FIELD OF THE INVENTION
[0001] The present disclosure generally relates to a pharmaceutically acceptable herbal preparationsubject to natural treatment and management of osteoporosis andrelated disorders. More specifically it relates to a pharmaceutically acceptable herbal preparationfor treating and preventing osteoporosis and the associated disorders. It further relates to processof preparing such herbal preparation.

BACKGROUND OF THE INVENTION
[0002] Background description includes information that may be useful in understanding the present invention. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, or that any publication specifically or implicitly referenced is prior art.
[0003] Osteoporosis is a condition characterized by a decrease in the density of bone, decreasing its strength and resulting in fragile bones. Osteoporosis literally leads to abnormally porous bone that is compressible like a sponge. As osteoporosis, or thinning bones, weakens the skeleton of the bone, it also results in frequent fractures (breaks) in the bones.
[0004] Risk factors for osteoporosis include genetics, lack of exercise, aging, lack of calcium and vitamin D, smoking, excessive alcohol consumption, low sex hormones, low body weight, history of rheumatoid arthritis, personal history of fracture as an adult, and family history of osteoporosis. Risk factors are also associated with aging, being female, after menopause and some medications. Asbone mass (bone density) decreases after 35 years of age, bone loss occurs more rapidly in women after menopause.
[0005] Prevention and treatment includeincreasing intake of calcium and vitamin D, magnesium, fluorides, and restriction of the amount of salt, caffeine, alcohol, and consumed animal protein, exercise, and osteoporosis medications. Prevention is likely to remain the most effective method of dealing with osteoporosis. Hormone replacement therapy, starting shortly after menopause, prevents rapid bone loss and leads to reduction in the fracture risk of up to 60%. Unfortunately, there are side effects and other risk factors associated with using estrogens in hormone replacement to prevent postmenopausal osteoporosis,e.g., breast cancer, osteonecrosis of the jaw, hypercalcemia, and hypertension.
[0006] Some chemical or pharmaceutical agents are known for promoting bone growth in humans. For example, WO9619246 describes a method for promoting bone growth in a human patient by (normally intermittent not continuous) administration of parathyroid hormone, PTH-related protein or an agonist for at least one month. In WO9619501, a pancreatic-derived factor inhibits the resorption of bone and stimulates bone cells to proliferate and increases the formation of bone.
[0007] Recently, a preparation and method for treatment of osteoporosis by transdermal drugs has been disclosed in US patent US9198931B2. But the effectiveness of transdermal drug delivery in osteoporosis remains questionable as per the case to case basis of osteoporosis. WO99/55351A discloses a herbal composition, characterized by the fact that it comprises dandelion root (Teraxaci radix) at 6-12 wt%.Although certain other preparations for treatment of osteoporosis are mentioned in prior arts, there is continual need of more efficient treatments with lesser or no side effects to the body.
[0008] Although these chemicals and pharmaceutical compounds have been proved effective for the treatment of bone disorders, it would be of interest to provide a safe and efficient nutritional way to promote bone growth and prevent or alleviate the symptoms of bone/joint disorders or precisely osteoporosis in mammals.In view of this, there remains an unmet need to develop alternative, safe, effective osteoporosis prevention measures and/or treatments that are cost effective and secure. There is a need for improved medicinal preparations for use in the treatment or regulation of such physiological and pathological conditions, without the adverse/toxic effects associated with conventional modes of treatment in such conditions. It is an object of the present invention to provide such a preparation.
[0009] Thus, the present invention provides a herbal preparation that can act as aneffective treatment or prophylaxis to manage the medical condition of osteoporosis and associated disorders.

OBJECTS OF THE INVENTION
[0010] Primary object of the present disclosure is to provide a herbal preparation for effective treatment and prevention of osteoporosis and related disorders.
[0011] Another object of the present disclosure is to provide a method of preparation of a herbal preparation for effective treatment and prevention of osteoporosis and related disorders.
[0012] Another object of the present disclosure is to provide a herbal preparation for effectivetreatment and prevention of osteoporosis and related disorders which is easy to formulate and is cost-effective.
[0013] Another object of the present disclosure is to provide a herbal preparation for effective treatment and prevention of osteoporosis and related disorders with minimalside-effects.
[0014] Another object of the present disclosure is to provide a herbal preparation that may promote human osteoblast cell proliferation and possess anti-osteoporotic potential.
