neurodegenerative
diseases and cognitive disorders, metabolic disorders including
insulin resistance, mood disorders, and anxiety disorders.
[0141] Ellagitannins (ETs) are polyphenols included within the so
called “hydrolyzable tannins” in which hexahydroxydiphenic acid
forms diesters with sugars (most often β-D-glucose). ETs can occur
as complex polymers reaching molecular weights up to 4000 and
higher. These polymers can be hydrolyzed with acids or bases to
yield ellagic acid (EA), which can be used indirectly to quantify
ETs. EA in turn is a source of additional metabolic products
including urolithins.
[0142] Many plant species containing ellagitannins have been used
for the treatment of diseases, particularly in Asia (Okuda et al.,
2009). These include Agrimonia pilosa (agrimoniin), Camelia
japonica (camelliatannin A), Cornus officinalis (cornussin A),
Geranium thunbergii (geraniin), Geum japonicum (gemin-A),
Liquidambar formosana (casuarictin), Mallotus japonicus
(mallotusinic acid), Oenothera erythrosepala (oenothein B), Punica
granatum (pomegranate) (granatin B), Rosa rugosa (rugosin), and
Terminalia chebula (chebulinic acid), among others. The main uses
of these medicinal plants have been associated to their
antioxidant, anti-diarrheic, anti-microbial, and immunomodulatory
activities.
[0143] Ellagitannins are also present in significant amounts in
many berries, including strawberries, red and black raspberries
(Zafrilla et al., 2001), blueberries, and blackberries.
Ellagitannins have also been found in apples, cherries,
cloudberries, cranberries, currants, grapes, lime, mango,
pineapple, pomegranate, prune, rhubarbs. Serrano et al. (2009) Mol
Nutr Food Res. 53:S310-29. The ellagitannin rubusuaviin C can be
isolated from the leaves of the Chinese sweet tea Rubus
suavissimus S. Lee. Ellagitannins have also been identified in
appreciable amounts in nuts, including walnuts (Fukuda et al.,
2003), pistachios, cashew nuts, chestnuts, oak acorns (Cantos et
al., 2003) pecans (Villarreal-Lozoya et al., 2007) and peanuts.
[0144] They are also abundant in pomegranates (Gil et al., 2000),
and muscadine grapes (Lee and Talcott, 2002) and are important
constituents of wood, particularly oak (Glabasnia and Hofmann,
2006). Ellagitannins can be incorporated into food products, such
as wines, and whiskey, through migration from wood to the food
matrix during different aging processes. Ellagic acid has also
been found in several types of honey and it has been proposed as a
floral marker for heather honey (Ferreres et al., 1996). Free
ellagic acid and different glycosidic derivatives are also present
in these food products, including glucosides, rhamnosides,
arabinosides and the corresponding acetyl esters (Zafrilla et al.,
2001).
[0145] A number of studies have shown that the ellagitannin
content of several food products can be quite high (Table 1). For
example, a glass of pomegranate juice (200 mL) can provide as much
as 1 g of ellagitannins and ellagic acid together, a raspberry
serving (100 g raspberries) around 300 mg, a strawberry serving 70
mg, and four walnuts some 400 mg of ellagitannins.
[0146] Representative dietary ellagitannins include punicalagin of
pomegranate, sanguiin-H-6 of strawberry and raspberry, and
pedunculagin of walnuts. All of these release ellagic acid upon
hydrolysis, although other metabolites can also be produced and
are distinctive of individual ellagitannins (e.g., gallagic and
ter-gallagic acids)…
Example 1
Preparation of Functional Extracts from Pomegranate
Compounds
[0336] The pomegranate extracts described in this application
containing specific molecules were prepared using an extraction
procedure based on adsorption of polyphenols in a standard polymer
adsorption-based column as described. For the preparation of the
extracts 31008 and 1108 derived from pomegranate juice,
pomegranates were juiced using a standard juicing and
manufacturing process and adsorbed onto a polymeric
chromatographic resin as pure juice. The resin Amberlite XAD-16
(Rohm & Haas) was packed into semi-preparative columns and
loaded with the extracted juice. The column was washed with water
to remove the sugars until completion (Brix levels were below
0.1%). The polyphenols were eluted with 100% ethanol. The
remaining ethanol was evaporated under vacuum to produce a
concentrated extract containing 4.5 g of total polyphenol per
liter as determined using the Folin assay for total polyphenol
content. Extract 1011 was prepared in a similar manner as extract
31008 and 1108, but the liquid extract was then spray dried
utilizing a spray dryer to produce a final powder extract.
Utilizing HPLC-MS for the identification of compounds, extract
31008, 1108, and 1011 were found to contain the molecules
punicalagin, punicalin, tellimagrandin, and pedunculagin.
[0337] The extract 71109 derived from the pomegranate husk was
prepared by manually separating the husk from the pomegranate
arils pulp, followed by pressing with a manual fruit press. To
extract the maximal amount of polyphenols, the cake/pomace of
pressed pomegranate parts were soaked in water consecutively for
several periods of time (5 minutes) in order to increase
extraction efficiency. The extracted pomegranate solution was
clarified by centrifugation before being adsorbed onto the
polymeric chromatographic Amberlite XAD-16 resin (Rohm &
Haas), packed in semi-preparative columns, and loaded with the
water extracted from pomegranate husk. The column was washed with
water to remove the sugars until completion (Brix levels were
below 0.1%). The polyphenols were eluted with 100% ethanol. The
remaining ethanol was evaporated under vacuum to produce a
concentrated extract containing 17.1 g of total polyphenol per
liter as determined using the Folin assay for total polyphenol
content. This technique is a modification of methods known in the
art as described by several published methods for purification of
polyphenols from various plants and berries. Tuck, K. L. and P. J.
Hayball (2002) “Major phenolic compounds in olive oil: metabolism
and health effects.” J Nutr Biochem 13(11):636-644; and Schieber,
A., P. Hilt, et al. (2003) “A new process for the combined
recovery of pectin and phenolic compounds from apple pomace.”
Innovative Food Sci. Emerging Technol. 4:99-107.
[0338] For the preparation of Extract 61109, an aqueous extract of
the pomegranate was fractionated utilizing centrifugal partition
chromatography. The isolation fractions were lyophylized to
produce extract 61109, highly enriched in punicalagin (>90%).
Purification of Punicalagin
Preparation of Extract
[0339] Extract from pomegranate was dissolved in 16 mL of the
organic/aqueous phase mixture (1:1) and filtered on a Teflon
filter (0.45 μm).
[0000] Separation of Punicalagin from Extract Using Centrifugal
Partition Chromatography
[0340] Separation of punicalagin from pomegranate extract was
achieved by utilizing Centrifugal Partition Chromatography CPC.
The CPC apparatus was a FCPC® 1000 apparatus provided by Kromaton
Technologies (Angers, France) that is fitted with a rotor of 1000
mL capacity. The solvents were pumped by a 4-way binary
high-pressure gradient pump. The samples were introduced into the
CPC column via a high pressure injection valve (Rheodyne) equipped
with a 20 mL sample loop. The effluent was monitored with a diode
array detection (DAD) detector equipped with a preparative flow
cell. Fractions were collected by a fraction collector. The
separation steps were conducted at room temperature.
[0341] To accomplish the extraction, the stationary phase was
first introduced into the column in the ascending mode without
rotating, and mobile phase was then pumped through the stationary
phase until an equilibrium stage was reached. Then, the rotation
speed was increased from 0 to 1000 rpm and the mobile phase was
pumped into the column at a flow-rate of 20 mL/min. After
injection of 10 g of pomegranate extract, fractions of 20 mL were
collected every minute. The content of the outgoing organic phase
was monitored by online UV absorbance measurement at λ=260 nm.
[0342] An elution-extrusion procedure was used to recover all the
compounds from the column: after a classical elution of 100 min,
the mobile phase was replaced by the stationary phase used as
mobile liquid, until all volume contained (1000 mL) was pushed out
the column. A fraction containing punicalagins (mixture of A and B
isomers) with 94-97% chromatographic purity was obtained between
51 and 63 minutes of elution, and a second fraction with a
chromatographic purity of 85-88% was obtained between 64 and 79
min.
Enhancing Autophagy or Increasing
Longevity by Administration of Urolithins or Precursors
Thereof
US2016000753
Disclosed are methods, compounds, and compositions useful for
increasing autophagy and promoting longevity. The methods,
compounds, and compositions relate to urolithins and urolithin
precursors and use thereof. Certain urolithins are represented by
Formula I, while certain urolithin precursors are represented by
Formula IV. The urolithin may be urolithin A, urolithin B,
urolithin C, or urolithin D. The urolithin precursor may be
ellagic acid or an ellagitannin. The methods include in vivo, ex
vivo, and in vitro uses of the compounds and compositions.
BACKGROUND
[0002] Autophagy is a lysosomal degradation pathway in both
animals and plants that is essential for development,
differentiation, homeostasis, and survival. In animals, autophagy
serves principally as an adaptive mechanism to protect organisms
against diverse pathologies, including infection, cancer,
neurodegeneration, heart disease, and aging. The repertoire of
routine housekeeping functions performed by autophagy includes
elimination of defective proteins and organelles, prevention of
the accumulation of abnormal protein aggregates, and elimination
of intracellular pathogens. The autophagy pathway is uniquely
capable of degrading entire organelles, such as mitochondria,
peroxisomes, and endoplasmic reticulum.
[0003] Multiple reports indicate that proteins required for
autophagy induction, such as sirtuin 1, have reduced expression in
aged tissues; levels of autophagy have been shown to diminish with
age. Reduced levels of autophagy have also been associated with
obesity, diabetes, cancer, neurodegenerative diseases,
cardiovascular disease, osteoarthritis, and age-related macular
degeneration.
[0004] A number of compounds that stimulate autophagy have been
identified, including rapamycin, resveratrol, metformin,
spermidine, and glucosamine.
[0005] Urolithins are ellagitannin- and ellagic acid-derived
metabolites produced, e.g., by mammalian colonic microflora,
including human colonic microflora. Urolithins are known to
exhibit anti-oxidant activity.
SUMMARY OF THE INVENTION
[0006] An aspect of the invention is a method of increasing
autophagy in an animal, comprising the step of administering to an
animal in need thereof an effective amount of a urolithin or a
precursor thereof, thereby increasing autophagy in the animal.
DETAILED DESCRIPTION OF THE INVENTION
Overview
[0193] Autophagy is a process by which cells degrade their own
components, recycling amino acids and other building blocks that
can be reused. Such degradation is performed by lysosomal acidic
hydrolases. It is a tightly regulated process that plays an
important role in normal cell growth, development, and
homeostasis, helping to maintain a balance between the synthesis,
degradation, and subsequent recycling of cellular products. It is
a major mechanism by which starving cells can reallocate nutrients
from less-essential processes to more essential processes.
[0194] During nutrient starvation, increased levels of autophagy
lead to the breakdown of non-vital components and the release of
nutrients, ensuring that vital processes can continue. Mutant
yeast cells that have a reduced autophagic capability rapidly
perish in nutrient-deficient conditions. A gene known as Atg7 has
been implicated in nutrient-mediated autophagy, and studies in
mice have shown that starvation-induced autophagy was impaired in
Atg7-deficient mice. Komatsu M et al. (2005) J Cell Biol.
169:425-434.
[0195] Autophagy degrades damaged organelles, cell membranes, and
proteins. The failure of autophagy is thought to be an important
factor in the accumulation of cell damage and, therefore, aging.
[0196] Three types of autophagy can be distinguished, depending on
the pathway along which cellular components are delivered to
lysosomes: macroautophagy, microautophagy, and chaperone-mediated
autophagy (CMA).
Macroautophagy
[0197] Macroautophagy involves the degradation of long-lived
proteins and whole cellular organelles through a multistep process
(FIG. 1). Macroautophagy begins with the formation of a
double-layered isolation membrane (phagophore) around the
molecules and/or organelles to be degraded. The phagophore engulfs
cytosolic components and seals around the content, forming an
autophagosome. Eventually, the autophagosome fuses with a
lysosome, evolving into an autophagolysosome (or autolysosome),
wherein lysosomal hydrolases digest the cargo. Microautophagy
involves the direct sequestration of cytosolic components through
invaginations or armlike projections of the lysosomal membrane.
Microautophagy may serve for the turnover of long-lived proteins;
however, the significance and regulation of this type of autophagy
remain poorly understood. Finally, chaperone-mediated autophagy is
a highly selective process devoted to the degradation of soluble
cytosolic proteins.
[0198] The microtubule-associated protein 1A/1B-light chain 3
(LC3), a mammalian homolog of the yeast Atg8, is a soluble protein
with a molecular mass of approximately 17 kDa which is distributed
ubiquitously in mammalian tissues and cultured cells. It is
processed immediately after its synthesis by Atg4B, a cysteine
protease, that exposes the C-terminal glycine residue (LC3-I).
During autophagy, autophagosomes engulf cytoplasmic components,
including cytosolic proteins and organelles. Concomitantly, a
cytosolic form of LC3 (LC3-I) is conjugated to
phosphatidylethanolamine (PE) to form and LC3-PE conjugate
(LC3-II), which is recruited to autophagosomal membranes (FIG. 1).
[0199] p62, also known as sequestosome-1, was identified as a
novel partner of the atypical protein kinase Cs (aPKCs) and is a
ubiquitiously expressed cellular protein. p62 is known to have
domains that interact with and bind to ubiquitinated proteins, and
it has been identified as a component of inclusion bodies observed
in human diseases, especially neurodegenerative diseases (e.g.,
Alzheimer's disease, Parkinson's disease, amyotrophic lateral
sclerosis) as well as in liver diseases. p62 also has been
identified as an LC3 interacting protein and it has been
demonstrated that an 11 amino acid sequence in the mouse p62
serves to recognize the LC3 protein. As seen in FIG. 1, LC3 binds
to p62 and transports it (and any ubiquitinated proteins or cell
components bound to it) into the autophagosome, where it is
degraded. Consequently, one of the hallmarks of autophagy is an
increase in the ratio of LC3-II/LC3-I with a concomitant decrease
in the level of cellular p62.
[0200] Of the three types of autophagy described, macroautophagy
is the best characterized in mammalian cells. Starvation is the
strongest stimulus of macroautophagy. During nutrient deprivation,
macroautophagy breaks down cellular components, generating amino
acids, fatty acids, and carbohydrates, which can be harnessed for
energy production and for the synthesis of essential cellular
molecules. Macroautophagy is also involved in specific cytosolic
rearrangements during embryogenesis and postnatal development.
Furthermore, macroautophagy is induced during viral or bacterial
infections, in hypoxia, and under various stress conditions,
including radiation exposure and increased reactive oxygen species
(ROS) generation. In these circumstances, macroautophagy is
essential for the maintenance of cell homeostasis by its promotion
of the removal of damaged components. Indeed, impairments in
macroautophagy induce premature aging and shorten the lifespan in
several organisms, including C. elegans, yeast, and Drosophila.
Hars E S et al. (2007) Autophagy 3:93-95; Matecic M et al. (2010)
PLoS Genet. 6:e1000921; Lee J H et al. (2010) Science
327:1223-1228. Conversely, upregulation of macroautophagy is
proposed to be a major mechanism underlying the lifespan-extending
properties of calorie restriction. Toth M L et al. (2008)
Autophagy 4:330-338; Morselli E et al. (2010) Cell Death Dis.
1:e10.
[0201] More than 35 Atg (AuTophaGy-related) proteins have been
identified in yeasts and mammals; however, the precise role each
Atg protein plays during autophagy is not yet fully established.
As illustrated in FIG. 1, the process of macroautophagy can be
divided into discrete steps, namely, induction and nucleation,
expansion, fusion, and degradation. The induction phase is
mediated by the ULK1-Atg13-FIP200 kinase complex. The regulation
of the nucleation stage, which consists of the recruitment of Atg
proteins to the phagophore assembly site, is not yet completely
understood. However, the vacuolar protein sorting-34 (Vps34), a
class III phosphatidylinositol-3-kinase (PI3K), is required for
this step. Vps34 associates with Beclin1, the mammalian homologue
of yeast Atg6, and subsequently recruits Atg14 and Vps15 (p150) to
the preautophagosomal structure. The elongation and expansion of
the phagophore membrane require two ubiquitin-like conjugation
systems involving Atg12 (conjugated to Atg5) and
Atg8/microtubule-associated protein 1 light chain-3 (LC3,
conjugated to phosphatidyl ethanolamine), along with other Atg
proteins such as Atg9 and Atg16. The fusion of the autophagosome
with a lysosome relies on canonical cellular fusion machinery that
consists of the Rab-SNARE (Soluble N-ethylmaleimide-sensitive
factor Attachment protein REceptor) system and requires the
presence of lysosomal membrane-associated protein-2 (LAMP-2) and
the UV radiation resistance-associated gene (UVRAG). Finally, the
digestion of the cargo is accomplished by lysosomal hydrolases,
followed by the transportation of degraded components into the
cytoplasm by lysosomal efflux transporters such as Atg22.
[0202] With regard to the regulation of macroautophagy, mTOR, the
mammalian target of rapamycin, is considered to be a major
checkpoint, linking the cellular nutritional state with the level
of ongoing autophagy. Under nutrient-rich conditions, mTOR is
active and inhibits the ULK1-Atg13-FIP200 complex required for the
induction of macroautophagy. Energy deprivation leads to mTOR
inactivation and stimulation of AMP-activated protein kinase
(AMPK), which both induce macroautophagy. AMPK functions as an
energy-sensing kinase and is activated by increases in the
cellular AMP to ATP ratio. Under such circumstances, AMPK promotes
autophagy by directly activating ULK1 and by relieving the
mTOR-mediated inhibition of macroautophagy.
[0203] Macroautophagy can be selectively directed toward the
removal of particular targets, e.g., peroxisomes (pexophagy),
endoplasmic reticulum (reticulophagy), intracellular lipids
(lipophagy), ribosomes (ribophagy), and intracellular pathogens
(xenopathy). Likewise, mitochondria can be selectively targeted
for degradation via macroautophagy (mitophagy).
Mitophagy: A Specialized Form of Macroautophagy
[0204] Mitophagy is a highly selective process that can promote
the elimination of dysfunctional or unnecessary mitochondria. Wang
K et al. (2011) Autophagy 7:297-300. The loss of mitochondrial
membrane potential (Δψm) represents a major trigger of mitophagy.
Indeed, laser-induced photo damage of selected mitochondria inside
living hepatocytes results in the rapid dissipation of Δψm,
followed by the quick removal of depolarized mitochondria through
mitophagy. In addition, oxidative damage can lead to the formation
of asymmetrical daughter mitochondria characterized by different
Δψm, with autophagy specifically targeting mitochondria with lower
Δψm. Apart from the degradation of damaged mitochondria under
stress conditions, mitophagy is essential for mitochondrial
turnover in the basal state and during cell differentiation, such
as the maturation of reticulocytes into mature red blood cells.
[0205] Investigations into the molecular regulation of mitophagy
have unveiled several mitophagy-specific proteins. Parkin and
Pink1 are believed to play important roles in the selective
degradation of damaged mitochondria, at least under certain
circumstances. Parkin is a cytosolic E3-ubiquitin ligase that is
selectively recruited to dysfunctional mitochondria and assists in
their removal by mitophagy. Narenda D (2008) J Cell Biol.
183:795-803. Pink1 is imported into healthy mitochondria through a
Δψm-dependent process and is degraded by the presenilin-associated
rhomboidlike (PARL) protease. Matsuda N et al. (2010) J Cell Biol.
189:211-221. The dissipation of Δψm results in the accumulation of
Pink1 on the mitochondrial surface, leading to the recruitment of
Parkin, which ubiquitinates outer membrane proteins, including the
voltage-dependent anion channel (VDAC). It is proposed that
ubiquitin-tagged mitochondria are targeted directly to autophagic
vacuoles through the interaction of ubiquitinated proteins with
the autophagosomal marker LC3 (Atg8). In addition, Parkin can
ubiquitinate the inner mitochondrial membrane and apoptosis
regulator protein B-cell lymphoma-2 (Bcl-2), thereby de-repressing
Beclin1.
[0206] Recent evidence also suggests that the opening of the
mitochondrial permeability transition pore (mPTP) may be required
for the selective removal of damaged mitochondria. Opening of the
mPTP causes a sudden increase of the inner membrane permeability
to solutes with molecular weight up to 1500 Da. This results in
mitochondrial depolarization, activation of the mitochondrial
ATPase (i.e., ATP synthase operating in reverse), and swelling and
rupture of the outer membrane. The loss of Δψm subsequent to
permeability transition targets individual mitochondria for
degradation. The loss of Δψm and the activation of macroautophagy
are prevented by cyclosporin A, an inhibitor of the mPTP component
cyclophilin D. Furthermore, starvation fails to induce
macroautophagy in cyclophilin D-deficient murine cardiomyocytes,
whereas autophagy is enhanced even under fed conditions in cardiac
cells from mice overexpressing cyclophilin D. The nicotinamide
adenine dinucleotide (NAD)-dependent deacetylase sirtuin-3 (SIRT3)
appears to be critically involved in the control of mPTP by
modulation of cyclophilin D.
[0207] Similar to the mPTP, the apoptotic proteins Bnip3 (Bcl-2
and adenovirus E1B 19-kDa-interacting protein-3) and Nix
(Nip3-like protein X) are thought to trigger selective mitophagy
through mitochondrial depolarization. Moreover, Bnip3 may induce
mitophagy by competitively disrupting the inhibitory interaction
between Bcl-2 and Beclin1. Finally, Nix associates with
mitochondrial membranes and directly interacts with LC3 (Atg8).
[0208] Although the molecular regulation of mitophagy has not yet
been completely elucidated, the mTOR/AMPK pathway is proposed to
be a major checkpoint. AMPK, in addition to stimulating
mitochondrial removal through autophagy, enhances the activity of
sirtuin-1 (SIRT1) and its downstream target PGC-1α, resulting in
stimulation of mitochondrial biogenesis. Hence, through the
activity of AMPK, mitophagy and mitochondrial biogenesis are
coordinately regulated, maintaining a healthy and functional pool
of mitochondria in the cell.
[0209] Lipophagy is a recently recognized alternative pathway of
lipid metabolism in which intracellular lipid droplet
triglycerides and cholesterol are taken up by autophagosomes and
delivered to lysosomes for degradation by acidic hydrolases,
thereby releasing free fatty acids. Lipophagy, therefore,
functions to regulate intracellular lipid stores, cellular levels
of free lipids, such as fatty acids, and energy homeostasis.
[0210] Xenophagy is a recently recognized mechanism of defense
against various types of intracellular pathogens, including
Mycobacterium tuberculosis, Salmonella typhimurium, Legionella
pneumophila, Brucella species, Chlamydia species, Coxiella
burnetti, Listeria monocytogenes, Shigella flexneri, Rickettsia
species, Mycobacterium marinum, Burkholderia species, and
Francisella tularensis.
[0211] Microautophagy involves lysosomes directly engulfing
cytoplasm by invagination, protrusion, or septation of the
lysosomal limiting membrane.
