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WO2002013840A1 - Essential oils and chemically related species for the treatment of increased bone resorption - Google Patents

Essential oils and chemically related species for the treatment of increased bone resorption Download PDF

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Publication number
WO2002013840A1
WO2002013840A1 PCT/CH2001/000395 CH0100395W WO0213840A1 WO 2002013840 A1 WO2002013840 A1 WO 2002013840A1 CH 0100395 W CH0100395 W CH 0100395W WO 0213840 A1 WO0213840 A1 WO 0213840A1
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WIPO (PCT)
Prior art keywords
monoterpene
essential oil
metabolite
chemically related
derived
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PCT/CH2001/000395
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French (fr)
Inventor
Roman MÜHLBAUER
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University Of Bern
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Publication date
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Priority to AU2001273759A priority Critical patent/AU2001273759A1/en
Publication of WO2002013840A1 publication Critical patent/WO2002013840A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease

Definitions

  • IPC International Patent Classification
  • the present invention relates to the use of cosmetical, nutritional or pharmaceutical compositions comprising essential oils extracted from plants, monoterpenes de- rived thereof, their metabolites and/or chemically related species that are useful in the treatment or prevention of diseases or conditions characterized by increased bone resorption, such as Paget's disease, tumor-induced bone disease or particularly osteoporosis.
  • Said compositions are particularly beneficial for mammals, i.e. humans and companion animals.
  • the invention further comprises a method for the preparation of said compositions.
  • Osteoporotic fractures besides causing suffering to the patient, are a major burden to health care as the direct expenditure for osteoporosis and associated fractures is around US$ 14 billion/year in the 'USA and exceeds US$ 10 billion/year .in Europe; novel strategies to prevent osteoporosis are therefore required.
  • This most common bone disorder can be generally defined as the reduction in the quantity of bone, either from the reduction in bone formation or the acceleration of bone resorption, in either event the result is a de- crease in the amount of skeletal tissue.
  • Osteoclasts bone resorbing cells
  • Osteoclasts are responsible for the excavation of a portion of bone during the resorption process. After resorption, osteoblasts (bone forming cells) appear, which then refill the resorbed portion with new bone.
  • estrogens, bisphosphonates and calcitonin are known as inhibitors of bone resorption.
  • One ac- cepted method for the treatment of postmenopausal osteoporosis is estrogen replacement therapy. Although therapy is generally successful, patient compliance with the therapy is low, primarily because estrogen treatment may produce undesirable side effects.
  • An ad- ditional method of treatment is the administration of a bisphosphonate compound, such as, for example, Fosamax® (Merck & Co. , Inc. ) .
  • Pine-oil baths are recommended to elderly people to reduce the back pain due to osteoporosis. Furthermore, the consumption of tea infusions made from dried leaves of sage is recommended to post-menopausal women suffering from hot flushes, as monoterpenes from sage are known to inhibit sweat production. Both procedures are recommended to alleviate the symptoms but not to treat the cause (R. Gladstar; Natural Health; 46-50 (1992) .
  • Yamaguchi et al. (Biosci. Biotechnol. Biochem., 63 (4), 731-735, 1999) isolated and characterized a single monoterpene and have been able to show that oral administration of said monoterpene "significantly suppressed the decrease in bone weight caused by ovariec- tomized mice". However, they have not been able to show the mechanism behind this suppressive effect caused by said monoterpene.
  • essential oils extracted from plants and their components are potent inhibitors of bone resorption, i.e. that the use of essential oils and monoterpenes inhibit the functionality of osteoclasts and/or influence the number of osteoclasts present, for osteoclasts are known to be responsible for the excavation of bone material during the resorption process.
  • the main aspect of the present invention is the use of essential oils extracted from plants and monoterpenes derived thereof, having an inhibitory effect on bone resorption and the use of said compounds at least in the preparation of a pharmaceutical, cos- metical or nutritional composition for the prophylaxis or treatment of a disease or condition characterized by increased bone resorption, such as Paget ' s disease, tumor-induced bone disease or particularly osteoporosis.
  • a disease or condition characterized by increased bone resorption such as Paget ' s disease, tumor-induced bone disease or particularly osteoporosis.
  • the invention further provides a method for the treatment or prophylaxis of a disease or condition which is characterized by increased bone resorption, such as Paget' s disease, tumor-induced bone disease or particularly osteoporosis, comprising the administration of a medicament or nutritional or cosmetical formulation to a human or other mammal, said medicament or nutritional formulation or cosmetics comprising essential oils extracted from plants and/or monoterpenes derived thereof, in an amount, which is effective for inhibiting bone resorption.
  • a disease or condition which is characterized by increased bone resorption, such as Paget' s disease, tumor-induced bone disease or particularly osteoporosis
  • the present invention also foresees the use of essential oils extracted from plants and monoterpenes de- rived thereof, having an inhibitory effect on bone resorption and the use of said compounds in the preparation of a pharmaceutical, cosmetical or nutritional composition for the treatment or prophylaxis of a dis- ease or condition characterized by increased bone resorption, such as Paget ' s disease, tumor-induced bone disease or particularly osteoporosis.
  • Osteoporosis as used herein includes osteoporosis in- prised by hormone deficiency (e.g. postmenopausal) and old age, as well as secondary Osteoporosis such as osteoporosis secondary to steroid treatment or secondary to malnutrition caused by anorexia nervosa.
  • hormone deficiency e.g. postmenopausal
  • secondary Osteoporosis such as osteoporosis secondary to steroid treatment or secondary to malnutrition caused by anorexia nervosa.
  • essential oil volatile oils are meant that occur in plants and in general give the plants their characteristic odors and flavors.
  • Essential oils are complex mixtures of organic compounds.
  • Essential oils may be derived from plants with the following processes:
  • o harvesting resin from plants, essential oils and oleoresins are then separated, o by mechanical pressure; in this method, the plant or different parts of a plant are pressed to push out the essential oil.
  • Hydro distillation also known as water distillation, is a process in which water and plant material are boiled together in a common tub.
  • Steam distillation uses dry steam to vaporize and extract the oil. Steam distillation is used by commercial ventures seeking to process large quantities of essential oils economically.
  • Solvent extraction uses organic solvents to extract both essential oils and oleoresins, which are then separated.
  • o Supercritical extraction is another form of solvent extraction in which carbon dioxide is used under extremely high pressure to extract both essential oils and oleoresins.
  • Essential oils can be extracted from different parts of a plant, such as leaves and tops, leaves and twigs, leaves and young branches, needles, flowers, flowering tops, flowering herbs, fruits, unripe fruits, dried ripe fruits, seeds, berries, root, rhizome, bulb, rind, wood, peel or entire plant.
  • a preferred group of essential oils can be harvested from plants from members of the following botanical families: Araceae, Aristolochiaceae, Bureraceae, Che- nopodiaceae, Compositae, Euphorbiaceae, Geraniaceae, Gramineae, Labiateae, Lauraceae, Magnoliaceae, Myristi- caceae, Myrtaceae, Pinaceae, Piperaceae, Rosaceae, Ru- taceae, Umbelliferae, Valerianaceae, Zingiberaceae.
  • the active ingredient which is an essential oil extracted from a plant such as cited in the following list ciclicaA" (Latin name in brackets), called oil of: Angelica (Angelica officinalis Moench.), Anise (Pimpi- nella anisum L., or Illicium verum Hook. Fil.), Anise- Japanese (Illicium anisatum L.), Asaru (Asaru cana- dense L.), Balm (Melissa officinalis L.), Basil (Ocimum basilicum L.), Bay (Pimenta (Myrcia) acris Kostel.), Bergamot (Citrus aurantium L., var. bergamia Wight &
  • Camphor Cinnamomum camphora T.
  • Nees & Eberm. ) Caraway (Carum carvi L.), Cardamom (Elettaria cardamomum Maton), Cascarilla (Croton elute- ria (L.) Sw. ) , Cedar Leaf (Thuja occidentalis L.), Cedar Wood (Juniperus virginiana L.), Celery (Apium gra- veolens L.), Chenopodium (Chenopodium ambrosioides L. var.
  • Geranium Pelargonium odora- tissimum Ait. and the like
  • Geranium-East Indian An- dropogon schoenanthus L. and the like
  • Ginger Zingi- ber officinale Roscoe
  • Hyssop Hyssopus officinalis L.
  • Juniper Juniper
  • Lavender Lavandu- la officinalis Chaix (L. vera DC.)
  • Lemon Ciitrus li- onium (L.) Risso (C. medica var.
  • Suitable methods of obtaining or chemically synthesi- zing monoterpenes are known in the art. Suitable monoterpenes are isolated from their natural sources by distillation of the plant matter with steam. They are volatile oils, less dense than water, and have normal boiling points in the range 150°-185° C (300°-365° F) . Purification is usually achieved by fractional distillation at reduced pressures or by regeneration from a crystalline derivative.
  • acyclic monoterpenes such as ocimen, myrcen, citronel- lal, linalool, geraniol, nerol, citronellol
  • monocyclic monoterpene such as ⁇ -Terpinen, Limonene, ⁇ -Phellandrene, ⁇ -Phellandrene, p-Cymene (p-Cymol)
  • Menthol, ⁇ -Terpineol, Terpinen-4-ol Carveol, Thymol, Carvacrol, Menthone, Piperitone, Piperitenone, Pule- gone, Carvone, Menthofurane, Cuminal, Safranal, 1,8- Cineol (Eukalyptole) , Ascaridol
  • bicyclic monoterpenes such as Sabinen, ⁇ -Thujen, Santen, ⁇ -Pinen
  • the amount of essential oil and/or monoterpenes to be supplied may vary within wide ranges, depending on the 10
  • Suitable nutritional compositions comprising the above mentioned essential oils or monoterpenes or a combination thereof represent a further object of the invention. They are characterized in that they comprise
  • the nutritional formulations of the invention may comprise nutritionally acceptable components such as vitamins, minerals, trace elements, fibers, flavors, pre- servatives, colorants, sweeteners, emulsifiers and the like.
  • the inventive nutritional formulations may be formulated and administered in any form suitable for enteral administration, for example oral administration or tube feeding.
  • the inventive nutritional compositions may be in form of a complete formula diet, such that, when used as sole nutrition source essentially all daily caloric nitrogen, fatty acids, vitamin, mineral and trace element requirements are met.
  • compositions as used herein comprises i.a:
  • Semi-luxuries including but not limited to baked products, biscuits and cakes, candies, cereal and/or fruit bars, chewing gums, chocolate compositions, confectionery products, dairy and dairy substitute foods, desserts, savoury snacks, yogurts and the like.
  • Food products which are likely to be classified as “functional foods” or “novel foods”, i.e. foods that are similar in appearance to conventional foods and are intended to be consumed as part of a normal diet or a supplement, but have been modified to physiological roles beyond the provision of simple nutrient require- ments.
