U.S. patent application number 14/660385 was filed with the patent office on 2016-06-09 for regulation of brain biogenic amines associated with depression by a formulation from botanical source.
The applicant listed for this patent is Anand Shah, B. Arvind Shah. Invention is credited to Aruna Agrawal, Govind Prasad Dubey, Rajesh Dubey, Shipra Dubey, Anand Shah, B. Arvind Shah.
Application Number | 20160158302 14/660385 |
Document ID | / |
Family ID | 56093283 |
Filed Date | 2016-06-09 |
United States Patent
Application |
20160158302 |
Kind Code |
A1 |
Dubey; Rajesh ; et
al. |
June 9, 2016 |
Regulation of Brain Biogenic Amines Associated with Depression by a
Formulation from Botanical Source
Abstract
Provided herein are anti depressant and anxiolytic herbal
formulations including: 225-450 mg of Nyctanthes arbortristis,
250-550 mg Ocimum tenuiflorum, 200-450 mg Hippophae salcifolia, and
optionally additives selected from minerals, vitamins, salt, filler
and binder. Also provided are methods of making such formulations
and methods of using such formulations for treating or improving
brain function.
Inventors: |
Dubey; Rajesh; (Brookline,
MA) ; Dubey; Shipra; (Brookline, MA) ; Dubey;
Govind Prasad; (Varanasi, IN) ; Agrawal; Aruna;
(Varanasi, IN) ; Shah; B. Arvind; (Chennai,
IN) ; Shah; Anand; (Chennai, IN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Shah; B. Arvind
Shah; Anand |
Chennai
Chennai |
|
IN
IN |
|
|
Family ID: |
56093283 |
Appl. No.: |
14/660385 |
Filed: |
March 17, 2015 |
Current U.S.
Class: |
424/745 |
Current CPC
Class: |
A61K 36/185 20130101;
A61K 36/63 20130101; A61K 45/06 20130101; A61K 36/63 20130101; A61K
36/53 20130101; A61K 36/53 20130101; A61K 2300/00 20130101; A61K
2300/00 20130101; A61K 36/185 20130101; A61K 2300/00 20130101 |
International
Class: |
A61K 36/63 20060101
A61K036/63; A61K 36/185 20060101 A61K036/185; A61K 45/06 20060101
A61K045/06; A61K 36/53 20060101 A61K036/53 |
Foreign Application Data
Date |
Code |
Application Number |
Dec 6, 2014 |
IN |
6161/CHE/2014 |
Claims
1. An anti depressant and anxiolytic herbal formulation comprising:
225-450 mg of Nyctanthes arbortristis; 250-550 mg Ocimum
tenuiflorum; 200-450 mg Hippophae salcifolia; and optionally
additives selected from minerals, vitamins, salt, filler and
binder.
2. The herbal formulation as claimed in claim 1, wherein the said
formulation comprises whole plant of Nyctanthes arbortristis, whole
plant of Ocimum tenuiflorum and only leaves and fruits of Hippophae
salcifolia.
3. The herbal formulation as claimed in claim 1 wherein the amount
of each of the components are: 325 mg Nyctanthes arbortristis; 350
mg of Ocimum tenuiflorum; 275 mg of Hippophae salcifolia.
4. A process for the preparation of a herbal formulation
comprising: grinding a portion of each of Nyctanthes arbortristis,
Ocimum tenuiflorum, and Hippophae salcifolia, to provide ground
herbal matter, subjecting the ground herbal matter to the step of
extraction using water to provide a: filtering the extracts to
provide a filtered product; subjecting the filtered product to the
step of distillation under vacuum; evaporating the water and drying
the filtered product under vacuum; and grounding and sieving the
filtered product.
5. The process as claimed in claim 4, wherein the temperature at
time of extraction is 70.degree.-80.degree. C. and the extraction
process lasts for a period of 3-4 hrs.
6. The process as claimed in claim 4, wherein the temperature of
distillation is 65.degree. C.
7. (canceled)
8. (canceled)
9. A method of treating depression in a patient comprising
administering to a patient in need thereof a formulation containing
extract of Nyctanthes arbortristis and Hippophae salciofolia in an
amount effective to promote sleep pattern and improve wake-cycle
and depression symptoms.
10. (canceled)
11. (canceled)
12. A method of treating depression in a patient comprising
administering to a patient in need thereof a formulation comprising
Ocimum tenuiflorum and Hippophae salcifolia in an amount effective
to ameliorate symptoms of depression in the patient.
13. The method of claim 12, wherein administering the formulation
improves brain functional impairment, maintains hippocampal volume,
and/or enhances neuronal plasticity in the patient.
14. The method of claim 12, wherein administering the formulation
improves sleep quality in the patient.
15. A method of treating depression in a patient comprising
administering to a patient in need thereof a formulation comprising
Nyctanthes arbortrist and Ocimum tenuiflorum in an amount effective
to ameliorate symptoms of depression in the patient.
16. The method of claim 15, wherein administering the formulation
reduces anxiety and/or phobic reactions in the patient.
17. The method of claim 15, wherein administering the formulation
improvise diffusion and hallucinations in the patient.
18. The method of claim 15, wherein administering the formulation
enhances immunity and/or improves fatigue in the patient.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application claims priority to Indian Patent
Application No. 6161/CHE/2014, filed on Dec. 6, 2014.
FIELD OF INVENTION
[0002] Present invention relates to the development of a plant
based formulation beneficial in the management of mild to moderate
depression. This preparation may be further advantageous if used
for the management of sleep disorders, behavioral abnormalities
like psychotic and neurotic behavior, cognitive dysfunction, etc.
The pharmacological activity of test formulation can be evaluated
on neurotransmitter biochemistry including biogenic amine like
dopamine, GABA, glutamate, 5-HT serotonin and also anxiety and
depression scores as these bio-markers are significantly involved
with depressive behavior. Melatonin is one of the most important
targeted markers involved with depression and having a role in
regulation of sleep. The activity of test formulation can be
assessed on melatonin receptors distributed in different part of
the brain and the drug can activate either the wake related
neuronal down stream pathways or promote sleep related
pathways.
BACKGROUND OF STUDY
[0003] Depression is a significant contributor to the major disease
burden affecting almost all the communities worldwide. Recently the
incidence is reported to be significantly high, mentioning that 350
million people are affected from depression. The World Mental
Health conducted a survey and reported that 1 in 20 people is
having an episode of depression. Depressive disorders often start
at a young age affecting their quality of life. Moreover,
depression often comes with symptoms of anxiety. These complaints
can become chronic or recurrent and lead to substantial impairment
of an individual's ability to take care of him- or herself in day
to day life. The worst part of depressive disorder is the
development of suicidal tendency which is reported around 9000
suicidal deaths every day.
