Title of Invention

HERBAL COMPOSITION FOR TREATMENT OF BONE METABELIC DISORDERS

Abstract The invention provides a novel synergistic herbal composition useful in the prevention and treatment of bone metabolic disorders, said herbal composition comprising plant parts of a calcinogenic plant together with powder of Emblica officinalis.
Full Text FIELD OF INVENTION
The present invention relates to a synergistic herbal composition for the treatment of bone metabolic disorders, said composition comprising plant parts of a calcinogenic plant together with Emblica officinalis.
BACKGROUND OF THE INVENTION:
Osteoporosis is a major health problem and important cause of morbidity worldwide, particularly, with advancing age in both men and women. Normal aging causes a decline in bone density that occurs to varying degrees in otherwise healthy people. This decline does not induce bone fractures.
Osteoporosis, on the other hand, causes a progressive marked reduction in bone mineral density that often results in pathological fractures, particularly of the vertebrae of the spine. It occurs more frequently in women than in men. Determinants of bone mass include external factors such as lifestyle, especially physical activity, exposure to sunlight and calcium intake
Osteoporosis is a disease characterized by low bone mass and microarchitectural deterioration of bone tissue, leading to enhanced bone fragility and risk of fracture. Bone is continuously removed by osteoclasts and replaced by osteoblasts in a process of remodelling. Although bone formation outstrips resorption in youth, accumulated small deficits from remodelling result in a gradual loss of bone mass after the third decade. Primary osteoporosis is therefore usually an
age-related disease. It can affect both sexes, though women are at greater risk because bone loss is accelerated to a variable degree after menopause. Osteoporosis can also be secondary to a number of diseases and drugs.
Treatment of Osteoporosis
In the patients with osteoporosis with or without fragility fractures, treatment involves supportive therapy and interventions to prevent further bone loss and reduce the risk of fractures.
For the treatment of Bone Metabolic Disorders the most commonly used drugs are Vitamin D, estrogen and anabolic steroids combined with calcium supplementation. Various other agents recommended in the treatment of Bone Metabolic Disorders include Sodium fluoride, bisphosphonstes, Calcitonin and Parathyroid hormone. However, treatment with fluoride and other agents like calcitonin has not been approved for general use. While many options are available for the treatment of osteoporosis, use of Vitamin D or its metabolites is a widely accepted modality of treatment. This modality also has a relevance in countries like India where the nutritional factors play an important role in the genesis of osteoporosis.
Vitamin D: Its role in humans
In humans, the UV rays of sunlight, with a wavelength of 290-315 nm, initiate the photo conversion of provitamin D3, 7-dehydro-cholesterol (7-DHC) to previtamin D3 in the skin. A heat induced isomerisation then converts pre-vitamin D3 to vitamin D3 over a period of days.
In addition to endogenous synthesis, Vitamin D3 is also absorbed from dietary sources. Maximum absorption of Vitamin D3 occurs in the jejenum. Following the formation in the skin or after absorption from the intestine, Vitamin D3 binds to Vitamin D binding protein (DBF) in plasms and gets transported to the liver.
Vitamin D3 derived either exogenously from the diet or endogenously from photobiogenesis in the skin, is converted into biologically active metabolite l,25cc-dihydroxy-cholecalciferol by two steps. The first metabolic step occurs in liver, where Vitamin D3 is hydroxylated to form 25-hydroxy vitamin D3. It is further hydroxylated in the kidney to form l,25a-dihydroxy-cholecalciferol. It is the physiologically active hormonal form of Vitamin D3.
The active form of Vitamin D3 has been reported to be very much sensitive to heat and light. It readily decomposes to 25-hydroxy vitamin D3in presence of light.
2. Description of the prior art
Since use of Vitamin D3 along with Calcium and Magnesium supplements is the preferred drug for treatment of Bone Metabolic Disorders, it has always been a topic of interest for researchers to explore an alternative herbal source of Vitamin D3. Earlier it was believed that animal kingdom is the only source of Vitamin D3. The presence of Vitamin D3 in plants has been considered to be very rare. Occurrence of 'Calcinosis' in grazing animals with symptoms similar to Vitamin D intoxication was the first clue which led to the discovery of Vitamin D3 metabolites in higher plants (Worker and Carrillo, 1967). Later calcinosis was reported from various parts of the world. The first calcinogenic plant identified was Solanum malacoxylon, which affected grazing animals in Argentina. Later, other plant species mostly belonging to family Solanaceae and Gramineae have been identified in most cases to cause this calcinotic disease. The causative agent in all the cases has been identified to be a substance having Vitamin D3-like activity. Plants in which presence of Vitamin D3 compounds or Vitamin D-like activity has been reported are Cestrum diurnum, Dactylis glomerata, Lycopersicon esculentum, Medicago saliva, Solanum malacoxylon, Solanum torvum, Solanum verbascifolium, Solanum melongena, Solanum tuberosum and other plants belonging to family Graminae and Solanaceae.
Herbal Source of Vitamin D in India:
T. P. Prema and N. Raghuramulu (1994) have reported the presence of Vitamin D metabolites in the leaves of Cestrum diurnum grown in Hyderabad. The plant material was tested for vitamin D-like activity in vitamin D-deficient rat and chick and also in rachitic rat models. Other calcinogenic plants identified by the authors are Lycopersicon esculentum, Solanum melongena, and Solanum tuberosum.
In all the citations the compound responsible for Vitamin D-like activity was isolated by Freeze- drying process which indicate that the compound is 'thermolabile in nature. Freeze-drying is a highly expensive method at commercial level, and stability of the active compound cannot be ascertained after drying of the plant material during further processing for preparation of a pharmaceutical dosage form. This could be a possible reason why a stable herbal
