HK1155675B - Compositions and methods for maintaining bone health or reducing bone loss - Google Patents

Description

Compositions and methods for maintaining bone health or reducing bone loss

Technical Field

The present invention relates to the use of a water soluble extract of rose hip for maintaining or improving bone health or bone strength and for treating or preventing bone disorders characterized by weakened or fragile bones. The present invention also relates to compositions comprising a water-soluble extract of rose hip for use in maintaining or improving bone health or bone strength and treating or preventing bone disorders characterized by weakened or fragile bones.

Background

Bone disorders are becoming more prevalent as the world population ages. These conditions, such as osteopenia and osteoporosis, are costly both financially and in terms of the loss of quality of life for the patient caused by the disease.

Bone comprises an extracellular protein matrix (osteoid) in which bone cells (osteocytes) are dispersed and a mineral component consisting of calcium salts and other minerals is located within the extracellular matrix. Bone undergoes remodeling by a resorption process, in which case the bone is degraded by osteoclasts and then replaced (reformed) by osteoblasts. Remodeling occurs to regulate calcium homeostasis, repair bone damaged by daily stress, and shape bone when growth or mechanical stress patterns change.

Osteoclasts degrade bone in a specific area and then undergo apoptosis. Osteoblasts remodel new bone and mediate its remineralization. During remineralization, some osteoblasts are encapsulated within calcified matter and then become osteoblasts.

Once the peak bone content has been reached in early adulthood, the resorption process is almost completely accompanied by reformation. However, as people age, the effective regulation of the remodeling system becomes weaker. With the onset of menopause in women, the two processes may become desynchronized, with resorption predominating. In addition, bone mineral density loss, bone microstructural destruction, and other changes that result in increased fracture risk with aging. The process of bone loss is often gradual and no obvious symptoms appear until the disease has progressed properly.

It is therefore important to maintain bone mass and density over time so that the bone does not become brittle with aging.

Osteopenia is a disease state in which bone mineral density is lower than normal and has been considered by some as a precursor to osteoporosis. However, not every person diagnosed with osteopenia will develop osteoporosis. Osteopenia is defined as a bone mineral density T-score as measured by dual energy X-ray absorption instrumentation (DXA) that is 1-2.5 standard deviations below the peak bone content (mean for 20 year old healthy women).

Osteoporosis is characterized by gradual thinning and weakening of bone, a condition that can result in weakening of the bone and increased risk of fracture without treatment. Osteoporosis is defined as bone mineral density below the peak bone content (average for 20 year old healthy women) by 2.5 standard deviations as measured by dual energy X-ray absorptiometry.

The pathology of osteopenia and osteoporosis is an imbalance between the process of bone degradation (resorption) and bone building (reformation) by osteoclasts and osteoblasts. In healthy individuals, the process of bone resorption and bone reformation is almost completely balanced. If this balance is disrupted for various reasons, i.e. menopause, drugs, etc., eventually bone degradation exceeds bone building and may progress to osteopenia or more severe osteoporosis.

Women are generally at higher risk of developing osteopenia and osteoporosis because of the reduced estrogen production after the climacteric period. However, elderly men and people suffering from certain hormonal disorders or long-term users of certain drugs are also prone to osteopenia and osteoporosis.

The treatment of bone disorders, like osteoporosis, is often unsatisfactory, resulting in patients suffering from side effects from the prescribed treatment methods for treating bone disorders.

Osteopenia is not often diagnosed and, if diagnosed, is not usually medically treated due to the cost and length of treatment time. Because osteopenic patients are often younger than people diagnosed with osteoporosis, they need to be treated for years. The cost/benefit ratio of such long-term treatment is unknown, and therefore osteopenia is generally not treated medically.

There are currently a number of drugs used for the prescribed treatment of osteoporosis. Bisphosphonates are often prescribed in cases where patients are identified as having osteoporosis. However, oral administration of the diphosphate is poorly absorbed and must be taken on an empty stomach. In addition, they are poorly tolerated in some people and are associated with esophagitis.

Other prescription drugs for osteoporosis such as teriparatide and strontium ranelate are also not well tolerated by some people. Teriparatide must be administered by injection and is not suitable for use in younger patients, who have previously been treated with radiation or who have suffered from Paget's disease. Strontium ranelate has fewer side effects than bisphosphonates but is associated with an increased risk of venous thromboembolism. In addition, strontium ranelate is taken up into the bone matrix to replace calcium, which results in a disproportionate increase in bone mineral density as measured by dual energy X-ray absorptometer scanning.

There is therefore a need to provide a treatment for maintaining or improving bone health which overcomes or ameliorates at least one of the problems associated with known treatments, or which at least provides the public with a useful choice.

Summary of The Invention

Thus, in a first aspect, the present invention relates to the use of a water-soluble extract of rose hip for the preparation of a composition for the treatment or prevention of bone disease states characterized by weakened or fragile bone.

In another aspect, the present invention relates to compositions comprising a water-soluble extract of rose hip for use in the treatment or prevention of bone disease states characterized by weakened or fragile bones.

In one embodiment, this composition further comprises one or more agents selected from the group consisting of calcium, magnesium, zinc, vitamin D, vitamin K, folic acid or folate, vitamin B6, and vitamin B12.

In another aspect, the present invention relates to a method for treating or preventing a bone condition characterized by weakened or fragile bone, the method comprising administering to an individual in need thereof a composition comprising an effective amount of a water-soluble extract of rosehip.

In another aspect, the present invention relates to a method for treating or preventing osteoporosis, the method comprising administering to an individual in need thereof a composition comprising an effective amount of a water-soluble extract of rosehip.

In another aspect, the present invention relates to a method for treating or preventing osteopenia, the method comprising administering to an individual in need thereof a composition comprising an effective amount of a water-soluble extract of rosehip.

In one embodiment, the subject is in need of maintained or increased bone formation, maintained or increased bone mineral density, maintained or increased bone content (including peak bone content), bone regeneration during fracture healing, reduced bone resorption, reduced bone loss, or maintained or increased bone strength.

The following embodiments may relate to any of the above aspects.

In one embodiment, the composition does not include any of the following: blueberry, blackberry, elderberry, cranberry, rosemary, clove, citrus medica, nettle root, artichoke, ganoderma, olive extract, green tea extract, grape seed extract, resveratrol, viniferin, aframomum melegueta, boswellia serrata extract, boswellia forte, ipriflavone, tocotrienols, evening primrose oil, INM-176, borage oil, krill oil, at least one type of xanthophyll (e.g., astaxanthin), green coffee extract, or ferulic acid.

In one embodiment, the disease state to be treated is osteoporosis or osteopenia.

In one embodiment, the composition maintains or increases bone formation, maintains or increases bone density, maintains or increases bone content, stimulates bone regeneration during fracture healing, reduces bone resorption, reduces bone loss, or maintains or increases bone strength.

In one embodiment, the composition is a food, a candy, a milk, a dairy product, a milk powder, a reconstituted milk, a fermented milk, a drinking yogurt, a set yogurt, a beverage, a food additive, a beverage additive, a dietary supplement, a nutraceutical, a medical food, a nutraceutical (nutraceutical), or a medicament.