  • the term "food products” is intended to cover the whole variety of foods and beverages, including but 12
  • yogurts ice creams, cheeses
  • baked products such as fresh or frozen bread, crisp bread sandwiches, biscuits and cakes, dairy and dairy substitute foods, desserts, confectionery products, edible oil compositions, spreads, cereal and/or fruit bars, breakfast cereals, savoury snacks, juices, soups, sauces and the like.
  • Dietary foods for special medical purposes which means a category of foods for particular nutritional uses, specially processed or formulated and intended for the dietary management of patients and to be used under medical supervision which are intended for exclusive or partial feeding of patients with a limited, impaired or disturbed capacity to take, digest, absorb, metabolize or excrete ordinary foodstuffs or certain nutrition requirements, whose dietary management cannot be achieved only by modification of the normal diet, by other foods for particular nutritional uses or by a combination of the two.
  • These foods are either nutritionally complete foods with a nutrient-adapted formulation specific for a disease, disorder or medical condition which, used in accordance with the manufacturer's instructions, may constitute the sole source of nourishment for the persons for whom they are intended, or they may be nutritionally incomplete foods with a nutrient-adapted formulation for a disease, disorder or medical condition which are not suitable to be used as the sole source of nourishment and are thus used as partial replacement or as a supplement to the patient's diet. 13
  • One mandatory active ingredient of a), b) and c) is an essential oil extracted from a plant and/or a monoterpene derived thereof or mixtures thereof.
  • Suitable nutritional compositions may be in liquid form or in solid form and comprise (in % by weight) for example, from approximately 0.01 % to 50 %, preferably from approximately 0.1 to approximately 25 %.
  • the invention further relates to pharmaceutical compositions in single dose unit form comprising
  • compositions as used herein comprises i.a. compositions for enteral administration, such as oral, nasal or rectal administration.
  • Suitable pharmaceutical compositions may be in liquid form or preferentially in solid form and comprise (in % by weight) for example, from approximately 0.001 % to 100 %, preferably from approximately 0.1 to approximately 50 % active ingredient.
  • the active ingredient is an essential oil extracted from a plant and/or a monoterpene derived thereof or mixtures thereof. It is also possible to have a mixture of two or more of said essential oils and monoterpenes.
  • compositions for enteral administration are, for example, those in single dose forms, such as dragees, tablets, capsules or sachets. They are prepared in a manner known per se, for example by means of conventional mixing (with suitable carriers), granulating, confectioning, dissolving or lyophilising proc- esses. 15
  • compositions comprise the active ingredients (that is an essential oil extracted from a plant and/or monoterpene derived thereof or mixtures thereof) together with one or more pharmaceutically acceptable carriers and optionally other therapeutic and/or prophylactic ingredients.
  • the carrier (s) must be acceptable in the sense of being compatible with the other ingredients of the formula and not deleterious to the recipient thereof.
  • compositions include those suitable for oral, topical (including dermal, buccal and sublin- gual), rectal and parenteral (including subcutaneous, intradermal, intramuscular and intravenous) , admini- stration as well as administration by naso-gastric tube.
  • the formulation may, where appropriate, be conveniently presented in discrete dosage units and may be prepared by any of the methods well known in the art of pharmacy. All methods include the step of bringing into association the active ingredients with liquid carriers or finely divided solid carriers or both and then, if necessary, shaping the product into the desired formulation.
  • compositions suitable for oral administration wherein the carrier is a solid are most preferably presented as unit dose formulations such as boluses, capsules or tablets each containing a predetermined amount of the active ingredients.
  • a tablet may be made by compression or moulding, optionally with one or more accessory ingredients. Compressed tablets may be 16
  • Capsules may be prepared by filling the active ingredients, either alone or in admixture with one or more accessory ingredients, into the capsule shells and then sealing them in the usual manner. Cachets are analogous to capsules wherein the active ingredients together with any accessory ingredient (s) are sealed in a rice paper envelope.
  • the active ingredients may also be formulated as dispersible granules, which may for example be suspended in water before administration, or sprinkled on food.
  • the granules may be packaged e.g. in a sachet.
  • Formulations suitable for oral administration wherein the carrier is a liquid may be presented as a solution or a suspension in an aqueous liquid or a non- aqueous liquid or as an oil-in-water liquid emulsion.
  • Formulations for oral administration include controlled release dosage forms e.g. tablets wherein the active ingredients are formulated in an appropriate release— controlling matrix, or are coated with a suitable release-controlling film. Such formulations may be par- ticularly convenient for prophylactic use. 17
  • the active ingredients may also be formulated as a solution or suspension suitable for administration via a naso-gastric tube.
  • compositions suitable for rectal administration wherein the carrier is a solid are most preferably presented as unit dose suppositories.
  • Suitable carriers include cocoa butter and other materials commonly used in the art.
  • the suppositories may be conven- iently formed by admixture of the active combination with the softened or melted carrier (s) followed by chilling and shaping in moulds.
  • compositions suitable for parenteral ad- ministration include sterile solutions or suspensions of the active combination in aqueous or oleaginous vehicles.
  • injectible preparations may be adapted for bolus injection or continuous infusion. Such preparations are conveniently presented in unit dose or multi-dose containers, which are sealed after introduction of the formulation until required for use.
  • the active ingredients may be in powder form, which are constituted with a suitable vehicle, such as sterile, pyrogen-free water, before use.
  • the active ingredients may also be formulated as a long- acting depot preparation, which may be administered by intramuscular injection or by implantation e.g. subcu- taneously or intramuscularly.
  • Depot preparations may include, for example, suitable polymeric or hydrophobic materials, or ion-exchange resins. Such long-acting formulations are particularly convenient for prophylactic use.
  • the pharmaceutical formulations for the various routes of administration described above may include, as appropriate one or more additional carrier ingredients such as diluents, buffers, flavouring agents, binders, surface active agents, thickeners, lubricants, preservatives (including anti- oxidants) and the like, and substances included for the purpose of rendering the formulation isotonic with the blood of the intended recipient.
  • additional carrier ingredients such as diluents, buffers, flavouring agents, binders, surface active agents, thickeners, lubricants, preservatives (including anti- oxidants) and the like, and substances included for the purpose of rendering the formulation isotonic with the blood of the intended recipient.
  • compositions suitable for veterinary use include those adapted for oral, parenteral, and intrarumenal administration.
  • compositions as used herein com- prises i.a. compositions for topical applications, that is, for applications of the active component through the skin.
  • Suitable cosmetical compositions may be in liquid form or in solid form and comprise (in % by weight) for example, from approximately 0.1 % to 100 %, preferably 19
  • the active ingredient is an essential oil extracted from a plant and/or a monoterpene derived thereof or mixtures thereof.
  • the invention further relates to cosmetical compositions in single dose unit form comprising
  • Cosmetical compositions for local topical administration are, for example, those in ointments, liniments or in liquid form. They are prepared in a manner known per 20
  • Suitable cosmetical compositions administrable through a large proportion of the body surface are, for example, bath products such as foam baths or therapeutic bath products that consist of a combination of the active ingredient with a base material prepared in a manner known per se.
  • the present invention relates to a process for preparing a cosmetical formulation containing an essential oil extracted from a plant and/or a monoterpene derived thereof or mixtures thereof, in the form of an ointment and its use.
  • Carrier materials may be aqueous solutions of alcohols, all types of emulsions, gels, foaming compositions and fatty carriers.
  • Use of one specific carrier selected from the group of the above mentioned substances allows formulation of various types of cosmetic preparations according to the invention, such as tonics, creams, balsams, cleaning milks etc. for daily body care as well as shampoos, hair balms, foaming bath compositi- ons, all with the addition of an essential oil extracted from a plant and/or a monoterpene derived thereof or mixtures thereof.
  • the above described nutritional, pharmaceutical and cosmetical composition are used for the treatment of mammals, i.e. humans and companion animals. 21
  • Figure lb Effect of bitter orange peel oil, DL-limonene, and trans-anethol on bone resorption. Applied dose: 100 mg/rat/day. For further details see description of Fig- ure la.
  • Bone resorption was assessed by the urinary excretion of [ 3 H]-Tetracycline from prelabeled rats, a model sen- sitive to inhibitors of bone resorption used clinically.
  • 3 H-labeled tetracycline [ 3 H]-Tc) is deposited in hard tissues during their formation. [ 3 H]-Tc is released when bone is resorbed, circulates in blood, is only poorly reutilized during bone turnover and is then excreted into urine where it can be assessed by counting the 3 H.
  • rats are injected subcutaneously from the first week of their life for 6 weeks with a solution containing 10 ⁇ Ci/ml of 7- [3H] (N) tetracycline (New England Nuclear, Boston, MA) dissolved in 0.15 M NaCl .
  • 7- [3H] (N) tetracycline New England Nuclear, Boston, MA
  • the rats are housed in individual metabolic cages.
  • 24-hour urine collections are started.
  • Dur- ing the first 10 days baseline bone resorption is assessed. Thereafter the rats are treated during 10 days.
  • 3 H in urine is determined by liquid scintillation counting. Aliquots of 1 ml urine are counted in 10 ml of Irga-Safe Plus TM scintillator (Packard International, Zurich, Switzerland) and the result (dpm) is multiplied by the urine volume.
  • the essential oil or monoterpene or combinations thereof of the invention are capable of considerably decreasing the cumulative 3 H-labeled tetracycline excretion in urine of rats which indicates a high inhibitory effect on bone resorption. Accordingly, the claimed nutritional, cosmetical and pharmaceutical compositions are useful for the treatment and prophylaxis of all kinds of diseases or conditions which are characterized by increased bone resorption, such as Paget' s disease, tumor-induced bone disease or particularly osteoporosis.
  • the inhibitory effect of the essential oil or monoterpene or combination thereof of the invention may also be assessed by an in vitro assay.
  • Osteoclasts are harvested from one day old rats, set- tied onto mammoth ivory as the mineral substrate and incubated for 24 hours at 37 C in a 5% CO ? /air atmosphere.
  • One 4x4 mm ivory slice is incubated per well of a 48-well plate in 250 ml of medium containing the onion extract. For each dose 8 slices are used.
  • the ef- feet of the- monoterpene borneol on osteoclast number and resorption pits is assessed by counting the number 25
  • Lanes 7, 8 and 25 of figure la show that low doses are effective when administered for 10 days, and, as shown on figure 2a, the curves of the 30 and 100 mg doses of pine oil appears to converge during the course of the administration of the modified diet, suggesting an accumulation of active components in some body compartment. Thus, when chronically used, lower doses may be effective.
  • Pine oil protects the animals from the loss of both trabecular BMD (by 70%) and total BMC (by 60%; Table 1).
  • the observed loss in total BMC is more than that which can be attributed to the trabecular bone compartment, as trabecular bone only contributes 8% to the bone mineral mass in the proximal metaphysis of rat 28
  • cis-verbenol inhibits osteoclast resorption activity, the parent compound ⁇ - pinene does, ' however, not. Therefore, the finding that the 3 main components of pine oil, ⁇ -pinene, ⁇ -pinene and bornylacetate, although effective in vivo, are ineffective in vitro, strongly suggests that they are me- tabolised to active compounds.