[0004] The clinical symptoms associated with depression are
depressed mood, loss of interest and enjoyment and increased early
fatigue. Based on symptoms a depressive episode can be categorized
as mild, moderate or severe. An individual with mild depressive
episode will have some difficulty in continuing with ordinary work
and social activities.
[0005] The burden of depression is 50 percent higher for females
than males (WHO 2008). Maternal depression may be a major risk
factor for poor growth in young children. Depression affects not
only this generation but also the next.
[0006] Disturbance of the sleep/wake cycle is one of the DSM-IV
diagnostic criteria for depression and people with major depression
commonly experience changes in sleep wake cycle regulation that are
seen as an abnormal total sleep duration, poor sleep efficiency
overwhelming rapid eye movement (REM) sleep and early morning
awakening.
[0007] A good number of depressive patients show regular changes in
the intensity of depressive mood during the day, with parallel
changes in anxiety symptoms that frequently accompany depression.
Further, in patients with unipolar and bipolar depression, evidence
has been provided for a phase advance of the temperature--REM sleep
cycle in relation to the rest/activity cycle, and melatonin
secretion studies of patients with depressive disorders.
[0008] A number of studies have investigated variants of genes that
control the circadian system for their association with mood
disorder and circadian related polymorphism in depressive
disorders.
[0009] Recently attention has been paid towards the investigations
of patho-physiology involved with affective disorders and their
efficient treatment. The drug being used for the management of
depression is based on modulation of nor-adrenergic and
serotonergic neurotransmitters mainly known as nor-adrenergic
reuptake inhibitors as well as MAO inhibitors. A significant
decrease in nor-adrenergic and serotonergic neuro-hormones are
reported which are regulated through intervention of
anti-depressant agents. As reported over activation of glutamate
and the decreased activity of GABA is responsible for mood
disorders. However, a long term treatment can only produce desired
results and moreover in most of the depression patient this
treatment is not much effective. Further, the workers have reported
that the anti-depressant drugs influence excitatory and inhibitory
neurotransmitter which contributes to their efficacy. Thus, this
type of activity may have better anti-depressant effect.
[0010] The treatment option consists of basic psychosocial support
combined with anti-depressant medication along with psychotherapy
such as cognitive behavior therapy, interpersonal psychotherapy or
problem solving treatment. Anti-depressant medications and brief
psychotherapy programs are effective. Anti-depressants can be a
very effective form of treatment for moderate to severe depression
but are not the first line treatment.
[0011] It is postulated that depression is the outcome of
deficiency of various amines which serve as neuro-transmitters in
the brain. The tricyclic agents are anti-cholinergic effects, may
cause postural hypertension and cardio-toxicity. Currently
available drugs are fluoxetine that are selective inhibitors of
5-HT re-uptake, MAO inhibitors which enhances the availability of
neuro-transmitters at synaptic cleft by inhibiting metabolism of
nor-epinephrine and 5-HT, and are less effective than tricyclic or
less effective in severe depression. Further, studies are there
showing that MAO inhibitors are more useful when depression is
associated with anxiety and phobic reactions. A number of studies
have indicated an increased level of platelet MAO in large series
of depressed patients. It is also pointed out that the
administration of tyrosine in the body increases the rates of brain
neuron synthesis of both dopamine and nor-epinephrine. A decreased
symptoms of depression after tyrosine administration is reported
which in turn indicates the decrease in the level of tyrosine and
increased MAO activity.
[0012] Anti-depressant drugs act through neurotransmitters in
circulation as well as brain tissues. It has been pointed out that
catecholamine and serotonin metabolism is impaired during mental
depression. Any synthetic medication beneficial in the management
of psychiatric illness produces incontinence, discomfort,
pathologic changes and some times danger to the patients.
[0013] However, the mechanism of action of anti-depressant drugs
must be considered in terms of its activity on melatonin receptors
(MT.sub.1 & MT.sub.2 receptors), distributed in the brain,
present in suprachiasmatic nucleus (SCN) of prefrontal cortex,
cerebellar cortex, hippocampus, basal ganglia, substantia nigra,
ventral segmental area, nucleus accumbens, etc. Another possible
mode of action of an anti-depressant drug is MT.sub.1 mediated
effects in SCN favor sleep initiation via the hypothalamic sleep
switch, a mechanism characterized by typical on-off responses. The
mechanism involved with this kind of sleep pattern is mainly
characterized by typical on-off responses. The mode of action
activates either wake-related neuronal downstream pathways or
promotes sleep-related pathways. Apart from sleep promotion,
melatonin receptors also appear to be involved in sedating and
anti-excitatory effects of melatonergic drugs. This type of
activity is mainly studied in relation to anti-convulsant role in
which the role of melatonin is assessed on .gamma.-amino-butyric
acid (GABA) transmission.
[0014] The activity of test formulation may be through its action
on GABA receptor complex that blocks GABA-mediated inhibition
indicating that the drug is mediated by GABA receptors, not via
5-HT receptor inhibition and effect could be related to
melatonergic action on the cholinergic system. Further, it is
proposed that the drug may have alerting action. It is explained as
5-HT receptors are concentrated in frontal cortex, amygdala,
hippocampus and cortico-limbic structures that are responsible in
the regulation of mood and cognition.
[0015] Due to global awareness, attention has been directed towards
the utilization of Ayurvedic plants in the prevention and
management of many physical and mental disorders. In the Ayurvedic
system of medicine, several drugs are known to influence the human
behavior. But the scientific evidence is lacking. Due to the
limitation of modern drugs, several plant based drugs have been
advocated for the prevention and management of depression and
associated behavioral changes. Ayurveda has listed a group of
plants acting on brain function i.e. sp Areca catechu (L). Bacopa
monniert (L), Celestrus paniculatus (L). Centella asiatica (L),
Curcuma longa (L), Nardostachys jatamansi (L), Withania somnifera
(L), Acorusm calamus, Argyreia speciosa, Clitoria ternatea,
Convolulus pluricaulis. Dioscorea bublifera. Glycyrrhiza glabra,
Jasminun sambac, Mucuna pruriens, Terminalia chebula, Tinosporu
cordifolia, Valeriana jatamansi, which are known to improve mental
performance, regulation of behavior and sleep pattern. Out of these
plants some of them i.e. Areca catechu (L), Bacopa monnieri (L),
Celestrus paniculatus (L), Centella asiatica (L), Curcuma longa
(L), Withania somnifera (L) are investigated for their cognitive
function amelioration property. Taking the lead from ancient text
we have investigated the anti-depressant property of plant based
formulation containing the aqueous extract of Nyctanhes
arbortristi, Ocimum tenuiflorum and Hippophae salcifolia on various
biomarkers involved with different degrees of depression.