formulation containing plant material as source of Vitamin D has not been developed so far.
The prior art lacks in any herbal composition for combatting the problem of treating bone disorders. Also, since vitamin D as found in plants is highly unstable, is lost in the process of handling or using the plant parts. Considering this background, the present invention is focussed to develop herbal composition useful in the treatment of bone disorders. Also, the invention provides cost effective technologies that can preserve Vitamin D3 metabolites in plant material as an alternative to Freeze Drying technology. Further, it is also envisaged that addition of certain herbal ingredients to any of the calcinogenic plants might cause a surprising potentiation of their therapeutic efficacy in osteoporosis. Therefore, after stabilizing the vitamin D, studies were also carried out to examine formulations with additive ingredients.
OBJECTS OF THE INVENTION
The main object of the present invention is to provide an effective herbal composition for the prevention and treatment of Bone Metabolic Disorders.
Another object of the invention is to develop a synergistic herbal composition having application in the prevention and treatment of Bone Metabolic Disorders.
Yet another object of the invention is to provide a process for the preparation of the synergistic composition.
Still another object is to provide a cost-effective process by which, the concentration of Vitamin D3 is preserved during post harvest processing in the calcinogenic plant material.
SUMMARY OF THE INVENTION
In accordance with the above and other objects, the invention provides a novel synergistic herbal composition useful in the prevention and treatment of bone metabolic disorders,
said herbal composition comprising plant parts of a calcinogenic plant together with a Emblica officinalis.
DETAILED DESCRIPTION OF THE INVENTION:
Thus, the invention provides a synergistic herbal composition for the treatment of bone metabolic disorders, said composition comprising plant parts of a calcinogenic plant together with powder of Emblica officinalis in the ratio of 1:1 to 1:7.
In an embodiment, the plant parts (of the calcinogenic plant) are freeze dried parts, organic extract of the parts or plant parts pretreated with a fixative agent or is a powder of the plant parts.
In another embodiment, the calcinogenic plants are selected from Cestrum diurnwn, lypopersicon esculentum, Solarium melongena, Dactylis glomerata, Medicago sativa, Solatium malacoxylon, Solanum torvum, Solanim verbasiflium and Solanum tuberosum. In particular, the leaves and tender twigs of Cestrum diurnum are utilized in the composition. Similarly, the leaves, twigs and fruits of other calcinogenic plants may be used. The calcinogenic plants contain vitamin D3 metabolites such as l,25a-dihydroxy-chplecalciferol as one of the active ingredients.
In yet another embodiment, fixative agent is obtained from the plant parts of Emblica officinalis or Triphala or Kushta. The powder of Emblica officinalis used in the composition is the dried powder of the fruit, freeze dried or spray dried extract of the fruits or freeze dried Emblica officinalis juice. Triphala extract comprises a combination of plant parts (fruits) of Emblica officinalis, Terminalia belerica and Terminalia chebula. Where the fixative agent is Kushta, the extract is obtained from the roots of the plant Saussurea lappa,
It is important to note that the composition of the invention may be formulated in different physical forms such as tablets or syrups etc as may be suitable for oral consumption. These compositions may be prepared in accordance with conventional methods known to a person skilled in the art. In case a tablet is intended to be prepared,
the powders of the calcinogenic plant parts and Emblica officinalis may be used. However, where the physical form of the composition is a liquid, the extracts of the calcinogenic plants and the fixative agents may be used. The extracts so used may be aqueous or organic. Needless to say, as in all herbal compositions, conventional additives as may be necessary may be added to the composition. In short, the intention of the applicants is to provide a herbal composition that may exist in different physical forms.
The dosage of the composition so prepared may vary depending on the patient or subject to whom administered and his pathological condition. In any case, the dosage recommended may comprise 450 to 50 mg of the Emblica officinalis and 400 to lOOmg of calcinogenic plant powder or an equivalent extract. The preferred dosage may comprise lOOmg of Emblica officinalis powder and lOOmg of calcinogenic plant powder per dose. The composition may also contain 325 mg of Emblica officinalis and lOOmg of calcinogenic plant powder per dose.
The herbal composition of the invention is a synergistic composition, exhibiting surprising and unexpected properties.
Another important aspect of the invention is that the fixative agent acts as a potentiating agent. In the sense that when the calcinogenic plant parts are used in isolation for treatment of bone disorders, the effect is observed to a certain degree, although not of significant interest. However, if the calcinogenic plant parts are treated with the fixative agent as illustrated in detail in this invention, the effect is surprisingly much more. This indicates that the fixative agent potentiates the activity of the calcinogenic plant parts. In other words, the composition of the invention is synergistic composition, wherein the ingredients as a combination exhibit surprising or unexpected properties. Further, the Applicant has found that the fixative agent is very selective in its activity in that the parts/extracts of all plants cannot be used as fixative agents.,Only a few plants as listed herein act as fixative agents. The selection of the appropriate fixative agent is a matter wherein the inventive ingenuity of the applicant is called for.
The invention also provides a process of treating calcinogenic plant parts comprising vitamin D3 metabolites, so as to preserve or stabilize the metabolites present therein, said process comprising the step of collecting the fresh plant parts of calcinogenic plants and treating the said parts with a fixative agent comprising extract of fruits selected from Emblica officinalis, Terminalia belerica, Terminalia chebula and roots of Saussurea lappa or mixtures thereof.