  • actin rings indicates polarisation of osteoclasts; in osteoclasts atta- ched to the bone surface, actin rings appear in the sealing zone surrounding the area to be resorbed (Lak- kakorpi, P.T. and Vanaanen , H.K., j. Bone Mineer. Res. 6, 817-826 (1991), Nakamura et al . , J. Bone Miner. Res. 11, 1873-1879 (1996); Suzuki, H. et al . , Endocrinology 137, 4686-4690 (1996). As shown in Fig. 3a, about 40 % of disaggregated osteoclasts attached to glass cover 30
  • Oil of cumin (oleum carvi, rect.), oil of eucalyptus (oleum eucalypti, >80% Ph. Eur.), oil of fennel (oleum foeniculi), oil of juniper berries (oleum juniperi e baccis, purum), bitter orange oil (oleum auratii ama- rum) , pine oil (oil of fir; oleum pini sibiricum) , dwarf-pine oil (oleum pini pumilionis) , oil of rosemary (oleum rosmarini, DAB) and sage oil (oleum salviae, Dalmation) were purchased from Carl Roth Inc., Reinach, Switzerland.
  • Sweet orange-peel oil (oleum auratii dulcis) was purchased from a local drugstore.
  • Thujone (mixture of » 60% ⁇ -isomere and «10% ⁇ - isomere) , eucalyptol (99%, purum), (+) -camphor (>99%), Borneol (>98%, mixture of isomers) , menthol (>98%, Chinese), (-) - ⁇ -pinene (>97%) and ⁇ -pinene, D,L-limonene (depur) were purchased from Carl Roth, Reinach, Switzerland, thymol (puriss.) from E. Merck, Darmstadt, Germany, (-) ⁇ -bornyl acetate (97%) and (S) -cis verbe- nol were purchased from Aldrich, Buchs, Switzerland.
  • the bisphosphonate clodronate (dichloromethylene)bis- phosphonate was administered by gavage 0.5 hours before food administration at the daily dose of 58.2 mg/kg body weight .
  • [ 3 H] -Tetracycline [ 3 H]- Tc) (Mtihlbauer, R.C. and Fleisch, H., Am. J. Physiol. 259, 679-689 (1990)).
  • [ 3 H]-Tc is deposited into bone and is released when bone is resorbed. After discon- tinuation of labelling, the rats were housed in metabolic cages and bone resorption was monitored by measuring the daily urinary [ 3 H] -excretion (Mtihlbauer, R.C. and Fleisch, H., Am. J. Physiol. 259, 679-689 (1990), Egger, CD. et al., J.Bone Miner. Res.
  • BMC and BMD was measured in the proximal metaphysis of the left tibia by quantitative computed tomography (XCT 34
  • a cross-section starting 5 mm distal to the joint space was evaluated, using a threshold for trabecular bone of 400 mg/cm 3 .
  • the peptide was measured in plasma using an immunoradi- ometric assay (IRMA) kit for rat osteocalcin (Immutop- ics, San Clemente, USA) .
  • IRMA immunoradi- ometric assay
  • Osteoclasts were isolated from femora and tibiae of 1-2 day old rats and settled for 40 minutes onto 4x4 mm ivory slices used as the mineral substrate. After washing off non-adherent cells they were incubated for 24 hours at 37°C in a 5% C0 2 /air atmosphere (Arnett, T.R. and Spowage M. Bone 18, 277-279 (1996); Jones, S.J. et al., Anat. Embryol. 170, 247-256 (1984)).

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Abstract

The present invention relates to cosmetical, nutritional or pharmaceutical compositions comprising essential oils extracted from plants and/or monoteerpenes derived thereof, their metabolites and chemically related species. Said essential oils and their monoterpenes are useful in the treatment or prevention of diseases or conditions characterized by increased bone resorption, such as Paget's disease, tumor-induced bone disease or particularly osteoporosis. The invention further comprises a method for the preparation of said compositions as well as their uses.

Description

H i /CH 01/00395
A. CLASSIFICATION OF SUBJECT MATTER
IPC 7 A61K35/78 A61P19/08
According to International Patent Classification (IPC) or to both national classification and IPC
B. FIELDS SEARCHED
Minimum documentation searched (classification system followed by classification symbols)
IPC 7 A61K
Documentation searched other than minimum documentation to the extent that such documents are included in the fields searched
Electronic data base consulted during the international search (name of data base and, where practical, search terms used)
EPO-Internal, WPI Data, PAJ, FSTA, BIOSIS, MEDLINE, PASCAL, LIFESCIENCES, CHEM ABS Data, CAB Data, EMBASE
C. DOCUMENTS CONSIDERED TO BE RELEVANT
Category ° Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No
WO 9850054 A (MUEHLBAUER ROMAN CONRAD)
12 November 1998 (1998-11-12)
WO 0020014 A (MUEHLBAUER ROMAN CONRAD)
13 April 2000 (2000-04-13)
WO 0020015 A (MUEHLBAUER ROMAN CONRAD) 13 April 2000 (2000-04-13)
D Further documents are listed in the continuation of box C Patent family members are listed in annex
' Special categories of cited documents
"T" later document published after the international filing date or priority date and not in conflict with the application but
'A1 document defining the general state of the art which is not cited to understand the principle or theory underlying the considered to be of particular relevance invention
Ε" earlier document but published on or after the international "X" document of particular relevance, the claimed invention filing date cannot be considered novel or cannot be considered to
'L- document which may throw doubts on priority claιm(s) or involve an inventive step when the document is taken alone which is cited to establish the publication date of another "Y" document of particular relevance, the claimed invention citation or other special reason (as specified) cannot be considered to involve an inventive step when the
O" document referring to an oral disclosure, use, exhibition or document is combined with one or more other such docuother means ments, such combination being obvious to a person skilled
'P" document published prior to the international filing date but in the art later than the priority date claimed "&' document member of the same patent family
Date of the actual completion of the international search Date of mailing of the international search report
27 November 2001 03/12/2001
Name and mailing address of the ISA Authorized officer
European Patent Office, P B 5818 Patentlaan 2 NL - 2280 HV Rljswijk Tel (+31-70) 340-2040, Tx 31 651 epo nl, Fax (+31-70) 3 0-3016 Rempp , G nformation on patent family members
F . /CH 01/00395
Patent document Publication Patent family Publication cited in search report date member(s) date
WO 9850054 12-11-1998 AT 208625 T 15-11- 2001 AU 721339 B2 29-06- 2000 AU 7761698 A 27-11- 1998 BR 9808748 A 11-07- 2000 CN 1255066 T 31-05- 2000 WO 9850054 Al 12-11- 1998 EP 0980250 Al 23-02- 2000 HU 0002971 A2 28-02-■2001 PL 336076 Al 05-06- 2000 ZA 9803764 A 06-11- 1998
W0 0020014 13-04-2000 AU 6088999 A 26-04-2000 WO 0020014 Al 13-04-2000 EP 1117416 Al 25-07-2001
W00020015 13-04-2000 AU 6332999 A 26-04-2000
WO 0020015 Al 13-04-2000
EP 1117418 Al 25-07-2001
US 2001012523 Al 09-08-2001
ESSENTIAL OILS AND CHEMICALLY RELATED SPECIES FOR THE TREATMENT OF INCREASED BONE RESORPTION
The present invention relates to the use of cosmetical, nutritional or pharmaceutical compositions comprising essential oils extracted from plants, monoterpenes de- rived thereof, their metabolites and/or chemically related species that are useful in the treatment or prevention of diseases or conditions characterized by increased bone resorption, such as Paget's disease, tumor-induced bone disease or particularly osteoporosis. Said compositions are particularly beneficial for mammals, i.e. humans and companion animals. The invention further comprises a method for the preparation of said compositions.
Background of the Invention
Bone mass in adult humans decreases with age, leading to an increase in the risk of fractures. Osteoporotic fractures, besides causing suffering to the patient, are a major burden to health care as the direct expenditure for osteoporosis and associated fractures is around US$ 14 billion/year in the 'USA and exceeds US$ 10 billion/year .in Europe; novel strategies to prevent osteoporosis are therefore required. This most common bone disorder can be generally defined as the reduction in the quantity of bone, either from the reduction in bone formation or the acceleration of bone resorption, in either event the result is a de- crease in the amount of skeletal tissue. Osteoclasts (bone resorbing cells) are responsible for the excavation of a portion of bone during the resorption process. After resorption, osteoblasts (bone forming cells) appear, which then refill the resorbed portion with new bone.
In young healthy adults, the rate at which the osteoclasts and osteoblasts are formed and operate maintains a balance between bone resorption and bone formation. After the menopause and/or as consequence of aging in both sexes, an imbalance in this remodeling process occurs, resulting in loss of bone. As imbalance continues over time, the reduction in bone mass and thus bone strength leads to fractures.
Many compositions and methods are described in the medical literature for the treatment of osteoporosis.
For example, estrogens, bisphosphonates and calcitonin are known as inhibitors of bone resorption. One ac- cepted method for the treatment of postmenopausal osteoporosis is estrogen replacement therapy. Although therapy is generally successful, patient compliance with the therapy is low, primarily because estrogen treatment may produce undesirable side effects. An ad- ditional method of treatment is the administration of a bisphosphonate compound, such as, for example, Fosamax® (Merck & Co. , Inc. ) .
Nevertheless, it would be preferable instead of treat- ing established osteoporosis with drugs, to prevent increased bone resorption with natural means. Natural means are generally much more acceptable to patients, which results in an increased compliance.
Pine-oil baths are recommended to elderly people to reduce the back pain due to osteoporosis. Furthermore, the consumption of tea infusions made from dried leaves of sage is recommended to post-menopausal women suffering from hot flushes, as monoterpenes from sage are known to inhibit sweat production. Both procedures are recommended to alleviate the symptoms but not to treat the cause (R. Gladstar; Natural Health; 46-50 (1992) .
Yamaguchi et al. (Biosci. Biotechnol. Biochem., 63 (4), 731-735, 1999) isolated and characterized a single monoterpene and have been able to show that oral administration of said monoterpene "significantly suppressed the decrease in bone weight caused by ovariec- tomized mice". However, they have not been able to show the mechanism behind this suppressive effect caused by said monoterpene.
Summary of the Invention
We have now surprisingly found that essential oils extracted from plants and their components (monoterpenes) are potent inhibitors of bone resorption, i.e. that the use of essential oils and monoterpenes inhibit the functionality of osteoclasts and/or influence the number of osteoclasts present, for osteoclasts are known to be responsible for the excavation of bone material during the resorption process.
Therefore, the main aspect of the present invention is the use of essential oils extracted from plants and monoterpenes derived thereof, having an inhibitory effect on bone resorption and the use of said compounds at least in the preparation of a pharmaceutical, cos- metical or nutritional composition for the prophylaxis or treatment of a disease or condition characterized by increased bone resorption, such as Paget ' s disease, tumor-induced bone disease or particularly osteoporosis..