Chaperone-Mediated Autophagy
[0212] Chaperone-mediated autophagy (CMA) concerns only those
proteins that have a consensus peptide sequence that can be
recognized by the binding of a hsc70-containing
chaperone/co-chaperone complex. The CMA substrate/chaperone
complex then moves to the lysosomes, where the CMA receptor
lysosome-associated membrane protein type-2a (LAMP-2A) recognizes
it. The protein is unfolded and translocated across the lysosome
membrane assisted by the lysosomal hsc70 on the other side. Thus,
CMA substrates are translocated across the lysosomal membrane on a
one-by-one basis, whereas in macroautophagy and microautophagy the
substrates are engulfed or sequestered in bulk. Moreover, CMA
degrades only certain proteins and not organelles.
Exemplary Therapeutic Indications for Increased Autophagy
[0213] Compounds, compositions, and methods of the invention can
be used to treat and prevent any of the following therapeutic
indications for increased autophagy.
[0214] Autophagy Protects Organisms from Metabolic Stress
[0215] Nutrient deprivation, growth factor depletion, and hypoxia
can induce metabolic stress leading to the induction of autophagy
and to the generation of free amino acids and fatty acids. These
can be recycled in a cell-autonomous fashion and be used for 1) de
novo synthesis of proteins important in the stress response, and
2) fueling the TCA cycle to maintain ATP function. The importance
of this process is demonstrated in the inability of mice and C.
elegans with deficiencies in the ATG proteins important for
autophagy to resist starvation. Thus, a critical role for
autophagy is the mobilization of intracellular energy resources to
meet cellular and organismal demand for metabolic substrates.
[0216] Induction of Autophagy for Treatment of the Heart
[0217] Cardiomyocyte function and survival rely critically on the
presence of basal levels of cardiomyocyte autophagy. Autophagic
recycling of damaged cellular components in nutrient-rich
conditions constitutes a major means of protein and organelle
quality control, ridding the cell of defective (e.g., misfolded or
oxidized) proteins and dysfunctional organelles. This fact is
highlighted by the observation that abrogation of autophagic
pathways in adult heart by conditional inactivation of either the
Atg5 or Atg7 genes triggers rapid-onset cardiac hypertrophy, left
ventricular dilation, and diminished cardiac output.
[0218] Danon disease, a condition marked by severe and progressive
myopathy, stems from defective fusion of autophagosomes with
lysosomes. In early cardiac development, Atg5 disruption provokes
in utero defects and embryonic lethality. At the other end of the
age spectrum, age-related declines in the efficiency of autophagic
clearance likely contribute to progressive accumulation of
defective proteins and organelles which ultimately lead to
functional deterioration over time. Normal aging is associated
with loss of cardiac function mainly due to impaired relaxation
during diastole. Varying formulations of caloric restriction (CR)
can prolong lifespan and improve LV diastolic function; the
underlying mechanisms are believed to be the induction of
autophagy. Together, these facts highlight the vital housekeeping
role for cardiomyocyte autophagy as a mechanism of protein and
organelle surveillance and quality control.
[0219] Autophagy can Improve Skeletal Muscle Function in
Setting of Muscular Atrophy
[0220] Skeletal muscle adapts its capacity to levels of load and
utilization. A central aspect of this adaption is the regulation
of fiber remodeling through degeneration or regeneration of muscle
fibers.
[0221] In the absence of muscle activity, muscular atrophy occurs,
resulting in decreased muscular capacity. This atrophy has been
shown to occur due to increased levels of oxidative stress in
disused muscle. Attenuation of this oxidative stress could lead to
decreased atrophy.
[0222] The autophagy process, and in particular mitophagy are
important in clearing damaged mitochondria and reducing the
effects of increased oxidative stress on muscle functional
capacity. Failure of the autophagy process has been shown to be an
important contributing factor to muscle disuse atrophy, by failing
to remove damaged mitochondria. This decrease in mitochondria
turnover leads to an accumulation of dysfunctional organs and
ensuing muscle damage.
[0223] Preserving Autophagy Function During Aging can Improve
Sarcopenia
[0224] Skeletal muscle atrophy and impaired muscle strength
represent an important health issue and may occur as a consequence
of immobilization, disuse, injury, starvation, and aging. In
particular, advanced age is ineluctably accompanied by the loss of
muscle mass and strength. This condition, known as sarcopenia of
aging, has significant effects on individual health and impacts
the severity of frailty. Moreover, poor muscular strength is
highly predictive of disability and mortality, and general
weakness often results in the loss of independent living, thereby
affecting individual quality of life and imposing a high burden on
healthcare expenditure. Aside from aging, skeletal muscle can
undergo significant atrophy following disuse.
[0225] Sarcopenia is characterized by a gradual loss of muscle
proteins. The size of stable post-mitotic tissues, such as
skeletal and cardiac muscles, is regulated by protein turnover,
and skeletal muscle is influenced by a balance between protein
synthesis and degradation and the turnover of contractile
proteins. A key factor influencing the development of sarcopenia
is the imbalance between the rates of protein synthesis and
degradation. Protein degradation in skeletal muscle cells is
essentially mediated by the activity of two highly conserved
pathways: the autophagic lysosomal pathway and the
ubiquitin-proteasome pathway.
[0226] Recent studies have shown that the impaired autophagy seen
in ATG7 null muscles is characterized by muscle atrophy, weakness,
and features of myofiber degeneration. Consequently, autophagy has
been found to be essential for myofiber maintenance and for the
clearance of damaged proteins and altered organelles.
[0227] Autophagy, which is activated when skeletal muscle is under
nutritional stress (such as metabolic stress), plays a role in the
catabolic condition and in the degradation of macromolecules and
organelles. Catabolic pathways are accelerated during exercise to
supply energy and substrates to the muscle for continuation of
contractions. It has been well established that the rates of amino
acid (relatively small) and glucose oxidation are increased during
endurance exercise, and increased energy consumption is likely
required to induce autophagy. It has been shown that autophagy is
required for myofiber maintenance and for the clearance of damaged
proteins and altered organelles.
[0228] Mild exercise has been shown to improve muscle function and
decrease the decline in muscle function observed in sarcopenia.
These positive benefits are at least in part due to an exercise
induced improvement in the autophagy process. In aging mice, the
autophagy proteins LC3-II, Beclin-1, ATG7, and MuRF-1
significantly decrease with age in muscle. However, mice
undergoing a training regimen during the aging process show a
significantly attenuated decrease in these autophagy proteins. In
overweight older women, mild exercise has been shown to increase
the transcript levels of the autophagy regulators LCB3, Atg7, and
LAMP-2 and thus improve the autophagy process. Thus, preservation
of autophagy may play an important role in skeletal myocyte
homeostasis and optimal mitochondrial turnover in aged muscle.
[0229] An age-related attenuation of autophagy has been shown and
results in a diminished efficiency of protein degradation and the
clearance of damaged organelles. A decrease in proteolytic
activity has been considered responsible, at least in part, for
the accumulation of damaged cellular components in almost all
tissues of aging organisms.
[0230] Improving Autophagy as a Therapeutic Target for Muscle
Degenerative Diseases
[0231] Muscular dystrophies are a group of genetic, hereditary
muscle diseases characterized by defects in muscle proteins. These
defects result in progressive skeletal muscle damage accompanied
by myofiber necrosis and chronic local inflammation, leading to
substitution of myofibers by connective and adipose tissue. In
Duchenne muscular dystrophy (DMD), the most severe form of these
diseases, the continuous and progressive skeletal muscle damage
leads to complete paralysis and death of patients, usually by
respiratory and/or cardiac failure.
[0232] The therapeutic protocols currently in use, based on
corticosteroid administration, provide some delay in the
progression of the disease, but they are associated with severe
side effects. Therapies that substitute corticosteroids or at
least may act as corticosteroid-sparing drugs are thus being
actively pursued, and biological mechanisms relevant to skeletal
muscle homoeostasis are explored, in order to identify new
targets.
[0233] Autophagy is emerging as an important process that limits
muscle damage. Inhibition/alteration of autophagy contributes to
myofiber degeneration leading to accumulation of abnormal
organelles. Mutations that inactivate Jumpy, a phosphatase that
counteracts the activation of VPS34 for autophagosome formation
and reduces autophagy, are associated with a centronuclear
myopathy. This observation suggests that unbalanced autophagy is
pathogenic in muscle degeneration. Likewise, hyperactivation of
Akt as a consequence of muscle-specific deletion of the mammalian
target of rapamycin (mTOR) leads to inhibition of autophagy and to
a muscle phenotype resembling the one observed in muscular
dystrophy. The validity of autophagy modulation as a therapeutic
strategy has been shown in a mouse model of Ulrich myopathy
characterized by defective autophagy and accumulation of
dysfunctional organelles. Forced reactivation of autophagy in
these animals yielded a beneficial therapeutic response.
[0234] In vivo and ex vivo analyses have shown that autophagy is
defective in both the human (DMD) and mouse (mdx) muscular
dystrophy and that such defect contributes to the pathogenesis of
the disease. Muscle biopsies from DMD patients have been shown to
have significantly lower levels of LC3 II and significant
accumulation of p62, a protein known to be incorporated into
autophagosomes and efficiently degraded, with respect to tissues
from control, non-affected individuals.
[0235] A low protein diet has been shown in mice to lead to a
prolonged induction of autophagy. In mice with DMD fed a low
protein diet, an induction of autophagy leads to an improvement
and management in the disease progression. Significant
improvements in muscle function have been observed with an
improvement of whole body tension, reduced muscle fibrosis,
decreased collagen disposition, reduced accumulation of damaged
organelles and reduced apoptosis of muscle fibers.
[0236] This demonstrates that induction of autophagy is an
important homoeostatic mechanism that is disrupted in dystrophic
muscles and indicates that novel therapeutic approaches aimed at
reactivating autophagy can serve as a valuable strategy to reduce
muscle damage in DMD.
[0237] Autophagy Protects the Liver from Oxidative Stress and
Disease
[0238] During liver diseases such as cancer and cirrhosis, the
liver can undergo tissue hypoxia. This process has been shown to
induce an autophagy process, which if inhibited resulted in
increased apoptosis of liver cells.
[0239] In α1-antitrypsin deficiency, the most common genetic cause
of human liver disease, there is significant chronic inflammation
and eventual carcinogenesis. In this disease, a point mutation
occurs in al-antitrypsin Z (ATZ) leading to improperly folding and
accumulation of aggregates. Deletion of ATG5 in hepatic cell lines
lead to an accumulation of the mutant ATZ protein, demonstrating
the important role for autophagy in reducing the impact of liver
disease.
[0240] Autophagy is Important in Limiting Ischemic
Reperfusion Injury
[0241] With advancing age, patients are more likely to acquire
primary and secondary hepatic malignancies that are amenable to
surgical resection and transplantation. Though the elderly
patients may be treated surgically, the aged liver has
significantly decreased reparative capacity following ischemia and
reperfusion injury associated with these operations.
[0242] Ischemic preconditioning is the only promising strategy for
improving the outcome of liver surgery, but its beneficial effects
are limited to young patients. To date, no therapeutic strategy
can suppress the age-dependent ischemia and reperfusion injury.
[0243] A reduction in autophagy has been observed in the old cells
subjected to a severe stress such as ischemia followed by
reperfusion. Studies have shown that by overexpression of
autophagy genes in aged livers of mice, autophagy was increased
and hepatocyte cell survival was increased after ischemia and
reperfusion. Consequently, defective autophagy has been shown to
be a causal mechanism for the age-dependent hepatic reperfusion
injury and that enhancement of autophagy has been demonstrated to
offer therapeutic benefit and reducing age-mediated liver ischemia
reperfusion injury.
[0244] Autophagy in Intestinal Epithelial Cells as a
Therapeutic Target
[0245] The intestinal epithelium interfaces directly with a
diverse community of bacteria that includes benign commensals,
opportunistic pathogens, and overt pathogens, and consequently is
the first line of defense against bacterial invasion of host
tissues. One means that the epithelial cells employ to defend
themselves includes secreting antimicrobial proteins.
Unfortunately, there are some intestinal pathogens, including
Salmonella tyhpimurium or opportunistically invasive commensal
bacteria, such as Enterococcus faecalis, which can avoid this
first line of defense and enter the epithelial cells.
[0246] Autophagy has been shown to be essential for the
recognition and degradation of intracellular pathogens, acting as
an innate barrier to infection. In cell culture, autophagy has
been shown to limit the replication of certain bacterial species.
[0247] It has been shown via genetic studies of inflammatory bowel
disease (IBD) that autophagy plays an important role in the
intestinal immune homeostasis. IBD is a chronic inflammatory
disease of the intestine that arises from dysregulated
interactions with resident microbiota.
[0248] Recently, it has been shown that polymorphisms in genes in
the autophagic pathway are linked to Crohn's disease (CD). Crohn's
disease is a chronic form of IBD that can affect any part of the
gastrointestinal system, but is usually found in the colon or
terminal ileum. The average onset is at 27 years of age in humans,
and is usually present throughout the normal lifespan of the
individual. It is characterized by severe colitis, strictures, and
perianal fistulas, typically requiring surgery.
[0249] The chronic inflammatory process characteristic of CD
requires the intensive interaction between intestinal epithelial
cells and immune competent cells. In CD, there is an exaggerated
immune response to the intestinal microbiota, characterized by an
abnormal increase in Th17 cells, which play a major role in
autoimmunity, and a down-regulation of Treg cells important for
controlling the immune response.
[0250] It has recently been shown that intestinal epithelial cell
autophagy is essential for mammalian intestinal defense against
invasive bacteria. Autophagy in the epithelial cells protects
against the dissemination of invasive bacteria. Following oral
infection with the invasive pathogen Salmonella typhimurium as
well as Enterococcus faecalis, mouse epithelial cells activate
autophagy as a consequence of exposure to these pathogens.
Autophagy was also shown to be critical to limit the
extra-intestinal spread of S. typhimurium. This indicates that
autophagy is a key epithelial cell-autonomous mechanism of
antibacterial defense that protects against dissemination of
intestinal bacteria.
[0251] The present invention provides the know-how to use
compounds that include urolithins and their precursors as
enhancers of autophagy for the administration to and the treatment
of individuals with inflammatory bowel disease (IBD) or Crohn's
disease (CD) and in need of increasing the levels of autophagy in
their in the epithelial cells of the intestine in order to treat
either IBD or CD.
[0252] Autophagy is Important in Aging Cardiac Muscle
[0253] The effects of autophagy induction on improved outcome for
ischemic injury and muscle maintenance makes it especially
relevant for cardiac muscle maintenance and protection from
injury. Cardiac muscle undergoes progressive decline in
mitochondrial function, similar to that observed in skeletal
muscle, resulting in an increase in reactive oxygen species, as
well as an increase in the accumulation of defective organelles.
The clearance of these damaged organelles by autophagy is
important for the maintenance of cardiac muscle function. As
autophagy decreases with age, promoting autophagy can serve to
protect cardiac muscle function.
[0254] Cardiac muscle is also strongly exposed to ischemic
episodes during cardiac infarcts. The level of cardiac muscle
damage that these ischemic episodes produce is strongly dependent
on the ability of the cells to mount an effective autophagy
response to clear damaged organelles. In aged animals, a defective
autophagy response leads to an increase in cardiac muscle damage
after ischemic events. Thus, promotion of autophagy during these
acute events could serve to protect cardiac muscle from damage.
[0255] Autophagy is Important in the Inflammatory Process
[0256] Due to the role of autophagy in clearing defective
organelles, a defect in this process leads to a buildup of
cellular debris and the induction of apoptosis. Autophagy also
plays an important role in defending the organism against
microbial pathogens by inducing their degradation. Additionally,
autophagy plays an important role in the trafficking events that
activate innate and adaptive immunity.
[0257] The autophagic removal of apoptotic corpses is critical for
preventing danger signals that could lead to an inflammation
response. In an impaired autophagy response, where apoptotic
clearance is not efficient the resulting induction of
inflammation, could overcome tolerance to self-antigens leading to
autoimmune diseases such as systemic lupus erythematosus. Thus,
induction of autophagy could serve to decrease inflammatory
responses and the development of autoimmune diseases.
[0258] Applications of Autophagy for Treatment of Disorders of
the Liver
[0259] A number of features of hepatocytes and the liver as a
whole make this organ particularly dependent on autophagy. The
liver is rather unique in its regenerative properties as while
hepatocytes are normally in a quiescent state, they retain the
ability to quickly enter the cell cycle when there is a loss of
liver tissue due to injury or surgical removal. The lack of cell
turnover makes hepatocytes particularly vulnerable to the effects
of impaired autophagy, as cells having long lives accumulate high
levels of damaged organelles, protein aggregates, etc. that are
normally cleared by autophagy. This leads to cellular injury and
potentially to transformation.
[0260] Hepatocellular Lipid Metabolism
[0261] The liver serves as the second largest repository of stored
lipids in the body after adipose tissue. Hepatocytes are a major
cellular storehouse for neutral lipids in the form of
triglycerides (TGs) and cholesterol esters contained in
specialized organelles termed lipid droplets (LD). Autophagy
mediates the breakdown of intracellular LD stores through the
process of lipophagy. This enables the hepatocytes to rapidly
mobilize their lipid stores in times of metabolic need. The loss
of hepatocyte autophagy leads to a marked increase in hepatic TG
and cholesterol content, indicating that lipophagy limits lipid
accumulation by the liver in vivo. Also, lipophagy controls
cellular energy homeostasis by providing free fatty acids (FFA)
from the breakdown of TGs, which subsequently drives mitochondrial
β-oxidation and cellular ATP generation. It has been shown that
the autophagosomal protein LC3, critical for autophagosome
membrane formation, associates with LDs.
[0262] Autophagy Protects Against Hepatic Diseases
[0263] SERPINA1/al-anti-trypsin deficiency (ATD) is the most
common genetic cause of human liver disease in children. This
disease is caused by homozygosity for the SERPINA1/al-antitrypsin
Z allele SERPINA1-Z, a point mutation, which renders the hepatic
secretory glycoprotein SERPINA1 prone to misfolding,
polymerization, and aggregation. The mutant SERPINA1-Z protein
accumulates in hepatocytes and the levels of SERPINA1 found in the
blood and body fluids are reduced to 10-15% of those normally
observed. Accumulation of mutant SERPINA1-Z in the endoplasmic
reticulum (ER) of hepatocytes leads to liver damage by a
gain-of-function. It has been shown that intracellular degradation
of SERPINA1-Z aggregates and polymers involves the autophagic
pathway.
[0264] The drug carbamazepine, known to induce autophagy, was
recently shown to be effective in cell based and mouse model of
ATD. Carbamazepine increases autophagic degradation of SERPINA1-Z
in cultured cells and when provided orally to the PiZ mouse model
of ATD, it reduced the hepatic load of SERPINA1-Z. Additionally,
inducing autophagy reduced hepatic fibrosis. Consequently, drugs
enhancing autophagy are attractive candidates for improving the
liver disease that develops in some patients with ATD.
[0265] The present invention provides the know-how to use
compounds that include urolithins and their precursors as
enhancers of autophagy for the treatment of individuals with ATD
and in need of increasing the levels of autophagy in their liver
and hepatocytes in order to reduce liver toxicity.
[0266] Autophagy Protects Against Nonalcoholic Fatty Liver
Disease
[0267] Nonalcoholic fatty liver disease (NAFLD) is an important
component of the metabolic syndrome together with obesity and
diabetes. NAFLD encompasses a spectrum of hepatic abnormalities
that range from simple fatty liver or steatosis, to fatty liver
with hepatocellular injury and inflammation, which is known as
nonalcoholic steatohepatitis (NASH). NAFLD is now the most
prevalent liver disease in the USA and accounts for about 75% of
all chronic liver diseases.
[0268] The most important role of autophagy in fatty liver disease
could be to regulate the process of excessive lipid accumulation.
In fact, mice with a hepatocyte-specific knockout of Atg7, a
protein required for autophagy, consuming a high-fat diet led to a
marked increase in liver TGs and cholesterol content, showing that
autophagy defects can induce hepatic steatosis. When considering
NASH, while its exact causes are unknown, free fatty acid
(FFA)-induced lipotoxicity has been implicated in the mechanisms
of hepatocellular injury of this disease. Evidence points to the
fact that hepatocyte autophagy renders the cells more resistant to
injury from FFA.
[0269] Autophagy is an attractive therapeutic target for the
treatment and prevention of both NAFLD and NASH. Therapeutic
intervention to increase autophagy may reverse not only the
hepatic manifestations of NAFLD, including hepatocellular
steatosis and injury, but also some of the underlying metabolic
abnormalities of the disease via its effects on insulin
resistance. Additionally, treatment by increasing autophagy may
prevent common end-stage complications of NAFLD, such as
hepatocellular carcinoma.
[0270] The present invention provides the know-how to use
compounds that include urolithins and their precursors as
enhancers of autophagy for the treatment of individuals with NAFLD
and in need of increasing the levels of autophagy in their liver
and hepatocytes in order to treat these conditions.
[0271] Autophagy Protects Against Alcoholic Liver Disease
[0272] Alcoholic liver disease (ALD) is a major cause of chronic
liver disease, and like NAFLD, has a wide spectrum of pathogenic
features, that range from steatosis to sever acute alcoholic
hepatitis, fibrosis, cirrhosis, and hepatocellular carcinoma.
[0273] Autophagy has been shown to play a role in treating ALD.
For example, induction of autophagy by administration of rapamycin
significantly suppresses acute alcohol-induced steatosis. Also, a
common feature of chronic alcohol abuse is the formation of
hepatic protein aggregates known as Mallory-Denk bodies, which are
cytosolic inclusion bodies enriched with Krt8/keratin 8 and Krt18
and proteins that include ubiquitin. Rapamycin treatment
significantly reduces the number of Mallory-Denk bodies in
proteasome inhibitor-treated KRT8 transgenic mice.
[0274] Consequently, enhancing hepatic autophagy is an attractive
target for improving alcohol-induced liver disease. The present
invention provides the know-how to use compounds that include
urolithins and their precursors as enhancers of autophagy for the
treatment of individuals with ALD and in need of increasing the
levels of autophagy in their liver and hepatocytes in order to
treat this condition.
[0275] Autophagy Protects Against Drug-Induced Liver Injury
[0276] Most drugs are metabolized and detoxified in the liver,
making the liver the principal target for drug damage. Liver
injury due to drugs is a common cause for the withdrawal of
approved drugs on the market, and it is thought that drug-induced
hepatotoxicity is responsible for more than half of acute cases of
liver failure. Acetaminophen, also known as paracetamol and
N-acetyl-p-aminophenol (APAP), is a widely used antipyretic and
analgesic drug and is also the most common source of severe
drug-induced hepatotoxicity. At therapeutic levels APAP is safe,
but overdosing leads to toxicity mainly due to its reactive
metabolite, N-acetyl-p-benzoquinone imine (NAPQI). NAPQI can
deplete hepatic stores of glutathione (GSH), an intracellular
antioxidant. Following the depletion of GSH, NAPQI is known to
react with cellular proteins as well as mitochondrial proteins to
form protein adducts. These APAP-induced mitochondrial protein
adducts can then lead to mitochondrial damage and subsequent
necrosis.
