A Synergistic Herbal Anxiolytic, Adaptogenic And Antistress Composition And A Process Thereof


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Abstract

The present disclosure relates to a synergistic herbal anxiolytic, adaptogenic and antistress composition and a process of its preparation. The composition comprises a synergistic mixture of extracts of Camellia sinensis (Green tea) leaves, fruits of Terminalia bellerica, fruits of Terminalia chebula, fruits of Emblica officinalis, roots of Withania somnifera and Ocimum sanctum leaves. All the extracts thus prepared are mixed thoroughly to obtain the final composition, which has potent anxiolytic, adaptogenic and antistress activity. The novelty of the disclosure is present in the careful selection of the ingredients and also in the blending ratios such that the resulting composition has maximum synergistic activity.

Information

Application ID 1108/CHE/2009
Invention Field TRADITIONAL KNOWLEDGE BIOTECHNOLOGY
Date of Application
Publication Number 15/2012

Applicants

Name Address Country Nationality
AMIT AGARWAL K & S PARTNERS INTELLECTUAL PROPERTY ATTORNEYS # 134, FIRST FLOOR, 60 FT. DOMLUR ROAD, INDIRANAGAR, BANGALORE 560 008, KARNATAKA, INDIA. India India

Inventors

Name Address Country Nationality
AMIT AGARWAL K & S PARTNERS INTELLECTUAL PROPERTY ATTORNEYS # 134, FIRST FLOOR, 60 FT. DOMLUR ROAD, INDIRANAGAR, BANGALORE 560 008, KARNATAKA, INDIA. India India

Specification

TECHNICAL FIELD:

The present disclosure relates to a herbal anxiolytic, adaptogenic and antistress composition. The composition is helpful to alleviate stress and to enhance host resistance to a variety of stressful conditions and infections. The embodiment of the disclosure resides in the novel composition of its constituents used in the preparation of such herbal composition.

The novelty of this composition lies not only in the selection of the ingredients but also in the proportion selected in such a manner as to exhibit maximum synergistic activity.

BACKGROUND:

Anxiety disorders account for 15-20% of medical clinic patients. Anxiety is defined as a subjective sense of unease, dread or foreboding. It can indicate a primary psychiatric condition or it can be a reaction of any other primary medical disease. Mental health clinicians recognize many distinct subtypes of anxiety disorders, and the prevalence of these is startling. Data collected from various surveys have found that at some point of time in their lives nearly one in four people will suffer from an anxiety disorder. The most common types are social and simple phobia (11% and 12%), Generalised anxiety disorder or chronic worry (5%) and panic disorder (3.5%) [Steven Fried man, Anxiety and anxiety disorders, source: http:// globalrph.healthology.com/ globalrph/ 16162.htm]. Its prevalence in India is 15-16/1000.

Unlike other psychiatric conditions such as schizophrenia or depression, anxiety is both a normal emotion and a psychiatric disorder. It commonly co-exists with other clinical conditions but can be diagnosed on the basis of symptoms and course of the disease. However, when the symptoms are disruptive and maladaptive enough to interfere with normal functioning of the individual, anxiety should be regarded as pathological requiring requisite therapy.

TYPES OF ANXIETY DISORDERS:

• Generalized anxiety disorder where there is persistent, excessive, unrealistic worry associated with other signs and symptoms.

• Panic disorder defined by the presence of recurrent panic attacks.

• Phobic disorder characterized by marked or persistent fear of objects or situations.

• Post traumatic stress disorder, where the anxiety develops after exposure to extreme trauma. It could be acute stress disorder or post traumatic stress disorder.

• Obsessive compulsive disorder which is characterized by obsessive thoughts and compulsive behaviors that impairs everyday functioning.

• Mixed anxiety depression where the pressure of symptoms of anxiety depression co-exists with sub-syndrome level.

INTRODUCTION TO STRESS:

Stress and strain have become synonymous in the modern life. Currently, pathological stress has become a common problem and one of the major health hazards of modern life. A number of factors ranging from emotional to physical like mental tension, fear, anger, excessive heat and cold, etc. can result in stress. Hence the importance of treatment of stress.

Constant strain or stress leads to irritability, anger, sleeplessness and other psychological disorder. Stress has been defined as a non-specific response made by the body to any kind of demand made upon it. Therefore it is a stereotyped response, irrespective of the nature of the stress-physical, infections, chemical or emotional. The demand refers to adaptive work of the body in order to maintain homeostasis.

Stress disturbs the chemical and hormonal equilibrium including the immune system, which leads to the development of many clinical disorders ranging from a simple anxiety neurosis to cancer. If the stress is extreme, unusual or long lasting, however, the normal mechanisms may not be efficient. In this case, the stress triggers a wide-ranging set of bodily changes, called the General Adaptation Syndrome (GAS).

STAGES AND FEATURES OF STRESS:

On response to noxious stimuli, any animal/human exhibits three phases as a response of GAS.

(1) Stage of alarm reaction: Alarm reaction is the starting response to an unexpected stressful stimulus. It lasts for few hours to few days. The subject or individual enters in to the next stage provided the stimulus is not too noxious.

(2) Stage of resistance: In which the subject adapts itself to the continuous exposure to the stress or various physiological changes.