The invention further provides a method for the treatment or prophylaxis of a disease or condition which is characterized by increased bone resorption, such as Paget' s disease, tumor-induced bone disease or particularly osteoporosis, comprising the administration of a medicament or nutritional or cosmetical formulation to a human or other mammal, said medicament or nutritional formulation or cosmetics comprising essential oils extracted from plants and/or monoterpenes derived thereof, in an amount, which is effective for inhibiting bone resorption.
The present invention also foresees the use of essential oils extracted from plants and monoterpenes de- rived thereof, having an inhibitory effect on bone resorption and the use of said compounds in the preparation of a pharmaceutical, cosmetical or nutritional composition for the treatment or prophylaxis of a dis- ease or condition characterized by increased bone resorption, such as Paget ' s disease, tumor-induced bone disease or particularly osteoporosis.
Osteoporosis as used herein includes osteoporosis in- duced by hormone deficiency (e.g. postmenopausal) and old age, as well as secondary Osteoporosis such as osteoporosis secondary to steroid treatment or secondary to malnutrition caused by anorexia nervosa.
By the term essential oil volatile oils are meant that occur in plants and in general give the plants their characteristic odors and flavors.
Essential oils are complex mixtures of organic compounds.
Suitable methods of obtaining essential oils of the plants mentioned below are known in the art. Essential oils may be derived from plants with the following processes:
o harvesting resin from plants, essential oils and oleoresins are then separated, o by mechanical pressure; in this method, the plant or different parts of a plant are pressed to push out the essential oil. o Hydro distillation, also known as water distillation, is a process in which water and plant material are boiled together in a common tub. o Steam distillation uses dry steam to vaporize and extract the oil. Steam distillation is used by commercial ventures seeking to process large quantities of essential oils economically. o Solvent extraction uses organic solvents to extract both essential oils and oleoresins, which are then separated. o Supercritical extraction is another form of solvent extraction in which carbon dioxide is used under extremely high pressure to extract both essential oils and oleoresins.
Essential oils can be extracted from different parts of a plant, such as leaves and tops, leaves and twigs, leaves and young branches, needles, flowers, flowering tops, flowering herbs, fruits, unripe fruits, dried ripe fruits, seeds, berries, root, rhizome, bulb, rind, wood, peel or entire plant.
A preferred group of essential oils can be harvested from plants from members of the following botanical families: Araceae, Aristolochiaceae, Bureraceae, Che- nopodiaceae, Compositae, Euphorbiaceae, Geraniaceae, Gramineae, Labiateae, Lauraceae, Magnoliaceae, Myristi- caceae, Myrtaceae, Pinaceae, Piperaceae, Rosaceae, Ru- taceae, Umbelliferae, Valerianaceae, Zingiberaceae. For use according to the present invention the active ingredient, which is an essential oil extracted from a plant such as cited in the following list „A" (Latin name in brackets), called oil of: Angelica (Angelica officinalis Moench.), Anise (Pimpi- nella anisum L., or Illicium verum Hook. Fil.), Anise- Japanese (Illicium anisatum L.), Asaru (Asaru cana- dense L.), Balm (Melissa officinalis L.), Basil (Ocimum basilicum L.), Bay (Pimenta (Myrcia) acris Kostel.), Bergamot (Citrus aurantium L., var. bergamia Wight &
Arn.), Bitter Orange (Citrus aurantium L.), Cajeput
(Melaleuca leucadendron L. and the like) , Calamus
(Acorus calamus L.), Camphor (Cinnamomum camphora T.
Nees & Eberm. ) , Caraway (Carum carvi L.), Cardamom (Elettaria cardamomum Maton), Cascarilla (Croton elute- ria (L.) Sw. ) , Cedar Leaf (Thuja occidentalis L.), Cedar Wood (Juniperus virginiana L.), Celery (Apium gra- veolens L.), Chenopodium (Chenopodium ambrosioides L. var. antihelmiticum (L.) Aellen) , Citronella (Cympopo- gon (Andropogon) nardus (L.) Rendle) , Cumin (Cuminum cyminum L.), Cypress (Cupressus sempervirens L.), Dwarf
• Pine Needles (Pinus montana Mill. (Pinus pumilio Haen- ke)., Eucalyptus (Eucalyptus globulus Labill and the like), Fennel (Foeniculum vulgare Mill.), Fir (Abies alba Mill. (A. picea Lindl . , A. pectinata DC), Fir- Siberian (Abies sibirica Ledeb.), Fleabane (Conyza ca- nadensis (L.) Cron. (Erigeron canadensis L., Leptilon canadense (L.)) Britt . ) , Geranium (Pelargonium odora- tissimum Ait. and the like), Geranium-East Indian (An- dropogon schoenanthus L. and the like), Ginger ( Zingi- ber officinale Roscoe) , Hyssop (Hyssopus officinalis L.), Juniper (Juniperus comunis L.), Lavender (Lavandu- la officinalis Chaix (L. vera DC.)), Lemon (Citrus li- onium (L.) Risso (C. medica var. limon L.), Lemon Grass (Cymbopogon (Andropogon) citratus (DC.) Stapf, and C. flexuosus (Nees) Stapf), Levant Wormseed (Artemisia marittima var. stech anniana Bess (A. pauciflo- ra Weber), Linaloe (Bursera delpechiana Poiss. and the like), Majoram (Origanum majorana L.), Myrtle (Myrtus comunis L.), Niaouli (Melaleuca viridiflora (Soland. ) Gaertn.), Nutmeg (Myristica fragrans Houtt.), Orange (Citrus aurantium var. sinensis L.), Orange flowers (Citrus aurantium var. sinensis L. and the like), Origanum (Origanum vulgare L.), Pennyroyal-American (He- deoma pulegioides (L.) Pers.), Pennyroyal-European (Mentha pulegium L.), Pepper (Piper nigrum L.), Peppermint (Mentha piperita L.), Pettigrain (Citrus vulgaris Rissop (C. bigaradia Loisel.), Pine Needles (Pinus syl- vestris L.), Poho (Mentha arvensis), Rose (Rosa gallica L. and the like), Rosemary (Rosmarinus officinalis L.), Sassafras (sassafras albidum (Nutt.) Nees (S. variifo- lium (Salisb.) Kuntze: S. officinale Nees & Eber . ) , Savin (Juniperus sabina L.), Spearmint (Mentha spicata L. (M. virdis L.), Spike (Lavandula spica L. (L. lati- folia Vill.), Sweet Bay (Laurus nobilis L. ) , Tansy (Ta- nacetum vulgare L.), Thyme (Thymus vulgaris L.), Turpentine (Pinus palustris Mill, and the like), Valerian (Valeriana officinalis L.), White Cedar (Thuja occiden- talis L.), Wormwood (Artemisia absinthium L.), Yarrow (Achillea millefolium L.). From a person skilled in the art it will be understood that the above list of essential oils extracted from plants comprises only a selection and does not claim completeness .
Suitable methods of obtaining or chemically synthesi- zing monoterpenes are known in the art. Suitable monoterpenes are isolated from their natural sources by distillation of the plant matter with steam. They are volatile oils, less dense than water, and have normal boiling points in the range 150°-185° C (300°-365° F) . Purification is usually achieved by fractional distillation at reduced pressures or by regeneration from a crystalline derivative.
The following is a list B of suitable monoterpenes: acyclic monoterpenes, such as ocimen, myrcen, citronel- lal, linalool, geraniol, nerol, citronellol; monocyclic monoterpene, such as α-Terpinen, Limonene, α-Phellandrene, β-Phellandrene, p-Cymene (p-Cymol) , Menthol, α-Terpineol, Terpinen-4-ol, Carveol, Thymol, Carvacrol, Menthone, Piperitone, Piperitenone, Pule- gone, Carvone, Menthofurane, Cuminal, Safranal, 1,8- Cineol (Eukalyptole) , Ascaridol; or bicyclic monoterpenes, such as Sabinen, α-Thujen, Santen, α-Pinene, β-Pinene, Camphene, Carene, Sabinol, Thujol, Thujone, Myrtenol, Borneol, Camphor, Fenchon. It will be understood by a person skilled in the art that the above list comprises only a selction of suitable monoterpenes.
The amount of essential oil and/or monoterpenes to be supplied may vary within wide ranges, depending on the 10
desired treatment, subject to be treated and his needs. Thus, where the subject to be treated is an adult person (typically of ca. 60 to 75 kg body weight), a satisfactory inhibitory effect on bone resorption is, in general obtained with compositions formulated to allow a daily administration of 0.1 to 20 grams essential oil or 0.1 to 10 grams monoterpenes or a combination thereof.
Nutritional compositions
Suitable nutritional compositions comprising the above mentioned essential oils or monoterpenes or a combination thereof represent a further object of the invention. They are characterized in that they comprise
a) at least one essential oil extracted from a plant cited in list "A" or mixures thereof;
or
b) at least one monoterpene cited in list „B" or mixtures thereof;
or
c) at least a mixture of at least one essential oil and one monoterpene. 11
The nutritional formulations of the invention may comprise nutritionally acceptable components such as vitamins, minerals, trace elements, fibers, flavors, pre- servatives, colorants, sweeteners, emulsifiers and the like. The inventive nutritional formulations may be formulated and administered in any form suitable for enteral administration, for example oral administration or tube feeding. The inventive nutritional compositions may be in form of a complete formula diet, such that, when used as sole nutrition source essentially all daily caloric nitrogen, fatty acids, vitamin, mineral and trace element requirements are met.
The term nutritional compositions as used herein comprises i.a:
a) . Semi-luxuries, including but not limited to baked products, biscuits and cakes, candies, cereal and/or fruit bars, chewing gums, chocolate compositions, confectionery products, dairy and dairy substitute foods, desserts, savoury snacks, yogurts and the like.
b.) Food products which are likely to be classified as "functional foods" or "novel foods", i.e. foods that are similar in appearance to conventional foods and are intended to be consumed as part of a normal diet or a supplement, but have been modified to physiological roles beyond the provision of simple nutrient require- ments. The term "food products" is intended to cover the whole variety of foods and beverages, including but 12
'not limited to yogurts, ice creams, cheeses, baked products such as fresh or frozen bread, crisp bread sandwiches, biscuits and cakes, dairy and dairy substitute foods, desserts, confectionery products, edible oil compositions, spreads, cereal and/or fruit bars, breakfast cereals, savoury snacks, juices, soups, sauces and the like.
c.) Dietary foods for special medical purposes (herein- after "medical nutrition") which means a category of foods for particular nutritional uses, specially processed or formulated and intended for the dietary management of patients and to be used under medical supervision which are intended for exclusive or partial feeding of patients with a limited, impaired or disturbed capacity to take, digest, absorb, metabolize or excrete ordinary foodstuffs or certain nutrition requirements, whose dietary management cannot be achieved only by modification of the normal diet, by other foods for particular nutritional uses or by a combination of the two. These foods are either nutritionally complete foods with a nutrient-adapted formulation specific for a disease, disorder or medical condition which, used in accordance with the manufacturer's instructions, may constitute the sole source of nourishment for the persons for whom they are intended, or they may be nutritionally incomplete foods with a nutrient-adapted formulation for a disease, disorder or medical condition which are not suitable to be used as the sole source of nourishment and are thus used as partial replacement or as a supplement to the patient's diet. 13
One mandatory active ingredient of a), b) and c) is an essential oil extracted from a plant and/or a monoterpene derived thereof or mixtures thereof.