[0277] When autophagy is enhanced with rapamycin, APAP-induced
necrosis is significantly inhibited, both in cultured primary
hepatocytes and in the livers of mice. Treatment with rapamycin
two hours after APAP administration has been seen to significantly
improve APAP-induced liver injury, even though APAP metabolism and
hepatic GSH depletion have already occurred. This is particularly
important as patients at risk for hepatotoxicity from an acute
APAP overdose do not receive medical care until they are past the
metabolic phase. Consequently, pharmacologic intervention
targeting an enhancement of autophagy holds a potential
therapeutic benefit for individuals with a risk of APAP
hepatotoxicity following an overdose.
[0278] The present invention provides the know how-to use
compounds that include urolithins and their precursors as
enhancers of autophagy for the treatment of individuals at risk of
hepatotoxicity due to drug side effects and in need of increasing
the levels of autophagy in their liver and hepatocytes in order to
treat or prevent the potential drug toxicity.
[0279] Autophagy is Important in Limiting Ischemia/Reperfusion
Injury
[0280] Ischemia/reperfusion (I/R) injury is a causal factor
contributing to morbidity and mortality. The vulnerability of the
liver to I/R injury is a major obstacle to liver resection and
transplantation surgery where reperfusion after sustained ischemia
is unavoidable during hepatectomy and vascular reconstruction.
Mitochondrial dysfunction is known to be one of the critical
downstream events that lead to I/R-mediated cell death.
[0281] Autophagy clears abnormal or dysfunctional mitochondria to
ensure an optimal cellular function and survival. With impaired or
insufficient mitophagy, cells accumulate damaged mitochondria,
which subsequently leads to uncontrolled ROS formation,
mitochondrial DNA mutation, energetic failure, and ultimately cell
death. Consequently, the failure of mitophagy to remove a small
number of damaged mitochondria during I/R can have a significant
impact on hepatocellular function and viability. Mitophagy is
essential for hepatic function and survival following I/R injury.
[0282] While minimizing I/R injury plays an important role in the
outcome of transplanted young livers, aged livers are even more
susceptible to negative impact of I/R injury. In the case of aged
livers, hepatocytes fail to respond to the I/R stress and
upregulate their endogenous protective autophagy response. Similar
to young livers following prolonged ischemia, aged livers after
short-term ischemia accumulate dysfunctional mitochondria, undergo
mitochondrial permeability transition, and lose their viability
soon after reperfusion.
[0283] Methods of enhancing autophagy, including pre-ischemia
nutrient depletion and over expression of pro-autophagy genes ATG7
or BECN1, lead to the suppression of the mitochondrial
permeability transition and increases hepatocyte survival
following reperfusion.
[0284] This indicates that treatments with agents that induce
autophagy in the liver will offer protection during a situation of
I/R and help to minimize cellular injury. Such treatments are
applicable in situations of the transplantation of both young and
aged livers. Treatments may involve: (i) pre-treatments of the
liver tissue ex vivo by perfusion of the liver with a solution
that contains an inducer of autophagy; (ii) treatment of the liver
donor with an autophagy inducer; or (iii) treatment of the liver
recipient prior to, during the operation and/or immediately after
the surgical intervention. Of course, these treatment modalities
may be applied individually or in any combination (for example: 1
and 2; 2 and 3; 1 and 3; 1, 2, and 3).
[0285] The present invention provides the know-how to use
compounds that include urolithins and their precursors as
enhancers of autophagy for the treatment of individuals and their
livers, that may be at risk of I/R injury. These compounds may be
provided orally or parenterally to the donor or recipient, or
provided in a preconditioning solution that may be applied to the
resected liver tissue.
[0286] Autophagy and Osteoarthritis
[0287] Osteoarthritis (OA) is the most common aging-related joint
pathology and is characterized by degradation of cartilage
extracellular matrix (ECM) and reduced cartilage cellularity.
Changes in the articular cartilage appear to be critical in OA
initiation and progression. Chondrocytes are the only cell
population of adult articular cartilage. The capacity of the adult
articular chondrocytes to regenerate the normal cartilage matrix
architecture is limited and declines with aging, due to cell death
and abnormal responsiveness to anabolic stimuli. Articular
cartilage is characterized by a very low rate of cell turnover and
it has been shown that autophagy play an important role in
chondrocyte cellular function and survival. In fact, autophagy is
a constitutively active and protective process for the maintenance
of cartilage homeostasis. Studies have shown both in joint aging
and OA in humans and in mice that there is a reduction in the
expression of autophagy regulators, which was accompanied by an
increase in chondrocyte apoptosis. Compromised autophagy is
thought to contribute to the development of OA. It has been shown
that treatment with the compound rapamycin, a known inducer of
autophagy, has been able to increase activation of LC3 in
cartilage in an animal model of OA and consequently reduce the
severity of articular cartilage degradation. In the present
invention, urolithins and their precursors have been shown to
increase the levels of autophagy in tissues following oral
consumption, making them ideal candidates for the treatment and
reduction of the severity of osteoarthritis in young and aging
humans and mammals.
[0288] Metabolic Syndrome, Diabetes, and Obesity
[0289] Compounds and methods of the invention are useful in the
treatment and prevention of metabolic syndrome, type 2 diabetes
mellitus, and obesity. As used herein, the term “metabolic
syndrome” refers to a combination of medical disorders that, when
occurring together, increase the risk of developing cardiovascular
disease and diabetes. It affects one in five people in the United
States and prevalence increases with age. Some studies have shown
the prevalence in the United States to be an estimated 25% of the
population. In accordance with the International Diabetes
Foundation consensus worldwide definition (2006), metabolic
syndrome is central obesity plus any two of the following:
Raised triglycerides: >150 mg/dL (1.7 mmol/L), or specific
treatment for this lipid abnormality;
Reduced HDL cholesterol: <40 mg/dL (1.03 mmol/L) in males,
<50 mg/dL (1.29 mmol/L) in females, or specific treatment for
this lipid abnormality;
Raised blood pressure: systolic BP>130 or diastolic BP>85 mm
Hg, or treatment of previously diagnosed hypertension; and
Raised fasting plasma glucose: (FPG)>100 mg/dL (5.6 mmol/L), or
previously diagnosed type 2 diabetes.
[0294] Autophagy and Neurodegenerative Diseases
[0295] In neurodegenerative diseases, brain tissue accumulates
autophagosomes, demonstrating an increase in autophagy, which in
model organisms has been shown to have a protective effect. It
plays an important role in clearing the misfolded proteins that
accumulate as a result of several neurodegenerative diseases.
These include proteins that have polyQ repeats as seen as in
Huntington's diseases and spinocerebellar ataxia, mutant
α-synucleins involved in Parkinson's as well as tau aggregates.
[0296] Knockdown of ATG genes important in autophagy in C. elegans
resulted in increased aggregate formation and toxicity of PolyQ
proteins. In Alzheimer's disease the autophagy process is impaired
as a result of a defect in autophagosomal maturation that could be
an important reason for aggregate accumulation. By contrast,
autophagy induction by rapamycin in both Drosophila and mouse
models of polyQ disease protected these animals from
neurotoxicity. These results demonstrate that autophagy induction
can have a protective role in neuronal cells against
neurodegeneration.
[0297] The development of neurodegenerative diseases in patients
implies that autophagy can reach a saturation point in which the
ability to degrade mutant protein aggregates is exceeded. Thus,
promotion of autophagy could help in delaying the onset of
neurodegeneration disease.
[0298] Cognitive Disorder
[0299] Compounds and methods of the invention are useful for
treating a cognitive disorder. As used herein, a cognitive
disorder refers to any condition that impairs cognitive function.
In one embodiment, “cognitive disorder” refers to any one or more
of delirium, dementia, learning disorder, attention deficit
disorder (ADD), and attention deficit hyperactivity disorder
(ADHD). In one embodiment, the cognitive disorder is a learning
disorder. In one embodiment, the cognitive disorder is attention
deficit disorder (ADD). In one embodiment, the cognitive disorder
is attention deficit hyperactivity disorder (ADHD).
[0300] Compounds and methods of the invention are useful for
improving cognitive function, even in the absence of a cognitive
disorder. As used herein, “cognitive function” refers to any
mental process that involves symbolic operations, e.g.,
perception, memory, attention, speech comprehension, speech
generation, reading comprehension, creation of imagery, learning,
and reasoning. In one embodiment, “cognitive function” refers to
any one or more of perception, memory, attention, and reasoning.
In one embodiment, “cognitive function” refers to memory.
[0301] Methods for measuring cognitive function are well known and
can include, for example, individual or battery tests for any
aspect of cognitive function. One such test is the Prudhoe
Cognitive Function Test. Margallo-Lana et al. (2003) J Intellect
Disability Res. 47:488-492. Another such test is the Mini Mental
State Exam (MMSE), which is designed to assess orientation to time
and place, registration, attention and calculation, recall,
language use and comprehension, repetition, and complex commands.
Folstein et al. (1975) J Psych Res. 12:189-198. Other tests useful
for measuring cognitive function include the Alzheimer Disease
Assessment Scale-Cognitive (ADAS-Cog) (Rosen et al. (1984) Am J
Psychiatry. 141(11):1356-64) and the Cambridge Neuropsychological
Test Automated Battery (CANTAB) (Robbins et al. (1994) Dementia.
5(5):266-81). Such tests can be used to assess cognitive function
in an objective manner, so that changes in cognitive function, for
example in response to treatment in accordance with methods of the
invention, can be measured and compared.
[0302] Protein Misfolding and Aggregation
[0303] These diseases and disorders, which are collectively
referred to herein as “amyloid-related diseases”, fall into two
main categories: (a) those which affect the brain and other parts
of the central nervous system; and (b) those which affect other
organs or tissues around the body.
[0304] Examples of amyloid-related diseases which fall under these
two categories are listed in the following two sections; however,
many other examples of rare, hereditary amyloid-related diseases
are known which are not included here, and additional forms of
amyloid-related disease are likely to be discovered in future.
[0305] Neurodegenerative Diseases Associated with Amyloidosis
[0306] Many different neurodegenerative diseases are associated
with the misfolding and aggregation of a specific protein or
peptide in a particular part of the brain, or elsewhere in the
central nervous system, depending on the specific disease.
Examples of such diseases follow.
[0307] Various forms of Alzheimer's disease (AD) as well as Down's
syndrome, hereditary cerebral hemorrhage with amyloidosis (HCHWA,
Dutch type), cerebral amyloid angiopathy, and possibly also mild
cognitive impairment and other forms of dementia are associated
with the aggregation of a 40/42-residue peptide called β-amyloid,
Aβ(1-40) or Aβ(1-42), which forms insoluble amyloid fibers and
plaques in the cerebral cortex, hippocampus or elsewhere in the
brain, depending on the specific disease. Alzheimer's disease is
also associated with the formation of neurofibrillary tangles by
aggregation of a hyperphosphorylated protein called tau, which
also occurs in frontotemporal dementia (Pick's disease).
[0308] Parkinson's disease (PD), dementia with Lewy bodies (DLB),
and multiple system atrophy (MSA) are associated with the
aggregation of a protein called α-synuclein, which results in the
formation of insoluble inclusions called Lewy bodies. Huntington's
disease (HD), spinal and bulbar muscular atrophy (SBMA, also known
as Kennedy's disease), dentatorubral pallidoluysian atrophy
(DRPLA), different forms of spinocerebellar ataxia (SCA, types 1,
2, 3, 6 and 7), and possibly several other inheritable
neurodegenerative diseases are associated with the aggregation of
various proteins and peptides that contain abnormally expanded
glutamine repeats (extended tracts of polyglutamine).
Creutzfeldt-Jakob disease (CJD), bovine spongiform encephalopathy
(BSE) in cows, scrapie in sheep, kuru,
Gerstmann-Straussler-Scheinker disease (GSS), fatal familial
insomnia, and possibly all other forms of transmissible
encephalopathy are associated with the self-propagating misfolding
and aggregation of prion proteins.
[0309] Amyotrophic lateral sclerosis (ALS), and possibly also some
other forms of motor neuron disease (MND) are associated with the
aggregation of a protein called superoxide dismutase.
[0310] Familial British dementia (FBD) and familial Danish
dementia (FDD), respectively, are associated with aggregation of
the ABri and ADan peptide sequences derived from the BRI protein.
[0311] Hereditary cerebral hemorrhage with amyloidosis (HCHWA,
Icelandic type) is associated with the aggregation of a protein
called cystatin C.
[0312] Systemic Diseases Associated with Amyloidosis
[0313] In addition to the neurodegenerative diseases listed above,
a wide variety of systemic ageing-related or degenerative diseases
are associated with the misfolding and aggregation of a particular
protein or peptide in various other tissues around the body (i.e.,
outside of the brain). Examples of such diseases follow.
[0314] Type II diabetes mellitus (also known as adult-onset
diabetes, or non-insulin dependent diabetes mellitus) is
associated with the aggregation of a 37-residue peptide called the
islet amyloid polypeptide (IAPP, or “amylin”), which forms
insoluble deposits that are associated with the progressive
destruction of insulin-producing β cells in the islets of
Langerhans within the pancreas.
[0315] Dialysis-related amyloidosis (DRA) and prostatic amyloid
are associated with the aggregation of a protein called
β2-microglobulin, either in bones, joints and tendons in DRA,
which develops during prolonged periods of hemodialysis, or within
the prostate in the case of prostatic amyloid.
[0316] Primary systemic amyloidosis, systemic AL amyloidosis and
myeloma-associated amyloidosis are associated with the aggregation
of immunoglobulin light chain (or in some cases immunoglobulin
heavy chain) into insoluble amyloid deposits, which gradually
accumulate in various major organs such as the liver, kidneys,
heart and gastrointestinal (GI) tract.
[0317] Reactive systemic AA amyloidosis, secondary systemic
amyloidosis, familial Mediterranean fever, and chronic
inflammatory disease are associated with the aggregation of serum
amyloid A protein, which forms insoluble amyloid deposits that
accumulate in major organs such as the liver, kidneys and splee.
Senile systemic amyloidosis (SSA), familial amyloid polyneuropathy
(FAP) and familial amyloid cardiomyopathy (FAC) are associated
with the misfolding and aggregation of different mutants of
transthyretin protein (TTR), which form insoluble inclusions in
various organs and tissues such as the heart (especially in FAC),
peripheral nerves (especially in FAP) and gastrointestinal (GI)
tract. Another form of familial amyloid polyneuropathy (FAP, type
II) is associated with the aggregation of apolipoprotein AI in the
peripheral nerves. Familial visceral amyloidosis and hereditary
non-neuropathic systemic amyloidosis are associated with
misfolding and aggregation of various mutants of lysozyme, which
form insoluble deposits in major organs such as the liver, kidneys
and spleen.
[0318] Finnish hereditary systemic amyloidosis is associated with
aggregation of a protein called gelsolin in the eyes (particularly
in the cornea).
[0319] Fibrinogen α-chain amyloidosis is associated with
aggregation of the fibrinogen A α-chain, which forms insoluble
amyloid deposits in various organs, such as the liver and kidneys.
[0320] Insulin-related amyloidosis occurs by the aggregation of
insulin at the site of injection in diabetics.
[0321] Medullary carcinoma of the thyroid is associated with the
aggregation of calcitonin in surrounding tissues.
[0322] Isolated atrial amyloidosis is associated with the
aggregation of atrial natriuretic peptide (ANP) in the heart.
[0323] Various forms of cataract are associated with the
aggregation of y-crystallin proteins in the lens of the eyes.
[0324] Autophagy and Endothelial Cell Function and Associated
Diseases
[0325] Endothelial Cell Dysfunction
[0326] Global endothelial cell dysfunction occurs in several
diverse diseases such as diabetes, hypertension, chronic kidney
disease, and atherosclerosis. In these diseases endothelial cell
dysfunction is thought to occur as a result of stress-induced
premature senencense (SIPS). SIPS is characterized by subverted
autophagy and lysosomal dysfunction, with the accumulation of
autolysosomal vacuoles.
[0327] Endothelial cell dysfunction also occurs as a result of
aging with an increased incidence of cardiovascular diseases. This
increase in cell dysfunction correlates with a decrease in
autophagy. In older humans, expression of autophagy markers in
arterial endothelial cells was impaired by 50% (P<0.05) and was
associated with a 30% (P<0.05) reduction in arterial
endothelium-dependent dilatation (EDD). Similarly, in C57BL/6
control mice aging was associated with a 40% decrease (P<0.05)
in arterial markers of autophagy and a 25% reduction (P<0.05)
in EDD, demonstrating that impaired autophagy is a cause of
age-related arterial dysfunction.
[0328] In old mice, treatment with the autophagy-enhancing agent
trehalose restored expression of autophagy markers, rescued
NO-mediated EDD by reducing oxidative stress, and normalized
inflammatory cytokine expression. The present invention provides
the know-how to use compounds that include urolithins and their
precursors as enhancers of autophagy for the treatment of
individuals having health conditions linked to endothelial cell
dysfunction and in need thereof.
[0329] Endothelial Cell Injury
[0330] Endothelial cell injury can occur as a result of disease
processes such as sickle cell anemia or thalassemia in which
pathologically high levels of heme and iron release can occur.
Severe skeletal muscle damage, as well as cardiac ischemia injury
results in the release of the heme protein, myoglobin, which also
results in endothelial cell injury. This damage to the vascular
endothelial cells can lead to vascular dysfunction and an increase
in cardiovascular complications. Endothelial cell injury caused by
heme toxicity is associated with a progressive decrease in
endothelial cell mitochondrial membrane potential, leading to
apoptosis.
[0331] Micro- and macro-vascular complications are commonly seen
in diabetic patients, and endothelial dysfunction contributes to
the development and progression of the complications. Abnormal
functions in endothelial cells lead to the increase in vascular
tension and atherosclerosis, followed by systemic hypertension as
well as increased incidence of ischemia and stroke in diabetic
patients. Mitochondrial dysfunction appears to be central to the
vascular endothelial dysfunction. Enhanced mitochondrial fission
and/or attenuated fusion leads to mitochondrial fragmentation and
disruption of the endothelial physiological function. Abnormal
mitochondrial biogenesis and disturbance of mitochondrial
autophagy increase the accumulation of damaged mitochondria, such
as irreversibly depolarized or leaky mitochondria, and facilitate
cell death. Augmented mitochondrial ROS production and
Ca<2+> overload in mitochondria not only cause the
maladaptive effect on the endothelial function, but also are
potentially detrimental to cell survival.
[0332] Endothelial cell injury can also result from cardiac
procedures such as angioplasty, bypass surgery, and valve
replacement. Upregulation of autophagy should lead to a reduction
in the injury to the associated endothelial cells.
[0333] Strategies that increase autophagy would have clear
therapeutic potential. The present invention provides the know-how
to use compounds that include urolithins and their precursors as
enhancers of autophagy for the treatment of individuals having
health conditions linked to endothelial cell injury resulting from
disease processes such as diabetes, sickle cell anemia, or
thalassemia, as well as protecting endothelial cells from the more
acute effects of severe muscle injury.
[0334] Autophagy and Cancer
[0335] Autophagy and cancer have similar regulatory pathways, with
several tumor suppression genes such as PTEN, TSC1 and TSC2
leading to the upstream inhibition of TOR signaling, leading to
the stimulation of autophagy. Additionally, the autophagy protein,
Beclin1 has been identified as a tumor suppressor deleted in many
human cancers. These results demonstrate that autophagy plays an
important role in tumor suppression.
[0336] Aging
[0337] By far the greatest risk factor for neurodegenerative
diseases, such as Alzheimer's disease (AD), Parkinson's disease
(PD), and amyotrophic lateral sclerosis (ALS), is aging.
Mitochondria have been thought to contribute to aging through the
accumulation of mitochondrial DNA (mtDNA) mutations and net
production of reactive oxygen species (ROS). Although most
mitochondrial proteins are encoded by the nuclear genome,
mitochondria contain many copies of their own DNA. Human mtDNA is
a circular molecule of 16,569 base pairs that encodes 13
polypeptide components of the respiratory chain, as well as the
rRNAs and tRNAs necessary to support intramitochondrial protein
synthesis using its own genetic code. Inherited mutations in mtDNA
are known to cause a variety of diseases, most of which affect the
brain and muscles—tissues with high energy requirements. It has
been hypothesized that somatic mtDNA mutations acquired during
aging contribute to the physiological decline that occurs with
aging and aging-related neurodegeneration. It is well established
that mtDNA accumulates mutations with aging, especially
large-scale deletions and point mutations. In the mtDNA control
region, point mutations at specific sites can accumulate to high
levels in certain tissues: T414G in cultured fibroblasts, A189G
and T408A in muscle, and C150T in white blood cells. However,
these control-region “hot spots” have not been observed in the
brain. Point mutations at individual nucleotides seem to occur at
low levels in the brain, although the overall level may be high.
Using a polymerase chain reaction (PCR)-cloning-sequencing
strategy, it was found that the average level of point mutations
in two protein-coding regions of brain mtDNA from elderly subjects
was ̃2 mutations per 10 kb. Noncoding regions, which may be under
less selection pressure, potentially accumulate between twice and
four times as many. The accumulation of these deletions and point
mutations with aging correlates with decline in mitochondrial
function. For example, a negative correlation has been found
between brain cytochrome oxidase activity and increased
point-mutation levels in a cytochrome oxidase gene (COI).
[0338] Net production of ROS is another important mechanism by
which mitochondria are thought to contribute to aging.
Mitochondria contain multiple electron carriers capable of
producing ROS, as well as an extensive network of antioxidant
defenses. Mitochondrial insults, including oxidative damage
itself, can cause an imbalance between ROS production and removal,
resulting in net ROS production. The importance to aging of net
mitochondrial ROS production is supported by observations that
enhancing mitochondrial antioxidant defenses can increase
longevity. In Drosophila, overexpression of the mitochondrial
antioxidant enzymes manganese superoxide dismutase (MnSOD) and
methionine sulfoxide reductase prolongs lifespan. This strategy is
most successful in short-lived strains of Drosophila, and has no
effect in already long-lived strains. However, it has recently
been shown that overexpression of catalase experimentally targeted
to mitochondria increased lifespan in an already long-lived mouse
strain.
[0339] Improving Activity During Aging
[0340] Activity in animals is driven largely by circadian rhythm
and is synchronized to the environment. Disruption of the
circadian rhythm or a desynchronization with the environment can
lead to increase in nighttime wakefulness or daytime naps.
[0341] Normal aging is accompanied by declining locomotor
activity, altered circadian rhythms, as well as altered sleep and
food intake patterns. These effects lead to a decrease in
alertness and vigilance decreases in the elderly, leading to an
increase in nighttime wakefulness, as well as an increase in
daytime naps. Activity patterns can also be disrupted in a similar
way by disease, such as Alzheimer's Disease.
[0342] Age-dependent changes in activity rhythms are also observed
in other animals, for example, rats, hamsters, mice and dogs, with
an increase in fragmentation and a decrease in synchronization
with the environment. These age-dependent circadian disruptions
have been linked to the degeneration of the suprachiasmatic
nucleus of the hypothalamus. Sleep disruption in both humans and
rodents have been shown to contribute to age-dependent cognitive
dysfunction.