BRIEF SUMMARY OF THE INVENTION
[0016] An object of present invention is to propose a plant based
anti-depressant drug for the management of mild to moderate
depression.
[0017] Another object of this invention is to propose a plant based
formulation having tricyclic or MAO inhibitory agent like
activity.
[0018] Still another object of this invention is to propose a plant
based formulation having potential to enhance melatonin
concentration and improve the sleep pattern along with anxiety in
depression cases.
[0019] Yet another object of this invention is to propose a plant
based formulation which can regulate the altered neurotransmitter
biochemistry in depression cases.
[0020] Further object of this invention is to propose a plant based
formulation which can manage the mood affected behavior of
depressive patients.
[0021] Still further object of this invention is to propose a plant
based formulation for the management of emotional, social and
cognitive functioning in depressive patients.
STATEMENT OF INVENTION
[0022] As per the present invention there is provided a novel plant
based formulation showing therapeutic benefits in the management of
mild to moderate depression through its potential therapeutic
effects via acting on GABA receptors not via 5-HT receptor
inhibition rather through its activity on melatonergic receptor in
the cholinergic system. The drug may also have effects on 5-HT
receptors accumulated in the frontal cortex, amygdala, hippocampus
and cortico-limbic structures responsible for regulation of mood
and cognition. It has been proposed that through circulatory
neurotransmitters as well as neurotransmitters present in brain
tissues, catecholamine and serotonin metabolism is impaired during
mental depression.
[0023] Recent data suggested that 350 million people are affected
from depression and out of 20 persons, 1 is having an episode of
depression. As it is well established that the ultimate goal of
anti-depressant treatment is the symptomatic and functional
recovery helps a return to normal daily life functioning. Giving
more importance to emotional, cognitive and social functioning in
day to day life of depressive patients impaired functioning may
have effect on a patient's life, therefore more alteration should
be given on functioning when assessing the response of treatment of
anti-depressant agent.
[0024] In the present study the pre-clinical safety and efficacy
profile suggested that test formulation may exert its
anti-depressant activity via its interaction with marker receptors
i.e. melatonin and 5-HT The test formulation is beneficial in
improving sleep and in desynchronizing the disturbed circadian
rhythms making the test drug as a very useful, safe and effective
anti-depressant agent. Thus the test formulation has potential to
improve emotional, cognitive and social impaired behavior
manifesting due to depressive behavior.
[0025] In fact depression is the leading cause of disease burden
for women in both low as well as high income countries. Looking
towards the global burden of depression which poses a substantial
public health challenge both at social and economic levels as well
as clinical levels, there are variety of evidence based therapeutic
agents which can effectively combat this disease burden. Based on
present study outcome it is proposed that treating depression
following test formulation treatment is of clinical significance,
affordable and also cost effective.
[0026] Based on the above approach of management of mild to
moderate depression, under this invention there is provided a
process for the preparation of present plant based formulation
beneficial in the management of depression as claimed in claim 1.
The test formulation comprising of aqueous extract of Nyctanthes
arbortristis (Parijat--whole plant), Ocimum tenuiflorum
(Van-Tulsi--whole plant) and Hippophae salcifolia
(Seabuckthorn--Leaves & Fruits) in effective doses determined
in pre-clinical studies as per GLP, GCP, ICH guidelines. The
extraction of plant material was done at 70.degree.-80.degree. C.
and maintaining the pH of solution between 4-6, separating the
active compound chromatographically present in each plant extract
by using TLC, HPLC and HPTLC supporting the molecular
characterization of plant extract by utilizing IR & NMR The
genetic bar coding of each plant was also done to confirm plant
species, before starting the extraction process.
BRIEF DESCRIPTION OF THE DRAWINGS
[0027] FIG. 1 shows a process for providing a formulation according
to one embodiment of the present invention.
DETAILED DESCRIPTION OF INVENTION
[0028] The aqueous extract of three medicinal plants found in the
Tribal area of Madhya Pradesh i.e. Nyctanthes arbortristis, Ocimum
tenuiflorum and Hippophae salcifolia the extract was prepared using
RO water and utilized for the development of present novel
formulation. The Water utilized for extraction was decontaminated
for any type of bacterial or abnormal growth by using reverse
osmosis plant. After extraction the active molecules were
identified and separated by HPLC, HPTLC and NMR methods. A
mechanism based studies on each plant extract as well as combined
formulation was carried out in various experimentally designed
models. Both toxicity as well as the efficacy profile were studied.
A dose dependent study was also conducted to determine the most
active and safe dose of each plant extract. The pharmacological and
biological activity of each plant extract selected to develop a new
formulation for the management of depression of varying etiology
was done through conducting mechanism based studies and the mode of
action by assessing their effect on various targets involved with
depression.
[0029] Under this procedure, the effect of test formulation was
evaluated for its 5-HT and MAO inhibitory role as well as its
activity as agonist of melatonin receptors in experimental studies.
Standardization of biologically active material is another
prerequisite for the determination of effective dose. Based on
documents it has been observed that no clinical data is available
to demonstrate the effects of this formulation on human behavior.
In our preliminary clinical trial this formulation has shown
beneficial effect in elevating the mood as well as improved sleep
in depressed patients; as the test formulation has shown regulation
of neurotransmitter system particular GABA & MAO activity along
with its activity on rhythmic release in melatonin content among
patients suffering from different degree of depression.
Extraction Procedure:
[0030] The shed dried raw material of Nyctanthes arbortristis
(whole plant), Ocimum tenuiflorum (whole plant) and Hippophae
salcifolia (Leaves & Fruits) were taken separately in corous
form for extraction. The determination of presence of specific
active molecules/active constituents as well as its quantification
was done by using IR & NMR. The extraction was done at the
temperature of 70.degree.-80.degree. C. and the pH of solution was
maintained at 4-6. The following procedure was adopted to isolate
the active compound, to evaluate and assess the biological activity
of test formulation and to develop a new drug entity having a
beneficial role in the management of mild to moderate depression.
(FIG. 1).
Steps--Procedure
[0031] High through-put-screening of plant species chromatographic
separation of active constituents is based on the following
steps--
[0032] According to this invention there is provided a plant based
formulation showing efficacy in the management of mild to moderate
depression through its activity on neurotransmitter and biogenic
pathways, behavior pattern and inducing sleep. The present test
formulation comprising the following ingredients--
TABLE-US-00001 Name of the plants Part used I. Nyctanthes
arbortristis Whole plant 2. Ocimum tenuiflorum Whole plant 3.
Hippophae salcifolia Leaves & Fruits
[0033] Preferably the aforesaid plant extracts are present in the
following dose range in the test formulation--
TABLE-US-00002 Name of the plants Dose I. Nyctanthes arbortristis
225-450 mg/day 2. Ocimum tenuiflorum 250-550 mg/day 3. Hippophae
salcifolia 200-450 mg/day
[0034] The formulation may also comprise known additives such as
minerals, vitamins, salt, filler (for capsulation or to prepare
syrup) and binders, if required to present in trace amount.
