HK1076009B - Composition for promotion of bone growth and maintenance of bone health - Google Patents

Description

Composition for promoting bone growth and maintaining bone health

The present invention relates to compositions for maintaining bone health or preventing, alleviating and/or treating bone disorders. The invention also relates to the use of the composition in the manufacture of a nutrient, supplement or medicament; and to methods of promoting bone growth or maintaining bone health comprising administering an effective amount of the composition.

Background

Healthy bone requires effective bone remodeling that involves a balance between bone formation and resorption. Most bone diseases are due to increased bone resorption, rendering the primary therapeutic goal to inhibit the increase in bone resorption, and therefore most agents developed to date are anti-resorptive. For example, estrogens inhibit the production of cytokines that promote osteoclast production and differentiation. SERMs (selective estrogen response modifiers) are being developed that benefit bone health while reducing the risk of hormonal side effects on breast or endometrial tissues. They are thought to act in bone by a similar mechanism to estrogens. Bisphosphonate compounds (e.g., alendronate, risedronate, etc.) concentrate in bone and are by far the most effective inhibitors of bone resorption. They inhibit a key enzymatic pathway that is required for osteoclast activity and osteoclast survival. Calcitonin is a polypeptide hormone that inhibits bone resorption by inhibiting osteoclast activity. Novel targets include blocking TNF receptor/ligand family members and their signaling pathways, in particular blocking RANK/RANKL, inhibiting bone-specific metalloproteinases such as cathepsin K or inhibiting specific kinases.

The development of therapeutic agents that stimulate bone formation lags behind the development of bone resorption therapeutic agents. Some chemicals or agents are known to promote bone growth in humans. For example, WO 9619246 describes a method of promoting bone growth in a patient by intermittent administration of parathyroid hormone, PTH-related protein or agonist for at least one month. In WO 9619501, factors derived from the pancreas inhibit bone resorption and stimulate bone cell proliferation and increase bone formation.

The major recent breakthrough was the demonstration of the role of bone morphogenetic protein 2(BMP-2) as a key factor in the stimulation of bone formation and the role of statins (potent cholesterol-lowering drugs through inhibition of cholesterol synthesis) in improving bone formation, which is mediated in part by the induction of BMP-2. Delivery of recombinant BMP-2 has been shown to induce bone or cartilage formation. In US 6150328, a method of inducing bone and cartilage formation is described, comprising administering a purified bone morphogenic protein produced by culturing cells transformed with DNA encoding BMP. Furthermore, WO 9711095 relates to the use of bone morphogenic protein compositions for the treatment of neural tumors and bone growth and wound healing. In addition to BMPs, growth factors such as insulin-like GF (IGF-1), transforming GF-beta (TGF-beta), Fibroblast GFs (FGFs) are under investigation for local treatment in fracture healing and bone defects. However, systemic administration is problematic due to the metabolism of the small intestine and also due to possible effects on other tissues. Gene therapy is being proposed as an option. This alternative approach is to target the regulation of osteoblast production by regulating their gene or protein expression by dietary or pharmaceutical regulators.

While these chemicals and pharmaceutical compounds have proven useful in the treatment of bone disorders, there remains interest in promoting bone growth and preventing, alleviating or treating bone/joint disorders in mammals by providing a safe, effective nutritional means.

Summary of The Invention

Accordingly, in a first aspect, the present invention provides a composition for preventing, alleviating and/or treating a bone disorder or maintaining bone health in a mammal, said composition comprising as an active ingredient an effective amount of at least one plant or plant extract selected to induce the expression of a bone morphogenic protein.

Notably, it has now been found that certain plants or plant extracts have the ability to promote bone growth by modulating endogenous growth factors in bone or cartilage tissue. They have the ability to induce bone morphogenic protein expression in vivo and in the local environment of bone, which has a positive effect on bone formation and repair, maintenance or prevention of bone health, alleviation and/or treatment of bone disorders.

The composition according to the invention can be used in the manufacture of a nutrient, supplement, therapeutic or pharmaceutical for humans or pets.