As stated earlier, the calcinogenic plant is selected from Cestrum diurnum, lypopersicon esculentum and Solatium melongena.
The fixative agent is extract of plants selected from Emblica officinalis or Triphala or Kushta. The powder of Emblica officinalis used in the composition is the dried powder of the fruit, freeze dried or spray dried extract of the fruits or freeze dried Emblica officinalis juice. If the fixative agent is Triphala extract, it comprises a combination of extracts of fruits of Emblica officinalis, Terminalia belerica and Terminalia chebula. And, where the fixative agent is Kushta, the extract obtained from the roots of the plant Saussurea lappa.
The step of 'treating' comprises immersing the plant parts of the calcinogenic plant in the freshly prepared fixative agent for a period of 40 to 60 minutes. In the alternative, it could mean spraying the calcinogenic plant parts with the fixative agent. All these operations are performed under shade as presence of sunlight may affect the stability of Vitamin D3.
The fixative agent is prepared by grinding the plant parts, boiling the powdered material with 8 times the amount of purified water till the quantity of mixture/extract is reduced to one-fourth, filtering the extract and allowing it to cool down to 20 to 30°C.
For best results, the plant parts of the calcinogenic plant are washed in water before treatment with the fixative agent.
Further, the invention provides a method for preparation of a synergistic herbal composition for treatment of bone disorders and stabilization of vitamin D3 metabolites in plants, said method comprising the steps of mixing powder of calcinogenic plant parts with a fixative agent in a ratio between 1: 1 to 1:7.
Particularly, the method comprises the steps of:
a. harvesting the plant parts of a calcinogenic plant at suitable period of time,
b. washing the leaves in water,
c. treatment of washed leaves with a fixative agent,
d. washing the treated leaves with water,
e. drying of treated and washed leaves under controlled conditions of light,
humidity and temperature,
f. grinding the dried leaves by suitable means, protected from light and high
temperatures,
g. freeze drying or extracting calcinogenic plant parts with organic solvents, and
h. mixing the powder obtained at step (f) or (g) with powder of Emblica
officianalis.
In the above method, the plant parts of step (a) are freeze dried parts, organic extracts of the parts or are plant parts pretreated with a fixative agent.
The calcinogenic plants are selected from Cestrum diurnum, lypopersicon esculentum, Solanum melongena, Dactylis glomerata, Medicago saliva, Solanum malacoxylon, Solarium torvum, Solanim verbasiflium and Solanum tuberosum. The powder of Cestrum diurnum is obtained from the leaves and tender twigs of the plant. In case of lypopersicon esculentum, Solanum melongena and other plants, the fruits or leaves of the plant are used.
As mentioned earlier, the fixative agent is extract of plants selected from Emblica officinalis or Triphala or Kushta. The powder of Emblica officinalis used in the composition as fixative agent, the dried powder of the fruit, freeze dried or spray dried extract of the fruits or freeze dried Emblica officinalis juice may be used. If the fixative agent is Triphala extract, it comprises a combination of extracts of fruits of Emblica
officinalis, Terminalia belerica and Terminalia chebula. And, where the fixative agent is Kushta, the extract obtained from the roots of the plant Saussurea lappa.
The dosage recommended may comprise 450 to 50 mg of the fixative agent and 400 to lOOmg of calcinogenic plant powder or an equivalent extract as stated above per dose. The preferred dosage may comprise lOOmg of Emblica officinalis powder and lOOmg of calcinogenic plant powder per dose.
The powder of Emblica officinalis is the dried powder of the fruit, freeze dried or spray dried extract of the fruits or freeze dried Emblica officinalis juice.
The invention also provides a method for preparation of a synergistic herbal composition for treatment of bone disorders and stabilization of vitamin D3 metabolites in plants, said method comprising the steps of:
a. Preparing an aqueous extract of fixative agents,
b. Boiling and cooling the extract of step (a),
c. Immersing the plant parts of Cestrum diurnum in the extract of step (b) for a
period of 40 to 120 min. after washing the plant parts in water,
d. Washing and drying the plant parts of Cestrum diurnum in shade, and
e. Preparing a herbal composition containing Cestrum diurnum.
In an embodiment, the extract of step (b) is cooled to 25 to 30°C. The calcinogenic plant and the fixative agents are the same as mentioned earlier.
To describe the invention in detail, leaves and tender twigs of Cestrum diurnum were collected from mature plants. Freshly collected leaves and tender twigs were immediately washed with water. To stabilize the Vitamin D3 metabolites, washed leaves and tender twigs were treated with freshly prepared herbal extracts (Quath) or solutions of various substances known to have antioxidant properties, for specified duration. The treated plant material was dried under controlled light, temperature and humidity conditions. The dried plant material thus dried was pulverized using a suitable grinder.
In a specific aspect of the invention, one of the components of the herbal composition is powdered dry fruits of Amla (Indian Gooseberry) preferably in a ratio from 1:1 to 1:5.
The present invention is the result of research with certain herbs, including but not limiting to Cestrum diurnum, which, upon treatment with different stabilizing agents, all determined to possess antioxidant properties, followed by drying unde.r .controlled conditions provides an effective herbal base for the treatment ot bone Metabolic Disorders.
The inventive steps for this invention can be divided under two major headings:
a) Stabilization of Vitamin D3 metabolites in the plant material
b) Development and assessment of a herbal based composition for
treatment of Bone Metabolic Disorders.