In one embodiment, the composition for use in the present invention further comprises one or more dairy ingredients. This dairy ingredient may be selected from the group comprising: formulated, powdered or fresh skim milk, reconstituted whole or skim milk powder, skim milk concentrate, skim milk retentate, condensed milk, ultrafiltrated milk retentate, milk protein concentrate, milk protein isolate, decalcified milk protein concentrate, low fat milk protein concentrate, casein, caseinate, milk fat, high CLA milk fat, cream, butter, anhydrous milk fat, buttermilk, cheese whey, hard milk fat fraction, soft milk fat fraction, sphingolipid fraction, milk fat globule membrane fraction, phospholipid fraction, complex lipid fraction, colostrum fraction, colostrum protein concentrate, whey, colostrum-derived immunoglobulin fraction, whey, lactoferrin, one or more lactoferrin fragments, whey protein isolate, whey protein concentrate, sweet whey, lactic acid whey, mineral acid whey, reconstituted whey powder, milk mineral, milk whey protein concentrate, milk whey, Components derived from any milk or colostrum-treated stream, components derived from retentate or permeate obtained by ultrafiltration or microfiltration of any milk or colostrum-treated stream, components derived from breakthrough or adsorption fractions obtained by chromatographic (including but not limited to ion and gel permeation chromatography) separation of any milk or colostrum-treated stream, extracts of any of these components including extracts prepared by multi-stage fractionation, differential crystallization, solvent fractionation, supercritical fractionation, near supercritical fractionation, distillation, centrifugal fractionation, fractionation with a modifier (e.g., soap or emulsifier), hydrolysates of any of these components, fractions of hydrolysates, and any combination of any two or more of these components, including combinations of hydrolyzed fractions, combinations of non-hydrolyzed fractions, and combinations of hydrolyzed and non-hydrolyzed fractions.

In one embodiment, the water-soluble extract of rose hips is administered in an amount of from about 1mg to about 2000mg/kg body weight/day, from about 100mg to about 1000mg/kg body weight/day, from about 50mg to about 500mg/kg body weight/day, or from about 0.05mg to about 300mg/kg body weight/day.

In one embodiment, the calcium is calcium or a calcium salt. In one embodiment, the magnesium is magnesium or a magnesium salt. In one embodiment, the zinc is zinc or a zinc salt. In one embodiment, vitamin D is vitamin D or a vitamin D derivative (including but not limited to vitamin D1[ photosterol (lamisterol) ], vitamin D2[ ergocalciferol ], vitamin D3[ cholecalciferol, 1, 25-dihydroxycholecalciferol ], vitamin D4[ dihydrotachysterol ], vitamin D5[ 7-dehydrositosterol ], or a vitamin D analog.

Salts useful herein include, but are not limited to, ammonium (NH)₄ ⁺) Boron, calcium, copper, iron (ferrous, Fe)²⁺And of trivalent iron, Fe³⁺) Magnesium, manganese, phosphorus, potassium, pyridine (C)₅H₅NH⁺) Quaternary ammonium (NR)₄ ⁺) Silicon, sodium, strontium and zinc salts or combinations thereof.

In one embodiment, the compositions useful herein further comprise a pharmaceutically acceptable carrier. In another embodiment, the composition is or is prepared as a food, beverage, food additive, beverage additive, dietary supplement, nutritional product, medical food, nutraceutical (nutraceutical), pharmaceutical, enteral or parenteral feeding product, or meal replacement. In one embodiment, this composition is in the form of a tablet, caplet, pill, hard or soft capsule, or lozenge. In one embodiment, such a composition is in the form of a cachet, an optional powder, granules, a suspension, an elixir, a liquid, or any other form that can be added to a food or beverage, including, for example, water, milk, or fruit juice. In one embodiment, such a composition further comprises one or more ingredients (such as antioxidants) that prevent or reduce degradation of the composition during storage or after administration. These compositions may include any edible consumer product capable of carrying a water-soluble extract of rose hips. Examples of suitable edible consumer products include aqueous products, baked goods, confectionery products including chocolate, gels, ice creams, reconstituted fruit products, snack bars, food bars, muesli bars (mueslibars), spreads, sauces, dairy products including yogurt and cheese, beverages including dairy and non-dairy based beverages, milk powders, sports supplements including dairy and non-dairy based sports supplements, fruit juices, food additives such as protein breads, and dietary supplement products including daily supplement tablets. Suitable nutraceutical compositions useful herein may be provided in similar forms.

In one embodiment, the composition comprises, or the method comprises administering, a formulation such as rose hip, calcium, magnesium, zinc, vitamin D, vitamin K, folic acid or folate, vitamin B6, vitamin B12, a dairy ingredient or pharmaceutical agent. In one embodiment, the compositions useful herein comprise, consist essentially of, or consist of at least about 0.01%, 0.02%, 0.05%, 0.07%, 0.1%, 0.2%, 0.5%, 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99%, 99.5%, 99.8%, or 99.9% by weight of one or more of these formulations, and useful ranges may be selected from any of these values (e.g., from about 0.1% to about 50%, from about 0.2% to about 50%, from about 0.5% to about 50%, from about 1% to about 50%, from about 5% to about 50%, from about 10% to about 50%, from about 15% to about 50%, from about 20% to about 50%, from about 25% to about 50%, from about 30% to about 50%, from about 35% to about 50%, from about 40% to about 50%, from about 45% to about 50%, about 0.1% to about 60%, about 0.2% to about 60%, about 0.5% to about 60%, about 1% to about 60%, about 5% to about 60%, about 10% to about 60%, about 15% to about 60%, about 20% to about 60%, about 25% to about 60%, about 30% to about 60%, about 35% to about 60%, about 40% to about 60%, about 45% to about 60%, about 0.1% to about 70%, about 0.2% to about 70%, about 0.5% to about 70%, about 1% to about 70%, about 0.5% to about 70%, about 10% to about 70%, about 15% to about 70%, about 20% to about 70%, about 25% to about 70%, about 30% to about 70%, about 5% to about 70%, about 40% to about 70%, about 45% to about 70%, about 0.1% to about 80%, about 0.2% to about 80%, about 0.5% to about 80%, about 1% to about 80%, about 80% to about 80%, about 10% to about 80%, about 20% to about 80%, about 25% to about 80%, about 30% to about 80%, about 35% to about 80%, about 40% to about 80%, about 45% to about 80%, about 0.1% to about 90%, about 0.2% to about 90%, about 0.5% to about 90%, about 1% to about 90%, about 5% to about 90%, about 10% to about 90%, about 15% to about 90%, about 20% to about 90%, about 25% to about 90%, about 30% to about 90%, about 35% to about 90%, about 40% to about 90%, about 45% to about 90%, about 0.1% to about 99%, about 0.2% to about 99%, about 0.5% to about 99%, about 1% to about 99%, about 5% to about 99%, about 10% to about 99%, about 15% to about 99%, about 20% to about 99%, about 25% to about 99%, about 30% to about 99%, about 35% to about 99%, about 40% to about 99%, and about 45% to about 99%).

The compositions useful herein are useful for treating or preventing bone disease states characterized by weakened or fragile bones. In one embodiment, the disease state to be treated is one in which it is desirable to maintain or increase bone content or bone strength or both. In one embodiment, the disease state is one in which reduced bone loss is desired. In one embodiment, the disease state is a disease state requiring bone cell proliferation, bone cell differentiation, bone cell mediated mineralization or a combination of any two or more of bone cell proliferation, bone cell differentiation and bone cell mediated mineralization. In one embodiment, the disease state is a disease state requiring any two or more of inhibition of bone resorption, increased bone formation, decreased bone loss, or a combination of inhibition of bone resorption, increased bone formation, and decreased bone loss. In one embodiment, the disease state is a disease state in which it is desirable to improve or maintain bone density, bone content, bone strength, or bone health or to improve or maintain a combination of any two or more of bone density, bone content, bone strength, and bone health. In one embodiment, the disease state is osteoporosis. In one embodiment, the disease state is osteopenia.