Administration of the food composition of the invention to an individual results in improved bone regeneration during fracture healing. The present composition further contributes to the inhibition of bone resorption. It helps to increase bone formation and bone mineral density during growth and optimize peak bone mass. Furthermore, the present food composition helps to reduce bone loss, in particular age-related bone loss in mammals. It therefore helps to maintain age-related bone mass and reduces the risk of osteoporosis.

In addition, it helps to build cartilage in mammals, prevent osteoarthritis in pets or humans, and thus make the individual more mobile or mobile.

In another aspect, the present invention relates to the use of a plant or plant extract selected for its ability to stimulate bone morphogenic proteins and/or inhibit bone resorption for the preparation of a composition for maintaining bone health and preventing, alleviating and/or treating bone disorders in a mammal.

In addition, the present invention provides a method for preventing, alleviating and/or treating bone diseases or maintaining bone health, comprising administering an effective amount of the above composition.

The present invention further provides a method of increasing bone formation, bone mineral density and optimizing peak bone mass during growth, treating or preventing osteoporosis, stimulating bone regeneration during fracture healing, comprising administering an effective amount of the above composition.

The present invention further relates to a method for the treatment, alleviation and/or prevention of osteoarthritis in pets and humans, comprising the step of feeding an individual a composition as described above.

Further provided is a method of reducing bone loss, particularly age-related bone loss, in a human or pet, comprising the step of feeding the individual the above composition.

Detailed Description

With respect to the first object of the present invention, the plant or plant extract according to the present invention comprises phytochemicals having anabolic potential by inducing bone morphogenic protein expression and may further be anti-resorptive.

In a preferred embodiment, the plant or plant extract is from any part of a plant source, such as a leaf, tuber, fruit, seed, root, caryopsis, embryo or cell culture. The plant or plant extract, depending on the plant source, may be in the form of a dried, freeze-dried extract of the leaves, roots and/or fruits, or a fresh plant, or a concentrated fraction obtained by inorganic or organic solvent extraction methods well known in the art.

Selecting a plant or plant extract having the ability to inhibit bone resorption and/or induce bone formation, in particular it may be selected from the group consisting of Lindera (Lindera), Artemisia (Artemisia), calamus (Acorus), rhodobryum (Carthamus), caraway (caraum), Cnidium (Cnidium), juniper (Amelanchier), Curcuma (Curcuma), Taraxacum (Taraxacum), Cyperus (cypeus), juniper (Juniperus), Prunus (Prunus), Iris (Iris), chicory (Cichorium), morus (Dodonaea vitta), Epimedium (emidium), Eriogonum, soybean (Soya), Mentha (Mentha), basil (ocim), thymus (thymus), Artemisia (anthurium), Plantago (Plantago), Mentha (bearberry), roseum (roseum), rosewood (rosewood), rosewood (rosewood), rose. It may also be a mushroom.

In a most preferred embodiment, it may be, for example, the aerial parts of Piper nigrum (Linderabenzein), Artemisia annua (Artemisia vulgaris), the rhizome of Acoruscalamus (Acoruscalamus), the seed or flower of Carthamus tinctorius, the fruit of Amelanchier ovatus (Amelanchier ovalis), the fruit of Amelanchier albolalis, the fruit of Amelanchier alboroides (Amelanchier alborolia), the root of Cichorium intybus, the root stem of Curcuma longa (Curcuma longa), the aerial parts of Epimedium brevicornum (Epimedium brevicornum), the aerial parts of Eriogonum giganteum, the leaves or roots of Taraxacum officinale (Taraxacum officinale) or roots of Taraxacum officinale (Cyperus officiana officinalis), the root of Rhizopus orientalis (Cyperus), the root of Iridium officinale (Iridium officinale) or the stem of Acacia, the fruit of Iridium officinale (Iridium officinale), the cultivated in the cell of Acacia, Iridium officinale (Iridium officinale), or the fruit of Acacia) of the plant, aerial parts of glochira caryophyllata (Ocimum gratissimum), aerial parts of Thymus species (Thymus sp.), aerial parts of Rhus glabra (Rhus glabra), anise (aneth), rosewood, fruits of Vitis vinifera (Vitis vinifera), aerial parts of Rosmarinus officinalis (Rosmarinus officinalis), aerial parts of Artemisia vulgaris (Tanacetum vulgare), Carum carvi, Plantago major (Plantago maior), aerial parts of Oxyden arboreum.