[0343] The increasing disruption of circadian rhythms is
accompanied by a gradual decline in motor activity with age in
several species, including humans, mice, monkeys, and dogs. Of
particular note, the daytime activity of senior dogs (>10 years
of age) declines as compared to young and middle-aged dogs. These
changes in activity can be monitored by devices intended to
measure activity, for example, by means of an accelerometer or by
employing motion sensing cameras.
[0344] Many of the disruptions seen in aging as a result of
decreased activity are also observed in younger populations where
cultural trends have resulted in decreased activity, accompanied
with increased caloric intake, leading to an obesity epidemic. The
resulting caloric imbalance has led to an increase in several
disease conditions such as type 2 diabetes, colon cancer, and
metabolic syndrome, as well as mental health issues. Several
prospective cohort studies and meta-analysis in humans have shown
that physical inactivity is associated with an elevated risk for
the development of metabolic syndrome, type 2 diabetes,
hypertension, coronary artery disease, stroke, and cardiovascular
disease.
[0345] Both humans and animals, particularly dogs, would benefit
from the present invention and its ability to improve activity
both during the youth and aging periods of life.
[0346] In one embodiment, the urolithin or precursor would
increase activity of the recipient, human or animal. In yet
another embodiment, the increase in activity is an increase by 1%
to 100%. For example, the activity may be increased by 1%, 2%, 3%,
4%, 5%, 6%, 7%, 8%, 9%, or 10%. In certain embodiments, the
increase in activity is an increase of 5-10%, 10-15%, 15-20%,
20-25%, 25-30%, 30-35%, 35-40%, 40-45%, 45-50%, 50-55%, 55-60%,
60-65%, 65-70%, 70-75%, 75-80%, 80-85%, 85-90%, 90-95%, and
95-100%.
[0347] In one embodiment, treatment by urolithin or a precursor
thereof would increase activity and lead to a reduction in risk of
metabolic syndrome. In one embodiment, treatment by urolithin or a
precursor thereof would increase activity and lead to a reduction
in risk of type 2 diabetes. In one embodiment, treatment by
urolithin or a precursor thereof would increase activity and lead
to a reduction in risk of hypertension, coronary artery disease,
stroke, and cardiovascular disease. In one embodiment, treatment
by urolithin or a precursor thereof would increase activity and
improve cognitive function.
[0348] Mood Disorders
[0349] Compounds and methods of the invention are useful for
treating a mood disorder (also known as an affective disorder). As
used herein, a “mood disorder” refers to a disturbance in
emotional state, such as is set forth in the Diagnostic and
Statistical Manual of Mental Disorders, published by the American
Psychiatric Association. Mood disorders include but are not
limited to major depression, postpartum depression, dysthymia, and
bipolar disorder. In one embodiment, the mood disorder is major
depression.
[0350] Compounds and methods of the invention are useful for
treating or preventing a stress-induced or stress-related mood
disorder….
[0351] Compounds and methods of the invention are useful for
treating an anxiety disorder. As used herein, an “anxiety
disorder” refers to a dysfunctional state of fear and anxiety,
e.g., fear and anxiety that is out of proportion to a stressful
situation or the anticipation of a stressful situation…
Process-Scale Synthesis of Urolithins
US2015183758
Disclosed is a method of preparing a urolithin, or an intermediate
or analog thereof, having a dibenzo[b,d]pyran-6-one core. The
method is especially advantageous for the large-scale preparation
of urolithins or intermediates or analogs thereof. The method may
optionally include the preparation of a urolithin, or an
intermediate or analog thereof, as a pharmaceutically acceptable
salt.
BACKGROUND
[0002] Pomegranate (Punica granatum) fruits have been used for
centuries in folk medicine. They are consumed fresh and as juice,
both of which are excellent sources of ellagitannins and ellagic
acid. Ellagitannins (ETs) are polymeric polyphenols abundant in
some fruits and nuts such as pomegranates, raspberries,
strawberries, black raspberries, walnuts and almonds. Despite
numerous reports of the biological properties and human health
benefits of ETs, knowledge of their bioavailability,
pharmacokinetics, disposition and metabolic fate in humans is
limited. Commercially-produced pomegranate juice contains
gallagyl-type ellagitannins, including punicalagin isomers
(1500-1900 mg/L), undefined hydrolyzable tannins (400-500 mg/L),
and ellagic acid and its glycosides (120-260 mg/L). Gil et al. J.
Agric. Food Chem. 2000, 48, 4581-4589. Punicalagins, ellagitannins
in which gallagic and ellagic acids are linked to a glucose
molecule, are abundant in pomegranate peel. Punicalagin isomers
and ellagic acid derivatives are not present in the aril juice,
but during industrial juice processing they are extracted from the
husk and membrane surrounding the arils and released in large
quantities into the juice. The fruit arils of pomegranates contain
other polyphenols, such as anthocyanins, responsible for the
fruit's bright ruby-red color. Ellagitannins belong to a group of
compounds known as hydrolyzable tannins, which release ellagic
acid (EA) upon hydrolysis.
[0003] Unfortunately, ellagitannins are typically poorly absorbed
by the human gut. However, a number of metabolites derived from
ellagitannins are absorbed by the human gut, including certain
metabolites ultimately formed in the gut by commensal
microorganisms (i.e., intestinal microflora). Ellagitannins
release ellagic acid under physiological conditions in vivo, and
ellagic acid is then gradually metabolized by the gut microflora
in the intestine to produce the urolithins. Once the metabolites
are absorbed, they are further metabolized to produce urolithin
glucuronides and/or sulfates. There is growing evidence that
urolithins have potent antioxidant, anticancer, and
anti-hyperproliferative activity. See US 2011/0065662; US
2012/0164243; and US 2014/0018415; all of which are incorporated
by reference.
[0004] Although urolithins are derived from ETs present in certain
foods (e.g., pomegranates), the consumption of these foods does
not always lead to sufficient bioavailability of the therapeutic
metabolites. Specifically, certain individuals, referred to herein
as non-producers, fail to produce detectable amounts of the
metabolites after consumption of ET-containing foods (e.g.,
pomegranate juice). Even among individuals who are producers,
there is a great deal of variation (from very low to very high) in
the amount of urolithin metabolites produced. Furthermore, any
FDA-approved therapeutic use of urolithins would require a
reliable and standard dosing regimen; that is, a known dose of a
fully-characterized compound or compounds. It would thus be
necessary to administer one or more selected urolithins directly
to patients in need thereof.
[0005] In light of the therapeutic promise of urolithin compounds,
a tremendous need exists for a safe, economical, reliable, and
scalable synthesis approach to the urolithins. A reliable source
of multi-kilo and commercial quantities of urolithin compounds
will allow their further clinical development, with the ultimate
goal of exploiting their full therapeutic potential.
SUMMARY OF THE INVENTION
[0006] One aspect of the present invention is a method for the
preparation of urolithin compounds or intermediates useful in
preparing such compounds or analogs thereof. Certain methods of
the present invention include a copper-catalyzed coupling of two
urolithin precursor fragments to form a coupling product (Method
A). Certain methods of the present invention include demethylating
one or more phenolic methoxy groups of a urolithin intermediate
(Method B). In certain embodiments, Methods A and B are performed
sequentially, but not necessarily in that order, to yield a
urolithin or analog thereof. The present invention is improved
over previous methods for producing the same or similar compounds
(e.g., in terms of cost, yield, purity of the resulting
product(s), catalyst loading, safety profile, reaction time,
temperature, or amount/type of solvent used)...
DETAILED DESCRIPTION
[0009] As mentioned above, ellagitannins generally are not
absorbed in the gut. Rather, they release ellagic acid (EA) in the
gut, which is only poorly absorbed in the stomach and small
intestine. EA is largely metabolized by unidentified bacteria in
the intestinal lumen to produce urolithins. Urolithins are
putative metabolites produced by human (or animal) gut microflora
from ellagic acid, punicalagin (PA), punicalin (PB),
tellimagrandin (TL), and other ellagitannins through a series of
chemical modifications grouped into several pathways, giving rise
to many known urolithins. In terms of chemical structure,
urolithins are dibenzopyran-6-one derivatives with varying
hydroxyl substitution patterns. The processing of ellagic acid
begins with the loss of one of the two lactones present in ellagic
acid (lactonase/decarboxylase activity), and is followed by
optional removal of one or more hydroxyl groups (dehydroxylase
activities) and optional further reactions including methylation
and glycosylation.
[0010] More specifically, microbial metabolism of ellagic acid
starts in the small intestine, and the first metabolites produced
retain four phenolic hydroxyls (urolithin D, four hydroxyl groups;
see FIG. 1); these are further metabolized along the intestinal
tract to remove hydroxyl units leading to urolithin C (three
hydroxyls), urolithin A (two hydroxyls) and B (one hydroxyl) in
the distal parts of the colon. The absorbed metabolites are
conjugated with glucuronic acid (one or two units), and/or
methylated to form methyl ethers (e.g., when ortho-dihydroxyl
groupings are present). Urolithin A and B conjugates are the main
metabolites detected in plasma and urine, although some trihydroxy
derivatives (hydroxyl-UA) or EA-dimethyl ether glucuronide have
also been detected in smaller amounts. The
tetrahydroxy-urolithins, trihydroxy-urolithins, and EA derivatives
generally are not detected in peripheral plasma, but they are
absorbed in the small intestine and they are transported to the
liver where they are further metabolized and excreted with bile to
the small intestine, establishing an enterohepatic circulation
that is responsible for the relatively long life of urolithins in
plasma and urine.
[0011] Over the last twenty years many papers have appeared on the
biosynthesis, isolation, and biological activity of tannins,
especially ellagitannins. Access to pure ellagitannins by
isolation from natural sources may be cumbersome and yield only
relatively small quantities of pure natural products. See, for
example, Okuda et al. (1982) Chem. Pharm. Bull. 30: 4230-4233;
Okuda et al. (1982) Chem. Pharm. Bull. 30: 4234-4236. Methods are
known for total synthesis of many ellagitannins. See, for example,
Khanbabaee, K., Strategies for the synthesis of ellagitannins, In:
Chemistry and Biology of Ellagitannins, Ed. S. Quideau, World
Scientific Publishing, Singapore, 2009, pp. 152-202, including
references cited therein.
[0012] The development of a process-scale synthesis of urolithins
required substantial innovation. A useful process-scale synthesis
must be efficient, cost-effective, and reproducible. Further, all
starting materials and reagents must be reliably available in
bulk, or able to be produced on site in a safe and economical
fashion. The exacting regulatory standards for low impurity levels
and overall safety of the process create additional challenges to
development.
[0013] An Ullmann coupling is frequently used to couple the two
phenyl rings present in all of the urolithin compounds.
Unfortunately, the coupling routinely gives rise to product that
is unacceptably contaminated with copper. The product also varies
in color from batch to batch, from yellow to dark purple. Some
residual copper may be removed by column chromatography; however,
in process scale syntheses it is highly desirable to avoid column
chromatography, due to its expense and large waste stream.
Remarkably, improvements were made to a problematic Ullmann
coupling. By drastically reducing the amount of copper catalyst,
the isolated Ullmann coupling product consistently contained <1
ppm residual copper, and was off-white to light yellow in color.
Moreover, the need for column chromatography was avoided.
[0014] A second improvement relates to a demethylation reaction.
As discussed further in the Examples, the hydroxyl groups present
in urolithins are often protected as methyl, ethyl, or alkyl
ethers. Functionalizing the hydroxyl groups as ethers also allows
access to a variety of more lipophilic and potentially
more-bioavailable urolithin analogs. To allow access to the
natural urolithins, demethylation/dealkylation of the ethers must
be performed. This transformation has frequently been accomplished
on similar substrates with BBr3 (boron tribromide), a chemical
reagent associated with various hazards and drawbacks. Remarkably,
it was discovered that the powerful Lewis acid AlCl3 (aluminum
trichloride) can bring about the desired transformation in greater
than 40%, greater than 50%, greater than 60%, greater than 70%,
greater than 80%, greater than 85%, or even greater than 88%
yield. Subsequent hydrolysis of the excess AlCl3, filtration, and
a recrystallization provided the pure demethylated product
containing <17 ppm aluminum...
[0061] In certain embodiments, the copper-containing catalyst is
selected from the group consisting of copper powder, copper-bronze
couple, CuSO4 hydrate, anhydrous CuSO4, Cu(acac)2, CuCl, CuCl2,
CuBr, CuBr2, CuI, Cu2O, CuO, CuOTf, CuCN, and mixtures thereof….
[0065] In certain embodiments, the amount of copper-containing
catalyst is at least a trace amount but less than 0.05 equivalents
relative to either formula II or formula III.
[0071] In certain embodiments, the amount of copper-containing
catalyst is at least a trace amount but less than 0.0001
(1×10<−4>) equivalents relative to either formula II or
formula III.
[0072] In certain embodiments, the aqueous alkaline solvent
comprises LiOH, NaOH, KOH, CsOH, Na2CO3, CaCO3, or Cs2CO3.
[0073] In certain embodiments, the aqueous alkaline solvent
comprises NaOH or KOH.
[0074] In certain embodiments, the method is conducted at a
temperature from about 20° C. to about 180° C.
&c...
PRODRUGS OF UROLITIHNS AND USES THEREOF
WO2015097231
The invention provides compounds of formula (I) or salts thereof,
wherein: A, B, C, D, W, X, Y and Z are as defined in the
specification, and at least one of A, B, C, D, W, X, Y and Z is
OR1; each R1 being independently H or C(=O)R2, and at least one R1
group being C(=O)R2; where each R2 is selected from: CHR3NHR4,
where R4 is H and R3 is a group selected from CH3, CH2CH(CH3)2,
CH(CH3)CH2CH3, CH2Ph, CH2-3-(1H-indole), CH2CH2SCH3, CH2OH,
CHOHCH3, CH2SH, CH2SeH and CH2PhpOH, wherein said R3 group can
optionally be substituted by one or more groups selected from
halogen, cyano, nitro, ORA or C1-C4 alkyl; or R3 and R4 together
with the C and N atoms to which they are attached form a
5-membered heteroalkyl ring, wherein said heteroalkyl ring can
optionally be substituted by one or more groups selected from
halogen, cyano, nitro, ORA or C1-C3 alkyl, wherein RA is C1-C4
alkyl optionally substituted with one or more halogen, cyano or
nitro groups. The compounds are effective pro-drugs for urolithins
and they enable the ready delivery of urolithins to the site in
the digestive tract where they can be absorbed into the body.
Urolithin A Urolithin B
Urolithins have been proposed as treatments of a variety of
conditions related to inadequate mitochondrial activity, including
obesity, reduced metabolic rate, metabolic syndrome, diabetes
mellitus, cardiovascular disease, hyperlipidemia,
neurodegenerative diseases, cognitive disorder, mood disorder,
stress, and anxiety disorder; for weight management, or to
increase muscle performance or mental performance. See
WO2012/088519 (Amazentis SA). In WO2007/127263 (The Regents of the
University of California), the use of urolithins for the treatment
of various neoplastic diseases is described.
International patent publication no. WO2014/004902 (derived from
application PCT/US2013/48310) discloses a method of increasing
autophagy, including specifically mitophagy, in a cell, comprising
contacting a cell with an effective amount of a urolithin or a
pharmaceutically acceptable salt thereof, thereby increasing
autophagy, including specifically mitophagy, in the cell.
Administration may be to a subject having a disease or condition
selected from metabolic stress, cardiovascular disease,
endothelial cell dysfunction, sarcopenia, muscle degenerative
disease, Duchenne muscular dystrophy, alcoholic liver disease,
nonalcoholic fatty liver disease, drug-induced liver injury, a 1
-antitrypsin deficiency, ischemia/reperfusion injury,
inflammation, aging of the skin, inflammatory bowel disease,
Crohn's disease, obesity, metabolic syndrome, type II diabetes
mellitus, hyperlipidemia, osteoarthritis, neurodegenerative
disease, Alzheimer's disease, Huntington's disease, Parkinson's
disease, amyotrophic lateral sclerosis, age-related macular
degeneration, mitochondrial diseases (including for example poor
growth, loss of muscle coordination, muscle weakness, visual
problems, hearing problems, heart disease, liver disease, kidney
disease, gastrointestinal disorders, respiratory disorders,
neurological problems, autonomic dysfunction sometimes learning
disabilities, and dementia as a result of mitochondrial disease),
muscle diseases; sporadic inclusion body myositis (sIBM), cancer,
cognitive disorder, stress, and mood disorder…
Treatments using compounds of the invention
As mentioned above, the invention provides compounds of formula
(I) or (la) or salts thereof for use in the treatment of a disease
or condition selected from the group consisting of metabolic
syndrome, reduced metabolic rate, metabolic stress, cardiovascular
disease, sarcopenia, muscle degenerative disease, Duchenne
muscular dystrophy, alcoholic liver disease, nonalcoholic fatty
liver disease (NAFLD), Nonalcoholic steatohepatitis (NASH),
drug-induced liver injury, drug-induced cravings, anaemia
disorders, a 1 -antitrypsin deficiency, ischemia/reperfusion
injury, inflammation, inflammatory bowel disease, Crohn's disease,
obesity, metabolic syndrome, type II diabetes mellitus,
hyperlipidemia, osteoarthritis, neurodegenerative disease,
Alzheimer's disease, Parkinson's disease, Huntington's disease,
anxiety disorder, ulceration, amyotrophic lateral sclerosis,
mitochondrial diseases (including for example poor growth, loss of
muscle coordination, muscle weakness, visual problems, hearing
problems, heart disease, liver disease, kidney disease,
gastrointestinal disorders, respiratory disorders, neurological
problems, autonomic dysfunction sometimes learning disabilities,
and dementia as a result of mitochondrial disease. Further
diseases related to mitochondrial dysfunction include: Diabetes
mellitus and deafness (DAD); Leber's hereditary optic neuropathy
(LHON); Leigh syndrome (subacute sclerosing encephalopathy);
neuropathy, ataxia, retinitis pigmentosa, and ptosis (NARP);
myoneurogenic gastrointestinal encephalopathy (MNGIE); Myoclonic
Epilepsy with Ragged Red Fibers (MERRF);
Mitochondrial myopathy, encephalomyopathy, lactic acidosis,
stroke- like symptoms (MELAS); and mtDNA depletion), sporadic
inclusion body myositis (sIBM), and cancer, cognitive disorder,
stress, and mood disorder; for improving cognitive function; for
weight management; or to increase muscle or mental performance.
The compounds of formula (I) or (la) or salts thereof are
particularly suitable for use in improving muscle function, muscle
strength endurance and muscle recovery.
In particular, the invention provides compounds of formula (I) or
(la) or salts thereof for use in the treatment of a disease or
condition selected from the group consisting of metabolic
syndrome, reduced metabolic rate, metabolic stress, cardiovascular
disease, sarcopenia, muscle degenerative disease, Duchenne
muscular dystrophy, alcoholic liver disease, nonalcoholic fatty
liver disease, drug-induced liver injury, drug-induced cravings,
anaemia disorders, a 1 -antitrypsin deficiency,
ischemia/reperfusion injury, inflammation, inflammatory bowel
disease, Crohn's disease, obesity, metabolic syndrome, type II
diabetes mellitus, hyperlipidemia, osteoarthritis,
neurodegenerative disease, Alzheimer's disease, Parkinson's
disease, anxiety disorder, ulceration, amyotrophic lateral
sclerosis, and cancer, cognitive disorder, stress, and mood
disorder; for improving cognitive function; for weight management;
or to increase muscle or mental performance.
The invention further provides compounds of formula (I) or (la) or
a salt thereof for use in the treatment of a disease or condition
selected from the group consisting of metabolic stress,
sarcopenia, muscle degenerative disease, Duchenne muscular
dystrophy, alcoholic liver disease, nonalcoholic fatty liver
disease, drug-induced liver injury, a 1 -antitrypsin deficiency,
ischemia/reperfusion injury, inflammatory bowel disease, Crohn's
disease, Alzheimer's disease, Parkinson's disease, amyotrophic
lateral sclerosis, and cancer. The invention further provides
compounds of the formula (I) or (la) or salts thereof for
increasing autophagy or mitophagy in a cell. For example, the
autophagy or mitophagy may be in embryonic stem cells, induced
pluripotent stem cells, adult stem cells, differentiated cells,
blood cells, hematopoietic cells, epithelial cells, exocrine
cells, endocrine cells, connective tissue cells, adipose cells,
bone cells, smooth muscle cells, striated muscle cells, nerve
cells, sensory cells, cardiac cells, hepatic cells, gastric cells,
intestinal cells, pulmonary cells, epidermal (i.e. skin) cells
(including keratinocytes and fibroblasts), kidney cells, and germ
cells. It may thus for example treat or prevent a disease or
condition selected from the group consisting of metabolic
syndrome, reduced metabolic rate, metabolic stress, cardiovascular
disease, sarcopenia, muscle degenerative disease, Duchenne
muscular dystrophy, alcoholic liver disease, nonalcoholic fatty
liver disease, drug-induced liver injury, drug-induced cravings,
anaemia disorders, a 1 -antitrypsin deficiency,
ischemia/reperfusion injury, inflammation, inflammatory bowel
disease, Crohn's disease, obesity, metabolic syndrome, type II
diabetes mellitus, hyperlipidemia, osteoarthritis,
neurodegenerative disease, Alzheimer's disease, Parkinson's
disease, anxiety disorder, ulceration, amyotrophic lateral
sclerosis, and cancer, cognitive disorder, stress, and mood
disorder; or it can assist with weight management, or increase
muscle or mental performance.
Amongst the neurodegenerative diseases, there may specifically be
mentioned AIDS dementia complex, Alzheimer's disease, amyotrophic
lateral sclerosis, adreno leukodystrophy, Alexander disease,
Alper's disease, ataxia telangiectasia, Batten disease, bovine
spongiform encephalopathy (BSE), Canavan disease, corticobasal
degeneration, Creutzfeldt- Jakob disease, dementia with Lewy
bodies, fatal familial insomnia, frontotemporal lobar
degeneration, Huntington's disease, Kennedy's disease, Krabbe
disease, Lyme disease, Machado-Joseph disease, multiple sclerosis,
multiple system atrophy, neuroacanthocytosis, Niemann-Pick
disease, Parkinson's disease, Pick's disease, primary lateral
sclerosis, progressive supranuclear palsy, Refsum disease,
Sandhoff disease, diffuse my elino clastic sclerosis,
spinocerebellar ataxia, subacute combined degeneration of spinal
cord, tabes dorsalis, Tay-Sachs disease, toxic encephalopathy,
transmissible spongiform encephalopathy, and wobbly hedgehog
syndrome. In one embodiment, the neurodegenerative disease is
selected from the group consisting of Alzheimer's disease,
amyotrophic lateral sclerosis, Huntington's disease, and
Parkinson's disease. In one embodiment, the neurodegenerative
disease is Alzheimer's disease. An aspect of the invention is in
improving cognitive function. In one embodiment, the cognitive
function is selected from the group consisting of perception,
memory, attention, speech comprehension, speech generation,
reading comprehension, creation of imagery, learning, and
reasoning. In one embodiment, the cognitive function is selected
from the group consisting of perception, memory, attention, and
reasoning. In one embodiment, the cognitive function is memory.