a) Stabilization material

of Vitamin D3 metabolites in the plant

Cultivation and collection of Cestrum diurnum leaves Plants of Cestrum diurnum were grown as per standard procedure. Leaves and tender twigs were collected in the month of December from sound, mature plants. The harvested twigs are then brought immediately to the shade, away from the reach of sunlight, and the leaves are picked carefully from the twigs, discarding the stems. Leaves are then washed with water to remove the dust and other foreign particles adhering to the surface.
Five different samples of the plant material were thus collected and dried in a Freeze Dryer. The dried material was analyzed and assayed for l,25a-dihydroxy-cholecalciferol content. The results are shown in
following table.
Table 1
(Table Removed)
As the value of l,25cc-dihydroxy-cholecalciferol was found to be nearly Img/kg, which is the reported value in fresh plant material, it is obvious that the natural concentration of l,25cc -dihydroxy-cholecalciferol is maintained in the plant material during Freeze Drying.
In order to see the effect of sunlight, six different samples of freshly collected and washed plant material were spread over paper under sunlight and dried for 4 consecutive days. The dried material was analyzed and assayed for l,25tf-dihydroxy-cholecalciferol content. The results are shown in following table.
Table 2

(Table Removed)
None of the samples dried under sunlight show the presence of l,25a-dihydroxy-cholecalciferol in the plant material. This indicates that the compound gets deteriorated during the course of drying.
Another set of experiment was designed to check the effect of sunlight. Six different samples of freshly collected and washed plant material were spread over paper and kept in a closed chamber protected from light. The chamber was provided with suitable means for circulation of air and kept at a temperature not exceeding 30°C. The plant material was thus dried for 4 days with intermittent raking after every 6 hours. The dried material was analyzed and assayed for 1.25cc-dihydroxy-cholecalciferol content. The results are shown in following table.
Table 3
(Table Removed)
Samples dried in shade at a controlled temperature show that the concentration of l,25a-dihydroxy-cholecalciferol is less in comparison to the Freeze-dried samples. This confirms the complete deterioration of l,25a-dihydroxy-cholecalciferol in sunlight. However, the natural concentration of l,25a-dihydroxy-cholecalciferol is not maintained during the drying process in dark conditions.
In accordance with the process of invention, a series of experiments were done for stabilization of the compound, l,25a-dihydroxy-cholecalciferol, in plant materials. For this purpose it was envisaged that use of certain herbal extracts for a pretreatment of freshly collected leaves of calcinogenic plants known for their vitamin D3 metabolites. A number of such herbal Fixative agents were evaluated in a series of experiments. As a result of these investigations, it was identified that Amla Extract. Triphala Extract and Kushta Extract work as effective Fixative agents. By such an unknown properties these extracts when used for a pretreatment of fresh leaves from herbs like but not limited to Cestrum diurnum to stabilize their active metabolites of vitamin D. The inventive steps also helped to standardize the duration of pretreatment.
Example 1
Following procedure was adopted to stabilize the Vitamin D3 metabolites in the plant material.
Treatment with Amla Extr.ict (Ouath)
The Amla Extract was prepared as per the conventional procedures. To elaborate dried ripe fruits of Amla (Emblica officinalis) were taken and ground to a coarse powder using Restz Grinder fitted with 5 mm sieve. Powdered material is boiled with eight times of Purified water till the quantity of Purified water 'is reduced to one fourth. Collected the decoction by straining -through muslin cloth. Allowed the decoction to cool down to room temperature (25°C - 30JC).
In order to optimize the duration of treatment of plant material, freshly collected plant material was washed with water and immersed in Amla Extract protected from sunlight at a temperature not exceeding 30°C. Samples were drawn after every 10 minutes till the material showed signs of degradation. The samples were immediately washed and dried in a closed chamber protected from light. Finally, they were analyzed for vitamin D3 content and water-soluble extractive value. The result of analysis are summarized as follows:
Table 4

(Table Removed)