In one embodiment, treating or preventing a bone disease state characterized by weakened or weakened bone comprises one or more of maintaining or increasing bone formation, maintaining or increasing bone mineral density, maintaining or increasing bone content (including peak bone content), treating or preventing osteoporosis, treating or preventing osteopenia, stimulating bone regeneration during fracture healing, reducing bone resorption, or increasing bone mass (e.g., as measured by fracture stress or bone strength).

The compositions useful herein are useful for administering to a subject in need thereof to treat a bone condition. In one embodiment, the subject is in need of increased bone content. In one embodiment, the subject is in need of treatment for a disease state that requires a reduction in bone loss. In one embodiment, the individual is in need of treatment for a disease state requiring any two or more combinations of bone cell proliferation, bone cell differentiation or bone cell mediated mineralization or bone cell proliferation, bone cell differentiation and bone cell mediated mineralization. In one embodiment, the subject is in need of treatment for a disease state requiring inhibition of bone resorption, increased bone formation, or decreased bone loss, or a combination of inhibition of bone resorption, increased bone formation, and decreased bone loss. In one embodiment, the subject is in need of treatment for a disease state that requires an improvement or maintenance of bone density, an improvement or maintenance of bone content, an improvement or maintenance of bone strength, or an improvement or maintenance of bone health, or a combination of any two or more of these effects.

In one embodiment, the subject is in need of reducing bone resorption or reducing osteoclast formation or both. Thus, in one embodiment, the invention relates to ameliorating, treating or preventing a disease state associated with net bone resorption or increased osteoclastogenesis.

In another embodiment, the individual is in need of increased bone formation or increased osteoblast proliferation, increased osteoblast differentiation, increased mineralization or a combination thereof. Thus, in this embodiment, the invention relates to the amelioration, treatment or prevention of a disease state associated with poor bone formation or reduced osteoblast proliferation, reduced differentiation and reduced mineralization or a combination thereof.

In one embodiment, treating or preventing a bone condition characterized by weakened or fragile bone comprises reducing bone loss, particularly bone loss associated with an increased age of the subject.

In one embodiment, the composition comprises, consists essentially of, or consists of about 0.1%, 0.5%, 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99%, 99.5%, 99.8%, or 99.9% by weight of one or more dairy ingredients selected from fresh whole milk, formulated whole milk or whole milk powder or milk derivatives, and useful ranges may be selected from any of these above values (e.g., about 0.1% to about 50%, about 0.2% to about 50%, about 0.5% to about 50%, about 1% to about 50%, about 5% to about 50%, about 10% to about 50%, about 15% to about 50%, about 20% to about 50%, about 25% to about 50%, about 30% to about 50%, about 35% to about 50%, from about 40% to about 50% and from about 45% to about 50%). The milk derivative is preferably selected from the group consisting of formulated, powdered or fresh skim milk, reconstituted whole or skim milk powder, skim milk concentrate, skim milk retentate, condensed milk, ultrafiltrated milk retentate, Milk Protein Concentrate (MPC), Milk Protein Isolate (MPI), decalcified Milk Protein Concentrate (MPC), low fat Milk Protein Concentrate (MPC), casein, caseinate, milk fat, high CLA milk fat, cream, butter, Anhydrous Milk Fat (AMF), buttermilk, cheese whey, hard milk fat fraction, soft milk fat fraction, sphingolipid fraction, milk fat globule membrane fraction, phospholipid fraction, complex lipid fraction, colostrum fraction, Colostrum Protein Concentrate (CPC), colostrum whey, colostrum-derived immunoglobulin fraction, whey, lactoferrin, one or more lactoferrin fragments, Whey Protein Isolate (WPI), Whey Protein Concentrate (WPC), Sweet whey, lactic whey, mineral acid whey, reconstituted whey powder, milk minerals, components derived from any milk or colostrum treatment stream, components derived from retentate or permeate obtained by ultrafiltration or microfiltration of any milk or colostrum treatment stream, components derived from breakthrough or adsorbed fractions obtained by chromatographic (including but not limited to ion and gel permeation chromatography) separation of any milk or colostrum treatment stream, extracts of any of these milk derivatives including extracts prepared by multistage fractionation, differential crystallization, solvent fractionation, supercritical fractionation, near supercritical fractionation, distillation, centrifugal fractionation, fractionation with fractionation of a modifier (such as soap or emulsifier), hydrolysates of any of these derivatives, fractions of hydrolysates and any combination of any two or more of these derivatives including combinations of hydrolyzed fractions, compositions comprising a mixture of hydrolyzed fractions, and compositions comprising a mixture of, Combinations of non-hydrolyzed moieties and combinations of hydrolyzed and non-hydrolyzed moieties.

Reference to a series of numbers disclosed herein (e.g., 1 to 10) is intended to also include all rational numbers within that range (e.g., 1, 1.1, 2, 3, 3.9, 4, 5, 6, 6.5, 7, 8, 9, and 10) and also include any range within that range (e.g., 2 to 8, 1.5 to 5.5, and 3.1 to 4.7). Accordingly, all subranges of all ranges disclosed herein are explicitly disclosed herein. These are only examples of what is specifically intended to be protected and all possible combinations of numerical values between the minimum and maximum values recited are to be considered to be expressly stated in this patent application in a similar manner.

In this specification, if reference is made to a patent specification, other external data, or other sources of information, this is generally for the purpose of providing a context for discussing the features of the invention. Unless specifically stated otherwise, reference to such external material is not to be construed as an admission that such document, or such source of information, in any jurisdiction, is prior art, forms part of the common general knowledge in the art. And (4) dividing.

The invention may also be said broadly to consist in the parts, elements and features referred to or indicated in the specification of the application, individually or collectively, in any or all combinations of two or more of the above parts, elements or features, and where specific integers are mentioned herein which have known equivalents in the art to which the invention relates, such known equivalents are deemed to be incorporated herein as if individually set forth.

Brief Description of Drawings

Fig. 1 is a graph showing the effect of rosehips on osteoblast differentiation (p ═ 0.0093) as compared to the control group, as indicated by the production of alkaline phosphatase.

Fig. 2 is a graph showing the effect of rose hip extract on osteoclastogenesis. Rose hip dose-dependent inhibition of osteoclastogenesis. Results compared to control p < 0.05.

Fig. 3 is a graph showing the fracture stress results of a right femoral three-point bending test. One group of rats underwent Sham surgery (Sham group (Sham); n-15) and were fed a control diet for 28 weeks. The other group was ovariectomized and fed either a control diet (OVX group; n ═ 15) or a treatment diet containing rose hip extract (Rosehip group; n ═ 15) for 28 weeks. Results are shown as mean ± SEM. The results show significant differences (p ═ 0.0105) between the sham and Ovariectomized (OVX) groups, whereas there were no significant differences between the sham and rosehip (p ═ 0.8450) or the Ovariectomized (OVX) and rosehip (p ═ 0.5754).