An aspect of the invention is in the treatment of cognitive
disorder. In one embodiment, the cognitive disorder is selected
from the group consisting of delirium, dementia, learning
disorder, attention deficit disorder (ADD), and attention deficit
hyperactivity disorder (ADHD). In one embodiment, the cognitive
disorder is a learning disorder. In one embodiment, the cognitive
disorder is attention deficit disorder (ADD). In one embodiment,
the cognitive disorder is attention deficit hyperactivity disorder
(ADHD).
An aspect of the invention is in the treatment of stress-induced
or stress-related cognitive deficit. An aspect of the invention is
in the treatment of a mood disorder. In one embodiment, the mood
disorder is selected from the group consisting of depression,
postpartum depression, dysthymia, and bipolar disorder. In one
embodiment, the mood disorder is depression. In one embodiment,
the mood disorder is dysthymia.
An aspect of the invention is in the treatment of stress-induced
or stress-related mood disorder, e.g., dysthymia. An aspect of the
invention is in the treatment of an anxiety disorder. In one
embodiment, the anxiety disorder is selected from the group
consisting of generalized anxiety disorder, panic disorder, panic
disorder with agoraphobia, agoraphobia, social anxiety disorder,
obsessive-compulsive disorder, and post-traumatic stress disorder.
In one embodiment, the anxiety disorder is generalized anxiety
disorder. In one embodiment, the anxiety disorder is
post-traumatic stress disorder. An aspect of the invention is in
the treatment of stress-induced or stress-related anxiety.
An aspect of the invention is in enhancing muscle performance. In
one embodiment, the muscle performance is selected from the group
consisting of strength, speed, endurance and recovery. In humans,
muscle function generally declines with age starting during the
third decade of life; the decline generally accelerates after age
65. An aspect of the invention is thus in maintaining muscle
performance during the aging process. The enhancement of muscle
performance may be as part of the use of the compounds in sports
nutrition, in aiding healthy aging (for example from age 45 to
65), and in slowing the rate of muscle decline in those aged over
65 (pre-frail)
An aspect of the invention is in the treatment of a muscle or
neuromuscular disease. In one embodiment, the muscle or
neuromuscular disease is a myopathy. Ine one embodiment, the
muscle or neuromuscular disease is sarcopenia. In one embodiment,
the muscle or neuromuscular disease is sporadic inclusion body
myositis (sIBM). In one embodiment, the muscle or neuromuscular
disease is a muscular dystrophy. In one embodiment, the muscle or
neuromuscular disease is Duchenne muscular dystrophy.
An aspect of the invention is in the treatment of mitochondrial
disease. For example, a subject may require treatment of poor
growth, loss of muscle coordination, muscle weakness, visual
problems, hearing problems, heart disease, liver disease, kidney
disease, gastrointestinal disorders, respiratory disorders,
neurological problems, autonomic dysfunction sometimes learning
disabilities, and dementia as a result of mitochondrial disease.
Further diseases related to mitochondrial dysfunction include:
Diabetes mellitus and deafness (DAD); Leber's hereditary optic
neuropathy (LHON); Leigh syndrome (subacute sclerosing
encephalopathy); neuropathy, ataxia, retinitis pigmentosa, and
ptosis (NARP); myoneurogenic gastrointestinal encephalopathy
(MNGIE); Myoclonic Epilepsy with Ragged Red Fibers (MERRF);
Mitochondrial myopathy, encephalomyopathy, lactic acidosis,
stroke-like symptoms (MELAS); and mtDNA depletion.
Amongst cancers, there can specifically be mentioned solid
tumours, for example prostate cancer, pancreatic cancer and colon
cancer.
The effective amount of the compound will vary depending upon the
manner of administration, the age, body weight, and general health
of the subject. Factors such as the disease state, age, and weight
of the subject may be important, and dosage regimens may be
adjusted to provide the optimum response. A therapeutically
effective amount of the compound may for example range from about
0.001 to 30 mg/kg body weight, preferably about 0.01 to 25 mg/kg
body weight, more preferably about 0.1 to 20 mg/kg body weight,
and even more preferably about 1 to 10 mg/kg, 2 to 9 mg/kg, 3 to 8
mg/kg, 4 to 7 mg/kg, or 5 to 6 mg/kg body weight. Treatment may be
by way of a single treatment or, preferably, can include a series
of treatments. In one example, a subject is treated with the
compound in the range of between about 0.1 to 20 mg/kg body
weight, once per week for between about 1 to 10 weeks, preferably
between 2 to 8 weeks, more preferably between about 3 to 7 weeks,
and even more preferably for about 4, 5, or 6 weeks. It will also
be appreciated that the effective dosage of the compound may
increase or decrease over the course of a particular treatment...
Pomegranate Patents
Extracts --
ORAL FORMULATIONS FOR COUNTERACTING EFFECTS OF AGING
US2016067291
An oral formulation as described herein can comprise
pomegranate extract, panax ginseng extract, and c. sinensis, where
each is present in an amount effective to counteract and/or
prevent effects of aging in a subject when administered to the
subject. The effects of aging can include age-related changes in
gene expression.
POMEGRANATE FRUIT POLYPHENOL COMPOSITION AND METHODS OF USE
AND MANUFACTURE THEREOF
US9056083
A pharmaceutical composition with an active pharmaceutical
ingredient including a pomegranate fruit polyphenol extract. The
pomegranate fruit polyphenol extract includes at least about 3%
combined punicalagin A and punicalagin B by weight, less than
about 5% ellagic acid and their derivatives by weight, and less
than about 1% anthocyanins by weight.
Extracting method for effective components of pomegranate
leaves and obtained product
CN105232594
The invention discloses an extracting method for effective
components of pomegranate leaves and an obtained product. The
method comprises the steps that pomegranate leaves are thoroughly
washed and kneaded with salt; the pomegranate leaves are dried
through microwaves at 40-50 DEG C after kneading until the water
content is lower than 10%; the dried pomegranate leaves are
refrigerated for 5-10 min at minus 15 DEGC and then refrigerated
for 20-30 min at minus 5 DEG C; the refrigerated pomegranate
leaves are put into a heating container, compacted, subjected to
heat preservation for 0.5-1 h at 50-60 DEG C and then heated and
boiled, distilled-off components are collected, and the
distilled-off components are aqueous extraction of the pomegranate
leaves. The method is simple, easy to implement, clean and
efficient, the pretreatment measures of kneading with salt,
microwave drying and low-temperature refrigerating are adopted,
more effective components can be extracted, and it is proved
through tests that the obtained aqueous extraction of the
pomegranate leaves has good antimicrobial and anti-inflammatory
effects and can serve as raw materials of anti-inflammation
medicine.
Preparation method of pomegranate extracting liquid
CN105147546
The invention provides a preparation method of pomegranate
extracting liquid. The preparation method comprises the following
steps: squeezing fresh whole pomegranates to obtain fresh
pomegranate juice and fruit residues; adding water into the
obtained fruit residues and carrying out continuous countercurrent
extraction to obtain pomegranate residue liquid; mixing the
obtained fresh pomegranate juice and the obtained pomegranate
residue liquid; filtering to obtain the pomegranate extracting
liquid. According to the preparation method provided by the
invention, a process of combining spiral squeezing and a packing
auger type continuous countercurrent extraction method is adopted,
and the processes of peeling off the pomegranates, processing and
the like are omitted, so that the comprehensive utilization time
of the pomegranates is shortened and the yield of effective
components is improved; the preparation method makes important
contributions to operation simplification, energy saving, cost
saving and improvement of the stability of the extracting liquid.
Preparation method of whole pomegranate seed powder
CN105053877
The invention discloses a preparation method of whole
pomegranate seed powder. Peeled mature pomegranate fruits are
processed into the whole pomegranate seed powder through the steps
of carefully selecting pomegranate fruits as raw materials,
performing color protection on whole pomegranate seeds, mashing
the whole pomegranate seeds after the color protection so as to
obtain pomegranate fruit paste, quenching and tempering the
pomegranate fruit paste, and spraying and drying the quenched and
tempered pomegranate fruit paste. The whole pomegranate seed
powder prepared by the preparation method disclosed by the
invention is rich in nutrition, high in retention rate of
bioactive components, good in reconstituability, short in
rehydration time, and favorable of storage and transportation;
besides, the shelf life of the whole pomegranate seed powder can
be prolonged; the defects of large loss of active components and
low utilization rate of raw materials existing in the fruit powder
which is prepared by spraying and drying fresh pomegranate juice
or concentrated juice thereof are overcome, and a new way for
processing and utilizing pomegranates is developed.
Cultivation --
Special pomegranate tree liquid organic fertilizer and
preparation method thereof
CN105237184
The present invention provides a special pomegranate tree
liquid organic fertilizer preparation method, which comprises:
dissolving potassium dihydrogen phosphate, potassium nitrate and
urea in pure water to achieve a saturated solution, and mixing to
obtain a solution A; dissolving zinc sulfate, active boron,
ferrous sulfate, manganese sulfate, copper sulfate and ammonium
molybdate in pure water to achieve a saturated solution, and
mixing to obtain a solution B; dissolving a surfactant and a
composite aid in pure water to achieve a saturated solution, and
mixing to obtain a solution C; dissolving amino acid original
powder and potassium fulvate in pure water to achieve a saturated
solution, and mixing to obtain a solution D; mixing the solution B
and the solution C to obtain a first mixed solution; mixing the
solution D and the solution A to obtain a second mixed solution;
and mixing the first mixed solution and the second mixed solution,
and adding potassium carbonate so as to obtain the special
pomegranate liquid organic fertilizer. The present invention
further provides the special pomegranate tree liquid organic
fertilizer prepared through the preparation method. The special
pomegranate tree liquid organic fertilizer of the present
invention has the simple and easy ratio adjustment advantage, and
is the green organic fertilizer.
Composite fertilizer for increasing pomegranate yield
CN105237091
The invention provides a composite fertilizer for increasing
pomegranate yield. The composite fertilizer comprises the
following effective components in parts by mass: 60-80 parts of an
organic fertilizer, 5-10 parts of urea, 10-15 parts of
monopotassium phosphate, 10-18 parts of calcium superphosphate and
15-25 parts of plant ash. Due to adoption of the composite
fertilizer, organic matters in soil can be supplemented,
micro-ecological environment can be improved, and the defect that
the fertilizer efficiency of an organic fertilizer is slow to take
into effect can be made up. Meanwhile the yield of crops can be
increased. As the organic fertilizer is cheap and easily
available, the cost of the composite fertilizer is also lowered.
Cultivation method of pomegranate trees and
alliumjaponicurn regel at hilly arid area and application
CN105052652
The invention, which belongs to the technical field of
agriculture growing, discloses a cultivation method of pomegranate
trees and alliumjaponicurn regel at a hilly arid area and
application. According to the method, a natural rain path is
improved and a cistern is built; a terraced filed at a hilly arid
area is modified into wave-shaped farmland; a hole bottom and the
ground are covered by plastic films and pomegranate trees are
planted in a seepage-proof and moisture preservation mode, and
alliumjaponicurn regel is planted also in an intercropping mode;
and then filed intercropping management is carried out on the
pomegranate trees and alliumjaponicurn regel. According to the
invention, the provided method is superior to the existing method
obviously in terms of cultivation management; the yield can be
improved; the gains can be increased on the condition that the
management cost is not increased; and objectives of water saving,
labor saving, and high yield and benefit increasing can be
achieved. The method and application are suitable for cultivation
of pomegranate trees and alliumjaponicurn regel at a hilly arid
area.
Method for cultivating pomegranate seedlings in
saline-alkali soil
CN105028107
The invention discloses a method for cultivating pomegranate
seedlings in saline-alkali soil. The method comprises three steps
for improving the saline-alkali soil, namely 1, phosphorus gypsum
powder is applied to the saline-alkali soil for improving the
saline-alkali soil; 2, organic fertilizer, mushroom residue and
oil cakes are applied to the saline-alkali soil for improving the
saline-alkali soil; 3, wild sweet potatoes are planted in the
saline-alkali soil for improving the saline-alkali soil. By means
of the method, the soil structure of the saline-alkali soil can be
improved effectively, the soil pH can be reduced effectively, the
saline-alkali soil is made to be suitable for the growth of the
pomegranate seedlings, and therefore the pomegranate seedlings can
be cultivated on the saline-alkali soil. The method has a
remarkable effect especially for improving the moderate
saline-alkali soil, the saline-alkali soil can be improved
completely, the pomegranate seedlings can be cultivated by
utilizing the saline-alkali soil, and the remarkable economic
benefit is obtained.
Pomegranate fresh-keeping method
CN104970096
The invention provides a pomegranate fresh-keeping method
which includes the steps of (1) collecting matured pomegranates,
selecting health fresh fruits of the pomegranates; (2) coating the
surface of the fresh fruits of the pomegranates with a film of a
chitosan solution being 0.5-1.0% in mass concentration; (3)
naturally drying the pomegranate fresh fruits coated with the
chitosan; (4) packaging the naturally-dried pomegranate fresh
fruits respectively into PE bags on which a plurality of holes are
formed; and (5) storing the pomegranates at 4-6 DEG C under the
relative humidity being 90-95%. The pomegranate fresh-keeping
method can prolong preservation time of the fresh fruits of the
pomegranates, allows off-season consumption, can increase the
economic benefit of the pomegranates, is simple in process, is
convenient to carry out, and is controllable in preservation
scale.
High yield cultivating method of pomegranates
CN104969814
The invention relates to the technical field of plant
planting, particularly to a high yield cultivating method of
pomegranates. The high yield cultivating method comprises the
following steps of: (1) preparing a seedling raising matrix; (2)
raising seedlings by cutting of short shoots, (3) transplanting
the seedlings; (4) managing the transplanted seedlings; and (5)
managing the seedlings at a fruiting period and seedlings after
pomegranate fruits are picked up, wherein a culturing matrix is
weighed in parts by weight: 30-50 parts of poultry dung, 20-40
parts of turf, 10-30 parts of distillers' grains, 12-24 parts of
vermiculite, 8-16 parts of perlite, and 2-5 parts of micro
elements. Carbendazol is used for disinfecting the seedling
raising matrix, and then the seedlings are raised, wherein the
consumption of the carbendazol is 90-120g/m<3>. Through the
high yield cultivating method of pomegranates, disclosed by the
invention, the survival rate is high, the growing trend of the
plants is quick, neat and consistent, the cycle of fruit setting
is short, the yield is high, and the economic returns are
considerable.
Leaf fertilizer special for pomegranate
CN104961607
he invention relates to a leaf fertilizer special for pomegranate.
The leaf fertilizer is composed of raw materials including, by
weight, 1-20 parts of isoleucine, 8-38 parts of proline, 1-18
parts methionine, 1-20 parts of cysteine, 8-30 parts of
asparaginate, 0.1-0.8 part of glutamine, 10-100 parts of water,
0.8-2 parts of salicylic acid, 1-28 parts of abscisic acid and
8-20 parts of ammonium bicarbonate. The leaf fertilizer promotes
crop growth, reduces the pesticide application amount, and is low
in raw material cost and capable of preventing insect disease.
Formula of organic inorganic high-efficiency and safe bulk
blended fertilizer specially used for pomegranate tree
CN104945036
The invention discloses a formula of an organic inorganic
high-efficiency and safe bulk blended fertilizer specially used
for a pomegranate tree. The formula is obtained by scientifically
preparing according to a 'wooden barrel law' principle of balanced
fertilization and a fertilizer requirement law that the
pomegranate tree needs nitrogen fertilizer all the time from
germination to a fruit swelling period and needs more phosphorus
and potassium from a flowering period to a harvesting period, when
every 1000kg of the bulk blended fertilizer is produced, 400kg of
urea, 200kg of monoammonium phosphate, 200kg of potassium
chloride, 150kg of organic fertilizer and 50kg of calcium,
magnesium and phosphorus fertilizer are used, stirred and blended,
so that the organic inorganic high-efficiency and safe bulk
blended fertilizer, specially used for the pomegranate tree,
meeting the requirements that ratio of three elements, namely
nitrogen, phosphorus and potassium is 23:9:12, total nutrient
accounts for 44%, medium trace element accounts for 4% and organic
matter, amino acid and humic acid account for 6% is prepared. The
formula of the organic inorganic high-efficiency and safe bulk
blended fertilizer specially used for the pomegranate tree
contains all the nutrients such as macro elements (nitrogen,
phosphorus and potassium), medium elements (calcium, magnesium and
sulphur) and trace elements (copper, iron, zinc, manganese,
molybdenum, chlorine and boron) as well as organic matter, amino
acid and humic acid and has the advantages of high efficiency,
safety, low cost and strong pertinence.
Seedling-raising substrate preventing pomegranate seedling
root rot
CN104926553
The invention provides a seedling-raising substrate preventing
pomegranate seedling root rot, and relates to the technical field
of pomegranate seedling raising. The substrate is prepared from
the raw materials of, by weight, 30 parts of sandy soil, 15 parts
of humus, 10 parts of wheat bran, 8 parts of soybean meal, 12
parts of corn flour, 1.5 parts of raw gypsum, 10 parts of
vermiculite, 12 parts of diatomaceous earth, 6 parts of plant ash,
18 parts of fermented mud, 5 parts of edible fungus strain, 3
parts of quispualis indica, 2.5 parts of cyrtomium fortunei, 3
parts of radix euphorbiae lantu, 7 parts of chinaberry seed, 15
parts of golden cypress, 10 parts of ash bark, 4 parts of alum, 6
parts of fructus cnidii, and 2 parts of honeysuckle flower. The
substrate has the advantages of high propagation speed, high
germination rate, and effective pest and disease inhibition
performance. With the substrate, pomegranate seedling root rot can
be reduced, and a good condition for the seedlings to thrive is
provided. With the substrate, transplanting survival rate is high.
The seedling-raising substrate has good breathability, and is
suitable for pomegranate seedling growth. The substrate is rich in
nutrients. With the substrate, pomegranate seedling growth speed
is high.
Bio-enzyme organic compound fertilizer for pomegranate
CN104926468
The invention relates to a bio-enzyme organic compound
fertilizer for pomegranate. The bio-enzyme organic compound
fertilizer is characterized by comprising the following components
in parts by weight: 16-20 parts of serine protease, 36-66 parts of
thiol protease, 16-31 parts of isoleucine, 16-31 parts of proline,
16-31 parts of aspartic protease, 16-31 parts of isoaspartic
protease, 2-6 parts of protease, 7-16 parts of vinyl ethyl ether,
7-16 parts of ethyl glycollate and 6-10 parts of solanum nigrum.
The bio-enzyme organic compound fertilizer has the advantages that
the ecological agriculture target is realized, and large-scale
agriculture is on the way to virtuous cycle.
Seed treating fluid for preventing pomegranate seedlings
from rotted roots
CN104926429
The invention provides a seed treating fluid for preventing
pomegranate seedlings from rotted roots, and relates to the
technical field of seedling growing of pomegranate. The seed
treating fluid is prepared from the following raw materials in
parts by weight: 1500g of fermentation liquor, 1.2mg of auxin IBA,
3g of radix stemonae, 2g of cyrtomium fortune, 1.5g of radix
euphorbiae lantu, 2.5g of Chinaberry seeds, 3g of golden cypress,
2g of roots of Chinese pulsatilla and 3.5g of honeysuckle flowers,
wherein the fermentation liquor is a liquid prepared by fermenting
45 parts of pig manure, 280 parts of water, 15 parts of corn
stalks and 15 parts of tobacco stems in a fermentation tank. The
seed treating fluid is high in reproduction speed, high in
germination rate, capable of effectively restraining plant
diseases and insect pests and reducing rotted root conditions of
the pomegranate seedlings and providing a condition for the
seedlings to grow strongly and healthily; the provided seedling
cultivation medium is good in air permeability, suitable for the
pomegranate seedlings to grow and rich in nutrient, so that the
pomegranate seedlings are high in growth speed; the treating fluid
for treating pomegranate seeds has germination accelerating effect
on seeds, and can be used for restraining the seed surfaces from
growing mildew and reducing occurrence of plant diseases and
insect pests, so that the survival rate of the pomegranate
seedlings is high.
Special long-acting slow-release fertilizer for pomegranate
CN104909921
The invention discloses a special long-acting
slow-release fertilizer for pomegranate, which is composed of the
following components in parts by weight: 6-10 parts of
diethylaminoethyl cellulose, 5-9 parts of naphthyl diisocyanate,
4-8 parts of butylated hydroxytoluene, 6-11 parts of
dimethylphenyl phosphate, 10-20 parts of polyethyleneglycol, 3-7
parts of BCG, 2-8 parts of Albizzia julibrissin, 1-5 parts of
borneol, 3-8 parts of red paeonia, 3-5 parts of reed root, 2-7
parts of asafetida, 6-8 parts of fried shallot, 8-14 parts of
alanine, 1-2 parts of serine, 1-4 parts of acyl peptide hydrolase
and 2-4 parts of slow-release agent. The special long-acting
slow-release fertilizer for pomegranate can supply nutritional
ingredients required by pomegranate for a long time, and enhances
the soil fertility due to the addition of the microbial inoculant
and organic fertilizer.
Cultivation method for pomegranate seedlings
CN104855214
The invention provides a cultivation method for pomegranate
seedlings, relating to the technical field of
pomegranate-seedling. The method comprises following steps:
selecting seeds fully and completely filled with grains, putting
and soaking the seeds into a treating fluid for 8 hours till the
seeds are completely soaked; sowing soaked seeds onto a seeding
bed. The seeding substrate laid on the seeding bed is
25-centimeter thick. Soil is covered to the thickness of 3
centimeters. Suppressing and watering operation are performed
after sowing. The depth of water soaking into the seeding
substrate is below the seeds for 2 to 3 centimeters. The
cultivation method for the pomegranate seedlings has following
beneficial effects: the cultivation method is featured by high in
reproduction speed and germination capacity and is capable of
effectively inhibiting diseases and insect pests and provides a
condition for thriving seedlings with high survive rate after
transplantation; the seeding substrate has fine air permeability
and is suitable for growth of growing-fast pomegranate seedlings;
the treating fluid used for treatment of pomegranate seeds has
pregermination effect upon the seeds; mycete of seed surfaces is
avoided; occurrences of diseases and insect pests are reduced; and
high survive rate of the pomegranate seedlings is obtained.
Pomegranate blooming-stage cultivation method
CN104429778
The invention provides a pomegranate blooming-stage
cultivation method which comprises the following steps: (1)
trimming flowers: clearing away bell-shaped flowers, keeping
tubular flowers and gourd-shaped flowers, arranging four foliage
branches around each fruit branch with bell-shaped flowers, and
spraying a growth regulator at an early blooming stage and a full
blooming stage; (2) trimming fruit: clearing away the third batch
of fruit, keeping the first batch of fruit and the second batch of
fruit, and clearing away and deeply burying diseased fruit, wormed
fruit and malformed fruit; (3) topdressing: in the full-bearing
period, applying urea to resulting trees and young trees, spraying
leaf fertilizer to leaves, and conducting watering, intertillage
for loosening the soil and weed uprooting after topdressing; (4)
trimming branches: girdling the branches which are more than 8 cm
in thickness, and cutting off too dense branches germinating in
spring and twigs germinating in summer; (5) pollination: releasing
honeybees at the early blooming stage and the full blooming stage,
gathering pollen-flying fruitless flowers every day, removing the
sepals and the petals of the fruitless flowers to expose anthers,
and point-pollinating open flower stigmas; (6) disease and pest
control. The pomegranate blooming-stage cultivation method can
improve the quality and the yield of fruit.