[0015] Another object of the present disclosure is to provide a herbal preparation for effective treatment and prevention of osteoporosis and related disorders as a primary medication.
[0016] Another object of the present disclosure is to provide a herbal preparation for effective treatment and prevention of osteoporosis and related disorders as a supplemental medication along with primary treatment.

SUMMARY OF THE INVENTION
[0017] The present disclosure generally relates to a pharmaceutically acceptable herbal preparation subject to natural treatment and management of osteoporosis and related disorders. More specifically it relates to a pharmaceutically acceptable herbal preparation for treating and preventing osteoporosis and the associated disorders. It further relates to process of preparing such herbal preparation.
[0018] In accordance with the present disclosure, the foregoing objectives and advantages have been readily achieved. It has now been found that some plants or plant extracts have a particular positive effect on bone formation and repair, on maintenance of bone health or prevention, alleviation and/or treatment of bone disorders including osteoporosis.The present disclosure provides a herbal preparation with potential anti-osteoporotic property comprising: Acacia nilotica herb, and at least one pharmaceutically acceptable excipient.
[0019] In an embodiment, said Acacia nilotica herb is present in the form of an extract in an amount ranging from about 10% to 30% by weight of the herbal preparation.
[0020] In an embodiment, said herbal preparation further comprises of herb Pergularia daemia in the form of an extract in an amount ranging from about 2% to 10% by weight of the herbal preparation.
[0021] In an embodiment of the present disclosure,said Acacia nilotica and Pergularia daemiaherbs comprise ofat least one of a rhizome, a root, a bud, a tender shoot, a leaf, a stem, a fruit, a bark, a flower, a seed,ora combination thereof.
[0022] In an embodiment, the herbal preparation comprises of at least a part of the herb, whole herb, an extract of the herb or a combination thereof.
[0023] In an embodiment of the present disclosure, said at least one pharmaceutically acceptable excipient is selected from a group consisting of a sweetening agent, a buffering agent, apreservative, a bulking agent, a binder, a disintegrant, a chelating agent, a glidant, aflavouring agent, a colouring agent, a sweetener, a tonicity agent, a carrier, a solvent, a co-solvent, or a combination thereof.
[0024] In an embodiment, said at least one pharmaceutically acceptable excipient isselected from agroup consisting of a whey protein, a soy protein, a casein protein,and a plant-based protein.
[0025] In an embodiment, the herbal preparation is formulated in a form selected from agroup consisting of a dry powder, a tablet, acapsule, a bolus,and a liquid.
[0026] In an embodiment of the present disclosure is provided a process of preparing a herbal preparation with potential anti-osteoporotic property, comprising the steps of:
- collecting parts of herb Acacia nilotica, preferably seeds of Acacia nilotica;
- washing the parts of herb Acacia niloticaand drying it until removal of moisture;
- grinding the dried parts of herb Acacia niloticaand sieving it to obtain a herbal powder with particle size of = 250 µm;
- extracting the herbal powder with a solvent to obtain a 10% to 30% herbal extract;
- concentrating the herbal extract to obtain a concentrate;
- formulating the concentrate in a suitable dosage form to obtain final herbal preparation.
[0027] In yet another embodiment of the present disclosure is provided a process of preparing a herbal preparation with potential anti-osteoporotic property, comprises the steps of:
- collecting parts of herb Acacia nilotica, preferably seeds of Acacia nilotica;
- washing the parts of herb Acacia niloticaand drying it until removal of moisture;
- grinding the dried parts of herb Acacia niloticaand sieving it to obtain a powder with particle size of = 250 µm;
- extracting the powder with a solvent to obtain a 10% to 30% Acacia nilotica extract;
- collecting part of herb Pergularia daemia, preferably leaves of Pergularia daemia, washing it, drying it until moisture removal and grinding it to obtain a fine powder with particle size of = 250 µm followed by extracting fine powder to obtain 2% to 10% extract;
- mixing the Acacia nilotica extract and the Pergularia daemiaextract through a blender to obtain a poly herbal extract;
- concentrating the poly herbal extract to obtain a concentrate;
- formulating the concentrate in a suitable dosage form to obtain final herbal preparation.
[0028] In an embodiment of the present disclosure, the solvent used for extraction is selected from a group consisting of water, alcohol, combination of water and alcohol in a ratio of 1:1 or any combination thereof.
[0029] In an embodiment of the present disclosure, said extraction step is performed at a temperature ranging from about 30 °C to 60 °C.