Fig. 4 is a graph showing the maximum load (maximum load) of the right femur. The maximum load is defined as the maximum force that the femur can withstand during the three-point bending test. 45 rats were fed the control diet for 3 weeks. After 3 weeks, one group of rats underwent sham surgery (sham group; n-15) and continued to be fed the control diet. The other 30 rats were ovariectomized. One group of ovariectomized rats was fed a control diet (OVX; n-15) and the other group was fed a diet supplemented with rosehips (rosehip group; n-15). Results are shown as mean ± SEM. The results show significant differences between the ovariectomized group (OVX) and the rosehip group (p ═ 0.035).

Fig. 5 is a graph showing a breaking load (break load). The breaking load is the force that fractures the femur, typically less than the maximum force. 45 rats were fed the control diet for 3 weeks. After 3 weeks, one group of rats underwent sham surgery (sham group; n-15) and continued to be fed the control diet. The other 30 rats were ovariectomized. One group of ovariectomized rats was fed a control diet (OVX; n-15) and the other group was fed a diet supplemented with rosehips (rosehip group; n-15). Results are shown as mean ± SEM. The results show significant differences between the ovariectomized group (OVX) and the rosehip group (p 0.030).

Fig. 6 is a graph showing the energy expended to fracture the right femur of a rat. 45 rats were fed the control diet for 3 weeks. After 3 weeks, one group of rats underwent sham surgery (sham group; n-15) and continued to be fed the control diet. The other 30 rats were ovariectomized. One group of ovariectomized rats was fed a control diet (OVX; n-15) and the other group was fed a diet supplemented with rosehips (rosehip group; n-15). Results are shown as mean ± SEM. The results show significant differences between the ovariectomized group (OVX) and the rosehip group (p 0.030).

Detailed Description

The present invention is based on the following findings: the water soluble extract of rose hip has positive effects in maintaining bone health and treating bone diseases such as osteoporosis or osteopenia.

1. Definition of

The term "comprising" as used in the specification and claims means "consisting at least in part of. When interpreting statements in this specification and claims which include that term, all features from each statement that follow that term need to be present, but other features can also be present. Related terms such as "comprise" and "comprise" are also to be interpreted in the same way.

The term "water-soluble extract of rose hips" refers to an extract produced by the following process: any food grade polar solvent such as water, acetic acid and alcohols (such as methanol, ethanol, propanol or butanol, for example) is used to extract the water soluble components from the rose hips. The solvent is preferably water.

The term "maintaining bone health" refers to maintaining the bones of an individual at a healthy density, for example, within ± 1 Standard Deviation (SD) of bone density of a 20 year old person, as defined by the World Health Organization (WHO) standards for dual energy X-ray absorptiometry (DEXA).

The term "weakened or fragile bone" refers to an individual having a bone density below healthy density, for example within ± 1SD of the bone density of a 20 year old person as defined by WHO's standard for DEXA.

An "effective amount" is an amount required to impart a therapeutic effect. Freirich et al (1966) describe dose correlations for animals and humans (in mg/square meter body surface). Body surface area can be roughly determined from the height and weight of the individual. See, for example, Scientific Tables, Geigy pharmaceuticals, Ardley, New York, 1970, 537. As recognized by one skilled in the art, effective dosages will also vary, depending on the route of administration, carrier use, species and individual genetic variation, and the like.

The term "oral administration" includes oral, buccal, enteral and intragastric administration.

The term "pharmaceutically acceptable carrier" means a carrier including, but not limited to, excipients, diluents, adjuvants or combinations thereof, which is capable of being administered to an individual as an ingredient of the composition of the present invention without diminishing the activity of such composition and which is not toxic when administered in a dosage sufficient to deliver an effective amount of the rose-hip extract. This formulation can be administered orally, nasally and topically.

An "individual" in connection with the present invention is an animal, preferably a mammal, more preferably a companion animal or a human of a mammal. Preferred companion animals include cats, dogs and horses.

The term "treatment" and its derivatives should be interpreted in their broadest possible context. This term should not be taken to mean that the individual is treated to full recovery. Thus, "treating" broadly includes ameliorating or preventing or ameliorating and preventing the onset of symptoms or the severity of a particular disease state; for example, preventing or ameliorating a reduction in bone density, preventing or reducing the risk of developing osteoporosis, preventing or reducing the risk of developing osteopenia, or preventing or ameliorating other disease symptoms. The term "treating" also broadly includes maintaining good bone health and building bone quality to prevent a disease or disorder.

2. Rose fruit

Rose hip, sometimes called rose hawthorn (rosehaw), is a fruit found on the rose plant (Rosa spp.) and contains several vitamins including high levels of vitamin C. Other vitamins found in rose hips are vitamins A, D and E. Rose hips also contain high levels of iron and some essential fatty acids and antioxidants.

Several rose species of fruit can be used to prepare the water-soluble extract of rose hips useful in the present invention. These species include, but are not limited to, canine rose (Rosa canina), Rosa dumali sp. boissieri, Rosa dumalia subsp. antalyensis, Rosa rugosa (Rosa villosa), Rosa pulverata, Rosa moyesii Rosa gallica, Rosa condata, Rosa rugosa, and Rosa pisiformis (Rosa pisiformis).

Possible rose-hip extraction processes are as follows: fresh rose hips are macerated or dried rose hips are powdered. The macerated or powdered rose hips are mixed with distilled water or other aqueous solution (e.g., buffer) in a ratio of 1: 5(w/w) to allow the water soluble components to be extracted from the rose hips. The aqueous solvent may be replaced one or more times, preferably three times, and each amount of solvent is allowed to remain in contact with the plant material for several hours, preferably 12, 24 or 48 hours, preferably with continuous stirring. After filtration or centrifugation of the total extract, the aqueous solution is then frozen or spray dried to form a powder, which can be used as the aqueous extract used herein.

This extract can be further isolated by standard methods of ion exchange chromatography or membrane separation or other such separation techniques known in the art. See, for example, IonExchange Chromatography & Chromatography of using, Principles and Methods, Amazonia Biosciences, Inc. 2004(Amersham Biosciences Limited 2004), Catalogue number 11-0004-21, version AA, http:// www.amersham.com.

3. Methods of treatment or prevention

The compositions and uses of the compositions described herein are used to treat or prevent bone disease states characterized by weakened or fragile bones, such as osteoporosis or osteopenia.

In one embodiment, the disease state to be treated is a disease state requiring increased bone content. In one embodiment, the disease state is one in which reduced bone loss is desired. In one embodiment, the disease state is a disease state requiring bone cell proliferation, bone cell differentiation or bone cell mediated mineralization or a combination of two or more thereof. In one embodiment, the disease state is one in which it is desirable to inhibit bone resorption, increase bone formation, or decrease bone loss, or a combination of two or more thereof. In one embodiment, the disease state is a disease state in which it is desirable to improve or maintain bone strength, bone density, or bone mass, or a combination of two or more thereof. In one embodiment, the disease state is osteoporosis. In one embodiment, the disease state is osteopenia.

4. Compositions useful in the invention

Described herein is a composition comprising, consisting essentially of, or consisting of a water-soluble extract of rose-hips and one or more ingredients selected from the group consisting of calcium, magnesium, zinc, vitamin D, vitamin K, folic acid or folate, vitamin B6, and vitamin B12.