(3) Stage of exhaustion: Further persistence of stress for a continuous length of time
eventually exhausts the subject or individual from its adaptability and thereby gives
way for manifestation of various physical disorders like insomnia, hypertension,
gastro dueodenal ulcers, allergic afflictions etc.

The effect of stress on an individual depends on the intensity of the stimulus, duration of the stressor and most important, the individual factors like genetic predisposition, age, sex, conditioning of the individual, etc.

Principle components of GAS are corticotropin releasing hormone and locus cereuleus norepinephrine systems. The stimulation of cells of hypothalamus causes the release of CRH (Corticotropin releasing hormone) which in turn releases ACTH (Adrenocorticotrophic hormone) from pituitary through which glucocorticoids and mineralocorticoids are released from the adrenal cortex. At the same time the nerve impulses from cortex also stimulate sympathetic centres in the spinal cord, which later bring about release of adrenaline and noradrenaline from adrenal medulla.

These two hormones are responsible for the ultimate sequel of persistent stress with systemic manifestations.

Stress has been shown experimentally to increase the levels of free oxygen radicals within the body that can aggravate the cardiovascular diseases, infections, diabetes etc.

PRESENTLY AVAILABLE THERAPIES FOR STRESS AND ANXIETY RELATED DISORDERS:

Benzodiazepines remain the anti-anxiety agents of choice because of their effectiveness and relative safety. However they are known to induce tolerance and physical dependence. This has led to the search for new and better anxiolytics. The newer agents reduce central serotonergic activity, like the 5-HT1A receptor agonist buspirone and the 5-HT3 antagonist ondansetron.

Pharmacological therapies include Benzodiazepines (BDZ) - Diazepam, Non BDZ -Zolpidem, Serotonin modulators (5HT1A - Buspirone, Reuptake inhibitors -Fluoxetine), Antihistamines - Hydroxyzine, p-Antagonist - Propranolol and others medicines

DRAWBACKS OF THE PRESENT ANXIOLYTIC THERAPIES:

Extended pharmacological actions, sedation, cognitive impairment, tolerance, dependence and withdrawal symptoms, appetite stimulation, weight gain, interaction with alcohol and toxic adverse drug reactions.

Non-pharmacological anxiolytic therapies include Psychotherapy, behavioural therapy (Transcendental meditation & Relaxation techniques) and Cognitive therapy.

ADAPTOGENS - A NEW CONCEPT

The term adaptogen was first coined by Brekhman I.I.; Dardymov I.V; [Reference may be made to Brekhman I.I.; Dardymov I.V New substances of plant origin which increase non specific resistance. Annual Reviews in Pharmacology ND Toxicology 1969, Vol.9, Page 419-428] an agent that causes a "State of Nonspecifically Increased Resistance"(NSIR). An adaptogen causes minimal alteration on physiological function of an organism; it is non-specific in action and is able to increase resistance to adverse influence of a wide range of factors of physical, chemical and biological nature. Further it has a normalizing action irrespective of the direction of the foregoing pathologic changes.

STATEMENT OF DISCLOSURE
Accordingly the present disclosure provides a synergistic herbal composition comprising extracts of Camellia sinensis, Terminalia bellerica, Terminalia chebula, Emblica officinalis, Withania somnifera and Ocimum sanctum, optionally along with pharmaceutically acceptable excipient(s); a process for preparation of synergistic herbal composition comprising extracts of Camellia sinensis, Terminalia bellerica, Terminalia chebula, Emblica officinalis, Withania somnifera and Ocimum sanctum optionally along with pharmaceutically acceptable excipients, said process comprising acts of a) obtaining extract from various parts of the Camellia sinensis, Terminalia bellerica, Terminalia chebula, Emblica officinalis, Withania somnifera and Ocimum sanctum; b) mixing the extracts; c) drying the mixed extract, and d) optionally adding excipients to obtain the synergistic herbal composition and a method for inhibiting release of corticotropin releasing hormone by providing synergistic herbal composition comprising extracts of Camellia sinensis, Terminalia bellerica, Terminalia chebula, Emblica officinalis, Withania somnifera and Ocimum sanctum optionally along with pharmaceutically acceptable excipients to a subject in need thereof.

DETAILED DESCRIPTION OF DISCLOSURE

The present disclosure is in relation to a synergistic herbal composition comprising extracts of Camellia sinensis, Terminalia bellerica, Terminalia chebula, Emblica officinalis, Withania somnifera and Ocimum sanctum, optionally along with pharmaceutically acceptable excipient(s).

In another embodiment of the present disclosure, concentration of Camellia sinensis extract is ranging from about 5% to about 40% w/w, concentration of Terminalia bellerica extract is ranging from about 1% to about 15% w/w, concentration of Terminalia chebula extract is ranging from about 1% to about 15% w/w, concentration of Emblica officinalis extract is ranging from about 1% to about 15% w/w, concentration of Withania somnifera extract is ranging from about 3% to about 30% w/w and concentration of Ocimum sanctum extract is ranging from about 3% to about 30% w/w.

In still another embodiment of the present disclosure, pharmaceutically acceptable excipient is selected from a group comprising binders, disintegrants, diluents, lubricants, plasticizers, permeation enhancers, solubilizers, preservatives and colouring agents.
In still another embodiment of the present disclosure, the binder is selected from a group comprising starch, pre-gelatinized starch and Dicalcium phosphate or any combination thereof.