[0035] Thus any known additive or supplement is added to prepare
the final formulation required in trace amount. Reference is made
here in capsule form. However, it would be apparent that the
preparation may also be in the form of syrup/tablet.
[0036] Preferably but without implying any limitation the present
preparation comprises the aqueous extract of following selected
plants in the following effective doses.
[0037] Preferably the aforesaid plants are present in the following
doses in the test formulation--
TABLE-US-00003 Name of the plants Dose I. Nyctanthes arbortristis
325 mg/day 2. Ocimum tenuiflorum 350 mg/day 3. Hippophae salcifolia
275 mg/day
Hypothesis:
[0038] Imbalance of GABA, glutamate and decreased amount of
platelet MAO B and disturbance in melatonin metabolism are
responsible for anxiety and depression. Several genes have been now
recognized to influence anxiety and depression. Similarly
regulation of various receptors like NMDA, AMPA, MT.sub.1 and
MT.sub.2 including glutamatergic, serotonergic, monoaminergic
receptors play a crucial role in the etiopathogenesis of
depression.
[0039] Some medicinal plants have shown significant effect in the
modulation of various neurotransmitter systems including GABA and
glutamate levels. Similarly some of plant extracts have shown
significant effect in enhancement of melatonin which regulates the
sleep pattern. Regulation of sleep patterns which is under the
control of clock gene may serve as a useful target for management
of depression.
[0040] The plants selected in the study contains various molecules
have shown a significant role in management of depression by
maintaining GABA-Glutamate homeostasis. Further, regulation of
platetet monoamine-oxidase activity also play important role in the
management of depression and as well as regulation of sleep
pattern.
[0041] In present invention combined effect of 3 plants containing
specific bio-molecules acts on above receptors has been
evaluated.
[0042] In order to prove above hypothesis, a novel process has been
developed by using various inventive steps to propose a new drug in
the management of mild to moderate depression.
1. Nyctanthes arbor-tristis (Night-flowering Jasmine) is a species
of Nyctanthes, belongs to family Oleaceae found all over the
country. The major chemical constituents present in various parts
of the plant is the leaves contain .beta.-sitosterol, flavanol
glycosides, oleanolic acid, ascorbic acid, lupeol, glycosides, etc.
The flowers mainly contain essential oils, carotenoids, glycosides.
The seeds contain linoleic, palmitic and a water soluble
polysaccharide. The bark contains glycosides and alkaloids. In
stems the major chemical constituents found are glycoside,
.beta.-sitosterol. However, the active constituents found in leaves
of the plant have activity like anti-inflammatory,
hepatoprotective, immunopotentiating and antioxidant. Flowers
mainly have activity like anti-inflammatory, anti-oxidant and
sedative. On the whole the pharmacological activity of this plant
is sedative, anti-inflammatory, anti-oxidant and immunomodulatory.
2. Hippophae salcifolia (Seabuckthorn)--This is high altitude plant
belongs to family Elaeagnaceae. Fruits and leaves have shown
medicinal property. Hippophae salcifolia is a rich source of
flavonoids, vitamins, proteins, amino acids, folic acid,
phytosterol, alpha-tocopherol and phenolic compounds. It has shown
anti-oxidant, immuno-modulatory, anti-inflammatory and homocysteine
lowering effects and uplifts the mental function. 3. Ocimum
tenuiflorum (Van Tulsi), This plant belongs to family Lamiaceae, it
has wild growth and available throughout the Eastern World Tropics.
Some of the main chemical constituents of this plant is eugenol,
oleanolic acid, ursolic acid, rosmarinic acid, etc. The main
pharmacological activity is COX-2 inhibitor, analgesic effect due
to high concentration of eugenol, anti-oxidant, repairs cell damage
due to exposure to radiation, anti-hyperlipidemic and
cardioprotective effects, promotes immune system function.
Rationale for Selection of Plants:
[0043] Based on various pre-clinical and clinical studies it has
been postulated that the active molecules present in three plants
regulates the GABA-Glutamate balance, inhibits the MAO activity and
regulates the Melatonin secretion. Thus the combined formulation is
beneficial in the management of depression.
EXAMPLE
Example-1
[0044] When the aqueous extract of Nyctanthes arbortristis (75
mg/kg), Ocimum tenuiflorum (75 mg/kg) and Hippophae salcifolia (50
mg/kg) was administered orally to animals of sleep deprivation
stress model exhibited its activity as 5-HT antagonist as well as
melatonin agonist effects suggesting improved sleep pattern as well
as its anti-depressant potentials. The drug is having 5-HT.sub.2c
receptor antagonist activity as it inhibits serotonin re-uptake by
acting on several neuroplasticity associated molecules in the
associated brain region. 5-HT antagonism will predominate on
melatonergic action.
Example-2
[0045] The aqueous extract of Ocimum tenuiflorum in the dose of 125
mg/kg and Hippophae salcifolia in the dose of 125 mg/kg mixed and
given to experimentally designed sleep deprived rats a significant
reduction in glutamate content was noticed along with increase in
GABA synthesis, suggesting the improved brain fimction impairment
in terms of anti-depressant activity of test drug. Overall the
neuronal plasticity was enhanced. The test formulation acted as
MT.sub.1 and MT.sub.2 agonist and maintained the hippocampal
volume,
Example-3
[0046] After determination of safety and efficacy of test
formulation in pre-clinical studies the test formulation was
utilized for human consumption. When the aqueous extract of
Nyctanthes arbortristis in the dose of 425 mg/day and Ocimum
tenuiflorum in the dose of 450 mg/day was mixed and given to
selected cases of mild to moderate depression, improvement in loss
of interest in various activities, loss of self esteem, loss
of energy and depressed mood was noticed through its activity on
neurotransmitter biochemistry.
Example-4
[0047] When the aqueous extract of Nyctanthes arbortristis in the
dose of 375 mg/day and Hippophae salcifolia in the dose of 400
mg/day was given to selected depression cases a very good response
in improving the functional impairment was noticed among depressive
patients. The psychomotor co-ordination/retardation of depressed
patients improved and a better cognitive function was recorded in
selected depression cases.
Example-5
[0048] When the aqueous extract of Nyctanthes arbortristis in the
dose of 450 mg/day and Ocimum tenuiflorum in the dose of 425 mg/day
orally administered to patients with depressive behavior
improvement in fatigue (energy) and general body immunity was
noticed. The early fatigue time enhanced significantly after
treatment with test formulation determined through Electronic
Fatigue Time Apparatus.