Suitable nutritional compositions may be in liquid form or in solid form and comprise (in % by weight) for example, from approximately 0.01 % to 50 %, preferably from approximately 0.1 to approximately 25 %.
They are prepared in a manner known per se, for example by means of conventional mixing (with suitable nutrient sources), granulating, confectioning, dissolving or ly- ophilising processes.
Pharmaceutical compositions
The invention further relates to pharmaceutical compositions in single dose unit form comprising
a) at least one essential oil extracted from a plant as cited in list "A" or mixtures thereof;
or
b) at least one monoterpene cited in list „B" or mixtures thereof; 14
or
c) at least a mixture of at least one essential oil and one monoterpene;
and
(d) a pharmaceutical acceptable carrier.
The term pharmaceutical compositions as used herein comprises i.a. compositions for enteral administration, such as oral, nasal or rectal administration. Suitable pharmaceutical compositions may be in liquid form or preferentially in solid form and comprise (in % by weight) for example, from approximately 0.001 % to 100 %, preferably from approximately 0.1 to approximately 50 % active ingredient.
The active ingredient is an essential oil extracted from a plant and/or a monoterpene derived thereof or mixtures thereof. It is also possible to have a mixture of two or more of said essential oils and monoterpenes.
Pharmaceutical compositions for enteral administration are, for example, those in single dose forms, such as dragees, tablets, capsules or sachets. They are prepared in a manner known per se, for example by means of conventional mixing (with suitable carriers), granulating, confectioning, dissolving or lyophilising proc- esses. 15
Pharmaceutical formulations comprise the active ingredients (that is an essential oil extracted from a plant and/or monoterpene derived thereof or mixtures thereof) together with one or more pharmaceutically acceptable carriers and optionally other therapeutic and/or prophylactic ingredients. The carrier (s) must be acceptable in the sense of being compatible with the other ingredients of the formula and not deleterious to the recipient thereof.
Pharmaceutical formulations include those suitable for oral, topical (including dermal, buccal and sublin- gual), rectal and parenteral (including subcutaneous, intradermal, intramuscular and intravenous) , admini- stration as well as administration by naso-gastric tube. The formulation may, where appropriate, be conveniently presented in discrete dosage units and may be prepared by any of the methods well known in the art of pharmacy. All methods include the step of bringing into association the active ingredients with liquid carriers or finely divided solid carriers or both and then, if necessary, shaping the product into the desired formulation.
Pharmaceutical formulations suitable for oral administration wherein the carrier is a solid are most preferably presented as unit dose formulations such as boluses, capsules or tablets each containing a predetermined amount of the active ingredients. A tablet may be made by compression or moulding, optionally with one or more accessory ingredients. Compressed tablets may be 16
prepared by compressing in a suitable machine the active compounds in a free-flowing form such as a powder or granules optionally mixed with a binder, lubricant, inert diluent, lubricating agent, surface-active agent or dispersing agent. Moulded tablets may be made by moulding an inert liquid diluent. Tablets may be optionally coated and if uncoated, may optionally be scored. Capsules may be prepared by filling the active ingredients, either alone or in admixture with one or more accessory ingredients, into the capsule shells and then sealing them in the usual manner. Cachets are analogous to capsules wherein the active ingredients together with any accessory ingredient (s) are sealed in a rice paper envelope. The active ingredients (that is an essential oil extracted from a plant and/or monoterpene derived thereof or mixtures thereof) may also be formulated as dispersible granules, which may for example be suspended in water before administration, or sprinkled on food. The granules may be packaged e.g. in a sachet. Formulations suitable for oral administration wherein the carrier is a liquid may be presented as a solution or a suspension in an aqueous liquid or a non- aqueous liquid or as an oil-in-water liquid emulsion.
Formulations for oral administration include controlled release dosage forms e.g. tablets wherein the active ingredients are formulated in an appropriate release— controlling matrix, or are coated with a suitable release-controlling film. Such formulations may be par- ticularly convenient for prophylactic use. 17
The active ingredients may also be formulated as a solution or suspension suitable for administration via a naso-gastric tube.
Pharmaceutical formulations suitable for rectal administration wherein the carrier is a solid are most preferably presented as unit dose suppositories. Suitable carriers include cocoa butter and other materials commonly used in the art. The suppositories may be conven- iently formed by admixture of the active combination with the softened or melted carrier (s) followed by chilling and shaping in moulds.
Pharmaceutical formulations suitable for parenteral ad- ministration include sterile solutions or suspensions of the active combination in aqueous or oleaginous vehicles. Injectible preparations may be adapted for bolus injection or continuous infusion. Such preparations are conveniently presented in unit dose or multi-dose containers, which are sealed after introduction of the formulation until required for use. Alternatively, the active ingredients may be in powder form, which are constituted with a suitable vehicle, such as sterile, pyrogen-free water, before use.
The active ingredients (that is an essential oil extracted from a plant and/or monoterpene derived thereof or mixtures thereof) may also be formulated as a long- acting depot preparation, which may be administered by intramuscular injection or by implantation e.g. subcu- taneously or intramuscularly. Depot preparations may include, for example, suitable polymeric or hydrophobic materials, or ion-exchange resins. Such long-acting formulations are particularly convenient for prophylactic use.
It should be understood that in addition to the aforementioned carrier ingredients the pharmaceutical formulations for the various routes of administration described above may include, as appropriate one or more additional carrier ingredients such as diluents, buffers, flavouring agents, binders, surface active agents, thickeners, lubricants, preservatives (including anti- oxidants) and the like, and substances included for the purpose of rendering the formulation isotonic with the blood of the intended recipient.
Compositions suitable for veterinary use include those adapted for oral, parenteral, and intrarumenal administration.
Cosmetical compositions
The term cosmetical compositions as used herein, com- prises i.a. compositions for topical applications, that is, for applications of the active component through the skin.
Suitable cosmetical compositions may be in liquid form or in solid form and comprise (in % by weight) for example, from approximately 0.1 % to 100 %, preferably 19
from approximately 5 to approximately 70 % active ingredient .
The active ingredient is an essential oil extracted from a plant and/or a monoterpene derived thereof or mixtures thereof.
The invention further relates to cosmetical compositions in single dose unit form comprising
a) at least one essential oil extracted from a plant cited in list „A" or mixtures thereof;
or
b) at least one monoterpene cited in list „B" or mixtures thereof;
or
c) at least a mixture of at least one essential oil and one monoterpene
and
(d) an acceptable carrier.
Cosmetical compositions for local topical administration are, for example, those in ointments, liniments or in liquid form. They are prepared in a manner known per 20
se, for example by means of conventional mixing (with suitable carriers) or dissolving processes.
Suitable cosmetical compositions administrable through a large proportion of the body surface are, for example, bath products such as foam baths or therapeutic bath products that consist of a combination of the active ingredient with a base material prepared in a manner known per se.
The present invention relates to a process for preparing a cosmetical formulation containing an essential oil extracted from a plant and/or a monoterpene derived thereof or mixtures thereof, in the form of an ointment and its use.
Carrier materials may be aqueous solutions of alcohols, all types of emulsions, gels, foaming compositions and fatty carriers. Use of one specific carrier selected from the group of the above mentioned substances allows formulation of various types of cosmetic preparations according to the invention, such as tonics, creams, balsams, cleaning milks etc. for daily body care as well as shampoos, hair balms, foaming bath compositi- ons, all with the addition of an essential oil extracted from a plant and/or a monoterpene derived thereof or mixtures thereof.
The above described nutritional, pharmaceutical and cosmetical composition are used for the treatment of mammals, i.e. humans and companion animals. 21
Brief Description of the Drawings :
Figure la: Effect of herbs, essential oils and monoterpenes on bone resorption. Doses are given as underlined numbers, lanes from 1-37 as bold numbers. The values of the treated groups (n=5) are given as the ratio treated/untreated control ± SEM. (Values below 1.0 in- dicate inhibition of bone resorption.) The 95% confidence interval (1.96 x SEM) of the control groups (n= 6) of each experiment is given as box. Mean values outside the 95% CI are significantly different from control (p<0.05) (Sachs, L. Angewandte Statistik, 90-91 (Springer Verlag, Berlin, Heidelberg, New York, 1974) . The significance of differences versus the hypothetical mean of 1.0 is indicated by the suffixes, + p< 0.05, ++ p< 0.01 and +++ p< 0.001.
Figure lb: Effect of bitter orange peel oil, DL-limonene, and trans-anethol on bone resorption. Applied dose: 100 mg/rat/day. For further details see description of Fig- ure la.
Figure 2 :
Time course of effects on bone resorption. Based on baseline bone resorption, rats are homogeneously as- signed to the groups (day -1) . At day 0 rats are switched for 10 days to the semipurified diet stimulat- 22
ing bone resorption (Mϋhlbauer, R.C. and Fleisch, H.,
J. bone Miner. Res. 11, Abstract #S395 (1996)). Values are given as mean ± SEM, n=6 for controls, n=5 for treated, a: control (closed circle), pine oil 30 mg/rat/day (open circle) and pine oil 100 mg/rat/day
(open triangle), b: control (closed circle), borneol
200 mg/rat/day (open diamond) , α-pinene 200 mg/rat/day
(open triangle) and clodronate 58 mg/kg body weight
(open square). ++ p<0.01, +++ p<0.001 indicates first day with significant difference vs. control.
Figure 3:
In vitro effects on osteoclasts. a: isolated rat osteoclasts were incubated with various concentrations of borneol (open circles), ImM α-pinene (closed circle) and 1 nM calcitonin (full square) for 3 h. The number of osteoclasts containing one or more actin rings is expressed as percentage of the total number of osteoclasts of 4 coverslips. b: after 3h preincubation, the osteoclasts were treated with borneol or calcitonin for various time periods. Control (closed circle) n=4-8 coverslips. 1 nM salmon calcitonin (open square) n=4-7. 1 mM borneol (open triangle) n=3-7. * p<0.05, ** p<0.01, *** p<0.001 indicates significance vs. control.
Detailed Description of the Invention
Bone resorption was assessed by the urinary excretion of [3H]-Tetracycline from prelabeled rats, a model sen- sitive to inhibitors of bone resorption used clinically. 23
3H-labeled tetracycline ([3H]-Tc) is deposited in hard tissues during their formation. [3H]-Tc is released when bone is resorbed, circulates in blood, is only poorly reutilized during bone turnover and is then excreted into urine where it can be assessed by counting the 3H.