It was observed that after 60 minutes of soaking in Amla Extract, colour of leaves changes from dark green to yellowish green. Also, separation of upper epidermal layer and a decrease in water-soluble extractive value was noted. A slight decrease in the concentration of Vitamin D3 was also noticed after 60 minutes. Therefore, the optimum duration of treatment for further studies was kept between 40 - 60 minutes.
In the next phase of the investigations different combinations containing the leaves of calcinogenic plants preferably Centrum din rn urn were prepared. These formulations were also tested for their toxicity and clinical effects. These investigations are summarized as phase 2 of the inventive steps.
Example 2
In order to assess the effect of other herbal extracts, a series of experiments were conducted. The duration of treatment was kept standard in each of the experiments; i.e. 40 - 60 minutes.
Treatment with Triphala Extract fQuath)
Dried ripe fruits of Amla (Emblica officinalis), Haritaki (Terminalia chebula) and Vibhitaki (Terminalia belerica), were taken in equal proportions (collectively known as Triphala) and ground to a coarse powder using Restz Grinder fitted with 5 mm sieve. Powdered material was boiled with eight times of Purified water till the quantity of Purified water was reduced to one fourth. Collected the decoction by straining through muslin cloth. Allowed the decoction to cool down to room temperature (25°C - 30°C).
Freshly collected plant material was washed and immediately soaked in to freshly prepared Triphala Extract for 60 minutes. The material was further processed as described earlier.
Treatment with Kushta Extract fQuath)
Dried roots of Kushta (Saussurea lappa) from were taken and ground to a coarse powder using Restz Grinder fitted with 5 mm sieve. Powdered material is boiled with eight times of Purified water till the quantity of Purified water is reduced to one fourth. Collected the decoction by straining through muslin cloth. Allowed the decoction to cool down to room temperature (25°C - 30°C).
Freshly collected plant material was washed and immediately soaked in to freshly prepared Kushta Extract for 60 minutes. The material was further processed as described earlier.
Treatment with Dashmool Extract fQuath)
Dried roots of Salaparni (Desmodium gangeticum), Prishniparni (Uraria lagopoides), Brihati (Solanum indicum), Kantkari (Solarium xantho carpum), Gokshura (Tribulus terrestris), Bilva (Aegle marmelos). Agnimantha (Premna integrifolia), Syonaka (Oroxylum indicum), Gambhari (Gmelina arborea) and Patala (Stereospermuni suaveolens), collectively known as Dashmool, were taken in equal proportions and ground to a coarse powder using Restz Grinder fitted with 5 mm sieve. Powdered material was boiled with eight times of Purified water till the quantity of Purified water was reduced to one fourth. Collected the decoction by straining through muslin cloth. Allowed the decoction to cool down to room temperature (25°C -30°C).
Freshly collected plant material was washed and immediately soaked in to freshly prepared Dashmool Extract for 60 minutes. The material was further processed as described earlier.
Treatment with Trikatu Extract CQiiath)
Dried fruits of Pippali (Piper long urn), Kali Mirch (Piper nig rum) and rhizomes of Sunthi (Zingiber officinalis), collectively known as Trikatu, were taken in equal proportions and ground to a coarse powder using Restz Grinder fitted with 5 mm sieve. Powdered material was boiled with eight times of Purified water till the quantity of Purified water was reduced to one fourth. Collected the decoction by straining through muslin cloth. Allowed the decoction to cool down to room temperature (25°C - 30°C).
Freshly collected plant material was washed and immediately soaked in to freshly prepared Trikatu Extract for 60 minutes. The material was further processed as described earlier.
Treatment with Ascorbic acid solution
Dissolved 300 grams of Ascorbic acid (M/s. S. D. Fine Chemicals) in 3 liter of Purified water with the help of mechanical stirrer at room temperature(25°C - 30°C).
Freshly collected plant material was washed and immediately soaked in to freshly prepared Ascorbic acid solution for 60 minutes. The material was further processed as described earlier.
Treatment with Citric acid solution
Dissolved about 300 grams of Citric acid Monohydrate (M/s. S. D. Fine Chemicals) in 3 liter of Purified water with the help of mechanical stirrer at room temperature(25°C - 30°C).
Freshly collected plant material was washed and immediately soaked in to freshly prepared Citric acid solution for 60 minutes. The material was further processed as described earlier.
The results obtained from analysis of plant material treated with
different Fixative agents and dried in a dark place as per the
procedure described in example 1 are summarized and compared in
following table:
Table 5