In still another embodiment of the present disclosure, the preservative is selected from a group comprising propyl paraben sodium, methyl paraben sodium and bronopol or any combination thereof, each having a concentration ranging from about 0.1% to about l%w/w.

In still another embodiment of the present disclosure, the pharmaceutically acceptable excipient is in a concentration ranging from about 40%w/w to about 70%w/w.
In still another embodiment of the present disclosure, said composition is formulated into dosage forms selected from a group comprising tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsion in hard or soft gel capsules, syrups, elixirs, phytoceuticals and neutraceuticals.

The present disclosure is also in relation to a process for preparation of synergistic herbal composition comprising extracts of Camellia sinensis, Terminalia bellerica, Terminalia chebula, Emblica officinalis, Withania somnifera and Ocimum sanctum optionally along with pharmaceutically acceptable excipients, said process comprising acts of

a) obtaining extract from various parts of the Camellia sinensis, Terminalia
bellerica, Terminalia chebula, Emblica officinalis, Withania somnifera and Ocimum
sanctum;

b) mixing the extracts;

c) drying the mixed extract, and

d) optionally adding excipients to obtain the synergistic herbal composition.

In still another embodiment of the present disclosure, the extracts are prepared in solvent selected from a group comprising water, organic solvent and a combination thereof.

In still another embodiment of the present disclosure, the organic solvent is selected from a group comprising methyl alcohol, ethyl alcohol, propyl alcohol, butyl alcohol and acetone.

In still another embodiment of the present disclosure, concentration of the Camellia sinensis extract is ranging from about 5% to about 40% w/w, concentration of the Terminalia bellerica extract is ranging from about 1% to about 15% w/w, concentration of the Terminalia chebula extract is ranging from about 1% to about 15% w/w, concentration of the Emblica officinalis extract is ranging from about 1% to about 15% w/w, concentration of the Withania somnifera extract is ranging from about 3% to about 30% w/w and concentration of the Ocimum sanctum extract is ranging from about 3% to about 30% w/w.

In still another embodiment of the present disclosure, the extracts of Camellia sinensis and Ocimum sanctum are obtained from leaves, extracts of Terminalia bellerica, Terminalia chebula and Emblica officinalis are obtained from fruits and the extract of Withania somnifera is obtained from roots.

In still another embodiment of the present disclosure, wherein the drying is carried out at a temperature ranging from about 50°C to about 60°C.
In still another embodiment of the present disclosure, wherein the drying is carried out under vacuum.

The present disclosure is also in relation to a method for inhibiting release of corticotropin releasing hormone by providing synergistic herbal composition comprising extracts of Camellia sinensis, Terminalia bellerica, Terminalia chebula, Emblica officinalis, Withania somnifera and Ocimum sanctum optionally along with pharmaceutically acceptable excipients to a subject in need thereof.

In still another embodiment of the present disclosure, the subject is an animal, including human being.

NEED FOR DEVELOPING HERBAL COMPOSITION HAVING ANXIOLYTIC, ADAPTOGENIC AND ANTISTRESS PROPERTIES

Ethnomedical literature as well as Ayurveda recommends several herbs as beneficial in relief from stress, boosting one's immune system for revitalization. In the search for new effective and safer anxiolytics, pharmacologists have delved in to nature and efforts are on to seek remedies for anxiety from the plant kingdom.

Many of the herbs, which are used in the Ayurvedic system of medicine as rejuvenators i.e. they retard the process of aging and protect cells from damage because of free radical scavenging activity. Ayurveda, the Indian system of traditional medicine recommends number of plants classified as "Rasayanas". These are said to be adaptogenic and help the individual in adjust to various circumstances. Therefore it was thought worthwhile to study a combination of such major plants.

The plants reported to have potent anxiolytic, adaptogenic and antistress effects were selected and screened to verify the published data then processed in an appropriate manner and blended together in a specific ratio in order to obtain the synergistic herbal composition of present disclosure.

Several undesirable side effects associated with the synthetic agents stimulated the search for alternative agents from the herbal segment.

The main objective of the present disclosure is therefore to provide a herbal anxiolytic, adaptogenic and antistress composition.

Another objective of the present disclosure is to provide a synergistic combination of the most potent herbs and other natural agents blended together in a specific ratio such that the resultant composition exhibits significant synergistic anxiolytic, adaptogenic and antistress activities.

Yet another objective of the present disclosure is to provide a process for the preparation of the herbal based anxiolytic, adaptogenic and antistress composition which alleviates stress and enhances host resistance to a variety of infections and stressful conditions.

The present disclosure is based on our experience and research about the medicinal properties of various herbs. In an attempt to develop a safe, effective herbal anxiolytic, adaptogenic and antistress composition, more than thirty Indian medicinal plants were selected for screening, based on their published reports both in modern and ethnomedical literature. The selected crude drugs were coarsely powdered and extracted / distilled with suitable solvents of varying polarity, using the conventional method of extraction / distillation. The extracts and distillates thus obtained were subjected to a series of in vitro bioassays relevant to anxiolytic and antistress properties. Based on the results of the in vitro assays, six different plant extracts were finally short listed. Various prototype combinations were prepared based on Ayurvedic principles and the scientific models of compatibility assessment. All the prototypes thus prepared were re-evaluated in the in vitro assays. Based on these results, the present composition was finalized.