The phytochemical may be genistein, daidzein, lactucin, lactucanthin (lactucopicrin), 3-deoxy-lactucin, geraniol or carvone.

The plants or plant extracts of the invention can be used to prepare food compositions. The composition may be in the form of a nutritionally balanced food or pet food, a dietary supplement, a therapeutic agent or a pharmaceutical composition.

The plant or plant extract can be used alone or in combination with other plants such as chicory, tea, cocoa, or in combination with other bioactive molecules such as antioxidants, fatty acids, prebiotic fibers, glucosamine, chondroitin sulfate.

In one embodiment, a food composition for human consumption is prepared. The composition can be a nutritionally complete formula, a dairy product, a chilled or ambient stable beverage, a soup, a dietary supplement, a dietary substitute, and a nutritional bar or confection.

In addition to the plant extract according to the invention, the nutritional formula may comprise a protein source. Preferably, dietary proteins are used as the protein source. The dietary protein may be any suitable dietary protein; for example, animal proteins (e.g., milk proteins, meat proteins, and egg proteins); vegetable proteins (such as soy protein, wheat protein, rice protein and pea protein); a mixture of free amino acids; or a combination thereof. Milk proteins such as casein, whey protein and soy protein are particularly preferred. The composition may also comprise a source of carbohydrates and a source of fat.

If the nutritional formula includes a fat source, the fat source preferably provides from about 5% to about 55% of the energy in the nutritional formula; such as providing about 20% to 50% energy. The lipids making up the fat source may be any suitable fat or fat mixture. Vegetable fats are particularly suitable; such as soybean oil, palm oil, coconut oil, safflower oil, sunflower oil, corn oil, canola oil, lecithin, etc. Animal fats such as milk fat may also be added if desired.

A carbohydrate source may be added to the nutritional formula. The carbohydrate source preferably provides about 40% to about 80% of the energy of the nutritional composition. Any suitable carbohydrate may be used, such as sucrose, lactose, glucose, fructose, corn syrup solids and maltodextrin, and mixtures thereof. Dietary fiber may also be added if desired. If carbohydrates are used, they preferably comprise about 5% of the energy of the nutritional formula. The dietary fibre may be from any suitable source including, for example, soy, pea, oat, pectin, guar gum, gum arabic and fructooligosaccharides. Suitable vitamins and minerals meeting the appropriate criteria may also be included in the nutritional formula.

If desired, one or more food grade emulsifiers may be incorporated into the nutritional formula; such as diacetyl tartaric acid esters of mono-and diglycerides, lecithin, and mono-and diglycerides. Suitable salts and stabilizers may likewise be included. The plant extract may also be combined with vitamins and minerals.

Preferably an enterally administrable nutritional composition; such as in the form of a powder, tablet, capsule, liquid concentrate, solid product, or drink. If it is desired to produce a powdered nutritional formula, the homogenised mixture is transferred to a suitable dryer such as a spray dryer or freeze dryer and converted to a powder.

In another embodiment, the nutritional composition comprises a milk-based cereal and comprises a probiotic formulation. Preferably, the milk-based cereal is an immature cereal that acts as a carrier for the probiotic formulation.

In another embodiment, a general food product may be fortified with at least one plant or plant extract of the present invention. Such as fermented milks, yoghurts, fresh cheeses, curd, sweets such as candies or sweet drinks, sweet bars, breakfast cereal flakes or bars, drinks, milk powders, soy products, non-dairy fermented products or nutritional supplements for clinical nutrition.

The amount of plant or plant extract in the composition may vary depending on the plant source and its use. In a preferred embodiment, the effective daily dose is at least about 1mg of active molecule per day, more preferably from 1mg to 200 mg of active molecule per day.