(Table Removed)
Amla Extract, Triphala Extract and Kushta Extract were found to be effective for stabilization of Vitamin D3 metabolites in the plant material, Amla Extract being the most effective one. Triphala Extract, Kushta Extract and Ascorbic acid Solution were found to have moderate action on stabilization of l,25a-dihydroxy-cholecalciferol. Trikatu Extract and Citric Acid Solution were found to have no activity on stabilization of l,25a-dihydroxy-cholecalciferol.
Example 3
In order to confirm the stabilization of Vitamin D3 metabolites in plant materials using Herbal Extracts, leaves and tender twigs of Lycopersicon esculentum and Solanum melongena were treated with Amla Extract and Kushta Extract. Treated and dried material was found to contain l,25a-dihydroxy-cholecalciferol at the same levels as reported in these plants.
From the above examples, it will be appreciated that the present invention provides an effective means of stabilizing Vitamin D3 metabolites in plant materials using Amla Extract or Triphala Extract or Ascorbic Acid Solution. The present invention provides a cost-effective method for maintaining the natural concentration of Vitamin D3 metabolites even after harvesting.
b) Development and assessment of herbal based composition for treatment of Bone Metabolic Disorders.
One of the calcingenic plants. Ct'xtrum diurnum reported to contain Vitamin D3 metabolites have been selected for studies under phase 2 of inventive steps. Freshly harvested plant material was first treated

with Amla Quath and dried as described earlier in Example 1. To evaluate the safety of plant material, acute toxicity studies were carried out on the dried plant material in Mice using both oral and intra-peritoneal routes of administration. The LD 50 value was found to be more than 2000 mg/kg of body weight. Based on the animal data on oral administration of the drug 'No Observed Effect Level' (NOEL) was noted to be 30 mg/kg of body weight.
As a result of all these studies it was possible to identify an effective herbal composition comprising of Amla Powder and the powder of Cestrum diurnum leaves which are pretreated as described in the inventive steps. The surprising benefits of pretreatment and combinations are illustrated in the foregoing examples.
Example 4
Following samples of capsules were prepared to evaluate and compare the efficacy of herbal compositions.
Sample A
Calcium carbonate powder is filled in to empty Hard Gelatin Capsules.
Each capsule contains:
Calcium carbonate 250 mg of elemental Calcium
Two capsules per day have been administered to 6 patients for 6 months duration. Bone Mineral Density was recorded before, after 3 months and after 6 months of treatment.
Sample B
Alfacalcidol powder is filled in to empty Hard Gelatin Capsules. Each capsule contains:
Alfacalcidol 0.125 |^g
Excipients q. s.
Two capsules per day have been administered to 7 patients for 6 months duration. Bone Mineral Density was recorded before, after 3 months and after 6 months of treatment.