[0030] In yet another embodiment of the present disclosure, said concentration step is performed at a reduced pressure and at a temperature rangingfrom about 40 °C to 60 °C.

DETAILED DESCRIPTION OF THE INVENTION
[0031] The following is a detailed description of embodiments of the disclosure. The embodiments are in such detail as to clearly communicate the disclosure. However, the amount of detail offered is not intended to limit the anticipated variations of embodiments; on the contrary, the intention is to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the present disclosure as defined by the appended claims.
[0032] Each of the appended claims defines a separate invention, which for infringement purposes is recognized as including equivalents to the various elements or limitations specified in the claims. Depending on the context, all references below to the “invention” may in some cases refer to certain specific embodiments only. In other cases it will be recognized that references to the “invention” will refer to subject matter recited in one or more, but not necessarily all, of the claims.
[0033] As used in the description herein and throughout the claims that follow, the meaning of “a,” “an,” and “the” includes plural reference unless the context clearly dictates otherwise. Also, as used in the description herein, the meaning of “in” includes “in” and “on” unless the context clearly dictates otherwise.
[0034] All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g. “such as”) provided with respect to certain embodiments herein is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention otherwise claimed. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the invention.
[0035] Various terms as used herein are shown below. To the extent a term used in a claim is not defined below, it should be given the broadest definition persons in the pertinent art have given that term as reflected in printed publications and issued patents at the time of filing.
[0036] Features that are described and/or illustrated with respect to one embodiment, aspects or implementations may be used in the same way or in a similar way in one or more other embodiments and/or in combination with or instead of the features of the other embodiments, aspects or implementations.
[0037] It should be emphasized that the term “comprises/comprising” when used in this specification is taken to specify the presence of stated features, integers, steps or components but does not preclude the presence or addition of one or more other features, integers, steps, components or groups thereof.
[0038] Herbal medicine has been used for centuries in rural areas by local healers, a number of substances used in modern medicine for the treatment of several diseases have originated from research on medicinal plants.
[0039] The term "herbal preparation", as used herein, intends to cover herbal preparation having therapeutic and prophylactic effect in treating osteoporosis.
[0040] The term “osteoporosis” includes all conditions related to low bone density caused through any of the earlier mentioned reasons.
[0041] It is also understood that the terms “collecting”, "washing", ‘grinding", "drying", "sieving" “roasting”, "extracting", “concentrating” used herein intend to cover all kinds of conventional methods used for the said purpose carried out at room temperature unless specified particularly herein.
[0042] The term "pharmaceutically acceptable" refers to molecular entities and preparations that are physiologically tolerable and do not typically produce an allergic or similarly untoward reaction, such as gastric upset, dizziness and the like, when administered to a human Preferably, as used herein, the term "pharmaceutically acceptable" means approved by a regulatory agency of the federal or a state government or listed in the U S Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.
[0043] The term "carrier" refers to a diluent, excipient, or vehicle with which the compound is administered Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. Water or solution saline solutions and aqueous dextrose and glycerol solutions are preferably employed as carriers, particularly for injectable solutions.
[0044] As used herein, "treatment", “management” and "treating" and the like generally mean obtaining a desired pharmacological and physiological effect. The effect may be prophylactic in terms of preventing or partially preventing a disease, symptom or condition thereof and/or may be therapeutic in terms of a partial or complete cure of a disease, condition, symptom or adverse effect attributed to the disease. The term "treatment" as used herein covers any treatment of a disease in a mammal, particularly a human, and includes: (a) preventing the disease from occurring in a subject which may be predisposed to the disease but has not yet been diagnosed as having it; (b) inhibiting the disease, i.e., arresting its development; or relieving the disease, i.e., causing regression of the disease and/or its symptoms or conditions.
[0045] The “patient” may be varied and includes a mammal such as a human or any other animal. When the patient is a human it may be an adult man or woman, child or infant.
[0046] The present disclosure generally relates to a pharmaceutically acceptable herbal preparation subject to natural treatment and management of osteoporosis and related disorders. More specifically, it relates to a pharmaceutically acceptable herbal preparation for treating and preventing osteoporosis and the associated disorders. It further relates to process of preparing such herbal preparation.
[0047] In accordance with the present disclosure, the foregoing objectives and advantages have been readily achieved. It has now been found that some plants or plant extracts have a particular positive effect on bone formation and repair, on maintenance of bone health or prevention, alleviation and/or treatment of bone disorders including osteoporosis.The present disclosure provides a herbal preparation with potential anti-osteoporotic property comprising: Acacia nilotica herb, and at least one pharmaceutically acceptable excipient.