Example-6
[0049] The Aqueous extract of Ocimum tenuiflorum in the dose of 375
mg/day and Hippophae salcifolia in the dose of 450 mg/day given in
combined form to diagnosed depressive patients the sleep duration
enhanced, sleep/wake cycle pattern suggested the improvement in
sleep and early morning wake including quality of every day
performance of the patients.
Example-7
[0050] When the aqueous extract of Nyctanthes arbortristis in the
dose of 375 mg/day, Ocimum tenuiflorum in the dose of 325 mg/day
and Hippophae salcifolia in the dose of 225 mg/day given in
combined form to depression cases a complete and sustained recovery
from depression, psychosocial impairments was noticed. The
anti-depressant activity of test drug is through the prevention of
breakdown like MAO inhibitor or reduction in re-uptake like
tricyclic agent.
Example-8
[0051] When the aqueous extract of Nyctanthes arbortristis in the
dose of 400 mg/day, Hippophae salcifolia in the dose of 375 mg/day
and Ocimum tenuiflorum in the dose of 150 mg/day orally given in
combined form to depression cases a significant decrease in
depression scores were noticed when evaluated on Beck Depression
Inventory Scale.
Example-9
[0052] When the aqueous extract of Nyctanthes arbortristis in the
dose of 425 mg/day, Hippophae salcifolia in the dose of 350 mg/day
and Ocimum tenuiflorum in the dose of 175 mg/day given to selected
depression patients the Hamilton Depression Rating Scale analysis
showed remarkable decrease in depression scores along with
reduction in anxiety scores also.
Example-10
[0053] When the aqueous extract of Nyctanthes arbortristis in the
dose of 325 mg/day, Ocimum tenuiflorum in the dose of 350 mg/day
and Hippophae salcifolia in the dose of 275 mg/day given to mild to
moderate depression patients most effective results were obtained.
The novel test formulation is an agonist of MT.sub.1 and MT.sub.2
melatonin receptors and an antagonist of 5-HT.sub.2c receptors. Its
anti-depressant activity is proven through its interaction with
both type of receptors. The psychosocial impairment increases the
severity of depressive symptoms, greatly improved social
functioning, improved personal and professional functioning among
patients with depression, resulting in improvement in routine
quality of physical and mental work performance.
[0054] The test formulation revealed disease remission, improved
functional impairment scores related to emotional, cognitive and
social factors. An improvement in loss of interest, decreased self
esteem, loss of energy, depressed mood was noticed which corrects
the patients ability to maintain relationship under influence of
test formulation treatment.
Regulation of Biogenic Amines Following Combined Formulation
Treatment:
[0055] The study is divided into four groups, 10 rats of Charles
foster strain were kept in each study group.
Group-I: In control group 10 rats were selected and kept on normal
saline for 7 days. On 7.sup.th days the animals were sacrificed and
various biogenic amines like 5-HT GABA & glutamate, melatonin
and MAO were estimated. After dissection of rat's brain the
biogenic amines were estimated in thalamus, hypothalamus and
cortex. Group-II: In this group sleep deprivation stress for 7 days
was given to all the 10 rats. After 7 days the rat's brain were
dissected and thalamus, hypo-thalamus and cortex were separated for
the measurement of brain biogenic amines. Group-III: In this group
the sleep deprivation stress was introduced along with the
hydro-alcoholic extract of three plants Nyctanthes arbortristis,
Ocimum tenuiflorum and Hippophae salcifolia The drug Nyctanthes
arbortrists (100 mg/kg bw./day), Ocimum tenuiflorum (50 mg/kg
bw./day), Hippophae salcifolia (50 mg/kg bw./day) was given in
combined form to all the 10 rats along with sleep deprivation
stress. On 7.sup.th day (after two hours of administration of test
drug) different parts of the rat's brain were dissected and 5-HT,
melatonin GABA. glutamate and histamine were measured. Group-IV:
The modern drug "ibrium" was given in the dose of 2 mg/kg/day for 7
days along with sleep deprivation stress. On 7.sup.th days the
animals were sacrifice and brain biogenic amines were estimated in
different parts of rat's brain.
[0056] The value obtained were compared with each other group
(Normal vs sleep deprivation stress and sleep deprivation stress vs
sleep deprivation stress+test formulation and sleep deprivation
stress vs modern drug treatment group. Apart from initial at 7 day
before sacrificing the animals circulating biogenic amines were
also measured in all the three groups.
Observation and Result:
[0057] As per the object of the present experimental study the
effect of test formulation containing the hydro-alcoholic extract
of three plants Nyctanthes arbortristis, Ocimum tenuiflorum and
Hippophae salcifolia in effective doses was orally induced to
animals following sleep deprivation stress. The beneficial role is
evaluated on various brain biogenic amines.
[0058] It is observed that 7 days sleep deprivation stress exerted
marked alteration on brain biogenic amines i.e. 5-HT, melatonin,
GABA, glutamate and also a biogenic amines like GABA, glutamate,
5-HT, MAO, histamine and histaminase levels, 5-HT level increased
markedly following sleep deprivation in all the three parts of
brain. The increase in the level is significantly less in test
formulation treated group following sleep deprivation stress (Table
1).
[0059] Melatonin and GABA levels were significantly decreased
following stress in comparison to normal control group. The test
formulation revealed its beneficial role in controlling the decline
of levels as decrease is significantly less in GABA content
following stress (Table 2-3).
[0060] The experimental rats following sleep deprivation stress
showed a significant rise in glutamate content in the different
parts of the brain. The 7 days treatment with test drug along with
stress the increase in the glutamate content is quite less than the
negative control group (Table 4).
[0061] The circulating brain biogenic amines GABA, glutamate, 5-HT,
histamine and histaminase significantly increased following sleep
deprivation stress where as MAO levels were decreased under stress.
All these altered levels of circulating biogenic amines regulated
under Ayurvedic test drug therapy. Decrease in MAO levels were
significantly less in treated groups than only sleep deprivation
stress groups of animals (Table5).
[0062] It is observed that the modern drug Librium exerted better
results on the neurochemical parameters but keeping the adverse
reaction and restricted application of modern synthetic chemicals
used for inducing sleep, the present Ayurvedic test formulation may
be a better choice of drug in the regulation of sleep patterns.
TABLE-US-00004 TABLE 1 Beneficial role of test formulation on 5-HT
level in experimental sleep deprivation stress model. 9 5-HT
(.mu.g/gm in wet tissue) Groups No. of Animals Hypothalamus
Thalamus Cortex Normal control* 10 1.685 .+-. 0.162 1.345 .+-.
0.295 0.852 .+-. 0.238 Sleep deprivation** Stress 10 2.876 .+-.
0.356 1.918 .+-. 0.306 1.476 .+-. 0.238 Sleep deprivation streess +
test formulation*** 10 2.196 .+-. 0.446 1.530 .+-. 0.285 1.145 .+-.