Based on these characteristics, a method for continual monitoring of bone resorption has been developed and validated, using the urinary excretion of [3H]-Tc from chronically prelabeled rats (Am J Physiol, 258, R 679-
R689, 1990; J Clin Invest, 95, 1933-1940, 1995). This method is sensitive to drugs used to inhibit bone re- sorption in patients suffering from osteoporosis (J
Bone Miner Res, 6,1003-1011, 1991; J Clin Invest, 95,
1933-1940, 1995) . Furthermore, with this method we found that foodstuffs such as some salads, herbs and vegetables inhibit bone resorption (Nature 401, 343- 344, 1999) .
In order to radioactively label the whole skeleton, rats are injected subcutaneously from the first week of their life for 6 weeks with a solution containing 10 μCi/ml of 7- [3H] (N) tetracycline (New England Nuclear, Boston, MA) dissolved in 0.15 M NaCl . One week after discontinuation of the [3H]-Tc injections the rats are housed in individual metabolic cages. After 10 days of adaptation, 24-hour urine collections are started. Dur- ing the first 10 days baseline bone resorption is assessed. Thereafter the rats are treated during 10 days. 24
3H in urine is determined by liquid scintillation counting. Aliquots of 1 ml urine are counted in 10 ml of Irga-Safe Plus™ scintillator (Packard International, Zurich, Switzerland) and the result (dpm) is multiplied by the urine volume.
Suitable experiments show that the essential oil or monoterpene or combinations thereof of the invention are capable of considerably decreasing the cumulative 3H-labeled tetracycline excretion in urine of rats which indicates a high inhibitory effect on bone resorption. Accordingly, the claimed nutritional, cosmetical and pharmaceutical compositions are useful for the treatment and prophylaxis of all kinds of diseases or conditions which are characterized by increased bone resorption, such as Paget' s disease, tumor-induced bone disease or particularly osteoporosis.
The inhibitory effect of the essential oil or monoterpene or combination thereof of the invention may also be assessed by an in vitro assay.
Osteoclasts are harvested from one day old rats, set- tied onto mammoth ivory as the mineral substrate and incubated for 24 hours at 37 C in a 5% CO?/air atmosphere. One 4x4 mm ivory slice is incubated per well of a 48-well plate in 250 ml of medium containing the onion extract. For each dose 8 slices are used. The ef- feet of the- monoterpene borneol on osteoclast number and resorption pits is assessed by counting the number 25
of tartrate-resistant acid phosphatase positive (TRAP+) multinucleated cells (MNC) and the number of resorption pits excavated into the ivory surface visualised by tangential illumination after gold spottering. Osteo- clast activity is given by the ratio of the number of resorption pits/the number of TRAP+ MNC. Figure la, shows that 1 g of powdered leaves from sage, rosemary and thyme mixed with the daily food inhibits bone resorption in the rat (lanes 1, 15 & 16) . The es- sential oils from sage and rosemary also inhibit bone resorption (lanes 3 & 31). This is also the case for pine oil, dwarf-pine oil and the related medicinal turpentine (lanes 17, 18 & 29) . Also oil of juniper and eucalyptus are inhibitory (lanes 14 and 30) . Sage oil, pine oil and turpentine (lanes 2-4, 21-24 and 28 & 29) inhibit bone resorption dose dependently; pine oil being the most potent established so far (lane 25) . Thus, 7 out of 10 essential oils investigated inhibit bone resorption in the rat.
In a further step the individual components of essen- tial oils were studied. A mixture of the 4 major monoterpenes occuring in 200 mg of sage oil (Wagner,
H., Pharmazeutische Biologie 5, Gustav Fischer Verlag, Stuttgart-New York (1993) ) (80 mg thujone, 24 mg eucalyptol, 14 mg camphor and 10 mg borneol), lane 5, is nearly as effective as 200 mg of sage oil (lane 10), suggesting that these 4 monoterpenes are the relevant components. When tested singly at the above doses, thu- jone, eucalyptol and camphor (lanes 6-8) are effective inhibitors. At the dose of 100 g/day, borneol appears 26
to be more potent than eucalyptol and camphor (lanes 11-13, ) . α-pinene, β-pinene and bornylacetate, the major components of the pine oil we have used are also potent inhibitors of resorption (lanes 19,20 & 37). This is also the case for menthol and thymol (lanes 26 & 27). Thus, 9 monoterpenes, major components of the above described active herbs or essential oils, inhibit bone resorption. A mixture of equal parts of these monoterpenes is also very active (lane 34), suggesting that their effect is additive..
A time course of the effect of 2 doses of pine oil, borneol, α-pinene and a bisphosphonate on bone resorption was investigated (Figure 2) . The first significant effect was detected 2 days after starting a diet con- taining 100 mg pine oil per day (panel a) . At the lower dose of 30 mg per day, a significant effect was reached after 4 days. Large doses of the monoterpenes borneol and α-pinene (200 mg/day) had a larger effect, but it did not reach significance earlier (panel b) . The time course for the other essential oils and monoterpenes reported in figures la and lb is similar (not shown) . The appearance of the effect is dose dependent, occurs earliest 2 days after beginning of treatment and is therefore different from the effect of the bisphospho- nate clodronate, a well known antiresorptive drug, which occurs after 4 days. The time course of the effect of essential oils and monoterpenes also appears to differ from that of two other inhibitors of bone resorption calcitonin (Mϋhlbauer, R.C. and Fleisch, H., J. Clin. Invest. 95, 1933-1940 (1995) and a vegetable 27
such as onion (Mϋhlbauer, R.C. and Li, F., Nature 401, 343-344 (1999).
To date neither the optimal schedule nor the minimal effective dose of these compounds required to inhibit bone resorption in preventive and curative models are known. Lanes 7, 8 and 25 of figure la show that low doses are effective when administered for 10 days, and, as shown on figure 2a, the curves of the 30 and 100 mg doses of pine oil appears to converge during the course of the administration of the modified diet, suggesting an accumulation of active components in some body compartment. Thus, when chronically used, lower doses may be effective.
As the incidence of osteoporosis in humans is much more frequent in women, bone turnover (resorption > formation) increasing after the menopause, an animal model, the ovariectomised (OVX) rat in which a marked decrease in trabecular bone mineral density (BMD) and bone mineral content (BMC) occurs (Wronski,T. J. , Walsh, C.C. & Ignaszewski,L.A.. Bone 7, 119-123 (1986)), has been used to study the effect of pine oil on trabecular BMD and BMC in the proximal metaphysis of the left tibia of the rat. Pine oil protects the animals from the loss of both trabecular BMD (by 70%) and total BMC (by 60%; Table 1). The observed loss in total BMC is more than that which can be attributed to the trabecular bone compartment, as trabecular bone only contributes 8% to the bone mineral mass in the proximal metaphysis of rat 28
tibia, suggesting that pine oil has some protective effect on the cortical shell as well.
As BMC and BMD were measured at one skeletal site only, we investigated whether pine oil affects whole body bone turnover by measuring osteocalcin (ng/ml plasma) at the end of the experiment described above. During the period of the experiment, osteocalcin increased from 13.51±1.09 (mean±SEM, n = 10 each) in sham oper- ated animals to 20.34±1.41 (p<0.01) in OVX rats. Osteocalcin in OVX rats receiving pine oil was 15.33±1.21 (p<0.05 vs. OVX). Thus, the addition of pine oil to the diet partially normalised whole body bone turnover in OVX rats.
The question whether monoterpenes directly affect bone cells led to the investigation of their effect on in vitro osteoclastic resorptive activity calculated as the ratio resorption pits/TRAP+ multinucleated cells. The resorptive activity decreased from 0.709±0.093 in control ivory slices (n=16) to 0.687+0.117 (n=8), 0.286±0.083 (n=8; p<0.01) and 0.106±0.044 (n=8; p<0.0001) in cultures to which thymol at concentrations of 0.01, 0.1 and 1.0 mM was added. Borneol added at concentrations of 0.1, 0.5 and 1.0 mM decreased resorptive activity from 0.748+0.105 (control n=16) to 0.346+0.155 (n=8), 0.154+0.063 (n=8; p<0.0001) and) and 0.154+0.105 (n=8; p<0.001) respectively. One mM camphor decreased osteoclastic resorption activity from 0.775+0.107 (control n=16) to 0.35+.0.062 (n=8; p<0.01). Thus, thymol, camphor and borneol at a dose of 1.0 mM 29
and borneol and thymol also at lower doses decrease osteoclastic resorption activity. The other monoterpenes α-pinene, β-pinene, menthol, eucalyptol, thujone and bornylacetate were not active in vitro at 1.0 mM (re- suits not shown) . As workers in saw-mills exposed to fumes containing α-pinene excrete the metabolite cis- verbenol in their urine (Erikson, K. and Levin, J.O., J. Chromatogr. B. Biomed. Appl . 67, 85-98 (1996)) this substance has also been investigated. The resorptive activity of osteoclasts decreased from 0.565±0.066 in control ivory slices (n=16) to 0.072+0.046 (n=8; p<0.0001) and 0.011+.0.011 (n=8; p<0.0001) in cultures to which cis-verbenol was added at the concentration of 1.0 and 2.0 mM. Thus, in vitro, cis-verbenol inhibits osteoclast resorption activity, the parent compound α- pinene does, 'however, not. Therefore, the finding that the 3 main components of pine oil, α-pinene, β-pinene and bornylacetate, although effective in vivo, are ineffective in vitro, strongly suggests that they are me- tabolised to active compounds.
The action of borneol on osteoclast function was then studied in more detail. Formation of actin rings indicates polarisation of osteoclasts; in osteoclasts atta- ched to the bone surface, actin rings appear in the sealing zone surrounding the area to be resorbed (Lak- kakorpi, P.T. and Vanaanen , H.K., j. Bone Mineer. Res. 6, 817-826 (1991), Nakamura et al . , J. Bone Miner. Res. 11, 1873-1879 (1996); Suzuki, H. et al . , Endocrinology 137, 4686-4690 (1996). As shown in Fig. 3a, about 40 % of disaggregated osteoclasts attached to glass cover 30
slips form actin rings. Borneol inhibits this formation dose dependently. α-pinene at a concentration of 1 mM has no effect whereas 1 nM calcitonin inhibits completely. As shown on Fig. 3b, the actin rings disappear rapidly after addition of 1 mM borneol and 1 nM calcitonin, respectively. The time course of the effect of borneol is similar to that of calcitonin indicating that borneol acts rapidly on mature osteoclasts inhibiting their polarisation (Suzuki, H. et al., Endocrino- logy 137, 4686-4690 (1996) .
These data clearly show that essential oils and monoterpenes positively affect bone metabolism in the rat. There is a long history of the use of such co - pounds in man in folk-medicine, for medical applications such as the relief of head and chest colds and in bathing. As these lipophilic compounds readily cross membranes they are well absorbed from the intestine, skin and the lung. The use of essential oils and monoterpenes could be an inexpensive way, which is not only nutrition based, to help to decrease the incidence of osteoporosis.