[0048] In an embodiment, said Acacia nilotica herb is present in the form of an extract in an amount ranging from about 10% to 30% by weight of the herbal preparation.
[0049] In an embodiment, said herbal preparation further comprises of herb Pergularia daemia in the form of an extract in an amount ranging from about2% to 10%by weight of the herbal preparation.
[0050] In an embodiment of the present disclosure, said Acacia nilotica and Pergularia daemiaherbs comprise ofat least one of a rhizome, a root, a bud, a tender shoot, a leaf, a stem, a fruit, a bark, a flower, a seed, or a combination thereof.
[0051] In an embodiment, the herbal preparation comprises of at least a part of the herb, whole herb, an extract of the herb or a combination thereof.
[0052] In an embodiment of the present disclosure, said at least one pharmaceutically acceptable excipient is selected from a group consisting of a sweetening agent, a buffering agent, apreservative, a bulking agent, a binder, a disintegrant, a chelating agent, a glidant, aflavouring agent, a colouring agent, a sweetener, a tonicity agent, a carrier, a solvent, a co-solvent, or a combination thereof.
[0053] In an embodiment, said at least one pharmaceutically acceptable excipient isselected from a group consisting of a whey protein, a soy protein, a casein protein,and a plant-based protein.
[0054] In an embodiment, the herbal preparation is formulated in a form selected from a group consisting of a dry powder, a tablet, acapsule, a decoction, a suspension, a syrup, a bolus, and a liquid.
[0055] In an embodiment of the present disclosure is provided a process of preparing a herbal preparation with potential anti-osteoporotic property, comprising the steps of:
- collecting parts of herb Acacia nilotica, preferably seeds of Acacia nilotica;
- washing the parts of herb Acacia nilotica and drying it until removal of moisture;
- grinding the dried parts of herb Acacia nilotica and sieving it to obtain a herbal powder with particle size of = 250 µm;
- extracting the herbal powder with a solvent to obtain a 10% to 30% herbal extract;
- concentrating the herbal extract to obtain a concentrate;
- formulating the concentrate in a suitable dosage form to obtain final herbal preparation.
[0056] In yet another embodiment of the present disclosure is provided a process of preparing a herbal preparation with potential anti-osteoporotic property, comprises the steps of:
- collecting parts of herb Acacia nilotica, preferably seeds of Acacia nilotica;
- washing the parts of herb Acacia nilotica and drying it until removal of moisture;
- grinding the dried parts of herb Acacia nilotica and sieving it to obtain a powder with particle size of = 250 µm;
- extracting the powder with a solvent to obtain a 10% to 30% Acacia nilotica extract;
- collecting part of herb Pergularia daemia, preferably leaves of Pergularia daemia, washing it, drying it until moisture removal and grinding it to obtain a fine powder with particle size of = 250 µm followed by extracting fine powder to obtain 2% to 10% extract;
- mixing the Acacia nilotica extract and the Pergularia daemiaextract through a blender to obtain a poly herbal extract;
- concentrating the poly herbal extract to obtain a concentrate;
- formulating the concentrate in a suitable dosage form to obtain final herbal preparation.
[0057] In an embodiment of the present disclosure, the solvent used for extraction is selected from a group consisting of water, alcohol, combination of water and alcohol in a ratio of 1:1 or any combination thereof.
[0058] In an embodiment of the present disclosure, said extraction step is performed at a temperature ranging from about 30 °C to 60 °C.
[0059] In yet another embodiment of the present disclosure, said concentration step is performed at a reduced pressure and at a temperature ranging from about 40 °C to 60 °C.
[0060] In an embodiment, the disclosed herbal preparation is capable of effectively managing osteoporosis and related conditions, with minimal side effects. The herbal preparation may be mass produced inexpensively and used in an easy, cost effective, environment friendly and productive way. The disclosure further provides a process of preparing the herbal preparation and demonstrates anti-osteoporotic potential of herbal preparation using human osteoblast like cell line (MG-63).
[0061] In an embodiment, the herbal preparation comprises of herb Acacia nilotica as active ingredient or a combination of one or more active ingredients extracted from said herb along with at least one pharmaceutically acceptable carrier.