Also described herein is a composition consisting essentially of a water-soluble extract of rose-hips and one or more ingredients selected from the group consisting of calcium, magnesium, zinc, vitamin D, folic acid, folate and vitamin B12, wherein such composition is formulated for simultaneous, separate or sequential administration of the water-soluble extract of rose-hips and the one or more ingredients.

Also described herein is a composition comprising a water-soluble extract of rose hips and one or more dairy ingredients.

The compositions useful herein include any composition capable of carrying a water-soluble extract of rose-hips, including any consumer product and any pharmaceutical product capable of carrying a water-soluble extract of rose-hips. The composition can be formulated as a food, beverage, food additive, beverage additive, dietary supplement, nutraceutical, medical food, nutraceutical, pharmaceutical or medicament. The composition of the invention is preferably formulated as a powder, liquid, food bar, spread, sauce, ointment, tablet or capsule. Suitable food and beverages include dairy and non-dairy food and beverages. In one embodiment, the composition is a milk powder, milk beverage, yogurt powder, yogurt drink, soy milk, acidified drink, ultra high temperature sterilized milk (UHT), pasteurized milk, butter, or cheese. Those skilled in the art can prepare appropriate formulations based on the techniques and teachings of this specification.

The nutraceutical composition for use in the present invention may be a dietary supplement (e.g., a capsule, mini-pack or tablet) or a food product (e.g., milk, juice, soft drink, herbal tea pack or candy). This composition can also include other nutrients such as proteins, carbohydrates, lipids, vitamins, minerals or amino acids. This composition can be in a form suitable for oral administration, such as a tablet, hard or soft capsule, aqueous or oily suspension, or syrup; or in a form suitable for parenteral use, such as an aqueous propylene glycol solution or a buffered aqueous solution. The amount of active ingredient in a nutraceutical composition depends to a large extent on the specific needs of the individual. This number is also different, as recognized by one skilled in the art, depending on the route of administration, species, genetic/physiological disposition and possible co-usage with other bone enhancers.

The food, food additive or food supplement comprising rose hip extract for use in the present invention includes any edible consumable product capable of carrying a water soluble plant extract. Examples of suitable edible consumer products include confectionery products, reconstituted fruit products, snack bars, muesli bars, bakery products, spreads, gravies, dairy products including yoghurts and cheeses, beverages including dairy-based and non-dairy-based beverages, milk powders, sports supplements including dairy-based and non-dairy-based sports supplements, food additives such as protein chips (sprinkles), and dietary supplement products including daily supplement tablets. Suitable nutraceutical compositions useful herein may be provided in a similar manner.

Compositions useful herein may be formulated to allow for administration to an individual by any selected route, including but not limited to oral or nasal administration.

Preferably, the compositions useful herein are administered orally.

It will be appreciated that the dosage, period of administration and conventional regimen of administration of the composition may vary from individual to individual, depending upon such variables as the severity of the individual's symptoms, the type of condition being treated, the mode of administration selected and the age, sex and general health of the individual. However, for example, the inventors contemplate administration of from about 1mg/kg body weight/day to about 2000mg/kg body weight/day rose hips, preferably from about 100mg/kg body weight/day to 1000mg/kg body weight/day rose hips or from about 50mg/kg body weight/day to about 500mg/kg body weight/day rose hips. In one embodiment, the inventors contemplate administration of from about 0.05mg/kg body weight/day to about 300mg/kg body weight/day rose hips.

Preferably, the compositions useful herein are administered at least once daily. Preferably the compositions useful herein are administered 2-3 times per day. Alternatively, the compositions useful herein may be administered once a week.

Preferably, the compositions useful herein are administered as a prophylaxis prior to the onset of menopause. Optionally, the compositions useful herein are administered after the onset of menopause to maintain or increase bone health or for the treatment or prevention of a bone condition.

It will be appreciated that one skilled in the art can, without undue experimentation, determine an effective dosage regimen (including daily dosages and timing of administration) for a given disease state in light of the skill in the art and the present disclosure.

The efficacy of the compositions useful in the present invention can be assessed in vitro and in vivo. See, for example, the examples below. Briefly, the compositions can be tested in vitro for their ability to promote osteoblast development and activity or to inhibit osteoclastogenesis, limit osteoclast activity or reduce osteoclast numbers. For in vivo studies, the composition can be administered to an animal (e.g., rat) and its effect on bone tissue can then be evaluated. Based on these results, an appropriate dosage range and route of administration can be determined.

The compositions useful herein may be used alone or in combination with one or more other therapeutic agents. The therapeutic agent can be a food, beverage, food additive, beverage additive, food ingredient, beverage ingredient, dietary supplement, nutritional product, medical food, nutraceutical, pharmaceutical, or medicament. The therapeutic agent is preferably effective to promote osteoblast development and activity, inhibit osteoclastogenesis, reduce osteoclast number, or limit osteoclast activity.

When used in combination with other therapeutic agents, the compositions for use herein and the other therapeutic agents may be administered simultaneously or sequentially.

Simultaneous administration includes administration of a single dosage form containing all the components or administration of separate dosage forms at substantially the same time. Continuous administration includes administration according to different schedules, and thus it is preferred that there is overlap in the time periods during which the compositions and other therapeutic agents useful herein are provided.

In one embodiment, the compositions useful herein include the following or are administered simultaneously or sequentially with the following: milk ingredients such as whey protein, whey protein fractions (including acidic or basic whey protein fractions or combinations thereof), glycomacropeptide, lactoferrin or functional lactoferrin variants or fragments, vitamin D derivatives, vitamin D analogs or calcium and its salts, or combinations thereof. Useful compositions comprising a milk component include, for example, a food, a beverage, a food additive, a beverage additive, a dietary supplement, a nutraceutical, a medical food, or a nutraceutical. Milk fractions enriched in these components may also be used.

It is to be understood that additional therapeutic agents as described above (both food-based therapeutic agents and pharmaceutical agents) may also be used in the methods of the invention, wherein they may be administered separately, simultaneously or sequentially with the compositions useful herein.

The compositions useful herein may also include other bone health agents such as calcium, fluoride, magnesium, zinc, vitamin a, folate or folic acid, or vitamin B12, vitamin B6, vitamin C, vitamin D derivatives (including but not limited to vitamin D (including vitamin D1[ photosterol ], vitamin D2[ ergocalciferol ], vitamin D3[ cholecalciferol, 1, 25-dihydroxycholecalciferol ], vitamin D4[ dihydrotachysterol ] and vitamin D5[ 7-dehydrositosterol ], and vitamin D analogs), vitamin E derivatives, vitamin E analogs, vitamin K derivatives, vitamin K analogs, vitamin K2, whey protein moieties (including acidic or basic whey protein moieties or combinations thereof), Glycomacropeptide, lactoferrin, functional lactoferrin variants, functional lactoferrin fragments, soy isoflavones, phytoestrogens, prebiotics or probiotics, or a combination of any two or more of these ingredients that can affect mineral reabsorption, polyunsaturated fatty acids, and combinations thereof.

The use of a pharmaceutical composition comprising rose-hips or components thereof is also within the scope of the present invention. This pharmaceutical composition can be used for the prevention and treatment of bone related disorders as described above. This pharmaceutical composition can also include an effective amount of other bone enhancers. Pharmaceutically acceptable carriers include solvents, dispersion media, coatings, antibacterial and antifungal agents, and isotonic and absorption delaying agents.