The present disclosure is a synergistic herbal anxiolytic, adaptogenic and antistress composition, comprising:

(a) taking the extract of Green Tea (Camellia sinensis) in the range of 5-40% w/w,

(b) taking the extracts of fruits of Terminalia chebula in the range of 1-15% w/w,

(c) taking the extracts of fruits of Terminalia bellerica in the range of 1-15% w/w,

(d) taking the extracts of fruits of Emblica officinalis in the range of 1-15% w/w,

(e) taking the root extract of Withania somnifera in the range of 3-30% w/w,

(f) taking the leaves extract of Ocimum sanctum in the range of 3-30%w/w,

Another embodiment of the present disclosure is that the extracts of the plants selected may be water or alcohol or acetone or a combination of any two extracts. The above composition of the present disclosure may also contain pharmaceutical excipients which are usually employed to prepare oral dosage forms like powder, tablets, capsules, syrups, liquid etc. The excipients such as starch, pre-gelatinized starch, Dicalcium phosphate or a mixture thereof may be used. The amount of excipients employed may range from 40-70%w/w.

The composition may also contain preservatives which may be selected from propyl paraben sodium, methyl paraben sodium or bronopol etc or a mixture thereof may be used. The amount of preservatives employed may range from 0.1 -l%w/w.

The present disclosure, which is an anxiolytic, adaptogenic and antistress composition, can be employed in any required dosage forms such as powder, liquid, tablet, capsule, syrup etc.

According to another feature of the disclosure there is provided a process for the preparation of a synergistic herbal anxiolytic, adaptogenic and antistress composition, comprising: mixing thoroughly,

1) taking extracts of Green Tea (Camellia sinensis) in the percentage of 5-40% w/w,

2) taking the extracts from the fruits of Terminalia chebula in the percentage of 1-15% w/w,

3) taking the extracts from the fruits of Terminalia bellerica in the percentage of 1-15% w/w,

4) taking the extracts from the fruits of Emblica officinalis in the percentage of 1-15% w/w,

5) taking the extract of roots of Withania somnifera (Ashwagandha) in the percentage of 3-30% w/w,

6) taking the extract of leaves of Ocimum sanctum (Tulsi) in the percentage of 3-30%w/w which are taken in a non-corrosive vessel and dried in an oven to eliminate the traces of volatile organic solvent impurities. Finally adding sufficient quantity of excipients along with preservatives and lubricants under constant mixing to obtain the desired dosage form.

It would be observed that the composition and the process of preparing the said composition of present disclosure is not a mere admixture of the ingredients used resulting in the aggregate properties of the ingredients employed. The composition is the result of the synergistic combination of the ingredients which has potent anxiolytic, adaptogenic and antistress activity. The disclosure is described in detail in the examples given below which are provided by way of illustration only and therefore should not be construed to limit the scope of the disclosure.

SCREENING OF PLANTS:

Amongst the several plant extracts / plant fractions tested, we found that the following plants gave interesting results in the DPPH scavenging assay. The results of these samples are shown in table-1.

Table no.l

Preparation of synergistic composition: Example 1

An amount of 120g (20%w/w) of Camellia sinensis, 60g(10%w/w) of Terminalia bellerica, 60g(12%w/w) of Terminalia chebula, 60g(10%w/w) of Emblica officinalis, 168g(28%w/w) of Withania somnifera, 132g(20%w/w) of Ocimum sanctum are mixed thoroughly in a non-corrosive vessel and dried in oven to eliminate the traces of volatile organic solvent impurities. To this mixture, 400g of a mixture of maize starch, pre-gelatinised starch and dicalcium phosphate (taken in equal quantities) is added as excipients and mixed thoroughly. The composition obtained is filled in hard gelatin capsules. Each capsule contains l00mg of active ingredients and can be used as a single or multiple doses as per the requirement for the treatment of stress and related disorders. This composition henceforth is referred to as NR-S1.

Example 2

An amount of 165g (30%w/w) of Camellia sinensis, 44g(8%w/w) of Terminalia bellerica, 44g (8%w/w) of Terminalia chebula, 44g (8%w/w) of Emblica officinalis, HOg (20%w/w) of Withania somnifera, 143g(26%w/w) of Ocimum sanctum are mixed thoroughly in a non-corrosive vessel and dried in oven to eliminate the traces of volatile organic solvent impurities. To this mixture, 450g of a mixture of maize starch, pre-gelatinised starch and dicalcium phosphate (taken in equal quantities) is added as excipients and mixed thoroughly. The composition obtained is filled in hard gelatin capsules. Each capsule contains 150mg of active ingredients and can be used as a single or multiple doses as per the requirement for the treatment of stress and related disorders. This composition henceforth is referred to as NR-S2.

Example 3

An amount of 60g (15%w/w) of Camellia sinensis, 56g(14%w/w) of Terminalia bellerica, 56g(14%w/w) of Terminalia chebula, 56g(14%w/w) of Emblica officinalis, 96g(24%w/w) of Withania somnifera, 76g(19%w/w) of Ocimum sanctum are mixed thoroughly in a non-corrosive vessel and dried in oven to eliminate the traces of volatile organic solvent impurities. To this mixture, 600g of a mixture of maize starch, pre-gelatinised starch and dicalcium phosphate (taken in equal quantities) is added as excipients and mixed thoroughly. The composition obtained is filled in hard gelatin capsules. Each capsule contains 200mg of active ingredients and can be used as a single or multiple doses as per the requirement for the treatment of stress and related disorders.