Cultivation method of polyploid pomegranate
CN103392606
The invention relates to a cultivation method of polyploid
pomegranate. The cultivation method comprises following steps:
selecting branched flower buds and branches which grow on the top
of a diploid plant, inducing the flower to generate polyploid
pollen by dropwise adding chemical materials into the flower buds,
carrying out hybridizations between induced male flowers and
normal female flowers or between induced female flowers and normal
male flowers so as to obtain polyploid hybrid fruits, culturing
the hybrid seeds in the hybrid fruits so as to obtain polyploid
plants, and culturing the polyploid plants in a tissue cultivation
mode so as to obtain polyploid pomegranate plants. The cultivation
method overcomes the shortage that a large amount of time is
consumed by the conventional chromosome reduplication methods. The
cultivation method utilizes the fact that the pollen and ovule
cells in the reproductive organs are haploids, namely 1n
chromosome, during the pomegranate blossom period, uses chemical
materials to process the pollen and ovule cells to induce
chromosome doubling, thus largely reduces the time of chromosome
doubling process, and has the advantages of available materials,
simple operation, and high success rate.
Preparation method for high-yield cultivation fertilizer
for pomegranate trees
CN102807427
The invention relates to a preparation method for high-yield
cultivation fertilizer for pomegranate trees. The high-yield
cultivation fertilizer consists of the following components in
percentage by weight: dry cow manure, dry chicken manure, dry duck
manure, phosphate fertilizer, zinc fertilizer, composite
fertilizer, bone meal, benomyl, procymidone, triazolone, Bordeaux
liquid, broad-spectrum bactericide and midothane wettable powder.
The high-yield cultivation fertilizer has low cost, readily
available materials and a simple preparation method. The growth
requirements of the pomegranate trees can be met by a large number
of beneficial microorganisms and organic fertilizer generated in
the fertilizer. The pomegranate has large volume, sufficient
weight, high growth speed, delicious taste and rich nutrients. The
yield of the pomegranate trees is increased and the defects that
the pomegranate has small volume, light weight and low growth
speed and is in illness easily are overcome.
Method for grafting pomegranate in summer
CN102763563
A method for grafting pomegranate in summer relates to the
technical field of pomegranate cultivation, and comprises the
steps of selecting appropriate rootstocks, cutting wood and cions,
cutting the rootstocks, grafting the scions and the rootstocks and
then binding membranes outside the well grafted cions and
rootstocks. The method provided by the invention has the benefits
that the restriction on the pomegranate grafting season is broken,
so that the pomegranate can be grafted and reproduced even in
summer, and the problem of difficulty in grafting the pomegranate
in summer is effectively solved. The method is simple to operate,
improves the survival rate and ensures both production increase
and income increase of the pomegranate.
Frame type cultivation method for pomegranates
CN102715098
The invention discloses a frame type cultivation method for
pomegranates and belongs to the field of fruit cultivation. The
method comprises adopting sandy loam to serve as a matrix of
nursery, enabling plant line spacing for cultivating fixed
planting to be 2m*3m, enabling one column to be erected between
every two pomegranate trees, respectively drawing steel wires with
the heights of 0.7-0.9m and 0.9-1.1m between adjacent columns,
enabling two steel wires which are 0.9-1.1m high to be
horizontally arranged and respectively distributed on two sides of
the columns. An arbitrary pomegranate can be arranged in a
quadrilateral area enclosed by a steel wire, branches can be bound
according to the need, the problem that the branches are bent due
to fruiting can be solved, or attractiveness of the pomegranates
is improved by binding modeling. The frame type cultivation method
is simple and practical, effectively controls growing density of
the pomegranates and guarantees sunshine supply.
Method for extracting natural flocculation active
ingredients from pomegranate bark and method for preparing
flocculating agent
CN105129948
The invention relates to a method for extracting natural
flocculation active ingredients from pomegranate bark and a method
for preparing a flocculating agent. The method for extracting the
natural flocculation active ingredients from the pomegranate bark
includes the steps that 1, the pretreated pomegranate bark and
ultra-pure water or distilled water or cooled boiling tap water
are mixed according to the mass-volume ratio of 1:33.33-1:2.78; 2,
particle residues in leaching liquor are filtered away to obtain a
water extracting solution of the pomegranate bark. The method for
preparing the flocculating agent by extracting the natural
flocculation active ingredients from the pomegranate bark includes
the steps that 1, the pretreated pomegranate bark and the
ultra-pure water or the distilled water or the cooled boiling tap
water are mixed according to the mass-volume ratio of
1:33.33-1:2.78; 2, the particle residues in the leaching liquor
are filtered away to obtain the water extracting solution of the
pomegranate bark; 3, 0.5-10 mL of the water extracting solution of
the pomegranate bark is taken and diluted to 30 mL through the
ultra-pure water or the distilled water or the cooled boiling tap
water, and then the final flocculating agent is obtained.
Cultivation process for organic pomegranate
CN102726268
The invention discloses a cultivation process for an organic
pomegranate. The cultivation process comprises field planting
preparing, field planting, field drying, trimming and pruning,
field managing, irrigating and filling water, bagging for
vegetables and fruits, and disease and insect controlling. The
cultivation process has a rational management specification, can
rationally number and record milpa, plants, fertilization,
irrigation, disease and insect control, pesticide usage time,
harvesting date, planting variety, planting number, planting
method, and the like; on the disease and insect control aspect,
agricultural control, physical control and biological control are
combined to effectively prevent diseases and insects, and a
self-made organic fertilizer is adopted to fertilize, so that the
fertilizer cost is saved, and the pollution of the organic
fertilizer material to the environment is also reduced.
DESCRIPTION
The present invention discloses a process for organic pomegranate
cultivation, including planting preparation, planting, will be
dry, plastic trim, field management, irrigation and drainage
water, fruit thinning and bagging pest control. The present
invention is a reasonable management practices in the cultivated
land, plants, fertilization, irrigation, pest prevention,
pesticide use of time and date of harvest, cultivation,
cultivation quantity, cultivation methods so that a reasonable
number is recorded; in pest control on, the principle of
agricultural control, physical and biological control combination,
can effectively control pests and diseases; the use of homemade
organic fertilizer in fertilization, saving fertilizer costs, but
also reduce the pollution of organic fertilizer raw materials to
the environment.
Background technique
Pomegranate pomegranate pomegranate families genus are deciduous
shrubs or small trees, crown plexiform natural round shape, Roots
brown, barren and more resistant to drought, fear of waterlogging,
water demand growth season very much. Ripe pomegranate rind color
Bright red or pink, often split, exposing the glistening
jewel-like as grain, sweet and juicy, very loved by the people.
Pomegranate cultivation but extensive management, low yields, such
as fertilizer can not keep up, trim or not to trim unreasonable,
Pest control is not timely unreasonable.
SUMMARY
Object of the present invention to overcome the deficiencies of
the prior art, there is provided a process for the organic
cultivation of pomegranate.
To achieve the above object, the present invention adopts the
following technical scheme:
An organic pomegranate cultivation techniques, including planting
preparation, planting, will be dry, plastic trim, field
management, Row irrigation, fruit thinning and bagging pest
control, in particular:
Planting Preparation: Setting line spacing and spacing according
to terrain, pull dig planting pits, and then mixing organic
fertilizer pit Soil, backfill compaction;
Colonization: straighten out the seeds, cut long roots, implanted
pit backfill and fine soil, light pinch back mention seed Fill, do
a tree plate along seed around, planting pouring enough water,
cover and cover;
Set dry: after spring planting of seedlings after germination of
new branches, cut the main branch, culture Celi branches, forming
crown;
Plastic trim: Maintain the natural ecological crown, layered
trimming, sawing off excess closely spaced branches, combined Pull
support, vertical pressure, the crown form a variety of forms;
Field management: implementation of number management and
fertilization management;
Irrigation and drainage water: Shupan ahead, irrigation, opened at
the top of the cover Shupan irrigation, irrigation cover On the
cover;
Thinning bagging: fruit be firmly secured, timely thinning,
thinning Been dense fruit, put the fruit fruit bags;
Pest control: the agricultural control, physical control and
biological pest control were combined.
Further technical solution is the spacing of 3 to 3.5 meters, the
spacing of 2 to 3 meters, the Planting hole 70 cm deep wide 60㎝,
if cultivated for the slopes, the planting of 100 to 120 per acre,
if cultivated land To the ground, then planted per acre from 50 to
70, the use of organic manure pit every 50 kg.
Further technical solution, said Shupan 40㎝ wide, 20 cm deep, the
covering comprises a plastic film, Straw in one or two.
Further technical solution is the second cut in the main branch
refers to Jude Biao cut in 30 to 40 centimeters Go to the main
branch, the branch training Celi 2-3.
Further technical solution is the use of weak individual
hierarchical pruning prune and cultivate young branches empty
chamber, Use topping foster long put crown, formed results Zhizu;
crown is too large, heavy shears retracted, do not cut in line
with scissors In addition, do not cut scissors yin yang, upright
branches cut, do not cut the level of its side branches, cut
overlapping interference sticks, do not cut the branches empty
chamber Principle; the variety of forms including hollow tower,
fan, umbrella shape.
Further technical solution is that the number of project
management including planting, the plants, fertilization,
irrigation Water, anti-pest, pesticide use time and date of
harvest, cultivation, cultivation quantity, cultivation methods.
Further technical solution is the fertilization management
projects include promoting fertilizers and reminders fruit
fertilizer, promote the Fertilizers pomegranate harvest in late
September, the organic fertilizer 800 ~ 1500kg / mu, homemade
bio-organic fertilizer 200~ 300kg / acre, the pro-fertilization
methods fertilizers are digging peripheral crown 40 to 50 cm wide
and deep radial Ditch, promoting fertilizers and soil pit mixed
uniformly applied into the trench, irrigation 2kg casing tramping;
the fertilizer in the fruit catalyst 5 In late May or early June
after the fruit firmly secured, reminders fruit fertilizer
applied, depending on the vigor fertilization, the results of the
amount of self-administered Health Organic fertilizer was 120 ~
200kg / acre.
Further technical solution, said irrigation per plant 150 ~ 200kg
/ times, 25 to 30 days watering once.
Further technical solution is the fruit firmly secured in late May
to mid-June on, the thinning Holding fruit leaves more than 300 to
400 leaves / fruit, fruit from 30 to 40 cm / fruit.
Further technical solution that includes the agricultural control
fertilization and irrigation, thinning, drainage groove, Pruning,
remove leaves and dig garden pests, including the use of physical
control of the black light trap phototaxis Moths, insect repellent
planting insecticidal chrysanthemum, including the protection of
natural enemies of the biological control, promote the use of
biological Pesticides, botanical pesticides, mineral pesticides
pest control...
Example 1
An organic pomegranate cultivation techniques, including planting
preparation, planting, will be dry, plastic trim, field
management, Row irrigation, fruit thinning and bagging pest
control, in particular:
Planting Preparation: Setting line spacing and spacing according
to terrain, pull dig planting pits, and then mixing organic
fertilizer pit Soil, backfill compaction;
Colonization: straighten out the seeds, cut long roots, implanted
pit backfill and fine soil, light pinch back mention seed Fill, do
a tree plate along seed around, planting pouring enough water,
cover and cover;
Set dry: after spring planting of seedlings after germination of
new branches, cut the main branch, culture Celi branches, forming
crown;
Plastic trim: Maintain the natural ecological crown, layered
trimming, sawing off excess closely spaced branches, combined Pull
support, vertical pressure, the crown form a variety of forms;
Field management: implementation of number management and
fertilization management;
Irrigation and drainage water: Shupan ahead, irrigation, opened at
the top of the cover Shupan irrigation, irrigation cover On the
cover;
Thinning bagging: fruit be firmly secured, timely thinning,
thinning Been dense fruit, put the fruit fruit bags;
Pest control: the agricultural control, physical control and
biological pest control were combined.
Further technical solution is the spacing of 3 m, the spacing is 2
meters, width of the planting pit 60Cm 70 cm deep, if cultivated
for the slopes, the 110 acre planted, if the ground for
cultivation, the per acre Planting 63, the organic fertilizer
(composting manure) pit per 50 kg.
Further technical solution, said Shupan 40㎝ wide, 20 cm deep, the
covering comprises a plastic film, Straw in one or two.
Further technical solution is to cut off the main branch of the
second refers to the main Jude Biao cut 30 centimeters Branch, the
branch training two Celi.
Further technical solution is the use of weak individual
hierarchical pruning prune and cultivate young branches empty
chamber, Use topping foster long put crown, formed results Zhizu;
crown is too large, heavy shears retracted, do not cut in line
with scissors In addition, do not cut scissors yin yang, upright
branches cut, do not cut the level of its side branches, cut
overlapping interference sticks, do not cut the branches empty
chamber Principle; the variety of forms including hollow tower,
fan, umbrella shape.
Further technical solution is that the number of project
management including planting, the plants, fertilization,
irrigation Water, anti-pest, pesticide use time and date of
harvest, cultivation, cultivation quantity, cultivation methods.
Further technical solution is the fertilization management
projects include promoting fertilizers and reminders fruit
fertilizer, promote the Fertilizers in late September after the
harvest pomegranate, organic manure (composting manure) 800kg /
acre, homemade bio Organic fertilizer 200kg / acre (10 kg or
organic fertilizer / plant, bio-organic fertilizer made 2 kg /
plant), the pro-flower Fertilization fertilizer is a peripheral
crown dig deep 40㎝ wide radial grooves, after promoting
fertilizers and soil mix evenly applied into the pit Trench,
irrigation 2kg casing tramping; the fertilizer is urging fruit in
late May or early June fruit firmly secured, Shi reminders fruit
fertilizer, depending on the vigor fertilization, the results of
the amount of applied homemade bio-organic fertilizer 120kg / acre
(or self-administered Health Of organic fertilizer 1.5kg /
strain).
Further technical solution, said irrigation per plant 150kg /
times, 25 days irrigation once.
Further technical solution is the fruit firmly secured in late May
to mid-June on, the thinning Holding more than 300 fruit leaves
leaf / fruit, fruit from 30 cm / fruit.
Further technical solution that includes the agricultural control
fertilization and irrigation, thinning, drainage groove, Pruning,
remove leaves and dig garden pests, including the use of physical
control of the black light trap phototaxis Moths, insect repellent
planting insecticidal chrysanthemum, including the protection of
natural enemies of the biological control, promote the use of
biological Pesticides, botanical pesticides, mineral pesticides
pest control.
Example 2
An organic pomegranate cultivation techniques, including planting
preparation, planting, will be dry, plastic trim, field
management, Row irrigation, fruit thinning and bagging pest
control, in particular:
Planting Preparation: Setting line spacing and spacing according
to terrain, pull dig planting pits, and then mixing organic
fertilizer pit Soil, backfill compaction;
Colonization: straighten out the seeds, cut long roots, implanted
pit backfill and fine soil, light pinch back mention seed Fill, do
a tree plate along seed around, planting pouring enough water,
cover and cover;
Set dry: after spring planting of seedlings after germination of
new branches, cut the main branch, culture Celi branches, forming
crown;
Plastic trim: Maintain the natural ecological crown, layered
trimming, sawing off excess closely spaced branches, combined Pull
support, vertical pressure, the crown form a variety of forms;
Field management: implementation of number management and
fertilization management;
Irrigation and drainage water: Shupan ahead, irrigation, opened at
the top of the cover Shupan irrigation, irrigation cover On the
cover;
Thinning bagging: fruit be firmly secured, timely thinning,
thinning Been dense fruit, put the fruit fruit bags;
Pest control: the agricultural control, physical control and
biological pest control were combined.
Further technical solution is the spacing of 3.5 m, the spacing is
3 m, the planting pit 60㎝ deep 70 cm wide, if cultivated for the
slopes, the 100 acre planted, if the ground for cultivation, each
Planting 50 acres, the organic fertilizer (composting manure) pit
per 50 kg.
Further technical solution, said Shupan 40㎝ wide, 20 cm deep, the
covering comprises a plastic film, Straw in one or two.
Further technical solution is to cut off the main branch of the
second refers to the main Jude Biao cut 40 centimeters Branch, the
branch training Celi 3.
Further technical solution is the use of weak individual
hierarchical pruning prune and cultivate young branches empty
chamber, Use topping foster long put crown, formed results Zhizu;
crown is too large, heavy shears retracted, do not cut in line
with scissors In addition, do not cut scissors yin yang, upright
branches cut, do not cut the level of its side branches, cut
overlapping interference sticks, do not cut the branches empty
chamber Principle; the variety of forms including hollow tower,
fan, umbrella shape.
Further technical solution is that the number of project
management including planting, the plants, fertilization,
irrigation Water, anti-pest, pesticide use time and date of
harvest, cultivation, cultivation quantity, cultivation methods.
Further technical solution is the fertilization management
projects include promoting fertilizers and reminders fruit
fertilizer, promote the Fertilizers in late September after the
harvest pomegranate, organic manure (composting manure) 1500kg /
mu, homemade bio Organic fertilizer 300kg / acre (15 kg or organic
fertilizer / plant, bio-organic fertilizer made 3 kg / plant), the
pro-flower Fertilization fertilizer is a peripheral crown dig deep
40㎝ wide radial grooves, after promoting fertilizers and soil mix
evenly applied into the pit Trench, irrigation 2kg casing
tramping; the fertilizer is urging fruit in late May or early June
fruit firmly secured, Shi reminders fruit fertilizer, depending on
the vigor fertilization, the results of the amount of applied
homemade bio-organic fertilizer 200kg / acre (or self-administered
Health Of organic fertilizer 2.5kg / strain).
Further technical solution, said irrigation per plant 200kg /
times, 30 days irrigation once.
Further technical solution is the fruit firmly secured in late May
to mid-June on, the thinning Holding more than 400 fruit leaves
leaf / fruit, fruit from 40 cm / fruit.
Further technical solution that includes the agricultural control
fertilization and irrigation, thinning, drainage groove, Pruning,
remove leaves and dig garden pests, including the use of physical
control of the black light trap phototaxis Moths, insect repellent
planting insecticidal chrysanthemum, including the protection of
natural enemies of the biological control, promote the use of
biological Pesticides, botanical pesticides, mineral pesticides
pest control.
Example 3
An organic pomegranate cultivation techniques, including planting
preparation, planting, will be dry, plastic trim, field
management, Row irrigation, fruit thinning and bagging pest
control, in particular:
Planting Preparation: Setting line spacing and spacing according
to terrain, pull dig planting pits, and then mixing organic
fertilizer pit Soil, backfill compaction;
Colonization: straighten out the seeds, cut long roots, implanted
pit backfill and fine soil, light pinch back mention seed Fill, do
a tree plate along seed around, planting pouring enough water,
cover and cover;
Set dry: after spring planting of seedlings after germination of
new branches, cut the main branch, culture Celi branches, forming
crown;
Plastic trim: Maintain the natural ecological crown, layered
trimming, sawing off excess closely spaced branches, combined Pull
support, vertical pressure, the crown form a variety of forms;
Field management: implementation of number management and
fertilization management;
Irrigation and drainage water: Shupan ahead, irrigation, opened at
the top of the cover Shupan irrigation, irrigation cover On the
cover;
Thinning bagging: fruit be firmly secured, timely thinning,
thinning Been dense fruit, put the fruit fruit bags;
Pest control: the agricultural control, physical control and
biological pest control were combined.
Further technical solution is the spacing of 3 m, the spacing is 2
meters, width of the planting pit 60Cm 70 cm deep, if cultivated
for the slopes, the 120 acre planted, if the ground for
cultivation, the per acre Planting, the organic fertilizer
(composting manure) pit per 50 kg.
Further technical solution, said Shupan 40㎝ wide, 20 cm deep, the
covering comprises a plastic film, Straw in one or two.
Further technical solution is to cut off the main branch of the
second refers to the main Jude Biao cut 30 centimeters Branch, the
branch training two Celi.
Further technical solution is the use of weak individual
hierarchical pruning prune and cultivate young branches empty
chamber, Use topping foster long put crown, formed results Zhizu;
crown is too large, heavy shears retracted, do not cut in line
with scissors In addition, do not cut scissors yin yang, upright
branches cut, do not cut the level of its side branches, cut
overlapping interference sticks, do not cut the branches empty
chamber Principle; the variety of forms including hollow tower,
fan, umbrella shape.
Further technical solution is that the number of project
management including planting, the plants, fertilization,
irrigation Water, anti-pest, pesticide use time and date of
harvest, cultivation, cultivation quantity, cultivation methods.
Further technical solution is the fertilization management
projects include promoting fertilizers and reminders fruit
fertilizer, promote the Fertilizers in late September after the
harvest pomegranate, organic manure (composting manure) 800kg /
acre, homemade bio Organic fertilizer 200kg / acre (10 kg or
organic fertilizer / plant, bio-organic fertilizer made 2 kg /
plant), the pro-flower Fertilization fertilizer is a peripheral
crown dig deep 40㎝ wide radial grooves, after promoting
fertilizers and soil mix evenly applied into the pit Trench,
irrigation 2kg casing tramping; the fertilizer is urging fruit in
late May or early June fruit firmly secured, Shi reminders fruit
fertilizer, depending on the vigor fertilization, the results of
the amount of applied homemade bio-organic fertilizer 120kg / acre
(or self-administered Health Of organic fertilizer 1.5kg /
strain).
Further technical solution, said irrigation per plant 150kg /
times, 25 days irrigation once.
Further technical solution is the fruit firmly secured in late May
to mid-June on, the thinning Holding more than 300 fruit leaves
leaf / fruit, fruit from 30 cm / fruit.
Further technical solution that includes the agricultural control
fertilization and irrigation, thinning, drainage groove, Pruning,
remove leaves and dig garden pests, including the use of physical
control of the black light trap phototaxis Moths, insect repellent
planting insecticidal chrysanthemum, including the protection of
natural enemies of the biological control, promote the use of
biological Pesticides, botanical pesticides, mineral pesticides
pest control.
Although the invention herein with reference to the explanatory
embodiment of the present invention has been described, but not
limited to System of the present invention, it should be
appreciated that those skilled in the art can design many other
modifications and embodiments, These modifications and embodiments
will fall within the scope and spirit of the principles disclosed
herein within. more specifically, In the present application
disclosure, the drawings and claims, the component parts of the
subject combination arrangement and / Or layout many variations
and modifications. In addition to variations and modifications in
the component parts and / or layout of For skilled in the art, the
use of the present invention will be apparent.
http://nhb.gov.in/report_files/pomegranate/POMEGRANATE.htm
POMEGRANATE PRODUCTION TECHNOLOGY
Agro-climatic requirements
Pomegranate grows well under semi-arid conditions and can be grown
upto an altitude of 500 m. above m.s.l.. It thrives well under
hot, dry summer and cold winter provided irrigation facilities are
available. The tree requires hot and dry climate during fruit
development and ripening. Pomegranate tree is deciduous in areas
of low winter temperature and an evergreen or partially deciduous
in tropical and sub-tropical conditions. It can tolerate frost to
a considerable extent in dormant stage, but is injured at
temperature below - 110 C.