The pharmaceutical compositions useful in the present invention may be formulated with an appropriate pharmaceutically acceptable carrier selected with regard to the intended route of administration and standard pharmaceutical practice. For example, the compositions of the present invention can be administered orally as a powder, lipid, tablet, effervescent or capsule. Suitable formulations may contain additional agents as required, including emulsifiers, antioxidants, flavoring or coloring agents, and may be adapted for immediate release, delayed release, modified release, sustained release, pulsed release or sustained release.

In a preferred embodiment, the composition used in the present invention is formulated for ingestion.

The capsule can contain any standard pharmaceutically acceptable material such as gelatin or cellulose. Tablets are formulated by compressing the active ingredient in admixture with a solid carrier and a lubricant according to conventional procedures. Examples of solid carriers include, but are not limited to, starch and sugar bentonite. The active ingredient can also be administered in the form of hard shell tablets or capsules containing a binder such as lactose or mannitol, conventional fillers and tableting agents.

The foaming sheet can be formulated using any conventional procedure and may comprise any suitable pharmaceutically acceptable foaming material. Examples of foaming substances include, but are not limited to, citric acid, malic acid, sodium bicarbonate, potassium bicarbonate, sodium carbonate, and potassium carbonate.

The compositions of the present invention can be formulated into dosage forms for different routes of administration using conventional methods. For example, it can be formulated into capsules, gel packs or tablets for oral administration.

Suitable pharmaceutical compositions may be formulated with suitable pharmaceutically acceptable excipients, diluents or carriers selected with regard to the intended dosage form and standard pharmaceutical manufacturing practice. Dosage forms useful herein can be administered orally as a powder, liquid, tablet or capsule. Suitable dosage forms may contain additional agents as needed, including emulsifiers, antioxidants, flavoring agents, or coloring agents. Dosage forms useful herein may be adapted for immediate release, delayed release, modified release, sustained release, pulsed release, or sustained release of the active ingredient.

Compositions comprising rose hips may be used for the treatment or prevention of skeletal disorders. Examples of such disorders include, but are not limited to, osteoporosis, hepatic osteodystrophy, osteomalacia, rickets, cystic fibrosing osteomyelitis, renal osteodystrophy, osteopetrosis, osteopenia, bone fibrogenesis deficiency, secondary hyperparathyroidism, hypoparathyroidism, hyperparathyroidism, chronic kidney disease, sarcoidosis, glucocorticoid-induced osteoporosis, idiopathic hypercalcemia, paget's disease, and osteogenesis imperfecta.

The rose-hip extract may be used alone or in combination with one or more other therapeutic agents, such as nutraceuticals, pharmaceuticals or medical foods. When used in combination with other therapeutic agents, the administration of the two formulations may be separate, simultaneous or sequential. Simultaneous administration includes administration of a single dosage form containing both formulations as well as administration of both formulations in separate dosage forms at substantially the same time. Continuous administration includes administration according to different schedules, such that there is preferably overlap in the time periods during which the two formulations are provided. Suitable formulations which can be administered with the compositions of the present invention include bone growth agents or bone disease treatment agents and other suitable formulations known in the art. Such formulations are preferably administered parenterally, preferably by intravenous, subcutaneous, intramuscular, intraperitoneal, intramedullary, epidural, intradermal, transdermal (topical), transmucosal, intraarticular and intrapleural administration, and oral, inhalation and rectal administration.

Suitable formulations capable of administration with the compositions useful herein include α v β 3 integrin receptor antagonists, antiestrogens or SERMs (selective estrogen receptor modulators) including, but not limited to, tamoxifen (tamoxifen), raloxifene (raloxifene), lasofoxifene (lasofoxifene), toremifene (toremifene), azoxifene (azoxifene), clomiphene (clomiphene), droloxifene (droloxifene), idoxifene (idoxifene), levomeloxifene (lerfeloxifene), zuclomiphene (zuclomiphene), enclomiphene (enclomiphene), nafoxidine (nafoxiden) and salts thereof, antiresorptive agents, bisphosphonates including, but not limited to, alendronate, clodronate (clodronate), telronate (irronethonate), irronate (dronate), irronedronate (irinoteconate), dronate (dronate), voronofenanate (dronate), dronate (dronate), dronate (dron, Palindronate (pilidronate), risedronate (risedronate), tiludronate (tiludronate), zoledronate (zoledronate) and pharmaceutically acceptable salts thereof, calcium receptor antagonists, calcium supplements, cathepsin K inhibitors, Dual Action Binders (DABAs) including, but not limited to, strontium ranelate, estrogens and estrogen derivatives including, but not limited to, 17 β -estradiol, estrone, conjugated estrogens, equine estrogens and 17 β -ethinyl estradiol, flavonoids, folic acid, folates, vitamin B12, osteogenesis promoters, osteoprotegerin, progesterone and progesterone derivatives including, but not limited to, norethindrone and medroxyprogesterone acetate, vacuolar atpase inhibitors, Vascular Endothelial Growth Factor (VEGF) antagonists, thiazolidinediones, calcitonin, protein kinase inhibitors, parathyroid hormone (PTH) analogs, Recombinant parathyroid hormone, growth hormone secretagogues, growth hormone releasing hormone, homocysteinemia insulin-like growth factor inhibitors, Bone Morphogenetic Protein (BMP), BMP antagonist inhibitors, prostaglandin derivatives, fibroblast growth factor, statins including but not limited to lovastatin, simvastatin, pravastatin, fluvastatin, atorvastatin, cerivastatin, rosuvastatin and pravastatin, calcium, fluoride, magnesium, zinc, calcium salt, fluoride salt, magnesium salt, zinc salt, vitamin a derivatives, vitamin a analogs, vitamin B6, vitamin C derivatives, vitamin C analogs, vitamin D derivatives including but not limited to vitamin D (which includes vitamin D1[ photosterol ], vitamin D2[ ergocalciferol ], vitamin D3[ cholecalciferol, 1, 25-dihydroxycholecalciferol ], vitamin D4[ dihydrotachysterol ] and vitamin D5[ 7-dehydrositosterol ], and vitamin D analogs), vitamin E derivatives, vitamin E analogs, vitamin K derivatives, vitamin K analogs, vitamin K2, whey protein fractions (which include acidic or basic whey protein fractions or combinations thereof), glycomacropeptide, lactoferrin, functional lactoferrin variants, functional lactoferrin fractions, milk fat, high CLA milk fat, milk fat fractions, soy isoflavones, phytoestrogens, prebiotics or probiotics, or combinations of any two or more of these agents, which may affect mineral absorption, polyunsaturated fatty acids, and combinations thereof, and other suitable agents known in the art.

Published international patent applications WO 03/082921 and WO 2007/043900 describe useful lactoferrin and lactoferrin fragments and hydrolysates and are incorporated herein by reference.

Published international patent application WO 2008/147228 describes fatty acids, fatty acid derivatives and milk fat compositions comprising high CLA milk fat and a milk fat fraction, which milk fat compositions are capable of treating or preventing bone conditions which benefit from reduced bone resorption, and which international patent application WO 2008/147228 is incorporated herein by reference.

In addition, it is contemplated that the compositions of the present invention may be formulated with additional active ingredients, which may be beneficial to an individual in a particular context. For example, therapeutic agents that target the same or different disease processes may be used.

The different aspects of the invention will now be illustrated in a non-limiting manner by the following examples.