This composition henceforth is referred to as NR-S3. Each of these three compositions (NR-S1 to NR-S3) are again tested and compared using the DPPH scavenging assay showed in the Table 2.

Table no.2

Based on above results the prototype encoded as NR-S3 was selected for further evaluation.

EFFICACY AND SAFETY STUDIES:

The selected prototype (NR-S3) was evaluated for efficacy using the following laboratory animal models.

1. Elevated plus maze

Anxiolytics tend to increase ambulatory behaviors of rodents placed in an unfamiliar environment where as anxiogenic agents tend to decrease this behaviors Elevated plus maze has been used extensively for evaluation of novel anxiolytic agents and to investigate psychological and neurochemical basis of anxiety Elevated plus maze is a wooden device in the shape of plus consisting of two closed arms and two open arms. Whenever the mouse is exposed to an elevated and open maze arm, it leads to an approach avoidance conflict which is considerably stronger than that evolved by experience to closed arm.

Mice were treated with NR-S1 and NR-S3 in the dose of 10 mg/kg body weight orally for 20 days. On 21st day the assay was carried out. The drug treated animals showed an overall increase in motor activity. However the time spent and number of entries into the open arm was more than closed arm in treated groups (7.0 + 1.66 and 22.13 ±6.26 Vs. 1.857 + 0.261 and 5.714 ± 0.865, p< 0.05 ). This establishes the anxiolytic property of NR-S3.

Table no.3

* P<0.05, significant value
NR-S3 was selected for further pharmacological evaluation using the following in-vivo assays:

1. Anti stress activity-Cold restrain induced ulcer.

2. Sedative activity

a. Potentiation of pentobarbitone sodium (PBS) induced sleep

b. Spontaneous motor activity using photoactometer

ANTISTRESS ACTIVITY:

2. Cold restrain induced ulcer

Benzodiazepine anxolytics are known to exhibit antistress activity as part of their behavioural profile. As such test parameter involving variety of stress situations are used to evaluate antistress activity.

Immobilization stress-induced gastric ulcerations increase the plasma corticosterone in rats and therefore is a useful model for evaluation of antistress activity

In this procedure albino rats were treated with NR-S3 orally at the dose of 10 mg /kg body weight in carboxy methylcellulose for a period of 20 days. Diazepam was used as a standard in the dose of 0.91mg/kg body weight for 20 days. On 21st day they were subjected to immobilization by a method of Sander (Sander et al, Gastroenterology, 1975, 68, 285) with some modification.

The rats were immobilized for 2 hours on a metal cage grill with the help of adhesive tapes and kept in refrigerator for 2 hours at 4°C. After 2 hours, they were sacrificed, the stomach was cut along the greater curvature and ulcer was graded as per the method of Williamson et al. (Williamson E et al, in "Pharmacological methods in Phytotherapy research" 1996, 1, Wiley & Johnson publications, New York , 40) as follows.

0 - No ulcer

1 - Gastric Hyperemia

2 - Erosion

3 - Frank ulcer

The tissue was then fixed in formalin and evaluated histopathologically. NR-S3 showed ulcer score of 1.625 + 0.565 compared to 2.625 + 0.532 in control. Further the Mucin content was also found to be increased statistically in NR-S3 treated group (2.50 ± 0.25 vs. 1.18 ± 0.09, p < 0.05). Diazepam showed 2.00 ± 0.327, which was also statistically significant when compared to the control.

Table no.4

P=<0.05 statistically very significant when compared to the control

Restrain stress in animals is shown to increase gastric motility and acidity without affecting the gastric mucosal blood flow (Yanos. S et al, Japanese Journal of Pharmacology, 1975, 28, 607; Kitagawa H et al, Gastroenterology, 1979, 77,298). It has also been shown that GABA or sodium valproate administration reduced the incidence of cold restrain ulcer (Bhargava K. P et al, British Journal of Pharmacology, 1985, 84, 619). Plant extracts, which increase the GABA level in the brain protected the rats against the cold restrain ulcer (Chaterjee et al Indian Journal of Experimental Biology, 1992, 30,889-891). Therefore it appears that NR-S3 is probably increasing the GABA levels in the brain.

3. SEDATIVE ACTIVITY:

Sedative activity is one of the properties of benzodiazepines used as anxiolytics.

Therefore this activity was undertaken. Sedative activity of NR-S3 was evaluated by

(a) Potentiation of pentobarbitone sodium (PBS) induced sleep

(b) Spontaneous motor activity using photoactometerfa)

POTENTIATION OF PENTOBARBITONE SODIUM (PBS) INDUCED SLEEP
Pentobarbitone sodium at the dose of 50 mg/kg mouse body weight was administered by the i.p. route and the time between the loss of righting reflex and the gain of righting reflex (narcosis or sleeping time) was noted down (Kasture S.B. et al, Indian Drugs, 1999, 36(9), 588-592)

In this study, 2 hours after oral drug administration (at the dose of 10 mg/kg mouse body weight of NR-S3 and 1.3 mg/kg body weight of Diazepam) PBS was given by i.p. route (50 mg/kg) and narcosis time was calculated. NR-S3 showed a mean Narcosis time of 130 ± 15.0 minutes when compared to the control (41.5 ± 9.8 mins) which was statistically significant, (p < 0.05)

Table no.5
* p ± 0.05 - Significant values

4. SEDATIVE ACTIVITY:

(b) SPONTANEOUS MOTOR ACTIVITY USING PHOTOMETER

Reduced motor activity is attributed to the central nervous system depression, as depression of locomotor activity is common to most anxiolytics. Hence evaluation of spontaneous motor activity is a good model for the evaluation of Anti-stress activity.