In one embodiment, a pharmaceutical composition comprising at least one of the above-described extracts or phytochemicals can be prepared in an amount sufficient to achieve the desired effect in an individual. The composition can be in the form of a tablet, liquid, capsule, soft capsule, paste, lozenge, gum or drinkable solution or emulsion, dry oral supplement, wet oral supplement. The pharmaceutical composition further comprises carriers and excipients suitable for delivering each active molecule of different nature to the target tissue. The kind of carrier/excipient and the amount thereof depends on the nature of the substance and the mode of drug delivery and/or administration. It will be appreciated that the skilled person, based on his own knowledge, may select the appropriate components and plant dosage forms.

The plants or plant extracts of the invention may be used to prepare pet food compositions. The composition may be administered as a supplement to the normal diet of the pet or as a nutritionally complete pet food component, and more preferably in a reduced calorie pet food. It may also be a pharmaceutical composition.

The plant or plant extract can be used alone or in combination with other plants such as chicory, tea, cocoa, or in combination with other bioactive molecules such as antioxidants, fatty acids, prebiotic fibers, glucosamine, chondroitin sulfate.

Preferably, the pet food composition comprises about 0.01 to 0.5 grams of dry plant per gram of dry pet food for a 15 kilogram dog; for a 15 kilogram dog, each gram of wet pet food contains about 0.001 to 0.1 grams of dry plant.

Nutritionally complete pet food according to the invention may be in a powdered, dry form, a therapeutic agent or a wet, cold or ambient stable pet food product. It can be frozen or provided as a stable product at ambient temperature. These pet foods may be produced in a manner well known in the art.

The pet food may also optionally contain prebiotics, probiotic microorganisms, or other active agents, such as long chain fatty acids. The amount of prebiotic in the pet food is preferably less than 10% by weight. For example, the prebiotic may be included at about 0.1% to about 5% by weight of the pet food. For pet foods using chicory as a prebiotic source, chicory may be included at about 0.5% to 10% by weight of the food mix; more preferably from about 1% to about 5% by weight.

If probiotic micro-organisms are used, the pet food preferably comprises about 10 per gram of pet food⁴To about 10¹⁰An individual cell of a probiotic microorganism; more preferably about 10 per gram⁶To about 10⁸Individual cells of a probiotic microorganism. The pet food may comprise from about 0.5% to about 20% by weight of the probiotic microorganism mixture; preferably from about 1% to about 6% by weight; such as from about 3% to about 6% by weight.

Minerals and vitamins can be added to the pet food to complete their nutrition if desired. In addition, various other ingredients such as sugar, salt, spices, seasonings, flavoring agents and the like may also be incorporated into the pet food as desired.

In another embodiment, a dietary supplement may be prepared to improve the quality of the pet food. As dietary supplements, they can be encapsulated, whether wet or dry, or can be provided in powder form and packaged with or separately from a main meal. By way of example, powders containing the extract of the invention may be packaged in powder form in sachets or in gels or fats or other suitable carriers. These individual packaging units may be provided with a main meal or in a multiple unit package for use with a main meal or therapeutic agent, depending on the requirements of the user.

The amount of pet food consumed by the pet to achieve a beneficial effect depends on the size of the pet, the type of pet, and the age of the pet. However, amounts of pet food that provide about 0.5 to 5 grams of dry plant per kilogram of body weight per day are generally sufficient for dogs and cats.

Administration of the above food or pet food composition to a human or animal results in improved bone regeneration during fracture healing. It helps to stimulate bone formation and bone mineral density during growth and optimize peak bone mass. In particular it may provide optimal bone growth in childhood. The food composition is useful for preventing bone loss, particularly bone loss associated with age or long term hospitalization in mammals. It reduces the risk of osteoporosis and improves recovery after fracture. In addition, it helps to build mammalian cartilage, preventing osteoarthritis in pets and humans, which results in better activity or mobility of the individual.

The following examples are given by way of illustration only and are not intended to limit the subject matter of the invention. All percentages are by weight unless otherwise indicated. The embodiments are preceded by the brief description of the figures.