Examples

Cell culture and assay Material

All cell culture materials were purchased from Invitrogen. The MC3T3-E1 preosteoblasts were obtained from the American Type Culture Collection (ATCC), Manassas, Va. Rosehip extract was purchased from Paninkret, Germany (Germany).

Example 1: osteoblast differentiation

MC3T3-E1/4 cells at 0.55X 10⁵Cells/ml were seeded in 48-well plates, 0.3ml per well. Each treatment in each experiment had quadruplicate wells, and the experiment was repeated three times to ensure consistency of results. The medium was α -MEM containing 10% FCS. Therapeutic agents are added 24 hours after initial plating to allow cells to adhere to the wells. After 3 days of culture, the medium was changed to α -MEM containing 10% FCS, 10mM β -glycerophosphate and 50 μ g/ml ascorbic acid, with or without the addition of a therapeutic agent. The culture medium was changed every 2-3 days. After 10 days of incubation, the medium was removed from the wells and the cells were washed with Phosphate Buffered Saline (PBS). Cells were incubated with 200. mu.l of alkaline phosphatase assay (0.05M p-nitrophenylphosphate in TBS, pH 9.5) for 1 hour at 37 ℃. Two samples (100 μ Ι) of detection reagent from each well were transferred to a 96-well plate and the absorbance was read at 405nm using an ELx808Ultra microplate reader (Bio-Tek Instruments inc., Vermont, USA). The cells were again washed with PBS, fixed with 1% formaldehyde for 15 minutes and the cell number was determined by crystal violet assay. Cells were incubated with 300 μ l of 1% crystal violet (dissolved in PBS) for 2 hours, then washed under running water for 15 minutes to remove excess dye and allowed to air dry. Mu.l/well of Triton-X100 (0.2% in double distilled water) was added to the dried plates, and the plates were then incubated at room temperature for 2 hours to dissolve the adsorbed dye. Two aliquots of 100. mu.l were removed from each well and transferred to 96-well plates. Absorbance was read at 550nm using an ELx808Ultra microplate reader (Bio-Tek Instruments Inc., Vermont, USA) and used0.2% Triton-X100 was used as a blank. The alkaline phosphatase activity on a per cell basis was determined by dividing the absorbance measurement at 405nm (measurement of p-nitrophenyl production or alkaline phosphatase activity) by the absorbance measurement at 550nm (from crystal violet assay, measurement of cell number). The generated index graph was used to represent alkaline phosphatase activity on a per cell basis. The results are shown in FIG. 1. Rose hip administration increased differentiation by 17% compared to control.

Example 2: osteoclast formation

Osteoclasts can be generated from the murine macrophage RAW264.7 cell line. RAW264.7 cells were seeded in 24-well plates containing coverslips and treated with murine RANK-L. The effect of compounds on osteoclastogenesis was examined by adding different concentrations of the compounds to the cell culture medium containing RANK-L. These cells were incubated for 5 days with medium change on day 3. Coverslips were then fixed and TRAP stained, followed by counterstaining with hematoxylin. Large multinucleated osteoclasts were quantified. Osteoclasts appear as large multinucleated cells stained with a purple color and can form even larger giant cells. The effect of the preparation on osteoclastogenesis was quantified by taking micrographs of three random areas on each coverslip and then counting the number of TRAP (+) cells with more than 3 nuclei and expressing this as the "number of TRAP (+) cells with more than 3 nuclei per unit area". The TRAP stain was also dissolved and measured colorimetrically. These results are shown as a graph. In the first step of in vitro assay confirmation, a cell density titration is performed.

To determine the optimal cell density of the seeded cells, 2X10 in cell culture medium containing 35ng/ml RANK-L³-6x10⁴Cells/ml seeded cells. From here 1.5x10 was chosen⁴The density of cells/ml was used for further plating. RANK-L titration (1ng/ml-35ng/ml) was then performed using the selected cell density. From these two results, 1.5x10 was chosen⁴Cell seeding density of cells/ml and RANK-L concentration of 15ng/ml were used as optimal conditions under which osteoclasts were observed in all subsequent in vitro assaysAnd (4) cell generation. The results are shown in fig. 2. Administration of rosehip resulted in a 29% reduction in osteoclast number compared to control. At 10ug/ml, no osteoclast-like cells were observed compared to the control. Cell viability was not affected.

Example 3: bone resorption

This study was designed to assess whether diets supplemented with rosehip in the OVX rat model could affect bone quality.

Animal(s) production

44 female rats of 5.5 months of age were fed on a casein-based diet for 2 weeks (weeks 2 to 0). At week 2, rats were subjected to an in vivo DEXA scan under anesthesia. At week 0, rats or sham surgery (sham group; n-15) or ovariectomy (n-30). Sham operated rats were anesthetized by isoflurane inhalation and an incision was made but the ovaries were kept intact. Ovaries were removed from the ovariectomized animals. Ovariectomized rats were randomly assigned to a control group (OVX) and fed a casein control diet, or to a treatment group (rosehip group) and fed 0.5% w/w rosehip extract on the control diet for 28 weeks.

Confirmation of successful ovariectomy was determined by measuring uterine weight. The two OVX (0.14 g; p < 0.0001) and rosehip (0.13 g; p < 0.0001) groups had significantly lower uterine weights than the sham (0.86g) group.

At week 28, rats were fasted overnight and then euthanized by exsanguination (cardiac puncture under anesthesia). Following cardiac puncture, animals were exposed to 100% CO₂. The bodies were dissected and the various bones were collected and fixed for further analysis.

Diet

The balanced semi-synthetic diet fed to animals was made up of 15% (w/w) casein (as caseinate), 5% (w/w) cellulose, 5% (w/w) corn oil, 0.5% (w/w) calcium, 62% (w/w) starch and added vitamins, minerals and amino acids to meet the requirements of AIN93M (National Research Council, 1995).

Mechanical properties of bone

The right femur was scraped clean of the meat attached to it and stored in phosphate buffer at-20 ℃. Prior to biomechanical testing, the bone was thawed. The length of the femur was measured using an electronic caliper. The midpoint was marked with a waterproof pen and the width and thickness of the femur was recorded at the midpoint. The femur was then kept at 23 ℃ to be at room temperature before and during testing. The femur was placed in a test fixture constructed for a three-point bending test. The distance between the support rods is a fixed length of 12 mm. The maximum load (N), stiffness (N mm) were measured using a Shimadzu Ezi-test texture analyzer (Shimadzu Ezi-test texture analyzer, Kyoto, Japan) with a constant deformation rate of 50mm min-1^-2) And energy (J). The results are shown in fig. 3. The amount of energy required to fracture the right femur in the three-point bending test was 118N/mm for the sham group, OVX and rosehip groups, respectively²、96N/mm²And 101N/mm². The sham group and OVX were significantly different (p ═ 0.0105). The rosehip group did not differ significantly from the sham or OVX group, but was intermediate in its effect of making the bone more elastic when fractured.

Example 4: bone resorption and mechanical Properties

This study was designed to evaluate whether diet supplemented with rosehip in OVX rat model could inhibit bone resorption or affect bone biochemical and mechanical properties.

Animal(s) production

45 female Sprague Dawley rats, about 5 months of age, were housed individually in shoe-box-like cages and were free to drink Milli-Q water. They were acclimated to this diet and their environment over a 3 week period, all fed a nutritionally adequate casein-based diet consisting of 14% (w/w) caseinate, 5% (w/w) cellulose, 5% (w/w) vitamins, 5% (w/w) minerals, 1.3% (w/w) calcium carbonate, 4% (w/w) corn oil, 5.7% (w/w) sucrose and 60% (w/w) wheat starch.