Sample C
Plant material prepared as per the procedure described in Example 3 is powdered and filled in to empty Hard Gelatin Capsules.
Each capsule contains:
Plant material (Treated) 100 mg
Excipients q. s.
Two capsules per day have been administered to 7 patients for 3 months duration. Bone Mineral Density was recorded before and after 3 months of treatment.
Sample D
Freshly harvested Plant material was taken and dried under shade at a temperature not exceeding 30°C for 4-5 days. Dried Plant material is powdered and filled in to empty Hard Gelatin Capsules.
Each capsule contains:
Plant material (Untreated) 100 mg
Excipients - q. s.
Two capsules per day have been administered to 7 patients for 3 months duration. Bone Mineral Density was recorded before and after 3 months of treatment.
Sample E
Plant material has been prepared as per the procedure described in Example 3 and powdered. The powdered Plant material was mixed with Dry Amla powder and filled in to empty Hard Gelatin Capsules.
Each capsule contains:
Plant material (Treated) 100 mg
Dried Amla Powder 450 ms
•o
Two capsules per day have been administered to 7 patients for 6 months duration. Bone Mineral Density was recorded before, after 3 months and after 6 months of treatment.

Effect of Sample A
(Table Removed)
Effect of Sample B

(Table Removed)
Effect of Sample C
(Table Removed)
Effect of Sample D

(Table Removed)
Effect of Sample E

(Table Removed)
The data obtained from' all the groups could be summarized as follows:


(Table Removed)
As seen in above tables, it is obvious that the treated plant material has a positive effect on Bone Mineral Density. Further, it has been established that 'the plant material when administered with Amla powder performs surprisingly better than Sample C which contained only Cestrum diurnum leaf powder.



WE CLAIM:
1. A synergistic herbal composition for the treatment of bone metabolic disorders, said
composition comprising calcinogenic plant parts and powder of fruits of Emblica
officinalis in the ratio of 1:1 to 1:7.
2. A composition as claimed in claim 1 wherein the plant parts of the calcinogenic plant
are freeze dried parts, organic extract of the plant parts or plant parts pretreated with a
fixative agent.
3. A composition as claimed in claim 1 wherein the calcinogenic plants are selected
from Cestrum diurnum, lypopersicon esculentum, Solanum melongena, Dactylis
glomerata, Medicago saliva, Solanum malacoxylon, Solanum torvum, Solanim
verbasiflium and Solanum tuberosum.
4. A composition as claimed in claim 1 wherein the fixative agent is extract of fruits of
Emblica officinalis.
5. A composition as claimed in claim 1 wherein the fixative agent is Triphala extract
comprising a combination of extracts of fruits of Emblica officinalis, Terminalia
belerica and Terminalia chebula.
6. A composition as claimed in claim 1 wherein the fixative agent is Kushta extract
obtained from the roots of the plant Saussurea lappa.
1. A composition as claimed in claim 1 wherein the plant parts of Cestrum diurnum are obtained from the leaves and tender twigs.
8. A composition as claimed in claim 1 comprising 450 to 50 mg of Emblica officinalis
powder and 400 to 50 mg of calcinogenic plant powder or an equivalent extract per
dose.
9. A composition as claimed in claim 1 comprising lOOmg of Emblica officinalis
powder and lOOmg of calcinogenic plant powder or an equivalent extract per dose.
10. A composition as claimed in claim 1 wherein the powder of Emblica officinalis is the
dried powder of the fruit, freeze dried or spray dried extract of the fruits or freeze
dried Emblica officinalis juice.
11. A composition as claimed in claim 1 wherein the calcinogenic plants contains vitamin
D3 metabolites such as l,25cc-dihydroxy-cholecalciferol as one of the active
ingredient.
12. A composition as claimed in claim 1 wherein the powder or the extract of the
calcinogenic plant is used.
13. A method for preparation of a synergistic herbal composition for treatment of bone
disorders and stabilization of vitamin D3 metabolites in plants, said method
comprising the steps of mixing powder or an extract of calcinogenic plant parts with
Emblica officinalis powder in a ratio between 1:1 to 1:7.
14. A method as claimed in claim 13, said method comprising the steps of:
a. harvesting the leaves of the calcinogenic plants at suitable period of time,
b. washing the leaves in water,
c. treatment of washed leaves with a fixative agent,
d. washing the treated leaves with water,
e. drying of treated and washed leaves under controlled conditions of light,
humidity and temperature,
f. grinding the dried leaves by suitable means, protected from light and high
temperatures,
g. freeze drying or extraction of calcinogenic plant parts with organic solvents,
and
h. mixing the product of step (f) or (g) with powder of Emblica officinalis.
15. A method as claimed in claim 13 or 14 wherein the plant parts of the calcinogenic
plant are freeze dried parts, organic extract of the parts or plant parts pretreated with
a fixative agent. '
16. A method as claimed in claim 13 or 14 wherein the calcinogenic plant is selected
from Cestrum diurnum, lypopersicon esciilentum, Solatium melongena, Dactylis
glomerata, Medicago saliva, Solatium malacoxylon, Solatium torvum, Solanim verbasiflium and Solarium tuberosum.
17. A method as claimed in claim 13 or 14 wherein the powder or the extract of the
calcinogenic plant is used.
18. A method as claimed in claim 13 or 14 wherein the fixative agent is extract of fruits
of Emblica officinalis.
19. A method as claimed in claim 13 or 14 wherein the fixative agent is Triphala extract
comprising a combination of extracts of fruits of Emblica officinalis, Terminalia
belerica and Terminalia chebula.
20. A method as claimed in claim 13 or 14 wherein the fixative agent is Kushta extract
obtained from the roots of the plant Saussurea lappa.
21. A method as claimed in claim 13 or 14 wherein the powder of Cestrum diurnum is
obtained from the leaves and tender twigs.
22. A method as claimed in claim 13 or 14 comprising 450 to 50 mg of Emblica
officinalis powder and 400 to 50 mg of calcinogenic plant powder or an equivalent
extract per dose.
23. A method as claimed in claim 13 or 14 comprising 325 mg of Emblica officinalis
powder and lOOmg of calcinogenic plant powder or an equivalent extract, per dose.
24. A method as claimed in claim 13 or 14 wherein the powder of Emblica officinalis is
the dried powder of the fruit, freeze dried or spray dried extract of the fruits or freeze
dried Emblica officinalis juice.
25. A herbal composition substantially as herein described with reference to the
examples.
26. A method for preparing a herbal composition substantially as herein described with
reference to the examples.