The Photoactometer consists of a box containing photoactive cells on two adjacent walls and two receivers on the other two sides. A reading is rewarded each time the beam of light is cut. Locomotion of the animal cuts the beam and readings are recorded for a period of 5 minutes (Gamaniel et al, Indian Drugs, 1998, 35(12), 740-743, Kasture et al
Indian Drugs, 1999, 36(9), 588 - 592).

NR-S3 was evaluated for its effect on spontaneous motor activity in mice at the doses of 10 mg/Kg body weight. Diazepam, used as the standard at the dose of 1.3mg/Kg (0.026mg/20 mg) showed 213.5 ± 22.1 number of movements for a period of 5 minutes when compared to 303.857 ±31.3 of control. NR-S3 decreased the motor activity to 219.6 ± 41.4 at 10 mg/kg. Both the standard and NR-S3 showed statistical significance.

Table no.6

*p<0.05, statistically significant when compared to the control.

5. TOXICITY STUDIES:

ORAL TOXICITY

Other toxicity studies that were done were on LD50 determination of NR-S3 as per the method of Ghosh M.N (1984), In: Fundamentals of experimental pharmacology-toxicity studies - Chapter 26, 2nd edition, 153-158 and repeat dose toxicity studies.

NR-S3 was tested in the doses of 300, 450, 675, 1012.75 and 1519.25mg/kg mouse body weight and the animals were observed for behavioural, autonomic and other changes once a day for 15 days after which the animals were sacrificed for histopathological changes. NR-S3 produced no clinical toxicity up to the dose of 1519.25 mg/kg. The LD50 of NR-S3 appears to be more than 1519.25 mg/kg mouse body weight.

6. TOXICITY STUDIES:

SUB ACUTE TOXICITY
Repeat dose toxicity studies are conducted to test the adverse effects of the drug on prolonged use. This is carried out to provide information on the possible health hazards likely to arise from repeated exposure over a relatively limited period of time

NR-S3 was administered initially at a dose of 10 mg/kg rat body weight for three days, which was graduated to 20 mg/kg for the next 3 days and again to 40 mg/kg for three weeks. The animals were observed daily for toxic signs and symptoms.

After 28 days haematological examinations were conducted for total leucocyte count and Haemoglobin percentage. Biochemical determinations were also done to investigate the major toxic effects in tissues like kidney and liver and the parameters included SGOT, SGPT, ALP, Bilirubin, Urea and Creatinine.

Gross observations, haematogical and biochemical investigations revealed significant changes in the TLC, GPT and ALP in the treated group when compared to control.

Table no.7
* p £ 0.05, statistically significant when compared to control

Conclusion: There has been an increase in TLC which could be due to the immunomodulatory property of one of the ingredients and reduction in SGPT and ALP does not seem to be pathological. Thus NR-S3 appears to be safe upto 40 mg/Kg. Rat body weight.

7. CLINICAL STUDIES:

Randomized double blind placebo controlled studies of NR-S3 in anxiety patients. NR-S3 has shown significant improvement in anxiety scores when compared to placebo treated group.

MATERIALS AND METHODS:

The study was carried out in collaboration with Psychopharmacology department at NIMHANS, Bangalore over a period of nine months.

STUDY DESIGN:

CASES:

The trial was a randomized, double blind placebo controlled study in patients diagnosed to have generalised anxiety disorder (GAD), mixed anxiety and depression (MAD) following Hamilton's anxiety score, patients global rating and rater's global rating by the investigator. Thirty nine patients of both sexes (22 males & 17 female patients) with either GAD or MAD, in the age group of 20-60 years, were initially
enrolled for the study based on suitable inclusion and exclusion criteria after obtaining
an informed written consent. Institutional human ethical committee approval was
taken.

CLINICAL EXAMINATION:

Patients enrolled for the study were subjected to Hamilton anxiety scores, Patients global rating and Rater's global rating on three occasions i.e. day zero, prior to initiating treatment and twice, after two weeks and four weeks during the treatment with trial medication.

DRUG TREATMENT SCHEDULES:

All the patients entering the study were randomly assigned treatment with either placebo or test medication NR-S3 capsules (Natural Remedies Pvt.Ltd.) for a period of one month. They were instructed to take two capsules of the study medications twice daily (each capsule containing l00mg Actives), orally with a glass of warm water after food in the morning and in the night, for a period of one month. Patients were advised to restrict themselves to the study medications (NR-S3 / Placebo) as treatment for anxiety with no other active treatment intervention during the study period except the use of rescue medications whenever their symptoms aggravated. They were instructed to mark the frequency of intake of the study medications on a calendar provided to them to ensure their daily compliance. In addition they were advised to enter the details of use of rescue medications. There was no restriction on the diet or routine activity of the patients during the treatment period.

PATIENT ASSESSMENT:

All the patients were assessed for Hamilton's anxiety score, Patients global rating and Rater's global rating on three occasions viz., at day 0 prior to initiating treatment, at 2nd week and 4th week.