0.372 Sleep deprivation stress + Librium (2 mg/kg)**** 10 1.942
.+-. 0.315 1.460 .+-. 0.218 1.093 .+-. 0.118 Comparison * vs ** P
< 0.001 P < 0.001 P < 0.001 ** vs *** P < 0.01 P <
0.02 P < 0.01 *** vs **** P < 0.05 P < 0.05 P <
0.05
TABLE-US-00005 TABLE 2 Beneficial role of test formulation on
melatonin level in experimental sleep deprivation stress model.
Melatonin (.mu.g/gm in wet tissue) Groups No. of Animals
Hypothalamus Thalamus Cortex Normal control* 10 11.67 .+-. 3.02
7.99 .+-. 3.46 9.86 .+-. 2.08 Sleep deprivation** Stress 10 5.91
.+-. 1.34 3.98 .+-. 2.01 4.73 .+-. 2.32 Sleep deprivation stress +
test formulation*** 10 9.33 .+-. 3.01 5.31 .+-. 2.38 6.25 .+-. 2.95
Sleep deprivation stress + Librium (2 mg/kg)**** 10 7.80 .+-. 1.02
5.95 .+-. 1.85 7.01 .+-. 1.46 Comparison * vs ** P < 0.001 P
< 0.001 P < 0.001 ** vs *** P < 0.01 P < 0.02 P <
0.01 *** vs **** P < 0.01 P < 0.05 P < 0.01
TABLE-US-00006 TABLE 3 Effect of test formulation on GABA content
in the brain following experimental sleep deprivation stress. GABA
(.mu.mol/gm in wet tissue) Groups No. of Animals Hypothalamus
Thalamus Cortex Normal control* 10 3.48 .+-. 0.50 2.17 .+-. 0.39
1.72 .+-. 0.42 Sleep deprivation** Stress 10 6.84 .+-. 1.39 5.04
.+-. 1.12 4.32 .+-. 0.77 Sleep deprivation stress + test
formulation*** 10 4.80 .+-. 1.22 3.75 .+-. 0.82 2.94 .+-. 0.82
Sleep deprivation stress + Librium (2 mg/kg)**** 10 4.68 .+-. 1.02
3.91 .+-. 0.58 2.62 .+-. 0.59 Comparison * vs ** P < 0.001 P
< 0.001 P < 0.001 ** vs *** P < 0.01 P < 0.02 P <
0.02 *** vs **** P < 0.05 P < 0.05 P < 0.05
TABLE-US-00007 TABLE 4 Effect of test formulation on glutamate
level following experimental sleep deprivation stress. Glutamate
(.mu.mol/gm in wet tissue) Groups No. of Animals Hypothalamus
Thalamus Cortex Normal control* 10 11.02 .+-. 2.98 13.06 .+-. 3.24
14.91 .+-. 2.64 Sleep deprivation** Stress 10 16.95 .+-. 2.56 18.04
.+-. 3.11 17.22 .+-. 4.01 Sleep deprivation stress + test
formulation*** 10 13.88 .+-. 2.80 13.82 .+-. 2.75 14.80 .+-. 2.73
Sleep deprivation stress + Librium (2 mg/kg)**** 10 12.66 .+-. 3.06
11.64 .+-. 3.01 13.97 .+-. 2.11 Comparison * vs ** P < 0.001 P
< 0.001 P < 0.001 ** vs *** P < 0.01 P < 0.02 P <
0.01 *** vs **** P < 0.05 P < 0.05 P < 0.05
TABLE-US-00008 TABLE 5 Beneficial role of test formulation on
circulating biogenic amines following experimental sleep
deprivation stress. GABA Glutamate 5-HT MAO Histamine Histaminase
Groups (.mu.mol/ml) (.mu.mol/ml) (.mu.g/ml) (nmol/m//hrs.)
(.mu.g/ml) (.mu.g/ml) Normal Control* 0.91 .+-. 0.22 0.79 .+-. 0.19
0.14 .+-. 0.10 3.90 .+-. 0.65 0.19 .+-. 0.05 30.20 .+-. 4.32 (N =
10) Sleep 1.68 .+-. 0.19 1.26 .+-. 0.12 0.58 .+-. 0.20 7.04 .+-.
1.22 0.51 .+-. 0.15 49.65 .+-. 5.30 deprivation** Stress (N = 10)
Sleep deprivation 1.22 .+-. 0.20 0.95 .+-. 0.15 0.30 .+-. 0.12 5.28
.+-. 1.30 0.32 .+-. 0.11 39.70 .+-. 3.38 stress + test
formulation*** (N = 10) Sleep deprivation 1.02 .+-. 0.18 1.11 .+-.
0.10 0.26 .+-. 0.15 4.72 .+-. 0.89 0.45 .+-. 0.10 35.46 .+-. 4.95
stress + Librium (2 mg/kg)**** (N = 10) Comparison * vs ** P <
0.001 P < 0.001 P < 0.001 P < 0.001 P < 0.001 P <
0.001 ** vs *** P < 0.01 P < 0.01 P < 0.01 P < 0.01 P
< 0.02 P < 0.001 *** vs **** P > 0.05 P > 0.05 P >
0.05 P > 0.05 P > 0.05 P > 0.05
Clinical Study
Material and Methods
[0063] In the present study 99 cases between the age group of 20 to
60 years presenting the symptoms of mild to moderate depression
were selected for clinical trial of test formulation containing
extract of Nyctanthes arbortristis, Ocimum tenuiflorum and
Hippophae salcifolia. The subjects were advised to take test
formulation twice in a day in two divided doses (475 mg morning and
475 mg in the evening). The level of mood was measured by Major
Depression Inventory (MDI) developed by Olsen L R et al (2003), the
fatigue and psychomotor performance was measured by Mental Fatigue
Apparatus and Rotary Pursuit Task. These tests and test battery are
recommended by Gupta and Dubey (1994) for the scientific and
objective assessment in the area of drug development. The
behavioral pattern was repeatedly assessed at the continuous
interval of one month for six months.
[0064] In control series (Sertraline treated) all the
investigations were repeated and compared with the treated
group.
Results
[0065] It is observed that specific clinical features such as lack
of interest, poor sleep, poor appetite, weight loss, and
psychomotor retardation showed considerable improvement after six
months of continuous administration of test formulation. In the
treated group significant improvement in clinical symptomatology,
after six months of treatment was noticed, whereas the
conventionally treated group did not exert such improvement in this
series (Table-1).
TABLE-US-00009 TABLE 1 Improvement in clinical symptomatology (in
percent) following test formulation treatment. Treated with Test
formulation Sertraline treated Clinical symptomatology Initial
After 6 months Initial After 6 months Lack of interest 97.82 92.20*
98.42 75.51.degree. Poor concentration 88.41 86.02* 92.73
82.90.degree. Pessimistic thinking 96.27 95.86* 95.90 87.32.degree.