Documents:

574-del-2001-abstract.pdf

574-del-2001-claims.pdf

574-del-2001-correspondence-others.pdf

574-del-2001-correspondence-po.pdf

574-del-2001-description (complete).pdf

574-del-2001-form-1.pdf

574-del-2001-form-19.pdf

574-del-2001-form-2.pdf

574-del-2001-form-26.pdf

574-del-2001-form-3.pdf

574-del-2001-form-5.pdf

574-del-2001-petition-138.pdf


Patent Number 209451
Indian Patent Application Number 574/DEL/2001
PG Journal Number 48/2007
Publication Date 30-Nov-2007
Grant Date 30-Aug-2007
Date of Filing 15-May-2001
Name of Patentee DABUR RESEARCH FOUNDATION
Applicant Address 22, SITE IV, SAHIBABAD, GHAZIABAD 201 010, INDIA.
Inventors:
# Inventor's Name Inventor's Address
1 DASALUKUNTE BHIMARAO ANANTA NARAYANA DABUR FOUDATION, 22, SITE IV, SAHIBABAD, GHAZIABAD 201 010, INDIA.
2 CHANDRA KANT KATIYAR DABUR FOUDATION, 22, SITE IV, SAHIBABAD, GHAZIABAD 201 010, INDIA.
3 NARASIMHA BABA BRINDAVANAM DABUR FOUDATION, 22, SITE IV, SAHIBABAD, GHAZIABAD 201 010, INDIA.
4 RAMESH KUMAR DUGGAL DABUR FOUDATION, 22, SITE IV, SAHIBABAD, GHAZIABAD 201 010, INDIA.
5 RAVI JAIN DABUR FOUDATION, 22, SITE IV, SAHIBABAD, GHAZIABAD 201 010, INDIA.
6 RAGHU RAMULU NAMALA DABUR FOUDATION, 22, SITE IV, SAHIBABAD, GHAZIABAD 201 010, INDIA.
7 AVINASH NARWARIA DABUR FOUDATION, 22, SITE IV, SAHIBABAD, GHAZIABAD 201 010, INDIA.
PCT International Classification Number A61K 035/78
PCT International Application Number N/A
PCT International Filing date
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 NA