Well drained, sandy loan to deep loamy or alluvial soils is
suitable for cultivation.
Varieties Cultivated
Important pomegranate varieties cultivated in India are Alandi or
Vadki, Dholka, Kandhari, Kabul, Muskati Red, Paper Shelled,
Spanish Ruby, Ganesh (GB I), G 137, P 23, P 26, Mridula, Aarakta,
Jyoti, Ruby, IIHR Selection, Yercaud 1 and Co 1.
Land Preparation
Land is prepared by ploughing, harrowing, leveling and removing
weeds.
Planting
Planting Material
Pomegranate is propagated vegetatively by cuttings, air layering
or gootee.
Planting season
Air layering is usually done during the rainy season and also in
November-December. Planting is usually done in spring
(February-March) and July-August in sub-tropical and tropical
regions respectively.
Spacing
High density planting is adopted in temperate regions. A spacing
of 5-6 m. in northern India and also in the plains of Deccan
plateau is usually followed. High density planting with a
spacing gives 2-2.5 times more yield than that obtained when the
normal planting distance of 5 X 5 m. is adopted. Farmers have
adopted a spacing of 2.5 X 4.5 m. Closer spacing increases disease
and pest incidence.
Planting Method
Square system of planting is mostly adopted. Planting distance is
decided on the basis of soil type and climate. A spacing of 4-5 m.
on marginal and very light soils is recommended.
Pits of 60 X 60 X 60 cm. size are dug (at a spacing of 5 cm. in
square system) about a month prior to planting and kept open under
the sun for a fortnight. About 50 g. of 5% BHC or carbaryl dust is
dusted on the bottom and sides of the pits as a pre-caution
against termites. The pits are filled with top soil mixed with 20
kg. farmyard manure and 1 kg. super phosphate. After filling the
pit, watering is done to allow soil to settle down. Cuttings/air
layers are then planted and staked. Irrigation is provided
immediately after planting.
Nutrition
The recommended fertilizer dose is 600-700 g. N, 200-250 g. P2O5
and 200-250 g. K2O /tree/year. Application of 10 kg. farmyard
manure and 75 g. ammonium sulphate to 5 year old tree annually is
adequate, whereas application of 50 kg. farmyard manure and 3.5
kg. oil cake or 1 kg. sulphate of ammonia prior to flowering is
ideal for healthy growth and fruiting. The time of
application is December/January for ambe bahar, May/June for Mrig
bahar and October/November for hasthe bahar.
The basal dose of farmyard manure @ 25-40 cart-loads /ha. besides
the recommended doses of N, P and K should be applied to
non-bearing trees in 3 split doses coinciding with growth of
flushes during January, June and September. Fruiting should be
encouraged from fourth year onwards. Nitrogenous fertilizer is
applied in two split doses starting at the time of first
irrigation after bahar treatment and next at 3 weeks interval,
whereas full dose of P and K should be applied at one time. These
should be applied in a shallow circular trench below tree canopy
not beyond a depth of 8-10 cm. After application, fertilizers are
covered with top soil and irrigated.
Irrigation
First irrigation is provided in case of mrig bahar crop in the
middle of May followed by regular irrigation till the monsoon sets
in. Weekly irrigation in summers and that during winters at
fortnightly intervals is recommended. The check basin system of
irrigation is usually followed.
Drip Irrigation
The average annual water requirement through drip irrigation is 20
cm. Drip irrigation helps to save 44% on irrigation and 64% when
sugarcane trash mulch is used. It also helps to increase the yield
by 30-35%.
Training
Plants are trained on a single stem or in multi-stem system. Since
the crops trained on single stem training system are more
susceptible to pests viz. stem borer and shoot hole borer, the
other system is more prevalent in the country.
Pruning
Pruning is not much required except for removal of ground suckers
, water shoots, cross branches , dead and diseased twigs and also
to give shape to the tree. A little thinning and pruning of old
spurs is done to encourage growth of new ones.
Inter-cropping
Inter-cropping with low growing vegetables, pulses or green manure
crops is beneficial. In arid regions, inter-cropping is possible
only during the rainy season, whereas winter vegetables are
feasible in irrigated areas.
Regulation of bearing
Pomegranate plants flower and provide fruits throughout the year
in central and southern India. Depending on patterns of
precipitation, flowering can be induced during June-July (mrig
bahar), September-October (hasta bahar) and January-February (ambe
bahar). In areas having assured rainfall where precipitation is
normally received in June and continues upto September, flowering
in June is advantageous; where monsoon normally starts in August,
flowering during August is beneficial. Areas having assured
irrigation potential during April-May, flowering during January
can be taken and where monsoon starts early and withdraws by
September induction of flowering in October is possible.
Considering comparable yields, prices and irrigation needs it is
recommended that October cropping could be substituted for January
flowering.
Plant Protection Measures
Insect Pests
Insect pests mostly observed are fruit borer, mealy bugs, aphids,
white fly and fruit sucking moths. Spraying with dimethoate ,
deltamethrin or malathion etc. depending upon the type of pest
infestation has been found to be effective in most cases.
Diseases
The main diseases reported are leaf spot and fruit rot.
Application of Mancozeb (2g./l.) during rainy season in case of
the former and application of Kavach (2g./l) and
Carbendazim/Thiophanate methyl/Baycor/Benomyl (1g./l.) during
September/October in case of the latter has been found to be
effective in most cases.
Disorders
Fruit cracking is a serious disorder. This physiological disorder
observed in young fruits is due to boron deficiency and that in
fully grown fruits is mainly due to moisture imbalances. Tolerant
varieties viz. Bedana Bose and Khog may be cultivated and in other
cases spraying with calcium hydroxide soon after fruit set has
been found to be beneficial.
Harvesting and Yield
Pomegranate being a non-climacteric fruit should be picked when
fully ripe. Pomegranate plants take 4-5 years to come into
bearing. Harvesting of immature or over mature fruits
affects the quality of the fruits. The fruits become ready for
picking 120-130 days after fruit set. The calyx at the distal end
of the fruit gets closed on maturity. At maturity, the fruits turn
yellowish-red and get suppressed on sides.
POST HARVEST MANAGEMENT
Grading
Fruits are graded on the basis of their weight, size and colour.
The various grades are super, king, queen and prince-sized.
Besides that, pomegranates are also graded into two grades- 12A
and 12 B. Fruits of 12-A grade are generally preferred in southern
and northern region.
Storage
Fruits can be stored in cold storage upto 2 months or 10 weeks at
a temperature of 50 C. Longer storage should be at 100 C and 95%
RH to avoid chilling injury and weight loss.
Packing
The size of packages varies according to the grade of the fruits.
Corrugated fibre board boxes are mostly used. In a single box, 4-5
queen sized fruits, 12 prince sized and some of 12-A and 12-B
grades may be packed. The white coloured boxes having 5 plies are
generally used for export purpose, whereas red-coloured ones
having 3 plies are used for domestic markets. The red coloured
boxes are cheaper than white coloured ones. The cut pieces of
waste paper are generally used as cushioning material.
http://homeguides.sfgate.com/pomegranate-tree-cultivation-58444.html
Pomegranate Tree Cultivation
by
Jill Kokemuller
The pomegranate (Punica granatum) is a deciduous fruiting plant
that grows as a tree or a shrub in U.S. Department of Agriculture
plant hardiness zones 7 through 11. Pomegranates grow about 10
feet tall in shrub form or 20 feet tall as a tree. They are summer
blooming, with red-orange flowers, round red fruit and dark green
foliage.
Soil and Sun
Pomegranates will grow in sand and heavy clay soils, although the
best results come from loamy soils. Sandy soils will decrease
fruit production, while clay soils will result in paler fruit.
Pomegranates are not tolerant of alkaline soils, needing a more
acidic soil pH between 5.5 and 7.0. Peat moss or sulfur will make
soil more acidic while dolomite lime will raise soil pH if your
soil is outside the optimal range. Full sun is necessary to enable
the plant to produce fruit.
Planting
Pomegranates should be planted as early as possible after the last
spring frost to give them time to harden for the winter. Work soil
before planting to loosen, and add amendments such as peat moss,
compost or other organic matter to increase moisture and
nutrients. Space shrubs 6 to 9 feet apart and trees 15 to 18 feet
apart. Planting holes should be at least twice the width of the
root ball and backfilled with the soil surface lower than the
crown of the tree or shrub.
Water and Fertilizer
Weekly watering will keep the soil moisture adequate for fruit
production. The trees need 50 to 60 inches of water per year.
During late summer and fall especially, keep soil consistently
moist, watering when the top inch of soil dries. Pomegranates are
drought tolerant, but without sufficient water may not have much
of a yield. Young trees need about 3 ounces of nitrogen applied in
late winter and early spring. When the tree matures, nitrogen
should be increased to 1/2 to 1 pound of nitrogen split between
the two application times. Pomegranates do not benefit from extra
potassium or phosphorus, although occasional zinc sprays may be
necessary.
Pruning
The pomegranate will naturally grow as a bushy shrub-type plant.
To train it into a tree form, choose the strongest trunk and
remove the others, pruning off any suckers that form. At planting,
cut plants back to 24 to 30 inches tall. Cut branches back 40
percent the following winter. In subsequent years, prune suckers
and water sprouts as needed. An annual winter pruning will keep
the tree or shrub neat and healthy. Remove dead, damaged or
diseased branches, and cut individual branches back to an
outfacing bud to shape the tree.
http://homeguides.sfgate.com/protecting-pomegranates-bugs-30843.html
Protecting Pomegranates From Bugs
by
Amanda Flanigan
A native of Iran and India, the pomegranate (Punica granatum) is a
shrub or small tree that grows in U.S. Department of Agriculture
Plant Hardiness Zones 7 through 10. Pomegranates produce
leather-skinned, unusually shaped fruit and showy blooms in hues
of orange and red. This sun-loving fruit is generally easy to care
for but may develop problems caused by bugs and insects. Reduce
pest damage by protecting the pomegranate from bugs.
Pomegranate Bugs
According to the University of California, aphids are a
widespread, serious problem affecting many plants including
pomegranates. Aphids considered the most damaging to the plant and
cotton aphids and pomegranate aphids, although they are usually
only damaging in heavy infestations. Mealy bugs can infest
pomegranates. These small, white-colored bugs have a cottonlike
appearance and suck the juices from the foliage. Other bugs that
can infest pomegranates are scales, whiteflies, leafrollers,
thrips, beetles and various insect larvae. Leaf wilting,
discoloration, curling, browning, yellowing and fruit damage are
all signs of pest infestation on pomegranates.
Treatment
Most pests that feed on or damage pomegranates can be controlled
without chemicals by introducing lady beetles, lacewings,
beneficial parasites and predatory insects. These beneficial bugs
eat the problematic pests and keep them under control. Applying
chemical pesticides can throw off the balance of beneficial bugs
and increase the numbers of unwanted bugs leading to a pest
problem. Chemical pesticides should be used only as a last resort.
Instead, use less toxic products, such as Neem oil, insecticidal
soaps and horticultural oil.
Prevention
If your pomegranate is healthy, it will handle pest and diseases
with few problems. Encourage a healthy pomegranate by keeping the
ground under the plant free from debris. Fallen plant matter --
leaves, twigs, fruit and branches -- invites unwanted pests,
fungus and diseases to the pomegranate and increases the
possibility of problems. A water-stressed plant is likely to
develop pest and disease problems, so water young pomegranates
during dry periods. After they are established, pomegranates
tolerate drought conditions. During the pomegranates’ first and
second springs, apply about 2 to 4 ounces of all-purpose nitrogen
fertilizer to the soil. After the second spring, fertilizer is not
generally needed. However, applying organic compost around the
pomegranate every year protects its roots and adds extra nutrients
to the soil.
Considerations
The pomegranate is generally a pest- and disease-free plant that
rarely sees anything more than cosmetic damage caused by pests.
However, hungry deer have been known to feed on the pomegranate
causing foliage and fruit damage. If deer are a potential problem,
use commercial deer deterrents that keep the creatures away from
the pomegranates without harming them. Water the pomegranates in
the morning hours to allow the foliage to dry before the sun sets
and decrease the chance of fungal diseases.
http://homeguides.sfgate.com/natural-insect-sprays-pomegranate-trees-54288.html
Natural Insect Sprays for Pomegranate
Trees
by
Amanda Flanigan
Producing brightly colored blooms and unusually shaped fruit,
pomegranate trees (Punica granatum) are a stunning addition to
your garden. These India and Iran native fruit trees grow in U.S.
Department of Agriculture plant hardiness zones 8 through 11. Like
other fruit trees, insects can attack the pomegranate, requiring
immediate action to reduce potential damage. Chemical pesticides
are a common tactic to combat these insects, but they can cause
more harm than good. These toxic insecticides kill the unwanted
pests as well as beneficial insects. This can lead to an even
worse infestation than before. Instead, use natural insect sprays
to protect the pomegranate tree from pests.
Pomegranate Insects
Aphids are a serious problem affecting pomegranates with
pomegranate aphids and cotton aphids being the two species causing
the most damage in heavy infestations, notes University of
California Division of Agriculture and Natural Resources. Other
insects that infest pomegranate trees include mealybugs, scale
insects, whiteflies, thrips, leafroller, beetles and the larvae of
various butterflies and moths. A common sign of an insect
infestation is discoloration, yellowing and wilting of the leaves
or damage to fruit.
Insecticidal Soap
Containing fatty acids which break down quickly and -- once dry --
losses the insect-controlling abilities, insecticidal soap is a
more natural option to pest control on pomegranate trees. It
controls an array of unwanted insects, including aphids, scales,
whiteflies and leafhoppers. For insecticidal soap to effectively
control pests, it must come into contact with the insects and
should only be applied when the temperatures are between 40 and 90
degrees Fahrenheit, on a calm day.
Homemade Oil Spray
A homemade oil spray consists of 6 tablespoons of canola oil mixed
with 1/4 cup of mild dish soap and 2 gallons of water. Never use
dish soap that contains fragrances, degreasers or bleach. This
natural insect spray controls various soft-bodied pests -- such as
mites, scales and aphids -- that feed on the leaves of the
pomegranate tree. Only use this spray when temperatures won’t
reach 85 F and thoroughly coat the tops and undersides of the
leaves with the concoction.
Bacterium Insecticide
Bacterium insecticide, such as Bacillus thuringiensis strain
Kurstaki, targets the larvae of butterflies and moths that feed on
various parts of the pomegranate tree. This nontoxic insecticide
is applied as a foliar spray and poisons the larvae when they
consume it, causing them to stop feeding. After a few days, the
larvae die of starvation. Bacillus thuringiensis doesn’t harm
predatory and beneficial insects and is safe to use around people
and animals. It is nontoxic to bees, birds and fish.
http://homeguides.sfgate.com/pomegranate-tree-problems-48471.html
Pomegranate Tree Problems
by
Teri Silver
Pomegranates (Punica granatum L.) include more than 500 known
cultivars and produce medium-sized, leathery fruits containing
tangy, juicy berries. Pomegranates grow in U.S. Department of
Agriculture plant hardiness zones 7 to 11. The trees are
relatively easy to grow and maintain, notes Clemson University
Extension, but they are susceptible to fruit rots, mold, pests and
wood damage. Pollination failure and inadequate sunlight may keep
pomegranate trees from producing flowers and fruits.
Molds
Gray mold (Botrytis cinerea) causes harvested pomegranates to
decay at a faster than normal rate. The pathogen develops spores
on flower petals, which remain in the blooms until the fruits have
ripened. It creates a gray coating of fungal spores that live in
the fruit tissue. Pomegranates that are stored in damp, humid
conditions exhibit the gray-coated surface as they decay.
Blue-green mold (Penicillium spp.) may develop on pomegranate
trees but it usually appears when fruits are stored. Symptoms are
wet areas on fruit skins and bluish-green powdery mold. Protecting
pomegranate fruits from gray or blue-green mold is not easy or
economical, notes University of California Extension, but removing
old fruit and dead branches will help reduce fungal spore
production.
Fruit Rots
Pomegranate trees are susceptible to rots caused by pathogens that
develop during flowering and fruit development. Alternaria fruit
rot (Alternaria alternate) grows inside the fruits, causing them
to become stunted and discolored. Rain and overly saturated soil
cause the fungus to grow within the fruit. Pathogens live on dead
plant and fruit debris during the tree’s dormancy. Aspergillus
fruit rot (Aspergillus niger) is similar to Alternaria -- the
fungus grows inside the flowering, growing fruits after rainfall.
Fruit skins become pale but not dramatically different. Insects
also damage diseased pomegranates; pest control may useful.
Pests
Pomegranate trees attract pomegranate butterflies (Virachola
isocrates), which deposit larvae on flower buds and fruits.
Caterpillars emerge, entering a bud through the flower’s calyx.
Borers can cause major destruction to the tree’s fruit crop. In
early summer, stem borers such as Pleuroplaconema or Ceuthospora
phyllosticta lay eggs. The larvae cause twig dieback by digging
into fruits and seeds -- trees may die if left untreated.
Pomegranate tree leaves also attract mealybugs, scale insects,
thrips, mites and whitefly. Deer enjoy chewing through foliage on
pomegranate trees. Sawdust at the base of the tree indicates
termite damage.
Leaf and Fruit Spots
Like many deciduous trees, pomegranate foliage develops leaf
blotch and fruit spot. Infected leaves are pale green or yellow
and have small reddish brown spots that turn to black. Premature
leaf drop is possible. Applying copper fungicide three times a
year helps to alleviate leaf spotting. Blight (Colletotrichum
gloesporioidesl, Pseudocercospora punicae, Curvularia lunata and
Cercospora punicae) creates water-soaked spots on pomegranate
foliage, causing leaves to drop prematurely. Useful fungal
treatments include Topsin-M, Sulfex, Difolatan and Dithane-M,
advises the National Horticulture Board. Cercospora fruit spot
(Cercospora spp.) features irregular brown to black lesions on
pomegranates. Although it is best to destroy diseased fruits,
Dithane or Captan may control the fungus, notes the NHB.
About Pomegranate Trees
Most pomegranate cultivars are deciduous trees that grow best in
full sun and well-draining soil with a pH of 5.5 to 7.0.
Fertilizing with a 10-10-10 mixture in March and July will help
fruit production. Pomegranate trees can tolerate cold snaps but
may be damaged if temperatures consistently dip below 10 degrees
Fahrenheit. Most cultivars are drought-resistant. Light pruning
encourages new growth, flowers and fruits. Most cultivars are
self-pollinating but some varieties must be cross-bred to produce
fruit. Planting suitable varieties close together helps trees
develop pollinated blooms.
http://homeguides.sfgate.com/kind-dormant-spray-can-spray-pomegranate-trees-55156.html
What Kind of Dormant Spray Can You
Spray on Pomegranate Trees?
by
Amanda Flanigan
Pomegranate trees (Punica granatum) are an interesting addition to
your backyard orchard, producing showy flowers and oddly-shaped
edible fruit, growing in U.S. Department of Agriculture plant
hardiness zones 7 through 10. Pomegranates benefit from sprays
which act as a protective cover applied during the dormant period
to eliminate diseases and insects that may be overwintering on the
tree and to protect the pomegranate tree from future insect,
fungal and bacterial infestations.
Horticultural Oil
Horticultural oil is made from refined petroleum and can be used
anytime throughout the year. Horticultural oil works by smothering
overwintering, soft-bodied pests. It controls scales, mealybugs,
aphids, mites and their eggs but doesn’t treat fungal or bacterial
diseases. Horticultural oil can also control certain caterpillar
species, such as leafrollers and tent caterpillars, that
overwinter as eggs. For horticultural to work effectively, the
insects must be entirely coated with the oil, so to ensure the
pests are controlled, you must apply a liberal amount of the spray
to the pomegranate trees. Most horticultural oils have a low
toxicity to people, mammals and beneficial predator insects and
dissipate quickly, leaving little to no residue.
Homemade Dormant Oil
Homemade dormant oil consists of inexpensive and less toxic
ingredients than commercial insecticides and allows you to adjust
the recipe depending on how much or how little oil spray you need.
In a large pot, bring one-half gallon of mineral oil, one-quarter
gallon of water and one-half pound of oil-based soap to a boil,
mixing continuously until the ingredients are thoroughly combined.
This concentrated solution is then diluted at a ratio of 1 part
solution to 20 parts water. Homemade dormant oil must be used
immediately, since the ingredients have a tendency to separate.
You should apply the homemade dormant oil on a day when
temperatures are above 40 degrees Fahrenheit, thoroughly coating
the pomegranate tree with the liquid.
Fixed Copper Fungicide
Applications of copper fungicide during the fruit tree’s dormant
period will help prevent the spread and reduce infection of
various fungal and bacterial diseases. Copper fungicides can
prevent future leaf spot and fruit spot infections when applied
during the pomegranate’s dormancy period. Depending on the disease
you are trying to prevent, several applications of the fungicide
may be required to protect newly emerged flowers and shoots or due
to an abundance of rain.
Considerations
Dormant sprays are applied when the pomegranate trees enter their
dormant stage after the tree has lost its leaves. Typically, fruit
trees are sprayed with dormant oil in late November until the bud
swell stage in February or March. Refrain from applying dormant
sprays after flower buds open, as doing so increases the chance of
damaging the pomegranate fruit. A University of California
Agriculture and Natural Resources Master Gardener's Tip Sheet
recommends applying dormant sprays after the rainy or foggy
weather ends and never apply during periods of rain, fog or during
freezing temperatures.
http://homeguides.sfgate.com/companion-plants-pomegranates-39139.html
Companion Plants for Pomegranates
by
Nicki Wolf
Pomegranates, a fruit popular in Spanish and Middle Eastern
cuisine, can add beauty and function to a garden. The tall, broad
tree grows in U.S. Department of Agriculture hardiness zones 7
through 10, and produces edible fruit in the fall and orange-red
flowers through the summer. You can help ensure healthy
pomegranate trees in your yard by including companion plants
nearby -- the right plantings attract beneficial insects, repel
predators and enhance the look of your trees.
Herbs
The pomegranate requires bees for pollination; without proper
pollination, your tree will not produce fruit. A variety of herbs
can attract bees, including dill, cilantro, parsley and mint.
Basil, thyme and summer savory also attract bees. It can be
especially helpful to allow herbs to flower, because introducing a
spectrum of color into your garden keeps bees returning to the
area.
Flowers
Planting a bed of lavender flowers near a pomegranate tree
attracts bees, as will beds with cosmos, coreopsis, zinnias and
sunflowers. Many flowers help repel insects harmful to
pomegranates as well. Aphids can inflict damage this fruit tree,
resulting in rotten spots on fruit, blossom drop and ideal
conditions for sooty mold infestation. Choose flowers that repel
aphids, such as nasturtiums. You may also opt for flowers that
attract beneficial insects that eat aphids, such as ladybugs and
lacewings. These insects are especially attracted to daisies and
Queen Anne's lace. Flowers may also be used to bring out the color
of pomegranates -- purple passion vine flowers contrast, and
jasmine's white flowers offset the pomegranate's orange-red
flowers.