FIG. 1: extracts from Piper hancei (P.E.740, 50. mu.g/ml)) or Cyperus rotundus (P.E.205, 10. mu.g/ml) were treated for 48 hours before measuring the expression of endogenous BMP-2mRNA in hPOB-tert cells by RT-PCR and showing that they stimulate BMP-2 3.8-fold and 2.8-fold, respectively, compared to controls. The assay uses lovastatin (0.5. mu.g/ml) as a positive control, which shows 2.5-fold induction of BMP-2 over the control.

FIG. 2: comparison of inhibition values from extracts from Ocimum gratissimum (738), Amelanchier aldehyda (734), Glycine max (768), Cyperus rotundus (205), Carthamus tinctorius (746) by calvaria assay (A) and pit assay (pit assay) (B).

Examples

Example 1: bone formation and resorption identification

1. Bone formation

91 extracts were screened, wherein the extracts for bone formation were screened by BMP-2 (bone morphogenetic protein) gene reporter assay and for bone resorption by calvaria assay. These 91 extracts corresponded to 30 different plants.

Materials and methods

● preparation of extracts for screening assays:

the ground plant material is defatted with hexane and then extracted with a mixture of alcohol and water, with different water percentages ranging from 10 to 90%, preferably with 50% water. The alcohol can be methanol or ethanol to obtain extract 1 a.

The remainder of the first extract is subjected to enzymatic hydrolysis by alpha and beta glycosidases. The enzyme may be replaced by acidic conditions. This operation can be carried out under mild conditions (room temperature) or by refluxing with different acid concentrations. The aqueous, hydrolyzed phase is extracted with an immiscible solvent, preferably ethyl acetate to obtain extract 2 a.

The extract may be dried, lyophilized or provided in liquid form.

In some cases, polyphenols can be discarded after treatment with polyvinylpolypyrrolidone (PVPP), avoiding artifacts of screening assays.

After preparation of the extracts, each extract was weighed, redissolved with dimethyl sulfoxide (DMSO) to a final concentration of 20mg/ml and stored in portions at-20 ℃. This was applied as a mother liquor and subsequently diluted with the medium used for each assay. A series of doses were tested in the assay system.

● bone formation assay

BMP-2 luciferase assay-the activity of the extracts was determined using 2T3 cells containing the BMP-2 promoter operably linked to a luciferase gene. An increase in luciferase activity in cell lysates reflects an increase in BMP-2 promoter activity. The extract was initially diluted to 100. mu.g/ml and then 1/2 was diluted to 0.2. mu.g/ml for assay. BMP-2 promoter activity was determined by measuring luciferase activity in cell extracts.

Table 113 plants gave significant positive results in stimulating BMP-2 expression

Latin name	Name of English	In part	Concentration of μ g/ml	Active extract/n
					Acorus calamus	Acorus calamus	Rhizome of rhizoid	5	MeOH/water/731
Amelanchier ovalis	Oval shape Amelanchier	(Fruit)	10	MeOH/water/219
					Artemisia vulgaris	Mugwort	Aerial parts	10	Ethyl acetate/225
Cyperus rotundus	Rhizoma Cyperi	Rhizome of rhizoid	10	Ethyl acetate/205
					Taraxacum officinalis	Medicine dandelion	Leaf of Chinese character	50	Ethyl acetate/750
Lindera benzoin	American mountain pepper	Aerial parts	50	Ethyl acetate/740
					Prunus persica	Peach shape	Seed of corn	25	Ethyl acetate/772
Glycine max	Soybean	Cell culture	50	Ethyl acetate/768
					Iris pallida	White iris	Tuber tuber	100	MeOH/water/239
Rosmarinus officianlis	Rosemary	Leaf of Chinese character	50	MeOH/water/2004 ethyl acetate/2005
					Carvi	Carum carvi	Seed of corn	25	Ethyl acetate/2074
Thyme	Thyme (Thymus vulgaris L.)	Leaf of Chinese character	25	Ethyl acetate/2067
					Mentha spicata	Mint	Leaf of Chinese character	100	Ethyl acetate/2072
Vitis vinifera	Grape	(Fruit)	100	Ethyl acetate/2069