Examples
Materials
Processing of herbs The locally grown herbs were carefully washed, air- dried and ground to a fine powder. 31
Essential oils
Oil of cumin (oleum carvi, rect.), oil of eucalyptus (oleum eucalypti, >80% Ph. Eur.), oil of fennel (oleum foeniculi), oil of juniper berries (oleum juniperi e baccis, purum), bitter orange oil (oleum auratii ama- rum) , pine oil (oil of fir; oleum pini sibiricum) , dwarf-pine oil (oleum pini pumilionis) , oil of rosemary (oleum rosmarini, DAB) and sage oil (oleum salviae, Dalmation) were purchased from Carl Roth Inc., Reinach, Switzerland.
Sweet orange-peel oil (oleum auratii dulcis) was purchased from a local drugstore.
Monoterpenes
Thujone (mixture of » 60% α-isomere and «10% β- isomere) , eucalyptol (99%, purum), (+) -camphor (>99%), Borneol (>98%, mixture of isomers) , menthol (>98%, Chinese), (-) -α-pinene (>97%) and β-pinene, D,L-limonene (depur) were purchased from Carl Roth, Reinach, Switzerland, thymol (puriss.) from E. Merck, Darmstadt, Germany, (-)α-bornyl acetate (97%) and (S) -cis verbe- nol were purchased from Aldrich, Buchs, Switzerland.
Example 1
Diets and treatments
Prior to the experiments, all rats had free access to tap water and were fed ad libitum pellets (Kliba 343, Kliba-Mϋhlen, Kaiseraugst, Switzerland) , containing 1.1% calcium (Ca) and 0.8% inorganic phosphate (P) . 32
Tetracycline model:
To avoid any interference from natural components of a
"normal" diet (Miihlbauer, R.C. & Fleisch,H J Bone Miner Res 11, S188, Abstract #S395 (1996) all rats were group-fed from the first day of treatment with a „semi- purified" diet with a similar composition to AIN 76 (American Institute of Nutrition (AIN) . J Nutr 107, 1340-1348 (1977) but containing 1.1% Ca and 1.2% P (SoDi 2160, ibid) . Powdered herbs, essential oils or monoterpenes were mixed with the wet food at the doses indicated. The batches of food sufficient for the duration of the experiment were aliquoted and kept frozen until use.
Osteoporosis model
Starting one week before ovariectomy all rats were group-fed a modified AIN-93M diet containing 0.28% Ca and 0.23% P (SoDi 2172, ibid) as wet food and deminer- alised water was given ad libitum. Pine oil was added as described above.
The bisphosphonate clodronate (dichloromethylene)bis- phosphonate was administered by gavage 0.5 hours before food administration at the daily dose of 58.2 mg/kg body weight .
Example 2
In vivo monitoring of bone resorption New-born male Wistar Hanlbm rats obtained from RCC, Fϋllinsdorf, Switzerland were injected from the first 33
week of life for 6 weeks with [3H] -Tetracycline ([3H]- Tc) (Mtihlbauer, R.C. and Fleisch, H., Am. J. Physiol. 259, 679-689 (1990)). [3H]-Tc is deposited into bone and is released when bone is resorbed. After discon- tinuation of labelling, the rats were housed in metabolic cages and bone resorption was monitored by measuring the daily urinary [3H] -excretion (Mtihlbauer, R.C. and Fleisch, H., Am. J. Physiol. 259, 679-689 (1990), Egger, CD. et al., J.Bone Miner. Res. 9, 1211-1219 (1994); Mϋhlbauer, R.C. and Fleisch, H.; J. Clin. Invest. 95, 1933-1940 (1995); Antic, V.N. et al., Calcif. Tissue Int. 58, 443-448 1996); Mϋhlbauer, R.C. and Li, F., Nature 402, 343-344 (1999)). After 10 days each of equilibration and baseline monitoring of bone resorp- tion, the dietary intervention was startedOsteoporosis modelOne year old retired breeder female Wistar Hanlbm rats obtained from RCC, Fϋllinsdorf, Switzerland were individually housed and carefully observed for 1 month. Animals with decreasing body weight and/or irregular food consumption were eliminated. One week before the dietary intervention all rats were switched to the ex- perimental diet without additions. 3 days before the dietary intervention baseline BMD and BMC was measured and the rats assigned homogeneously to the 3 groups. OVX and sham operation were performed 1-2 days prior to the dietary intervention.
Assessment of bone „mass"
BMC and BMD was measured in the proximal metaphysis of the left tibia by quantitative computed tomography (XCT 34
Research SA, software version 5.40, Stratec Medizin- technik, Pforzheim, Germany), as previously described (Mtihlbauer, R.C. , Schenk,R.K., Chen,D., Lehto- Axtelius,D. & Hakanson,R., Calcif Tissue Int 62, 323- 326 (1998); Li,F. & Mϋhlbauer, R.C. ; J Bone Miner Res 14, 1457-1465 (1999) .
A cross-section starting 5 mm distal to the joint space was evaluated, using a threshold for trabecular bone of 400 mg/cm3.
Measurement of osteocalcin
The peptide was measured in plasma using an immunoradi- ometric assay (IRMA) kit for rat osteocalcin (Immutop- ics, San Clemente, USA) .
Example 3
In vitro assessment of osteoclast activity Osteoclasts were isolated from femora and tibiae of 1-2 day old rats and settled for 40 minutes onto 4x4 mm ivory slices used as the mineral substrate. After washing off non-adherent cells they were incubated for 24 hours at 37°C in a 5% C02 /air atmosphere (Arnett, T.R. and Spowage M. Bone 18, 277-279 (1996); Jones, S.J. et al., Anat. Embryol. 170, 247-256 (1984)). Individual slices were incubated in 48-well tissue culture plates in MEM Earle's salt solution containing 15 mM bicarbonate, 10% fetal bovine serum (FBS) and the test sub- stances. For each dose 8 slices were used. After fixation cells were stained for tartrate-resistant acid-

Claims

35phosphatase (TRAP) (kit 386-A, Sigma, Buchs, Switzerland) . Osteoclasts were counted blinded as the number of TRAP positive (TRAP+) multinucleated cells (MNC) . After removing the cells the slices were gold spottered and the resorption pits counted blinded (Vitte, C. et al., Endocriniolgy 137, 2324-2333 (1996)).Example 4Actin ring in isolated osteoclastsOsteoclasts were isolated as above. After adherence for 1 h to glass cover slips, precoated over night with 30% FBS diluted in saline, non-adherent cells were washed off, and the osteoclasts incubated at 37°C in air in Medium 199 containing Hank's salt and 20 mM HEPES buffer, pH 7.2, and 10% FBS and the compounds to be tested (Amano et al., Calcif. Tissue Int. 57, 367-370 (1995)). After 3 h, the cells were fixed for 5 min at room tem- perature with 3% paraformaldehyde in phosphate buffered saline without Ca-2+ and Mg2+ (PBS), washed 3 times with• PBS and permeabilized with 0.5% Triton X-100 in PBS for5 min. Thereafter, the cells were incubated with Phal- loidin conjugated with rhodamin (Molecular Probes, JURO, Luzern, Switzerland) for 40 min. After washing with PBS and embedding, the fluorescent actin rings were counted blinded. In the time course experiment (Fig. 3b) , the osteoclasts free of non adhering cells, were preincubated for 3h and then incubated with the test compound. After various time periods the cells were fixed and treated as described above. 36Table 1 Effect of pine oil on bone "mass" in aged ova- riectomized rats a = p<0.05, b = p<0.001 indicates significance of the slope of the mean regression line calculated from the individual regression for each rat during the 9 weeks of treatment vs. zero hypothesis. * = p<0.05 indicates significance of OVX + pine oil vs OVX. ""pO.Ol and """"pO.OOl indicates significance of OVX vs. sham. 37Claims
1. Use of an essential oil, a monoterpene derived thereof, a metabolite and/or a chemically related species in the preparation of a pharmaceutical, cosmetical or nutritional composition for the treatment or prophylaxis of a disease or condition involving bone resorp- tion, such as Paget' s disease, tumor-induced bone disease or osteoporosis in mammals, wherein said essential oil or monoterpene inhibits the activity of cells responsible for bone resorption.
2. The use of an essential oil, a monoterpene derived thereof, a metabolite and/or a chemically related species according to claim 1, wherein the treated mammal is a human or companion animal.
3. The use of an essential oil, a monoterpene derived thereof, a metabolite and/or a chemically related spe- cies according to claims 1 or 2, wherein said cells responsible for bone resorption are osteoclasts.
4. The use of an essential oil, a monoterpene derived thereof, a metabolite and/or a chemically related species according to claim 3, wherein said essential oil or monoterpene inhibits polarisation of said osteoclasts. 38
5. The use of an essential oil, a monoterpene derived thereof, a metabolite and/or a chemically related species according to claims 3 or 4, wherein said essential oil or monoterpene inhibits polarisation of said osteo- clasts by inhibiting formation of actin rings.
6. The use of an essential oil, a monoterpene derived thereof, a metabolite and/or a chemically related species according to claim 1, wherein said essential oil or monoterpene influences the number of osteclasts.