[0062] Inan embodiment, the herbal preparation further comprises of herb Pergularia daemiaor a combination of one or more active ingredients extracted from said herb.
[0063] In an embodiment of the present disclosure, said herbal preparation comprises precise refined parts of herbs in an amount pharmaceutically effective as an active ingredient, demonstrating its ability to manage and reduce problems of osteoporosis and contribution in increasing bone density.
[0064] In an embodiment of the present disclosure, said herbal preparation may also comprise of any chemical or functional equivalents or horticultural or botanical equivalents of the parts of herbs utilized.
[0065] In an embodiment of the present disclosure, said herbal preparation comprising refined parts of herbs may also contain a blend of extracts or refined parts of other medicinal herbs to enhance the efficacy of said herbal preparation in treatment and prophylaxis of osteoporosis and associated disorders.
[0066] In a preferred embodiment of the present discloure, said herbal preparationincludes refined parts of Acacia nilotica, preferably including (but not limited to) seeds of plant Acacia nilotica. However, a person skilled in the art may appreciate that any plant part or awhole plant may also be used for preparing said herbal preparation for management and reduction of osteoporosis and associated disorders.
[0067] In anembodiment of the present disclosure, said herbal preparation comprising refined parts ofPergularia daemiamay include whole parts of the herb.
[0068] For the purpose stated in present disclosure, the herbsAcacia nilotica andPergularia daemiaare collected from surroundings of Ganganagar, Rajasthan, India.
[0069] In yet another embodiment, said herbal preparation based on plant parts of Acacia nilotica can be formulated in various pharmaceutical formulations with active ingredient of said plant, wherein said pharmaceutical formulations include (but not limits to) nano-formulations, encapsulations, or any kind of enteral formulations.
[0070] In a preferred embodiment of the present disclosure, said herbal preparation may be formulated in various enteral dosage forms selected from a group consisting of powder, capsule, tablet, bolus, solution, decoction, suspension, syrup or like.
[0071] Said pharmaceutically acceptable excipients may be selected from a wide range of excipients acceptable for utilization in herbal formulations including but not limited to a sweetening agent, a buffering agent, a preservative, a bulking agent, a binder, a disintegrant, a chelating agent, a glidant, a flavouring agent, a colouring agent, a sweetener, a tonicity agent, a carrier, a solvent, a co-solvent, and a combination thereof.
[0072] In a preferred embodiment, the pharmaceutically acceptable excipients include at least one of the following: a sweetening agent/diluent in a range preferably, 0.03 to 0.7 % by weight, a buffering agent in a range preferably 0.03 % to 0.07% by weight, a preservative in a range preferably 0.05 % to 0.8 % by weight, a bulking agent in a range preferably 10 % to 25 % by weight, a binder in a range preferably 2 % to 9 % by weight, a disintegrant in a range preferably 0.5 to 4 % by weight, co-solvents in a range preferably 1 % to 10 % by weight, a glidant in a range preferably 0.1 to 0.5 % by weight, depending upon the form of herbal preparation.
[0073] In an embodiment, said suitable necessary solvents can beselected from a wide range of solvents including polar, non-polar and semi-polar solvents. In a preferred embodiment, said suitable solvent may be water or an alcohol or a hydro-alcohol or combination thereof. In another preferred embodiment, said suitable solvent can be water or a combination of water and water miscible solvents.
[0074] Inanother embodiment, said suitable solvent may be selected from agroup consisting of water, acetone, dimethyl isosorbide, glycols, propylene glycol, ethyl alcohol, cetyl alcohol, glyceryl stearate, isopropyl alcohol, diethylamine, glyceryloleate, glycerine, myristyl alcohol, gelatin, simple syrup, cyclodextrin, polyvinyl pyrrolidone (Povidone), benzyl alcohol, glycerin, propylene glycol, ethanol, sorbitol, maltitol, xylitol, inositol, mannitol, invert sugars, sorbitol and the like or a combination thereof.
[0075] In an embodiment, the solvent used for the extraction of herb may include a hydro-alcohol solvent. Preferably, the hydro-alcohol solvent can include 5% water and 95% alcohol, by volume. Preferred alcohol solvent can be selected from the group consisting of methanol, ethanol, isopropanol and butanol. In yet another preferred embodiment, the solvent may include a mixture of water and ethanol in ratio of 1:1.
[0076] In an embodiment, the at least one carrier may be selected from agroup consisting of a whey protein, a soy protein, a casein protein, a plant-based blend, or other protein powders.