Early fatiguability 98.01 91.12* 97.45 78.26.degree. Poor sleep
98.04 79.41* 96.34 74.35.degree. Poor appetite 58.89 50.03* 61.33
33.78.degree. Weight loss 42.01 40.32* 43.81 17.35.degree.
Psychomotor retardation 74.66 78.09* 79.11 57.35.degree. *No
improvement in percent; .degree.Improvement in percent
[0066] The alpha brain wave which indicates the relaxed state of
mind also showed significant improvement in the treated group. The
initial value of alpha wave significantly increased after six
months treatment. However, the control group did not show any
change in the alpha pattern (Table 2).
TABLE-US-00010 TABLE 2 Effect of test formulation on alpha brain
wave among minor depressive cases Comparison Alpha Brain Wave (Hz)
Initial Vs After 2 After 4 After 6 After 6 Groups Sample size
Initial months month months months Sertraline 41 6.18 .+-. 2.03
6.31 .+-. 2.41 5.93 .+-. 2.18 5.76 .+-. 1.93 p > 0.05 treated
Test 58 5.92 .+-. 2.47 7.56 .+-. 2.91 9.15 .+-. 3.23 10.38 .+-.
3.19 P < 0.05 formulation treated
[0067] The mental fatigue level also showed considerable
improvement after six months of treatment in comparison to control
group. The initial value of mental fatigue level significantly
increased after six months in the treated group (Table 3).
TABLE-US-00011 TABLE 3 Effect of test formulation on mental fatigue
level among minor depressive cases Comparison Mental fatigue level
(Hz) Initial Vs After 2 After 4 After 6 After 6 Groups Sample size
Initial months months months months Sertraline 41 18.22 .+-. 3.17
19.68 .+-. 4.02 19.02 .+-. 3.72 18.45 .+-. 2.11 P > 0.05 treated
Test 58 17.86 .+-. 3.16 21.24 .+-. 2.90 23.10 .+-. 3.62 24.98 .+-.
3.87 P > 0.05 formulation treated
[0068] The rotatory-pursuit task which measures psychomotor
performance showed significant improvement after six months of
treatment of test formulation in minor depressive cases. The task
performance considerably improved in the treated group as errors
reduced significantly after six months of therapy in comparison to
the control group (Table 4).
TABLE-US-00012 TABLE 4 Effect of test formulation on psychomotor
coordination among minor depressive cases Comparison Rotatory
pursuit task (Hz) Initial Vs After 2 After 4 After 6 After 6 Groups
Sample size Initial months months months months Sertraline 41 28.39
.+-. 3.74 28.03 .+-. 3.58 27.47 .+-. 2.14 26.52 .+-. 3.46 P >
0.05 treated Test 58 30.13 .+-. 4.57 28.42 .+-. 4.73 25.98 .+-.
5.13 24.78 .+-. 5.02 P > 0.05 formulation treated
TABLE-US-00013 TABLE 5 Changes in anxiety level and mental fatigue
time following oral administration of test formulation in
depressive disorder cases. Anxiety level Comparison Mental fatigue
Comparison (score) (initial vs (Time) (initial vs Treatment Sample
After 6 after 6 After 6 after 6 groups size Initial months months)
Initial month months) Sertraline 41 56.5 .+-. 4.78 58.5 .+-. 3.35
<0.05 94.5 .+-. 12.85 90.3 .+-. 10.78 >0.05 treated Test 58
61.40 .+-. 3.11 54.82 .+-. 3.79 <0.05 86.6 .+-. 14.87 70.77 .+-.
12.85 <0.05 formulation treated
TABLE-US-00014 TABLE 6 Change in attention and alpha frequency
after oral administration of test formulation in depression
patients. Attention span Comparison Alpha Comparison (Score)
(Initial vs frequency (Hz) (initial vs Clinical After 6 after 6
After 6 after 6 groups Initial months months) Initial months
months) Sertraline treated 10.49 .+-. 2.78 9.32 .+-. 1.64 >0.05
6.42 .+-. 1.08 7.92 .+-. 0.73 >0.05 (N = 41) Test formulation
8.73 .+-. 2.45 12.68 .+-. 2.01 <0.05 5.98 .+-. 1.22 8.12 .+-.
2.06 <0.05 treated (N = 58)
TABLE-US-00015 TABLE 7 Effect of test formulation on GABA and
glutamate levels in depression cases. GABA (.mu.mol/ml) Comparison
Glutamate (.mu.mol/ml) Comparison Clinical After 6 (initial vs
after After (initial vs after 6 groups Initial months 6 months)
Initial 6 months months) Sertraline treated 1.070 .+-. 0.385 1.410
.+-. 0.475 >0.05 0.419 .+-. 0.094 0.385 .+-. 0.092 >0.05 (N =
41) Test formulation 1.64 .+-. 0.421 1.02 .+-. 0.211 <0.05 0.502
.+-. 0.101 0.386 .+-. 0.099 <0.05 treated (N = 58)
TABLE-US-00016 TABLE 8 Changes in platelet MAO activity following
oral administration of test formulation in depressive disorder
cases. Platelet MAO (nmol/ml/hr) Comparison Glutamate (nmol/mg/hr)
Comparison Clinical After 6 (initial vs after After 6 (initial vs
after 6 groups Initial months 6 months) Initial months months)
Sertraline treated 5.04 .+-. 1.62 5.98 .+-. 1.35 >0.05 3.14 .+-.
0.08 4.23 .+-. 1.01 >0.05 (N = 41) Test formulation 6.11 .+-.
2.04 4.21 .+-. 0.77 <0.05 4.10 .+-. 0.85 2.87 .+-. 0.31 <0.05
treated (N = 58)
TABLE-US-00017 TABLE 9 Changes in epinephrine and norepinephrine
level under influence of test formulation in depressive cases.
Epinephrine Comparison Norepinephrine Comparison ng/ml (initial vs
ng/nl (initial vs Clinical After 6 after 6 After 6 after 6 groups
Initial months months) Initial months months) Depression C 5.32
.+-. 1.308 6.42 .+-. 1.928 >0.05 8.5 .+-. 1.83 9.82 .+-. 1.78
>0.05 (N = 41) T 6.20 .+-. 1.378 4.40 .+-. 1.038 <0.05 7.5
.+-. 2.85 6.20 .+-. 1.132 >0.05 (N = 58)
TABLE-US-00018 TABLE 10 Decrease in interleukin-1.beta.
concentration following test formulation treatment among mild to
moderate depressive patients. Comparison IL-1.beta. (.mu.g/ml)
Initial Vs After 2 After 4 After 6 After 6 Groups Sample size
Initial months months months months Sertraline 41 2.098 .+-. 0.624
1.886 .+-. 0.312 1.923 .+-. 0.228 1.790 .+-. 0.312 P > 0.05
treated Test 58 2.194 .+-. 0.921 1.773 .+-. 0.624 1.462 .+-. 0.558
1.398 .+-. 0.486 P > 0.05 formulation treated
TABLE-US-00019 TABLE 11 Effect of test formulation on TNF-.alpha.
among mild to moderate depressive patients. TNF-.alpha. (ng/mL)
Comparison Sample After 2 After 4 After 6 Initial Vs After 6 Groups
size Initial month months months months Sertraline 41 4.82 .+-.