DROPOUTS:

Four patients dropped out of each group at the 2 week follow-up. In the placebo group, two patients dropped out because of inefficacy of treatment, and 2 dropped out for unknown reasons. In the NR-S3 group, 1 patient dropped out because of inefficacy of treatment, and 3 patients dropped out because of unknown reasons.

STATISTICS:

The mean values of investigations were subjected to statistical analysis of significance using t test for quantitative variables with homogenous variances, Mann- Whitney test for quantitative variables with heterogenous variances and Chi square test (with Yates' continuity correction)

RESULTS:

Table no.8: Sociodemographic and clinical description of patients randomised to receive placebo (n=16) or (n=23)*

*Values are range, mean (standard deviation) for quantitative variables and frequency counts for qualitative.

** t - test for qualitative variables with homogeneous variances, Mann- Whitney test for quantitative variables with heterogenous variances and Chi square test (with Yates' continuity correction) for qualitative variables.

Table no.9: Adverse effects in placebo and groups
Table no.10: Hamilton anxiety ratings and global ratings at baseline, week 2 and
week 4

A two way repeated measures multivariate analysis of variance of these data yielded the following results:

HAMILTON ANXIETY SCORES:

The main effect for groups was significant (F = 5.80, df = 1,37, P = 0.021), indicating that the anxiety scores were significantly lower in the NR-S3 group than in the placebo group. The main effect for time was significant (Pilla's trace = 0.41, F = 12.48, df = 2,36, P < 0.001), indicating that there was significant improvement in both groups across time. Post hoc analysis showed that the improvement took place only between baseline and week 2. The group vs time interaction was significant (Pillai's trace = 0.16, F = 3.49, df = 2,36, P = 0.041), indicating that the NR-S3 group improved significantly more than did the placebo group.

PATIENTS' GLOBAL RATING SCORES:

The main effect for groups was significant (F = 5.68, df = 1,37, P = 0.022), indicating that the anxiety scores were significantly lower in the NR-S3 group than in the placebo group. The main effect for time was significant (Pillai's trace = 0.35, F = 9.64, df = 2,36, P < 0.001), indicating that there was significant improvement in both groups across time.

Post hoc analysis showed that the improvement took place only between baseline and week 2. The group Vs time interaction was not significant (Pillai's trace = 0.10, F = 2.06, df = 2,36, P = 0.14), indicating that the two groups improved comparably in this global ratings across time.

RATER'S GLOBAL RATING SCORE:

The main effect for groups was significant (F = 3.67, df = 1,37, P = 0.063), indicating that the anxiety scores, overall did not differ significantly between the two groups. The main effect for time was significant (Pillai's trace = 0.26, F = 6.41, df = 2,36, P = 0.004), indicating that there was significant improvement in both groups across time. Post hoc analysis showed that the improvement took place only between baseline and week 2. The group Vs time interaction was not significant (Pillai's trace = 0.11, F = 2.27, df = 2,36, P = 0.12), indicating that the NR-S3 group improved significantly more than did the placebo group.

A subgroup analysis using the paired t test was conducted to assess whether the decline in the scores in the NR-S3 group was significant between weeks 2 and 4. The results are presented below:

• Hamilton's rating : t = 3.54, df= 22, P = 0.002

• Patients' global rating : t = 3.02, df = 22, P = 0.006

• Rater's global rating : t = 2.86, df= 22, P = 0.009

Thus cases receiving NR-S3 continued to improve significantly between weeks 2 and 4. When response was defined as a final (week 4) Hamilton's score that was 10 or less, 3 out of 16 (18.8 %) placebo-treated patients were classified as responders, and 10 of 23 (43.5%) of NR-S3 treated patients were classified as responders. The advantage for NR-S3 was statistically significant. (Chi square = 4.11, df = 1, P = 0.04).

Dose determination:

Based on the above studies a dose of 400 mg per man per day for a duration of 10 to 12 weeks was finalized however it was noted that the product is capable of eliciting desirable effects ranging from 100mg to 2000mg per man per day. Similarly the duration may be varied from 4 weeks up to 24 weeks.

It would be very clear and undoubtedly confirmed that the composition of the present disclosure which is the result of specific and unique combination of the specific ingredients, which combination imparts unique synergistic anxiolytic, adaptogenic and antistress activity to the resulting composition, thereby imparting the novelty to the composition.

Advantages of the present disclosure

• It has got antistress, antioxidant, immunomodulatory and adaptogenic properties.

• Synergistic combination of scientifically validated most potent herbs like
Withania somnifera, Ocimum sanctum, Camellia sinensis, Terminalia chebula, Terminalia bellerica and Emblica officinalis.

• It alleviates stress and enhances host resistance to a variety of infections and stressful situation.

• The composition is capable of scavenging free radicals and minimizing its after effects.

• It increases the adaptability of the body against stress by regulating the functions of the adrenal glands and increasing its non-specific resistance to combat any environmental variations and various untoward influences.

I CLAIM

1) A synergistic herbal composition comprising extracts of Camellia sinensis, Terminalia bellerica, Terminalia chebula, Emblica officinalis, Withania somnifera and Ocimum sanctum, optionally along with pharmaceutically acceptable excipient(s).