[0077] In an embodiment, the herbal preparation may be administered to a patient in morning on empty stomach.
[0078] In an embodiment,said herbal preparation in active dose may be utilized as primary medicament or herbal supplement for effective treatment and prophylaxis of osteoporosis or associated low bone density disorders with minimalside-effects.
[0079] Said herbal preparation and combination is under extensive clinical trialsfor demonstrating its ability to manage and reduce osteoporosis effects and other associateddisorders and increase bone density.
[0080] In an embodiment, said concentrating of extract implies to obtain a concentrate of extract that may be obtained through any concentrating method or means known to a person skilled in the art including, but not limited to, heating on a water bath, heating in an oven, performing evaporation in a evaporator including rotary evaporator, etc.
[0081] In an embodiment, said formulation of extract obtained in effective amount as active dose in to at least one dosage form including an oral dosage form through methods known in the art.
Examples
[0082] The present disclosure is further explained in the form of following examples. However, it is to be understood that the foregoing examples are merely illustrative and are not to be taken as limitations upon the scope of the invention. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art. Such changes and modifications may be made without departing from the scope of the invention.
Example 1
[0083] Seeds of herb Acacia nilotica were taken, washed properly and dried under shade. Fully dried seeds were grinded to obtain a fine powder. The raw herbal powder wasstored in a dry place and wasused to effectively treat and prevent osteoporosis and related bone disorders in following dosage: 5 g of powder was given with warm cow/goat milk twice a day after food for 15-20 days. For children (< 10 year): the dosage given was 2.5 g for effective treatment and prevention of osteoporosis and related conditions.
Example 2
[0084] Seeds of Acacia nilotica herb were collectedand washed under flowing water. Washed parts of herb were dried until total removal of moisture and grinded to obtain a coarse powder with particle size of = 250 µm. The fine coarse powder in an amount about 7g to 15 g obtained was then subjected to extraction with water and ethanol in a ratio of 1:1 and at a temperature of 40 to 50°C. After number of rounds of extraction, 15% to 20% herbal extract obtained was then concentrated in a rotary evaporator at reduced pressure and temperature within 40 to 60°C. The concentrate obtained was then formulated into a100 ml syrup. The formulated syrup was given to a patient in an amount of 5ml in single or divided doses per day for at least 10-15 days for effective treatment and prevention of osteoporosis and related conditions.
Example 3
[0085] Seeds of Acacia nilotica herb were collected, preferably including seeds of Acacia nilotica and washed under flowing water. Washed parts of herb were dried until total removal of moisture and grinded to obtain a coarse powder with particle size of = 250 µm. The fine coarse powder in an amount of about 7g to 15g was then subjected to extraction with water and ethanol in a ratio of 1:1 and at a temperature of 40°c to 50°C. After number of rounds of extraction, 15% to 20% Acacia nilotica extract was obtained. The extract was then concentrated in a rotary evaporator at reduced pressure and temperature within 40°c to 60°C. Similarly,leaves of Pergularia daemiaherb were collected, washed and dried until total removal of moisture and grinded to obtain a coarse powder with particle size of = 250 µm. The fine coarse powder in an amount about 2g to 5g was then subjected to extraction in similar extraction method to obtain a 2% to 10% Pergularia daemiaextract. The Acacia nilotica extract and Pergularia daemiaextract werethen concentrated in a rotary evaporator at reduced pressure and temperature within 40°C to 60°C. The concentrate of poly herbal extract was then mixed in blender properly and formulated into a syrup. The formulated syrup was given to a patient in 5ml of single of divided doses per day for at least 10-15 days for effective treatment and prevention of osteoporosis and related conditions.
Example 4
[0086] Evaluation of anti-osteoporotic potential of herbal preparation using human osteoblast like cell line (MG-63)
[0087] Materials and Methods
[0088] Test System:
Human bone osteosarcoma cell line MG-63 was used as test system in present evaluation study. The details are following:
Cell Line: MG-63 (Human bone osteosarcoma)
Organism: Homo sapiens, human
Tissue: Bone
Disease: osteosarcoma
Growth Medium for routine culture: DMEM supplemented with 10% FBS
Growth Medium during experiments: Phenol free DMEM supplemented with 10% CD-FBS
Antibiotics: Penicillin (100U/ml), Streptomycin (100µg/ml)
Growth conditions: 37°c, 5% CO2 and 95%humidity
Sub-culturing medium: Trypsin (0.2%) and EDTA (0.02%)
[0089] Test Groups
Group I:The herbal preparation as mentioned in example 1 is weighed and dissolved in DMSO to attain 200 mg//ml stock solution, which is further diluted in DMEM to achieve non-cytotoxic weight/volume concentrations for subsequent treatment of cells.