1.06 3.71 .+-. 1.22 3.85 .+-. 1.12 4.14 .+-. 1.06 P > 0.05
treated Test 58 5.13 .+-. 1.06 4.63 .+-. 1.02 4.19 .+-. 0.98 3.58
.+-. 0.88 P < 0.01 formulation treated
TABLE-US-00020 TABLE 12 Effect of test formulation on TNF-.alpha.
among mild to moderate depressive patients. DHA (g/100 g of total
fatty acid) Comparison Sample After 2 After 4 After 6 Initial Vs
After 6 Groups size Initial months months months months Sertraline
41 1.096 .+-. 0.246 1.082 .+-. 0.193 1.920 1.980 .+-. 0.318 P >
0.05 treated 0.214 Test 58 1.047 .+-. 0.106 1.284 .+-. 0.293 1.886
.+-. 0.421 2.011 .+-. 0.398 P < 0.01 formulation treated
TABLE-US-00021 TABLE 13 Decrease in muscle action potential
following test formulation treatment in depression cases Occipito
frontalis muscle action potential Comparison Sample After 2 After 4
After 6 Initial Vs After 6 Groups size Initial month months months
months Sertraline 41 43.78 .+-. 17.23 41.75 .+-. 15.25 42.82 .+-.
11.64 40.86 .+-. 16.22 P > 0.02 treated Test 58 57.13 .+-. 21.45
45.87 .+-. 15.90 37.22 .+-. 20.42 33.49 .+-. 13.25 P < 0.001
formulation treated
TABLE-US-00022 TABLE 14 Delta activation following test formulation
treatment in depression cases Delta Wave (Hz) Comparison Sample
After 2 After 4 After 6 Initial Vs After 6 Groups size Initial
month months months months Sertraline 41 0.68 .+-. 0.09 0.94 .+-.
0.12 1.06 .+-. 0.11 1.02 .+-. 0.10 P > 0.05 treated Test 58 0.71
.+-. 0.13 1.22 .+-. 0.09 1.45 .+-. 0.13 1.64 .+-. 0.16 P < 0.001
formulation treated
TABLE-US-00023 TABLE 15 Increase in Neuro-peptide-.gamma. content
following test formulation treatment in depressed patients
Neuro-peptide-.gamma. (pg/ml) Comparison Sample After 2 After 4
After 6 Initial Vs After 6 Groups size Initial month months months
months Sertraline 24 129.68 .+-. 17.22 148.90 .+-. 30.25 151.89
.+-. 29.87 156.04 .+-. 13.77 P < 0.05 treated Test 36 133.80
.+-. 14.02 168.92 .+-. 27.25 192.45 .+-. 26.87 228.93 .+-. 38.04 P
< 0.01 formulation treated
TABLE-US-00024 TABLE 16 Table shows improvement in sleep pattern
following test formulation treatment in depressive patients Before
treatment After 6 months treatment Sleep Total Sleep Sleep Total
Sleep No. of period Sleep Latency period Sleep Latency Groups cases
(min) (min) (min) (min) (min) (min) Sertraline 60 445.90 .+-. 20.11
234.98 .+-. 35.80 13.62 .+-. 2.41 438.35 .+-. 30.78 201.94 .+-.
39.73 14.98 .+-. 3.17 treated Test 82 438.73 .+-. 31.64 211.06 .+-.
23.73 14.04 .+-. 2.73 439.74 .+-. 25.73 314.97 .+-. 36.82 11.79
.+-. 2.45 formulation treated
TABLE-US-00025 TABLE 17 Effect of test formulation on sleep pattern
recorded in terms of different stages among depressive patients
Before treatment After 6 months treatment Stage- Stage- Stage-
Stage- Stage- Stage- Stage-I II III IV Stage-I II III IV (Time
(Time (Time (Time (Time (Time (Time (Time No. of in in in in in in
in in Groups cases min) min) min) min) min) min) min) min)
Sertraline 60 179.32 .+-. 21.74 45.82 .+-. 6.11 18.22 .+-. 3.97
Could 162.98 .+-. 42.74 46.22 .+-. 9.31 15.82 .+-. 4.73 Could
treated not reach not reach Test 82 174.93 .+-. 14.73 48.20 .+-.
7.11 20.68 .+-. 6.02 Could 178.93 .+-. 21.73 34.79 .+-. 5.11 16.22
.+-. 3.97 Could formulation not reach not treated reach
[0069] The neuro-chemical assessment exhibited modulation of
altered neuro-transmitters and biogenic amines after treatment with
test formulation in selected depression cases (Table 5-9). The
inflammatory cytokines IL-1.beta. and TNF-.alpha. decreased
significantly in test formulation treated group indicating
beneficial role of test formulation in reducing the severity as
well as frequency of depressive episodes among mild to moderate
depressive patients (Table-10 & 11). There is an increase in
DHA content after oral administration of test formulation for 6
months (Table-12). Table-13 shows a marked decrease in occipito
frontalis muscle action potentials following test formulation
treatment indicating a decrease in anxiety and tension among
depressed persons. Similarly the test formulation also exerted
delta activation after treatment in depressed patients suggesting
improvement in the sleep pattern of the patients (Table-14).
Neuro-peptide-.gamma. is the 36-amino acid which affects anxiety,
stress, pain, circadian rhythm and also associated various
psychiatric disorders particularly depression of varying degree.
The test formulation exerted significant increase in NP-.gamma.
content in the brain suggesting improvement in anxiety, stress and
depressive behavior of the depression patients (Table-15). The
effect of test formulation on sleep pattern measured by recording
on polysomnographic studies indicated improvement in sleep period,
total sleep, sleep latency and various stages of sleep. Though none
of the cases could achieve the stage four. The results obtained in
the test formulation treatment group were compared with the
conventional drug treated group and exerted significant difference
when the conventional treated group was compared with the test
formulation treated group (Table-16-17). The drug was found safe
and effective after 6 months of continuous oral administration to
depressed patients. From the results, it is evident that the oral
administration of the test formulation is an effective and well
tolerated drug for the management of mild to moderate
depression.
* * * * *