2) The synergistic herbal composition as claimed in claim 1, wherein concentration of Camellia sinensis extract is ranging from about 5% to about 40% w/w, concentration of Terminalia bellerica extract is ranging from about 1% to about 15% w/w, concentration of Terminalia chebula extract is ranging from about 1% to about 15% w/w, concentration of Emblica officinalis extract is ranging from about 1% to about 15% w/w, concentration of Withania somnifera extract is ranging from about 3% to about 30% w/w and concentration of Ocimum sanctum extract is ranging from about 3% to about 30% w/w.

3) The synergistic herbal composition as claimed in claim 1, wherein said pharmaceutically acceptable excipient is selected from a group comprising binders, disintegrants, diluents, lubricants, plasticizers, permeation enhancers, solubilizers, preservatives and colouring agents.

4) The synergistic herbal composition as claimed in claim 3, wherein the binder is selected from a group comprising starch, pre-gelatinized starch and Dicalcium phosphate or any combination thereof.

5) The synergistic herbal composition as claimed in claim 3, wherein the preservative is selected from a group comprising propyl paraben sodium, methyl paraben sodium and bronopol or any combination thereof, each having a concentration ranging from about 0.1% to about l%w/w.

6) The synergistic herbal composition as claimed in claim 3, wherein the pharmaceutically acceptable excipient is in a concentration ranging from about 40%w/w to about 70%w/w.

7) The synergistic herbal composition as claimed in claim 1, wherein said composition is formulated into dosage forms selected from a group comprising tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsion in hard or soft gel capsules, syrups, elixirs, phytoceuticals and neutraceuticals.

8) A process for preparation of synergistic herbal composition comprising extracts of Camellia sinensis, Terminalia bellerica, Terminalia chebula, Emblica officinalis, Withania somnifera and Ocimum sanctum optionally along with pharmaceutically acceptable excipients, said process comprising acts of

a) obtaining extract from various parts of the Camellia sinensis, Terminalia bellerica, Terminalia chebula, Emblica officinalis, Withania somnifera and Ocimum sanctum;

b) mixing the extracts;

c) drying the mixed extract, and

d) optionally adding excipients to obtain the synergistic herbal composition.

9) The process as claimed in claim 8, wherein the extracts are prepared in solvent
selected from a group comprising water, organic solvent and a combination thereof.

10) The process as claimed in claim 8, wherein the organic solvent is selected from a group comprising methyl alcohol, ethyl alcohol, propyl alcohol, butyl alcohol and acetone.

11) The process for preparation of synergistic herbal composition as claimed in claim 8, wherein concentration of the Camellia sinensis extract is ranging from about 5% to about 40% w/w, concentration of the Terminalia bellerica extract is ranging from about 1% to about 15% w/w, concentration of the Terminalia chebula extract is ranging from about 1% to about 15% w/w, concentration of the Emblica officinalis extract is ranging from about 1% to about 15% w/w, concentration of the Withania somnifera extract is ranging from about 3% to about 30% w/w and concentration of the Ocimum sanctum extract is ranging from about 3% to about 30% w/w.

12) The process for preparation of synergistic herbal composition as claimed in claim 11, wherein the extracts of Camellia sinensis and Ocimum sanctum are obtained from leaves, extracts of Terminalia bellerica, Terminalia chebula and Emblica officinalis are obtained from fruits and the extract of Withania somnifera is obtained from roots.

13) The process for preparation of synergistic herbal composition as claimed in claim 8, wherein the drying is carried out at a temperature ranging from about 50°C to about 60°C.

14) The process for preparation of synergistic herbal composition as claimed in claim 8, wherein the drying is carried out under vacuum.

15) A method for inhibiting release of corticotropin releasing hormone by providing synergistic herbal composition comprising extracts of Camellia sinensis, Terminalia bellerica, Terminalia chebula, Emblica officinalis, Withania somnifera and Ocimum sanctum optionally along with pharmaceutically acceptable excipients to a subject in need thereof.

16) The method as claimed in claim 15, wherein the subject is an animal, including human being.

Documents

Name Date
1108-CHE-2009 CORRESPONDENCE 25-08-2009.pdf 2009-08-25
1108-che-2009 abstract 11-05-2010.pdf 2010-05-11
1108-CHE-2009 POWER OF ATTORNEY. 25-08-2009.pdf 2009-08-25
1108-CHE-2009 FORM-1 25-08-2009.pdf 2009-08-25
1108-che-2009 form-5 11-05-2010.pdf 2010-05-11
1108-che-2009 form-3 11-05-2010.pdf 2010-05-11
1108-che-2009 form-2 11-05-2010.pdf 2010-05-11
1108-che-2009 correspondence others 11-05-2010.pdf 2010-05-11
Form-3.pdf 2011-09-03
1108-che-2009 claims 11-05-2010.pdf 2010-05-11
Form-5.pdf 2011-09-03
1108-che-2009 form-1 11-05-2010.pdf 2010-05-11
1108-CHE-2009 FORM-13 13-08-2012.pdf 2012-08-13
1108-CHE-2009 CORRESPONDENCE OTHERS 13-08-2012.pdf 2012-08-13
1108-che-2009 description(complete) 11-05-2010.pdf 2010-05-11
Form-1.pdf 2011-09-03
1108-CHE-2009 FORM-1 13-08-2012.pdf 2012-08-13

Orders

Applicant Section Controller Decision Date URL