Group II: ß-Estradiol was taken as Positive control.
Group III: Untreated.
Culture and maintenance of cell line
[0090] MG-63 cell line was maintained under conditions as described and was sub-cultured by trypsinization followed by splitting the cell suspension into fresh flasks and supplementing with fresh cell growth medium. Three days before the start of the experiment, the growth medium was replaced with fresh phenol-free DMEM supplemented with 10% CD-FBS and 1% penicillin-streptomycin.
[0091] MG-63 cells were processed to determine non-cytotoxic concentrations of herbal preparation and effect on alkaline phosphatase enzyme activity.
RESULTS
Non cytotoxic concentrations of herbal preparation:
[0092] The non-cytotoxic concentrations were determined by viability assay using MTT. The herbal preparation as mentioned in example 1 is diluted in a concentration range of 0.005mg/ml – 10mg/ml (weight/volume) in serum free phenol free DMEM. The table1 shows the viability of MG-63 cells when treated with Group I with respect to untreated cells.
Table 1: Non-cytotoxic dose determination of Group I
Concentration Group I
0.005mg/ml 109.63
0.01mg/ml 88.27
0.02mg/ml 102.96
0.05mg/ml 108.48
0.1mg/ml 109.25
0.2mg/ml 119.92
0.5mg/ml 131.17
1mg/ml 151.57
5mg/ml 112.15

Effect of herbal preparation on cellular proliferation and alkaline phosphatase activity
[0093] The dose was considered non-toxic if the viability was more than 70%. Based on said result the doses for ALP activity assessment were selected for Group I as 0.1mg/ml-5mg/ml. Table 2 shows the effect of herbal preparation on cellular proliferation and alkaline phosphatase activity.
Table 2 Effect of herbal preparation on cellular proliferation and alkaline phosphatase activity
Test Groups Concentrations Cellular Proliferation Percentage ALP Activity
Group I 5mg/ml 112.15 572.29
1mg/ml 151.57 419.28
0.5mg/ml 131.17 135.54
0.2mg/ml 119.92 125.30
0.1mg/ml 109.25 106.63
Group II 100nm 107.543 145.78
50nm 94.726 112.05
25nm 116.689 130.12
10nm 125.501 138.55
Group III - 100 100

Conclusion
[0094] Under the objective of evaluating the effect of herbal preparation on cellular proliferation and ALP activity as a marker of anti-osteoporotic potential, the herbal preparation as taken under Group I led to increase in cellular proliferation and ALP activity at all tested concentrations and therefore it is concluded that the herbal preparation disclosed is effective as anti-osteoporotic medicament.
[0095] Accordingly, the disclosure is intended to embrace all such alternatives, modifications and variations as may fall within the spirit and scope of the present disclosure. The foregoing descriptions of specific embodiments of the present disclosure have been presented for purposes of illustration and description. They are not intended to be exhaustive or to limit the disclosure to the precise forms disclosed, and obviously many modifications and variations are possible in light of the above teaching. The embodiments were chosen and described in order to best explain the principles of the disclosure and its practical application, to thereby enable others skilled in the art to best utilize the invention and various embodiments with various modifications as are suited to the particular use contemplated. It is understood that various omissions, substitutions of equivalents are contemplated as circumstance may suggest or render expedient, but is intended to cover the application or implementation without departing from the spirit or scope of the claims of the present disclosure.

ADVANTAGES OF THE INVENTION
[0096] The present disclosure provides for a herbal based preparation for effective treatment and prevention of osteoporosis and related disorders.
[0097] The present disclosure provides for a method of preparation of said herbal based preparation for effective treatment and prevention of osteoporosis and related disorders.
[0098] The present disclosure provides fora herbal preparation for effective treatment and prevention of osteoporosis and related disorders which is easy to formulate and is cost-effective.
[0099] The present disclosure provides for a herbal preparation for effective treatment and prevention of osteoporosis and related disorders with minimal side-effects.
[00100] The present disclosure provides for a herbal preparation that may promote human osteoblast cell proliferation and possess anti-osteoporotic potential.