Vegetables
There are several vegetables you can plant near a pomegranate tree
that make good companions and help you maintain a truly edible
garden. Fennel and leaf celery draw ladybugs and lacewings, both
of which will eat the aphids that might damage your pomegranate
tree.
Fruit
Melons and berries attract bees, another insect beneficial for the
pomegranate. Planting a fruit garden near your tree helps ensure
proper pollination. Opt for a variegated Calamondin orange as a
companion plant to the pomegranate for a visually stunning and
edible fruit orchard. The orange blossoms also help to attract
bees.
http://homeguides.sfgate.com/kind-soil-pomegranates-40936.html
What Kind of Soil for Pomegranates?
by
Richard Corrigan
Pomegranate (Punica granatum L.) grows as a large shrub or small
tree, and is known for its red, pulpy fruit. Though it is native
to arid parts of Asia and the Middle East, pomegranates can also
be grown in the United States, where they thrive in U.S.
Department of Agriculture plant hardiness zones 8 to 10. Though
they are highly adaptable, but planting pomegranates in the right
type of soil is essential to a healthy fruit crop.
Soil Types
Pomegranates are able to adapt to a variety of soils, ranging from
acidic sandy loam to alkaline calcareous soils and everything in
between. In India, they have been known to grow in rocky gravel.
The only soil that will not support pomegranates is heavy clay,
because soils with excessive clay tend to present drainage
problems.
Irrigation and Drainage
Drainage is one of the most important things to consider when you
choose a location to grow pomegranate. Pomegranates demand
well-drained soil, and they will not fare well in a location where
the ground becomes wet and waterlogged. Irrigation is usually not
necessary in most areas because pomegranates can withstand
significant periods of drought. Fruiting can be diminished in
drought conditions, so watering may be helpful if you are growing
pomegranate primarily for its fruit.
Soil pH
Neutral to slightly acidic soil is best for pomegranate. They will
still survive under considerably more acidic or alkaline
conditions, but a pH range of 5.5 to 7.0 is best. If necessary,
you can raise the pH of your soil by adding ground agricultural
limestone, or lower it by incorporating some form of sulfur. A
soil test kit, available at most garden stores, will tell you the
pH as well as the nutrient content of your soil.
Fertilization
Pomegranates are fairly well adapted to growing in poor-quality
soils, but moderate nutrient levels are better for fruit
production. You can fertilize your trees with a small application
of some form of nitrogen fertilizer during their first two
springs. Fertilizer is usually not necessary after that, though
adult trees can still benefit from a fresh layer of organic mulch
applied annually.
http://homeguides.sfgate.com/protect-pomegranates-squirrels-28618.html
How to Protect Pomegranates From
Squirrels
by
B. Sinclair
Pomegranates are easy to grow and flourish in U.S. Department of
Agriculture hardiness zones 7 through 10. Grown as a shrub or
small tree, the plant produces nearly round fruit with red,
leathery, tough skin that protects sacs filled with juicy, edible
ruby seeds. Although relatively free of insect and pest problems,
squirrels can pose a problem by eating the fruit before you have a
chance to harvest it. Take protective measures to ensure you’ll
have a chance to savor the thirst-quenching pomegranate fruit.
Cover the fruit with sandwich-size paper bags and seal with a
twist tie. Check the fruit every few days for ripening.
Pomegranates mature five to seven months after blooming. Harvest
when the fruit makes a metallic sound when tapped.
Cover the pomegranates with netting made of 1/4- to 1/2-inch mesh
plastic. For pomegranates grown as shrubs, build a frame out of
PVC piping around the plant and lay the netting over it so that it
doesn’t actually touch the shrub. Drape the netting over
pomegranate trees and tie it around the trunk of the tree so
squirrels can’t get inside.
Cut trees back and keep them away from roof overhangs, telephone
lines or anything else squirrels can use to hop on your
pomegranates. Also, consider letting your dog have full run of the
yard. They are usually a good deterrent to squirrels.
http://homeguides.sfgate.com/grow-pomegranate-trees-containers-43843.html
How to Grow Pomegranate Trees in
Containers
by
Diane Watkins
Pomegranate trees (Punica granatum) are especially suited for
growing in containers. The dwarf trees are easier to care for than
a full-size tree, while still producing a good harvest of fruit.
Full-size trees grow up to 20 feet tall. Dwarf varieties such as
"State Fair" grow to be about 5 feet tall and "Nana" is only 2 to
3 feet tall when fully mature. They are suitable for outdoor
growing in U.S. Department of Agriculture plant hardiness zones 7b
to 10. Bring them indoors for the winter in colder climates.
Expect your tree to produce fruit the second or third year after
planting.
Allow the soil to dry slightly before watering your pomegranate
tree. Water thoroughly, saturating the soil. Allow excess water to
draing through the drain holes.
Fertilize the pomegranate tree in November, February and May with
an ammonium sulfate fertilizer. Apply approximately 1 ounce per
foot of the tree's height around the edge of the pot, keeping the
fertilizer away from the trunk.
Prune the tree to remove dead and diseased branches and branches
that rub or cross fruit or other branches. Shape the tree to a
pleasing size and shape. Remove excess suckers, leaving a few only
where you want new branches.
Place the pot in a place that gets direct sun for most of the day.
Bring the tree indoors whenever temperatures drop below 40 degrees
Fahrenheit. While trees in the ground can withstand temperatures
much lower, container-grown trees are more susceptible to cold
damage.
Dust the tree with sulfur in June and again in July if mites are
present. Control aphids by spraying them with soapy water.
Bring the container indoors in the fall, increasing the time spent
indoors by one or two hours daily. Transitioning the tree this way
gives it time to adjust to lower light levels. Reverse the process
in the spring, bringing it outdoors for a longer period each day.
Check the tree every spring for signs that the roots are becoming
pot-bound. Repot the tree in a larger pot before the roots have a
chance to out grow the pot. Place 1 inch or more of organic
potting mix in the bottom of a pot that is at least 2 inches
larger than the original container. Remove the pomegranate from
its container and place it in the center of the pot. Fill in with
soil around the outside. Tamp the soil down, making sure the trunk
remains exposed.
http://ressources.ciheam.org/om/pdf/a42/00600264.pdf
Cultivation of Pomegranate
by
A Blumenfeld
SUMMARY -- This paper describes various aspects of
pomegranate cultivation. Tradition, consumption and production
target, recommended soil, fertilisation ...
http://www.yourarticlelibrary.com/cultivation/pomegranate-plant-cultivation-in-india/24715/
Pomegranate Plant Cultivation in India
By
Samiksha S
Botany Name: Punica Granatum L
Family: Punicaeae
Pomegranate is one of the esteemed table fruit. Its fruits are
known for their sweetness and fine blend of acidity.
Its juice is cool and refreshing with medicinal value. The hardy
nature of pomegranate makes it the choicest kitchen garden fruit.
Highly acidic fruits with 7-8% acidity are used to make ‘anardana’
(daru).
Origin:
Pomegranate is native to Iran. It is commercially cultivated in
Afgharustan, Baluchistan, Iraq, Spain, Russia, India, Pakistan,
Myanmar, Chiria and Japan.
Area and Production:
In India, pomegranate production is 8.07 lacs MT from an area of
109.2 thousand hactare and Maharashtra state contributed a share
of 75.09 per cent to the total India production of pomegranate.
The major growing states one Maharashtra, Kaniataka, Gujarat,
Andhra Pradesh. Chhatisgarh, Himachal Pradesh, Nagaland, Orissa,
Rajasthan and Tamil Nadu.
Uses:
The sweet fruits are used as table purpose and acidic for making
‘anardana’ a condiment used for ‘chatru’ making. Juice is prepared
and preserved for use. Wine can be prepared from sweet coloured
juice. Sodium citrate/citric acid is prepared from very acidic
fruits. Pomegranate shoot bark is used for body slimming.
The root or rind (peel) of the fruit is used to control dysentery,
diarrhoea and worm killing in the intestines. Every part of the
pomegranate plant is used for one or the other human disease. Dyes
are prepared from flower petals. The edible portion (grains) of
fruit contain upto 78 percent water, 5% fibre, 1.6% protein,
16-18% carbohydrates (sugars). Each 100 gram of fruit flesh may
contain calcium 10 mg, phosphorus 70 mg. and sufficient quantity
of iron, riboflavin and vitamin C.
Botany:
It grows as a shrub but can be trained on modified leader system
as a small tree. Pomegranate is evergreen as well as deciduous.
Shrub trained plants remain smaller than single stemmed trained as
trees. It is hardy tree and can live over 40 years. The leaves
have small petioles and are oval to lanceolate in shape. The
shoots have thorns which originate deep from the wood. It bears
very beautiful red coloured flowers.
Flowers may be sohtary, axillary or appear in clusters on short
spurs. The calyx is persistent and tubular with 5-7 petals
inserted in calyx. Ovary have many locules. Pomegranate fruit is
special fruit botanically. The edible portion is aril juicy
covering of seed. Both types of seed are present is different
cultivars. The hard seeded and soft seeded cultivars.
Soil and Climate:
Pomegranate thrives well in semiarid conditions. It can adapt wide
range of soil and climatic conditions. Deep loam to sandy loam
soils are considered ideal. It can tolerate alkaline/ saline soils
with 9.0 pH having lower EC than 0.5 mm hos/cm. It can grow in
light soils but with assured irrigation. It requires hot and dry
summers with cool winters.
It is tolerant to frost and freeze fairly well very high
temperature in summers and too low in winters encourage fruit
cracking. Pomegranate have both types of cultivars, i.e. some are
deciduous in winter and others are evergreen. The tree requires
hot and dry climate for the production of high quality
pomegranates. The evergreen cultivars do not shed leaves in very
cold winters.
Cultivars:
Mostly pomegranate cultivars originated as seedling selections.
Some promising cultivars have also been developed through
controlled hybridization. The recommended cultivars are area
specific for example Kandhari and Nabha for north plains,
Jaloreseedless and Jodhpur white in Rajasthan, Velludu in Tamil
Nadu, Dholka in Gujrat, Basse-in-seedless, Bhagawa for Karnataka
and Phule Arakta, Ganesh in Maharashtra.
Some of the promising cultivars are discussed below :
Ganesh: A seedling selection from Alandi made by Dr. G.S. Cheema
at Pune. Plants bear profusly and regularly. It is evergreen bush
and precocious. Its fruits are of medium size, with yellow
coloured peel with pink blush. Average fruit size 350 g. Arils are
white, transparent with little pink tings. Seeds are soft and can
be eaten along with arils. The juice percentage on the basis of
grain weight is 80% or more. The TSS of juice varies from 13% to
16% with very Uttle acidity (0.3 to 0.5%). Tree can bear Ambe,
Mrig and Hast bahars. One can opt for any bahar as per requirement
of the area.
Under North Indian conditions, it is beneficial to take
Ambe-bahar, which ripens in August. To improve the fruit size,
flower buds appearing after 15th April should be manually removed.
Fruit yield is 12-15 tonnes per hectare.
Kandhari: Trees are deciduous, vigorous and upright growing. It is
regular bearer with good yield per tree. It bears only Ambebahar
(April-May flowering). Fruits are of big size with yellow coloured
peel and red splash. Average fruit weight is 370 g Arils are light
pinkish to deep pinkish, with semihard seeds, depending upon the
dry weather and prevailing temperature. The grains have 77 percent
juice on the basis of weight of grains. The juice have 13-15% TSS
and well blended acidity (0.5 to 0.6%).
To get well-sized fruits of high quality, remove the flower bud
appearing after 25th of April manually. There is a considerable
variation in yield from area to area. However, on an average it
can yield 10-12 tonnes per hectare.
Nabha: Tree growth is just like Kandhari and is also deciduous in
nature. The rind of the fruit remain yellow in colour. The fruit
size becomes larger than kandhari if thinning is done. The average
fruit weight is 350 g. The grains are of white colour, with hard
seeds. The grains have 75 percent juice with 13% TSS and 0.5%
acidity. Yield 10 tonnes per hectare.
Assam Local: Under North Indian conditions tree grows upto 3 m
high and 1.5 m spread. Tree is healthy. The fruits remain free
from fungal/bacterial attack during the rainy season. It also bear
only one crop. Average fruit weight is 220 g. The arils are small,
hence juice percentage on the basis of weight of grains in 47 with
15% TSS and 0.6% acidity. Average yield per hectare only 7 tonnes.
Jyothi: Trees are dwarf and evergreen. Under North Indian
conditions also it bear three crops, i.e. Ambe, Mrig and Hast
bahars. Due to very low temperature in January the fruits of mrig
bahar crack. Therefore it is beneficial to get only ambe bahar.
Fruits remains small in size. Average fruit weight is 200 g. The
rind develops a typical red colour with pinkish-reddish coloured
arils. The juice percentage is 75 on the basis of grains weight.
The juice have 17% TSS with very low acidity 0.3%. Average yield 8
tonnes per hectare (ambebahar only).
P.A.U Selection: It originated as a chance seedling from Ganesh
and was evaluated along with other thirty cultivars at Ludhiana.
The plants are as vigorous as Ganesh and evergreen. The flesh
characters resemble Ganesh, but seeds are little hard. It bears
big-sized fruits like kandhari. Average fruit weight 450 g. Juice
percentage on the basis of grain weight is 71% with 13% TSS and
0.7 acidity. Average yield 8 tonnes per hectare.
Mridula: It is a soft seeded evergreen cultivar and it is cross
between Gul Shah Red x Ganesh. It also bear three crops, but it is
better to get Ambe-bahar only. The plants remain dwarf than
Ganesh. The fruits have red coloured rind. The grains are also
blood red in colour. If thinning is done in Ambe-bahar the average
fruit weight is 240 grams. The juice is 78 percent on the basis of
grain weight the TSS of juice 17-18% with only 0.3% acidity.
Basse-in-Seedless: As the name shows the grains are seedless. The
plants are as vigorous as Ganesh and are evergreen in nature. The
fruits are prone to fungal/bacterial attack during rains. The
average fruit weight in ambebahar is 340g. The grains are light
pinkish as of Ganesh. The juice is 80 percent with 14% TSS and
0.4% acidity.
Ruby: The plants are slow growing, evergreen and bear all the
three crops. It is a hybrid from a 3 way cross between Ganesh X
Kabul X Yercand and Gulsha Rose Pink. It bears red coloured fruits
of small size with red coloured arils. Average fruit weight is 200
g. Juice percentage 80 with 15% TSS and 0.3% acidity.
Some other important cultivars are, G-137 a clonal selection from
Ganesh; P-23 and P-26 seedling selections from Muscat; IIHR
selection, it is a selection from open pollinated seedlings;
Yercand-1 and Co-1.
Propagation:
Pomegranate should not be propagated through seeds, due to cross
pollination, considerable variation occurs in the seedlings. Some
workers say that pre-conditioning of shoots during June-July by
girdling or etiolation increases the level of root promoting
cofactors, which help in rooting of cuttings. The maturity of
shoots used for cuttings plays a great role in the rooting of
cuttings. For getting healthy and precocious plants, pomegranate
should be propagated through hard wood cuttings.
One season old cuttings should be prepared during first week of
December. The length of cuttings should be 20-25 cms. Treat the
lower ends of these cuttings with 100 ppm of IBA (Indole butyric
acid) solution for 24 hours, before planting in the nursery beds
for rooting. After 10 to 15 days of planting, the beds may be
applied 5 litres of chloropyriphos solution @ 10 ml/L. to one
square meter of the bed. Repeat this treatment 20 days after to
control white ants attack. Chloropyriphos solution should be
applied with a shower over the cuttings slowly so that it reaches
the rootzone apply light irrigation after the treatment.
The cuttings should be irrigated lightly at an interval of 15-10
days from December to March and 7 days after wards. Aphid attacks
the newly emerging foliage, which should be controlled by spraying
Rogor @ 2 ml/L. of water or Nuvacuron (monocrotophos) @ 1 ml/L.
More than 80 percent cuttings root and produce healthy plants.
Deciduous cultivars root better than the evergreens. To improve
rooting of cuttings in evergreen cultivars take only one crop,
i.e. ambebahar remove flowers from June onward.
Planting:
Pomegranate should be planted on square system of planting at a
distance of 5m x 5m apart. Ever green cultivars may be planted at
4m x 4m distance. Prepare 1 meter diameter and 1 m deep pits one
month prior to planting of pomegranate.
Thus one need 400 to 625 plants for planting an hectare. The best
time for planting of pomegranate is December-January. The
evergreen cultivars should be planted in December. Young plants
are attacked by white ants in the pits also. So apply one litre
solution of chloropyriphos @ 10 ml/L. of water after two months of
planting to each plant.
Training:
Normally no importance is given to the training of pomegranate
plants. Pomegranate is left as such to grow as a bush which is not
desirable. Plants can be trained as single stemed tree (e.g.
Kandhari etc.) and 2 to 4 stemed bushes (evergreen). Multi stemed
trained plants create problems later on as the stems inter mingle
with each other.
Pomegranates should be trained as single stemed to get strong
scaffold system. No branch should be allowed to develop from
ground level to 30 cm of trunk height. Head back the leader at 1 m
height to force the scaffolds to develop. Select only 4-5 well
placed scaffolds on all sides of the main trunk.
If the plants are trained on two or three stems, then the plant it
self develop a balanced scaffold system. Remove only intermingling
branches from the stems. It has been recorded that deciduous
cultivars perform better as single stemed trees and evergreens
with 2 to 3 stems.
Pruning:
Pomegranate do not require annual pruning. The fruit is borne on
short spurs as well as in leaf axils and shoot apex. Remove
criss-crossing and dried branches. Some branches may be headed
back during December by removing 30 percent of the growth to
encourage fresh growth.
Crop Regulation:
Pomegranate is a precocious bearer. Plants start bearing in the
3rd year of planting. It continues to bear for 30 years. The
evergreen cultivars flower through out the year and bear three
crops. To obtain good quality of fruits and high yield. Select
only one bahar as suited to the area. Remove the flowers manually
for rest the bahars. Deciduous cultivars only bear Ambe bahar
(flowering April-May). The evergreens flower in Ambe bahar as well
as Mrig bahar (July) and Hast bahar (October-November).
The inflorescence is cyme, due to abscission of side buds and
persistence of central flower, flowering appears to be solitary.
Keep only one fruit at a place to get well-sized fruits. The
flowers should be manually removed after mid-April when sufficient
number of fruits have already set. On a mature tree select 80-100
fruits only.
Pollination:
Pomegranate bears three types of flowers i.e. pure male with
rudimentary ovary; hermaphrodite with medium style and
hermaphrodite with well developed style. The percentage of these
flowers varies from cultivar to cultivar and bahar to bahar. There
is no shortage of pollen and pollinating agents. Sufficient number
of fruits set from hermaphrodite (pin) flowers. Pollen is
available at noon and stigma remains receptive for 2-3 days. Both
cross and self pollination take place.
Irrigation:
Apply irrigation just after transplanting the plants in December.
Light and frequent irrigations should be given at an interval of
20 to 10 days from January to May and at weekly interval from May
to end July. No irrigation may be given if rains come in. Increase
the interval of irrigation. After the harvest of fruit in August
September. The interval of irrigation may be a month. Deciduous
cultivars may not be applied any irrigation during
December-January when leaf fall starts.
Intercropping:
Pomegranate have short juvenile period. Plants are also planted
closely and remain bushy and spreading in nature, hence, no
intercrop should be grown. When planting distance is more than 4m
x 4m then intercrops can be grown for the first two to three
years. Growing of vegetables and pulses should be preferred over
rabi crops and fodder crops. If wheat is to be grown prepare
separate irrigation system for irrigation to pomegranate plants
during March-April.
Manuring and Fertilization:
Manuring should be done as per nature of evergreen or deciduous.
If crop is not regulated then trees bear two crops and evergreen
three crops Accordingly, fertilizer requirements increase as ^e
crops in a year. As it has been recommended to take e-bahar in
North India, to get good quality of fruit, fertilizer doses may be
applied.
Add farm yard manure, super phosphate and muriate of potash in
December-January. Mix well in plant basins. Urea should be split
into two parts. Apply half in February at sprouting and half in
May after fruit set.
However, different fertilizer doses are given in different areas.
In general a higher dose has been recommended in literature i.e.
1.5 kg urea, 1 kg superphosphate and 400 grams of muriate of
potash per mature plant. This dose may hold good where more than
one crop per year is taken.
Weed Control:
Pomegranate is closely planted, hence use of a tractor to
inter-cultivate is not desirable. Weeds should be manually removed
by hoeing the basins once in December at the time of
fertilization and then again in May. The left out places may have
some weeds/grass growing. If need be spray Grammoxone (paraquat) @
6 ml/L of water during July. Keep the nozzle of spray close to the
weed growth to avoid drift of the chemical.
Harvesting and Marketing:
Pomegranate is non climacteric fruit hence should be harvested
when fully ripe. Pomegranate fruits become ready for harvesting
after 4-5 months of fruit set. As the flowering continues for over
two months, in the same way fruit ripening and harvesting also
continue for a month or so. The fruits are harvested at full
maturity, when rind has developed a typical colour of the
respective cultivar. At this stage calyx at the tips dries up.
Fruits should be harvested with the help of secateurs.
Grading:
Fruits are graded on the basis of fruit weight and size. For big
markets following grades are followed:
Super sized : Weight more than 750g, withoutspots.
King sized : 500g – 750g without spots.
Queen sized : 400g – 500g without spots.
Prince sized 300g – 400g well coloured.
12-A: 250 – 300g without spots.
12-B : 250 – 300g with some spots.
Marketing:
Fruits are packed as per their grade. CFB boxes are used for
packaging and fruits are cushioned with paper strips/cut pieces.
The fruit is transported to the markets. For small markets fruits
are packed in wooden boxes of 5 to 10 kg size.
The suitable temperature for cold storage of pomegranate fruits
ranges between 5-10°C, with 90-95% relative humidity. Fruits
stored at lower temperatures 0-3°C develop chilling injury.
Physiological Disorder:
Fruit Cracking:
Fruit cracking usually occurs in fruits of mrig and hast bahars.
It is more serious in arid zone. Cracked fruits get affected by
fungii and fruits become unfit for consumption. Mrig bahar fruits
crack due to wide variation in humidity and soil moisture.
Prolonged drought in ambe-habar causes peel hardening and when
rain comes or irrigation is applied the arils sweet and peel
cracks. Some cultivars are more prone to fruit cracking. Hast
bahar fruits crack due to low temperature in December and January.
Cracking can be controlled by supplying irrigation at regular
intervals, this will reduce the fluctuations in soil moisture.
Planting of wind break around the orchard also help in the
reduction of fruit cracking. Spray of borax @ 0.1 to 0.2 percent,
spray of GAj 30-40 days before fruit ripening @ 15-20 ppm checks
cracking of fruits. Cultivars Ganesh, Kandhari and PAU selection
show little cracking in ambe bahar fruits.
Internal Break Down of Arils:
Blackening or dis-colouration of arils in ripened fruits is a
serious malady throughout the pomegranate growing areas. All
cultivars are affected by this disorder. No cause or remedial
measure can be suggested at this stage. The fruit should be
harvested at ripening stage without keeping it on the tree for
more time than required.