None of the diets were supplemented with vitamin C and the vitamin mix did not contain any vitamin C. The rose hips added to their diets provided about 0.08% (w/w) vitamin C.

After 3 weeks, 30 animals were ovariectomized and 15 additional sham surgeries were performed to simulate ovariectomy without actual removal of the ovaries. Thereafter, randomly selected 15 ovariectomized rats and 15 sham operated animals were fed a standard casein based diet, while another 15 ovariectomized animals received a basal diet supplemented with 2% rose hip extract. After 24 weeks, animals were euthanized as described in example 3, their right femurs were removed and prepared for bone biomechanical testing using the three-point fracture test protocol as described in example 3. Results were compared by one-way analysis of variance (1-way ANOVA).

Biochemical analysis

Biomechanical analysis was performed on OVX, sham surgery and right femurs of rose hip group animals. The maximum load, breaking load and energy were calculated using a Shimadzu Ezi-test texture analyser (Kyoto, Japan). The maximum load is defined as the maximum force recorded applied to the femur during the three-point bending test. The fracture load is defined as the force that fractures the bone during the three-point bending test. This force is measured by a load cell. The device fractures the bone and collects and records the force and displacement. Plotting this yields a force-displacement curve that clearly identifies the maximum and breaking loads. The software automatically identifies these points.

Energy is a measure of the energy expended to fracture the bone and is also calculated using a Shimadzu Ezi-test texture analyser (Kyoto, Japan), using the recorded forces and known points at the fracture and maximum force measurements.

These results show that bone fractures from animals fed 2% rose hip extract require more force/energy than OVX animals, and comparable force/energy to sham animals. The maximum load is shown in fig. 4, the breaking load is shown in fig. 5 and the energy is shown in fig. 6.

Industrial applications

The invention has utility in maintaining bone health or improving an imbalance in bone remodeling that results in bone loss.

The composition can be used as a food, beverage, food additive, beverage additive, dietary supplement, nutraceutical, medical food, nutraceutical, pharmaceutical or medicament.

It will be appreciated by persons skilled in the art that the above description has been provided by way of example only and is not limiting of the invention.

Reference to the literature

Freireich EJ，Gehan EA，Rail DP，Schmidt LH，Skipper HE(1966)Quantitative comparison of toxicity to anticancer agents in mouse，rat，hamster，dog，monkey and man.Cancer Chemother Rep 50：219-244Gillies，R.Didier，N.，Denton，M.Determination of cell number inmonolayer cultures，Analytical Biochemistry，1986，159：109-113)Green JH，Booth C，Bunning R.Post prandial metabolic responses to milkenriched with milk calcium are different from responses to milk enrichedwith calcium carbonate.Asia Pacific J Clin Nutr 2003；12：109-19Murray T.Calcium nutrition and osteoporosis.CMAJ 1996；155：935-9.National Research Council.1995.Nutrient requirements of laboratoryanimals.4th Edition.National Academic Press.Washington，DC.Zikan V，Roubal P，Haast T，Stepan JJ.“Acute effects of calcium carbonateand milk on the calcium-parathyroid axis and bone resorption in healthywomen.”In：Bruckhardt P，Dawson-Hughes B，Heaney RP eds.NutritionalAspects of Osteoporosis：Proceedings of the 4^th International SymposiumLausanne，May 2000.Academic Press，London，2001.131-140.

Claims (17)

1. Use of a water soluble extract of rose hip for the preparation of a composition for the treatment or prevention of a bone condition characterized by weakened or brittle bone, wherein the bone condition is treatable or preventable by reducing bone resorption, reducing osteoclast formation, increasing bone formation, increasing osteoblast proliferation, increasing osteoblast differentiation or increasing mineralization or a combination thereof.

2. The use of claim 1, wherein the composition maintains or increases bone formation, maintains or increases bone mineral density, maintains or increases bone content, stimulates bone regeneration during fracture healing, reduces bone resorption, reduces bone loss, or maintains or increases bone strength.

3. The use of claim 1, wherein the disease state is osteoporosis or osteopenia.

4. The use of claim 3, wherein the disease state is osteoporosis.

5. The use according to claim 3, wherein the disease state is osteopenia.

6. The use of any one of claims 1 to 5, wherein the composition provides from 1mg to 2000mg/kg body weight/day rose hip.

7. The use of any one of claims 1 to 5, wherein the composition provides 100mg to 1000mg/kg body weight/day rose hip.

8. The use of any one of claims 1 to 5, wherein the composition provides 50mg to 500mg/kg body weight/day rose hip.

9. The use of any one of claims 1 to 5, wherein the composition provides from 0.05mg to 300mg/kg body weight/day rose hip.

10. Use according to any one of claims 1 to 5, wherein the composition further comprises one or more dairy ingredients.

11. Use according to claim 10, wherein the dairy ingredient is selected from the group comprising: formulated, powdered or fresh skim milk, reconstituted whole or skim milk powder, skim milk concentrate, skim milk retentate, condensed milk, ultrafiltrated milk retentate, milk protein concentrate, milk protein isolate, decalcified milk protein concentrate, low fat milk protein concentrate, casein, caseinate, milk fat, high CLA milk fat, cream, butter, anhydrous milk fat, buttermilk, cheese whey, hard milk fat fraction, soft milk fat fraction, sphingolipid fraction, milk fat globule membrane fraction, phospholipid fraction, complex lipid fraction, colostrum fraction, colostrum protein concentrate, whey, colostrum-derived immunoglobulin fraction, whey, lactoferrin, one or more lactoferrin fragments, whey protein isolate, whey protein concentrate, sweet whey, lactic acid whey, mineral acid whey, reconstituted whey powder, milk mineral, milk whey protein concentrate, milk whey, A component derived from any one of the milk or colostrum-treated streams, a component derived from a retentate or permeate obtained by ultrafiltration or microfiltration of any one of the milk or colostrum-treated streams, a component derived from a breakthrough or adsorption fraction obtained by chromatographic separation of any one of the milk or colostrum-treated streams, an extract of any one of these components including extracts prepared by multi-stage fractionation, differential crystallization, solvent fractionation, supercritical fractionation, near supercritical fractionation, distillation, centrifugal fractionation, fractionation with a modifier, a hydrolysate of any one of these components, a fraction of the hydrolysate, and any combination of any two or more of these components, including combinations of hydrolyzed fractions, combinations of non-hydrolyzed fractions, and combinations of hydrolyzed and non-hydrolyzed fractions.

12. The use of claim 11, wherein the chromatography comprises ion or gel permeation chromatography.

13. The use of claim 11, wherein the modifying agent comprises a soap or an emulsifier.

14. The use of any one of claims 1 to 5, wherein the composition further comprises one or more agents selected from the group consisting of calcium, magnesium, zinc, vitamin D, vitamin K, folic acid or folate, vitamin B6, and vitamin B12.

15. Use according to any one of claims 1 to 5, wherein the composition is a food or food additive.

16. Use according to any one of claims 1 to 5, wherein the composition is a beverage or a beverage additive.

17. The use of any one of claims 1 to 5, wherein the composition is a dietary supplement, a nutritional product, a medical food or a medicament.