[00101] The present disclosure provides for a herbal preparation for effective treatment and prevention of osteoporosis and related disorders as a primary medication.
[00102] The present disclosure provides for a herbal preparation for effective treatment and prevention of osteoporosis and related disorders as a supplemental medication along with primary treatment.

CLAIMS:
1. A herbal preparation with anti-osteoporotic property comprising:
a pharmaceutically effective amount of Acacia niloticaherb; and
at least one pharmaceutically acceptable excipient.
2. The herbal preparation as claimed in claim 1, wherein said Acacia nilotica herb comprises at least a part of the herb,whole herb, an extract of the herb or a combination thereof.
3. The herbal preparation as claimed in claim 1, wherein said Acacia nilotica herb is present in the form of an extract in an amount ranging from about 10% to 30% by weight of the herbal preparation.
4. The herbal composition as claimed in claim 1, wherein said Acacia nilotica herb comprises ofat least one of a rhizome, a root, a bud, a tender shoot, a leaf, a stem, a fruit, a bark, a flower, a seed,ora combination thereof.
5. The herbal preparation as claimed in claim 1, wherein said herbal preparation further comprises of herb Pergularia daemia in the form of an extract in an amount rangingfrom about2% to 10%by weight of the herbal preparation.
6. The herbal preparation as claimed in claim 5, wherein said Pergularia daemia herb comprises of at least a part of the herb,whole herb, an extract of the herb ora combination thereof.
7. The herbal composition as claimed in claim 5, wherein Pergularia daemiaherb comprises ofat least one of a rhizome, a root, a bud, a tender shoot, a leaf, a stem, a fruit, a bark, a flower, a seed,ora combination thereof.
8. The herbal preparation as claimed in claim 1, wherein said at least one pharmaceutically acceptable excipient is selected from a group consisting of a sweetening agent, a buffering agent, apreservative, a bulking agent, a binder, a disintegrant, a chelating agent, a glidant, aflavouring agent, a colouring agent, a sweetener, a tonicity agent, a carrier, a solvent, a co-solvent, or a combination thereof.
9. The herbal preparation as claimed in claim 1, wherein said at least one pharmaceutically acceptable excipientisselected from agroup consisting of a whey protein, a soy protein, a casein protein,and a plant-based protein.
10. The herbal preparation as claimed in claim 1, wherein said herbal preparation is formulated in a form selected from agroup consisting of a dry powder, a tablet, acapsule, a bolus ,and a liquid.
11. A process of preparing a herbal preparation with anti-osteoporotic property, comprises the steps of:
- collecting parts of herb Acacia nilotica, preferably seeds of Acacia nilotica;
- washing the parts of herb Acacia niloticaand drying it until removal of moisture;
- grinding the dried parts of herb Acacia niloticaand sieving it to obtain a herbal powder with particle size of = 250 µm;
- extracting the herbal powder with a solvent to obtain a 10% to 30% herbal extract;
- concentrating the herbal extract to obtain a concentrate;
- formulating the concentrate in a suitable dosage form to obtain final herbal preparation.
12. A process of preparing a herbal preparation with anti-osteoporotic property, comprises the steps of:
- collecting parts of herb Acacia nilotica, preferably seeds of Acacia nilotica;
- washing the parts of herb Acacia niloticaand drying it until removal of moisture;
- grinding the dried parts of herb and sieving it to obtain a powder with particle size of = 250 µm;
- extracting the powder with a solvent to obtain a 10% to 30% Acacia niloticaextract;
- collecting part of herb Pergularia daemia, preferably leaves of Pergularia daemia, washing it, drying it until moisture removal and grinding it to obtain a fine powder with particle size of = 250 µm followed by extracting fine powder to obtain 2% to 10% extract;
- mixing the Acacia nilotica extract and thePergularia daemiaextract through a blender to obtain a poly herbal extract;
- concentrating the poly herbal extract to obtain a concentrate;
- formulating the concentrate in a suitable dosage form to obtain final herbal preparation.
13. The process as claimed in claim 11 and 12, wherein the solvent used for extraction is water, alcohol, combination of water and alcohol in a ratio of 1:1 or any combination thereof.
14. The process as claimed in claim 11 and 12, wherein extracting step is performed at a temperature ranging from about 30 °C to 60 °C.
15. The process as claimed in claim 11 and 12, wherein concentrating step is performed at a reduced pressure and at a temperature ranging 40 °C to 60 °C.