Examples of novel preparations and subfractions thereof from the same plant that stimulate BMP-2 are: active extract/n of Piper hancei^OEthyl acetate/740/2059; active subfraction/n^O2060 herba Taraxaci, active extract/n^OEthyl acetate/750/2034; active subfraction/n^O2035 rhizoma Cyperi, active extract/n^OEthyl acetate/205/2011; active subfraction/n^O2012、2013 rhizoma Iridis Tectori, active extract/n^OMeOH/water/239; active subfraction/n^O760/762/2021、2022

The subfractions are prepared by fractionation on a reverse phase silica gel column eluting with solvents of different polarity. Genistein and daidzein (10) in pure soybean isoflavones^-6M) stimulates BMP-2 and estradiol does not stimulate BMP-2.

The induction of BMP-2 appears not to be limited to estrogen-like activity as it is stimulated by phytoestrogens rather than by estrogens themselves. This suggests that the activity of phytoestrogens (such as genistein and daidzein) can be mediated through non-estrogenic mechanisms. BMP-2 promoter activity was not stimulated by estradiol, and thus estrogenicity of the plant compound was not required for this assay.

Plant and method for producing the same	Active extract number	Concentration (μ g/ml)
			Soybean	Ethyl acetate/2001	10、50
Rosemary	MeOH/water/2004	10、50
			Rosemary	Ethyl acetate/2005	10
Rhizoma Cyperi	Subfraction/2012	10、50
			White iris	Subfraction/2022	10
Thyme (Thymus vulgaris L.)	Ethyl acetate/2067	10
			Carum carvi	Ethyl acetate/2074	10

Examples of bone formation in calvarial organ cultures

In Science 286: 1946-1949(1999) describe methods of use. The extracts were evaluated in a 4 day trial using an in vitro neonatal rat calvaria assay. The bone was incubated with the extract for the entire 4 days. Bone formation was evaluated histologically.

2. Bone resorption, calvaria assay

The extracts were evaluated for IL-1 inhibition in a neonatal bone resorption assay (10)^-10M) ability to stimulate bone resorption. The ability of each extract to inhibit bone resorption at 10. mu.g/ml was evaluated. The extract prepared in example 1 was evaluated for IL-1 inhibition in a neonatal bone resorption assay (10)^-10M) ability to stimulate bone resorption. The ability of each extract to inhibit bone resorption at 10. mu.g/ml was evaluated.

Table 2 lists positive plant extracts:

latin name	Name of English	In part	Active extract/n
				Amelanchier alnifolia	Tong Di	(Fruit)	Ethyl acetate/734
Ocimum gratissimum	Species of Ocimum basilicum	Leaf of Chinese character	MeOH/HO/737
				Ocimum gratissimum	Species of Ocimum basilicum	Leaf of Chinese character	Ethyl acetate/738
Carthamus tinctorius	Safflower	Seed of corn	Ethyl acetate/746
				Cyperus rotundus	Rhizoma Cyperi	Rhizome of rhizoid	Ethyl acetate/205
Glycine max	Soybean	Cell culture	768

The following plants were active in the bone resorption assay: amelanchier aldehyda, Ocimum gratissimum, flos Carthami, and semen glycines. Cyperus rotundus inhibits bone resorption and induces BMP-2.

Example 2: effect of plant extracts on the expression of endogenous BMP-2 in human osteoblasts

The BMP-2-induced positive plants of example 1 were further tested for their ability to induce endogenous BMP-2 expression in the human periosteum/preosteoblast cell line hPOB-tert. This test on osteoblasts confirmed the results shown in example 1.

For example, hPOB-tert cells treated with extracts of Piper hancei (extract 740, 50. mu.g/ml) and Cyperus rotundus (extract 205, 10. mu.g/ml) stimulated BMP-2 expression 3.8 and 2.8 times that of controls for 48 hours (see FIG. 1). This assay was validated using lovastatin (0.5 μ g/ml) as a positive control showing that it induced 2.5 times more BMP-2 than the control.