7. The use of an essential oil, a monoterpene derived thereof, a metabolite and/or a chemically related species according to any of the preceeding claims wherein the essential oil is harvested from a plant selected from the group consistising of Angelica (Angelica officinalis Moench.), Anise (Pimpinella anisum L., or Illicium verum Hook. Fil.), Anise-Japanese (Illicium anisa- tum L.), Asarum (Asarum canadense L.), Balm (Melissa officinalis L.), Basil (Ocimum basilicum L.), Bay (Pi- menta (Myrcia) acris Kostel . ) , Bergamot ( Citrus aurantium L., var. bergamia Wight & Arn.), Bitter Orange (Citrus aurantium L.), Cajeput (Melaleuca leucadendron L. and the like), Calamus (Acorus calamus L. ) , Camphor (Cinnamomum camphora T. Nees & Eberm. ) , Caraway (Carum carvi L.), Cardamom (Elettaria cardamomum Maton) , Ca- scarilla (Croton eluteria (L. ) Sw.), Cedar Leaf (Thuja occidentalis L.), Cedar Wood (Juniperus virginiana L.), Celery (Apium graveolens L.), Chenopodium (Chenopodium ambrosioides L. var. antihelmiticum (L.) Aellen), Ci- tronella (Cympopogon (Andropogon) nardus (L.) Rendle), 39
Cumin (Cuminum cyminum L.), Cypress (Cupressus semper- virens L.), Dwarf Pine Needles (Pinus montana Mill. (Pinus pumilio Haenke) , Eucalyptus (Eucalyptus globulus Labill and the like), Fennel (Foeniculum vulgare Mill.), Fir (Abies alba Mill. (A. picea Lindl., A. pec- tinata DC), Fir-Siberian (Abies sibirica Ledeb.), Fle- abane (Conyza canadensis (L.) Cron. (Erigeron canaden- sis L., Leptilon canadense (L.)) Britt . ) , Geranium (Pelargonium odoratissimum Ait. and the like), Geranium- East Indian (Andropogon schoenanthus L. and the like), Ginger ( Zingiber officinale Roscoe) , Hyssop (Hyssopus officinalis L.), Juniper (Juniperus comunis L.), Lavender (Lavandula officinalis Chaix (L. vera DC.)), Lemon (Citrus limonium (L.) Risso (C. medica var. limon L.), Lemon Grass (Cymbopogon (Andropogon) citratus (DC.) Stapf, and C. flexuosus (Nees) Stapf) , Levant Wormseed (Artemisia marittima var. stechmanniana Bess (A. pau- ciflora Weber), Linalσe (Bursera delpechiana Poiss. and the like), Majoram (Origanum majorana L.), Myrtle (Myr- tus comunis L.), Niaouli (Melaleuca viridiflora (So- land.) Gaertn.), Nutmeg (Myristica fragrans Houtt . ) , Orange (Citrus aurantium var. sinensis L.), Orange flowers (Citrus aurantium var. sinensis L. and the like), Origanum (Origanum vulgare L.), Pennyroyal-American (Hedeoma pulegioides (L.) Pers.), Pennyroyal-European (Mentha pulegium L.), Pepper (Piper nigrum L.), Peppermint (Mentha piperita L.), Pettigrain (Citrus vulgaris Rissop (C. bigaradia Loisel.), Pine Needles (Pinus syl- vestris L.), Poho (Mentha arvensis), Rose (Rosa gallica L. and the like), Rosemary (Rosmarinus officinalis L.), Sassafras (sassafras albidum (Nutt.) Nees (S. variifo- 40
lium (Salisb.) Kuntze: S. officinale Nees & Eberm. ) , Savin (Juniperus sabina L.), Spearmint (Mentha spicata L. (M. virdis L.), Spike (Lavandula spica L. (L. lati- folia Vill.), Sweet Bay (Laurus nobilis L.), Tansy (Ta- nacetum vulgare L.), Thyme (Thymus vulgaris L.), Turpentine (Pinus palustris Mill, and the like), Valerian (Valeriana officinalis L.), White Cedar (Thuja occiden- talis L.), Wormwood (Artemisia absinthium L.), and Yarrow (Achillea millefolium L.) or a mixture thereof.
8. The use of an essential oil, a monoterpene derived thereof, a metabolite and/or a chemically related species according to claim 7, wherein the essential oil is selected from the group consisting of, sage, rosemary, thyme, pine, dwarf-pine, turpentine, juniper and eucalyptus oil, bitter orange peel oil or mixtures thereof.
9. The use of a monoterpene or a metabolite according to any of the preceeding claims wherein the monoterpene is selected from the group consisting of ocimen, myr- cen, citronellal, linalool, geraniol, nerol, citronel- lol; monocyclic monoterpene, such as alpha-Terpinen, Li- monene, alpha-Phellandren, beta-Phellandren, p-Cymen (p-Cymol), Menthol, alpha-Terpineol, Terpinen-4-ol, Carveol, Thymol, Carvacrol, Menthon, Piperiton, Piperitenon, Pulegon, Carvon, Menthofuran, Cuminal, Sa- franal, 1,8-Cineol (Eukalyptol) , Ascaridol; or bicyclic monoterpenes, such as Sabinen, alpha- Thujen, Santen, alpha-Pinen, beta-Pinen, Camphen, 41
Caren, Sabinol, Thujol, Thujon, Myrtenol, Borneol, Cam- pher, and Fenchon or a mixture thereof.
10. The use of a monoterpene or a metabolite according to claim 9, wherein the monoterpene is selected from the group consisting of thujone, eucalyptol, camphor, borneol, α-pinene, β-pinene, bornylacetate and limonene or mixtures thereof.
11. The use of an essential oil, a monoterpene and/or metabolite derived thereof according to any of claims 7 to 10, wherein at least one essential oil according to claims 7 and 8 and at least one monoterpene or metabolite according to claims 9 and 10 is combined.
12. The use of an essential oil, a monoterpene derived thereof, a metabolite and/or a chemically related species according to any of the preceeding claims for the preparation of a nutritional composition comprising a) at least one essential oil extracted from a plant named in claim 5 or mixtures thereof; b) at least one monoterpene named in claim 7 or mixtures thereof; or c) a mixture of at least one essential oil and at least one monoterpene.
13. The use of an essential oil, a monoterpene derived thereof, a metabolite and/or a chemically related species according to claim 12, wherein the nutritional composition comprises 0.01% (w/w) to 50% (w/w), pre- 42
ferably 0.1 to 25% of said essential oil or monoterpene or mixtures thereof.
14. The use of an essential oil, a monoterpene de- rived thereof, a metabolite and/or a chemically related species according to claims 12 or 13, wherein the nutritional composition comprises suitable nutrient sources.
15. The use of an essential oil, a monoterpene derived thereof, a metabolite and/or a chemically related species according to any of the preceeding claims for the preparation of a pharmaceutical composition comprising a) at least one essential oil extracted from a plant named in claim 5 or mixtures thereof; b) at least one monoterpene named in claim 7 or mixtures thereof; c) a mixture of at least one essential oil and at least one monoterpene; and d) a pharmaceutical acceptable carrier.
16. The use of an essential oil, a monoterpene derived thereof, a metabolite and/or a chemically related species according to claim 15, wherein the pharmaceutical composition comprises 0.001% (w/w) to 100% (w/w), preferably 0,1 to 50% of said essential oil or monoterpene or mixtures thereof.
17. The use of an essential oil, a monoterpene derived thereof, a metabolite or a chemically related 43
species according to claims 15 and 16, wherein the pharmaceutical composition is prepared to be suitable for oral, topical, rectal and parenteral administration as well as by naso-gastric tubes.
18. The use of an essential oil, a monoterpene derived thereof, a metabolite and/or a chemically related species according to any of the preceeding claims for the preparation of a cosmetical composition comprising a) at least one essential oil extracted from a plant named in claim 5 or mixtures thereof; b) at least one monoterpene named in claim 7 or mixtures thereof; or c) a mixture of at least one essential oil and at least one monoterpene.
19. The use of an essential oil, a monoterpene derived thereof, a metabolite and/or a chemically related species according to claim 18, wherein the cosmetical composition comprises 0.01% (w/w) to 100% (w/w), preferably 5 to 70% (w/w) of said essential oil or mono- terpene or mixtures thereof.
20. The use of an essential oil, a monoterpene de- rived thereof, a metabolite and/or a chemically related species according to claims 18 or 19, wherein the cosmetical composition is suitable for topical applications comprising ointments, liniments or liquids.
21. A method for the preparation of a pharmaceutical, cosmetical or nutritional composition for the treatment
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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003053167A1 (en) * 2001-12-11 2003-07-03 Societe Des Produits Nestle S.A. Composition for promotion of bone growth and maintenance of bone health
FR2869230A1 (en) * 2004-04-23 2005-10-28 Alessio Patrizia D COMPOSITION FOR PREVENTING OR TREATING CELL DEGENERATION WITH AT LEAST ONE MOLECULE CAPABLE OF MAINTAINING THE REVERSIBILITY OF ADHESION MOLECULE EXPRESSION AND POLYMERIZATION OF ACTIN FIBERS
WO2012029148A1 (en) 2010-09-01 2012-03-08 株式会社オステオファーマ Freeze-dried recombinant human bone morphogenetic protein-2 preparation
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CN108601806A (en) * 2016-01-27 2018-09-28 西贝柳斯有限公司 Plant composition and application method with the Sage for extending sequential life activities

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998050054A1 (en) * 1997-05-06 1998-11-12 Muehlbauer Roman Conrad Plant extracts for the treatment of increase bone resorption
WO2000020014A1 (en) * 1998-10-05 2000-04-13 Muehlbauer Roman Conrad Plant extracts for the treatment of increased bone resorption
WO2000020015A1 (en) * 1998-10-05 2000-04-13 Muehlbauer Roman Conrad Plant extracts for the treatment of increased bone resorption

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998050054A1 (en) * 1997-05-06 1998-11-12 Muehlbauer Roman Conrad Plant extracts for the treatment of increase bone resorption
WO2000020014A1 (en) * 1998-10-05 2000-04-13 Muehlbauer Roman Conrad Plant extracts for the treatment of increased bone resorption
WO2000020015A1 (en) * 1998-10-05 2000-04-13 Muehlbauer Roman Conrad Plant extracts for the treatment of increased bone resorption

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2263480A1 (en) * 2001-12-11 2010-12-22 Société des Produits Nestlé S.A. Composition for promotion of bone growth and maintenance of bone health, comprising Mentha extracts
EP1325681A1 (en) * 2001-12-11 2003-07-09 Société des Produits Nestlé S.A. Composition for promotion of bone growth and maintenance of bone health
EP2272382A1 (en) * 2001-12-11 2011-01-12 Société des Produits Nestlé S.A. Composition for promotion of bone growth and maintenance of bone health, comprising soybean or soybean extracts
WO2003053167A1 (en) * 2001-12-11 2003-07-03 Societe Des Produits Nestle S.A. Composition for promotion of bone growth and maintenance of bone health
AU2002366734B2 (en) * 2001-12-11 2008-12-11 Societe Des Produits Nestle S.A. Composition for promotion of bone growth and maintenance of bone health
WO2005105074A3 (en) * 2004-04-23 2006-01-12 Alessio Patrizia D Composition for treating or preventing cell degeneration using at least one molecule capable of maintaining adhesion molecule expression reversibility and vascular endothelium actin fibre polymerisation
FR2869230A1 (en) * 2004-04-23 2005-10-28 Alessio Patrizia D COMPOSITION FOR PREVENTING OR TREATING CELL DEGENERATION WITH AT LEAST ONE MOLECULE CAPABLE OF MAINTAINING THE REVERSIBILITY OF ADHESION MOLECULE EXPRESSION AND POLYMERIZATION OF ACTIN FIBERS
US8912230B2 (en) 2004-04-23 2014-12-16 Aisa Therapeutics Method for treating cell degeneration using at least one molecule capable of inhibiting adhesion molecule expression and vascular endothelium actin fibre polymerization
WO2012029148A1 (en) 2010-09-01 2012-03-08 株式会社オステオファーマ Freeze-dried recombinant human bone morphogenetic protein-2 preparation
CN102688294A (en) * 2012-06-13 2012-09-26 浙江农林大学 Illicium henryi-contained anti-virus ointment for external use and preparation method and application thereof
CN102935119A (en) * 2012-11-21 2013-02-20 吉林鑫水科技开发有限公司 Method for preparing ointment for treating lumbar disc herniation
CN108601806A (en) * 2016-01-27 2018-09-28 西贝柳斯有限公司 Plant composition and application method with the Sage for extending sequential life activities
EP3407904B1 (en) * 2016-01-27 2021-08-25 Sibelius Limited Botanical compositions of salvia officinalis with activity in extending chronological lifespan and uses thereof

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