After the cells grew into monolayers, the cells were incubated with lovastatin at 0.05. mu.g/ml or with plant extracts. Total RNA was extracted using TRIzol reagent (Gibco). Mu.g of RNA was reverse transcribed using the first strand cDNA Synthesis kit (Boehringer). The BMP-2cDNA sequence was amplified for 35 cycles using specific oligonucleotide primers (5 ': TTGCGGCTGCTCAGCATGTT; 3': CATCTTGCATCTGTTCTCGGAA) at an annealing temperature of 55 ℃. The PCR products were separated by agarose gel electrophoresis and detected by staining with ethidium bromide. Quantification was performed using NIH imaging software and results were corrected using actin as the housekeeping gene.

The results are shown in FIG. 1.

Bone resorption

Plant extracts positive in the calvaria assay were tested in a second assay of bone resorption, a sink assay with rabbit bone mixed cells cultured on bovine bone sections (Tezuka K et al, 1992, biochem. Biophys. Res. Commun.186 (2): 911-7 and Loget F. et al, 2000, biochem. Biophys. Res. Commun.268 (3): 899-903). The absorption pits were observed by TRAP (tartrate-resistant acid phosphatase) staining. Positive cells were observed and counted.

FIG. 2 shows a comparison of the activity measured on 10. mu.g/ml extract by the two assay systems

Example 3: dry pet food

The feed mixture consisted of about 58% corn by weight, about 5.5% corn bran by weight, about 22% chicken feed by weight, 2.5% dried chicory, about 10% cyperus rotundus tuber, with the remainder consisting of salt, vitamins and minerals.

The feed mixture is charged to a preconditioner and moistened. The wet feed was then charged to an extruder-cooker and gelatinized. The gum base leaving the extruder is extruded through a die and pressed out. The extrudate is cut into suitable dog-fed pieces, dried at 110 ℃ for about 20 minutes, and cooled to form pieces.

The dry diet has a positive effect on bone and cartilage health and increases dog mobility.

Example 4: wet canned pet food with supplement

A mixture was prepared from 73% poultry carcass, pig lung and beef liver (ground), 16% wheat flour, 2% dye, vitamins and inorganic salts. The mixture is emulsified at 12 ℃ and pressed into pudding form, and then cooked at 90 ℃. Cooled to 30 ℃ and cut into large pieces. 45% of these chunks were mixed with 55% of a flavor made of 98% water, 1% dye and 1% guar gum. Loading into tin plate and sterilizing at 125 deg.C for 40 min.

As a supplement to be mixed with the pet food before feeding, it is contained in an additional package (sachet) containing 25 grams of powdered cyperus rotundus aerial parts for addition to the daily food. The corresponding amount for the pet is about 25 grams per day and this may be supplied as a supplement to the can (e.g., on top of the can).

Claims (12)

1. Food composition for the prevention, alleviation and/or treatment of bone diseases and maintenance of bone health in humans and pets, which composition comprises as active ingredient an effective amount of at least one plant or plant extract selected from the group consisting of the aerial parts of Lindera glauca (Lindera grandiflora), the aerial parts of Artemisia vulgaris (Artemisia vulgaris), the root-bark of Acorus calamus (Acorus calamus), the seed or flower of Carthamus tinctorius, the fruit of amellaria ovata (amelanchi ovalis), the fruit of Amelanchier alborophylla (Amelanchier alpinia), the root of chicory (cichoriumlinegybus), the root-bark of Curcuma longa (Curcuma longa), the aerial parts of epimedium brevicornum (epimeddium), the aerial parts of erigeronum, the aerial parts of erigiosum giganteum, the aerial parts of Taraxacum (Taraxacum), the root-bark of Taraxacum officinale (Taraxacum grandiflorum), the root-bark of berberis (Taraxacum grandiflorum), the root-bark of berberidactylophora (Iris officinalis), the root of berberidactyloides (berberis) or the root of berberidaceae (berberidaceae) as, the root of a alba (berberidactylicalis), the root of a mange, the root of a mangifera (mangifera) or the root of a alba (mang, Root-like stems of white Iris (Iris pallida) or yellow Iris (Iris tectorum), fruits of juniper juniperi (Juniperus communis), seeds of peaches (Prunus persica), aerial parts of spearmint (Mentha spicata), aerial parts of caryophylli flos (Ocimum gratissima), aerial parts of thyme species (Thymus sp.), aerial parts of euphorbia glabra (Rhus glabra), aerial parts of fennel (aneth), rhododendron, fruits of grapes (Vitis vinifera), aerial parts of rosemary (Rosmarinus officinalis), aerial parts of artemisia vulgaris (Tanacetum vulgare), aerial parts of caraway (caraway), Plantago asiatica, aerial parts of oxygorum regolium, aerial parts of caraway (tanarium vulgare), phytolacum carvium carvi, phytolacca sativum, lettuce-lettuce extracts or lettuce extracts containing phytochemicals such as alopecurone, lactuca sativa, lettuce.

2. The composition according to claim 1, wherein the plant or plant extract further inhibits bone resorption.

3. The composition of claim 1 or 2 in the form of a nutritionally balanced food or pet food, a dietary supplement, a therapeutic agent or a pharmaceutical composition.

4. Use of a composition according to claim 1 or 2 for the preparation of a food product for assisting bone regeneration during fracture healing, for increasing bone formation and bone mineral density during growth, and for optimizing peak bone mass or reducing bone loss in humans or pets.

5. The use according to claim 4, wherein the bone loss is age-related bone loss.

6. Use according to claim 4 or 5, wherein the prepared food product is useful for building cartilage in humans or pets.

7. Use according to claim 4 or 5, wherein the food product is prepared to help prevent osteoarthritis in humans or pets.

8. Use of a plant or plant extract comprising phytochemicals having the ability to stimulate bone morphogenetic proteins and/or inhibit bone resorption, wherein the plant or plant extract is selected from the group consisting of aerial parts of Lindera glauca (Lindera benzone), aerial parts of Artemisia annua (Artemisia vulgaris), root-tuber of Acorus calamus (Acorus calamus), seeds or flowers of Carthamus tinctorius, fruit of Amelanchier ovalicalis, fruit of Amelanchier japonica (Amelanchier orientalis), root of Amelanchier japonica (Amelanchier nifolila), root of Cichorium intybus (Cichorium intybus), root-tuber of Curcuma longa (Curcuma longa), root-tuber of Epimedium brevicornum (Epimedium brevicornum), aerial part of Nudiferum, root-tuber of Epimedium officinale (Epimedium officinale), root-tuber of Taraxacum officinale (Acronatum), root-tuber of Moraceae, Mongolian officinalis (Acronella officinalis) or petiolus food composition for the prevention, alleviation and/or maintenance of bone health in a human or companion animals, cell cultures of Iris alba (Iris pallida), the rhizome of Iris germanica (Iris germanica), Iris alba (Iris pallida) or Iris yellow Iris (Iris pseudoculus), the fruit of Juniperus communis (Juniperus eomulus), the seed of Prunus persica (Prunus persica), the aerial parts of Mentha spicata (Mentha spicata), the aerial parts of Ocimum gratissimum (Ocimum gratissimum), the aerial parts of Thymus vulgaris (Thymus sp.), the aerial parts of Rhus glabra (Rhus glaberba), the aerial parts of Foeniculum vulgare (anethoides), the fruit of Rhododendron, Vitis vinifera (Vittiferae), the aerial parts of Rosmarinus officinalis (Rosmarinus officinalis), the aerial parts of Artemisia vulgaris (Tanacituturn), the aerial parts of Carum carvacarin (Carrageenan), the aerial parts of Lactusson sativum, Lactuca sativa indica (Lactuca sativa), the aerial parts of Lactuca sativa, Lactuca indica (Lactuca sativa) or Lactuca sativa, the said plant (Lactuca sativa indica), the said plant variety Lactuca sativum.

9. Use according to claim 8, wherein the composition is as defined in any one of claims 1 to 3.

10. Use according to claim 8 or 9, wherein the plant or plant extract is used alone or in combination with other plants, or in combination with other bioactive molecules.

11. Use according to claim 10, wherein the other bioactive molecule is an antioxidant, a fatty acid, a prebiotic fibre, glucosamine, chondroitin sulphate.

12. Use according to claim 10, wherein the other plant is chicory, tea, cocoa.