TR202005078A2 - DEVELOPMENT OF MICROEMULSION FORMULATION FROM ROSMARINUS OFFICINALIS ESSENTIAL OIL AND THE ANTIFUNGAL EFFECT OF THESE FORMULATIONS - Google Patents
DEVELOPMENT OF MICROEMULSION FORMULATION FROM ROSMARINUS OFFICINALIS ESSENTIAL OIL AND THE ANTIFUNGAL EFFECT OF THESE FORMULATIONS Download PDFInfo
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- TR202005078A2 TR202005078A2 TR2020/05078A TR202005078A TR202005078A2 TR 202005078 A2 TR202005078 A2 TR 202005078A2 TR 2020/05078 A TR2020/05078 A TR 2020/05078A TR 202005078 A TR202005078 A TR 202005078A TR 202005078 A2 TR202005078 A2 TR 202005078A2
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- Turkey
- Prior art keywords
- essential oil
- microemulsion
- agent
- pharmaceutical
- rosmarinus officinalis
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Classifications
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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
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
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- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
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- Mycology (AREA)
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Abstract
Buluş, antimikrobiyal ve antifungal özelliğine sahip, Rosmarinus officinalis uçucu yağını içeren bir farmasötik veya nutrasötik preparat/ ajan ile ilgilidir. Yenilik olarak bahsedilen farmasötik veya nutrasötik preparat/ ajan mikroemülsiyon formunda sağlanmaktadır.The invention relates to a pharmaceutical or nutraceutical preparation / agent containing the essential oil of Rosmarinus officinalis, which has antimicrobial and antifungal properties. The novel pharmaceutical or nutraceutical preparation / agent is provided in the form of a microemulsion.
Description
TARIFNAME ROSMARINUS OFFICINALIS uçucu YAGINDAN MIKROEMÜLSIYON FORMULASYONUNUN GELISTIRILMESI ve BU FORMÜLASYONLARIN ANTIFUNGAL ETKISI TEKNIK ALAN Bulus, antimikrobiyal ve antifungal özelligine sahip Rosmarinus officinalis uçucu yagini içeren bir farmasötik veya nutrasötik ajan elde edilmesi ile ilgilidir. ÖNCEKI TEKNIK Günümüzde bitkisel ürünler gibi dogal kaynaklar hastaliklarin tedavisinde oldukça popüler hale gelmistir. Bitkilerden elde edilen ürünler ve fitokimyasallar geçmisten günümüze farmasötik alanda kullanilan preparatlarin içeriginde yer almaktadir. DESCRIPTION MICROEMULSION FROM ROSMARINUS OFFICINALIS ESSENTIAL OIL IMPROVEMENT OF THE FORMULATION AND THESE FORMULATIONS ANTIFUNGAL EFFECT TECHNICAL FIELD The invention is based on Rosmarinus officinalis volatile product with antimicrobial and antifungal properties. It relates to obtaining a pharmaceutical or nutraceutical agent containing oil. PRIOR ART Today, natural resources such as herbal products are very important in the treatment of diseases. has become popular. Plant-derived products and phytochemicals from the past It is included in the content of the preparations used in the pharmaceutical field today.
Bitkisel ekstreler ve dogal maddelerden hastaliklara karsi etken madde elde etme alaninda çok sayida çalisma gerçeklestirilmekte ve Rosmarinus officinalis bitkisel ürünlerinin konvansiyonel olarak antibakteriyel, antiviral ve antifungal özellikleri açisindan tercih edildigi bilinmektedir. Ancak hastaliklara karsi antimikrobiyal etkileri arttirilmis ve daha etkili yeni preparat/ajanlarin gelistirilmesi gerekmektedir. Obtaining active substances against diseases from herbal extracts and natural substances Numerous studies are carried out in the field of Rosmarinus officinalis herbal medicine. antibacterial, antiviral and antifungal properties of its products conventionally It is known that it is preferred in terms of However, its antimicrobial effects against diseases Increased and more effective new preparations/agents need to be developed.
Kandidiyazis maya benzeri bir fungus tarafindan olusturulan bir enfeksiyondur. Candidiasis is an infection caused by a yeast-like fungus.
Bütün dünyada görülme prevalansi artmaktadir. Candida türleri mukokutanöz tutulumdan herhangi bir organi etkileyebilecek invaziv hastaliga kadar degisen genis spektrumda enfeksiyonlara yol açmaktadir. Bu enfeksiyonlar orofaringeal ve özofagal gibi mukozal tutulumdan, siklikla immünsüpresif hastalarda gözlenen daha yaygin ve derin yerlesimli internal organ tutulumlarina kadar degiskenlik gösterebilmektedir. Bu kadar genis spektrumdaki hastaliklarda genis tedavi yöntemleri gereklidir. Bu enfeksiyonlar için topikal ve sistemik tedavi yöntemleri ve profilaktik tedavi rejimleri bulunmaktadir. Candida albicans insan patojeni olan önemli birfungustur. Bununla birlikte bitki hastaliklari da tarimsal alanda son derece önemli bir faktördür ve farkli genuslara ait fitopatojenik funguslar sayisiz mahsulü enfekte ederek tarimda ekonomik kayiplara neden olmaktadir. Its prevalence is increasing all over the world. Candida species mucocutaneous wide-ranging disease ranging from involvement to invasive disease that can affect any organ spectrum of infections. These infections are oropharyngeal and from mucosal involvement, such as esophageal, more frequently observed in immunosuppressed patients. variability up to extensive and deeply located internal organ involvement can show. Broad treatment for such a broad spectrum of diseases methods are required. Topical and systemic treatment methods for these infections and There are prophylactic treatment regimens. Candida albicans is a human pathogen It is an important fungus. However, plant diseases are also extremely common in the agricultural field. is an important factor and phytopathogenic fungi belonging to different genera produce numerous crops. causes economic losses in agriculture by infecting
Son yillarda tüm dünyada çiftçiler, bitki hastaliklarini kontrol etmekte geleneksel olarak kullanilan fungisitlerin etkinliginde azalma oldugunu bildirmislerdir. Ayni zamanda fungisitlerin uygun olmayan kullanimlari da fungal dirençliliginin artmasina neden olmus ve güncel olarak kullanilan fungisitler, benzimidazol ve dikarbomozitler gibi konvensiyonel antifungallere karsi dirençlilik gelismistir. Bu dirençliligin üstesinden gelmek için güncel kontrol stratejilerinin yerine geçebilecek yeni antifungal ajanlarin kesfedilmesi son derece önem arz etmektedir. Bu nedenlerle önemli bitki patojenleri olan Fusarium moniliforme, Fusarium culmorum, Botrytis cinerea gibi funguslar ve Candida albicans basta olmak üzere mikrobiyal patojenlere karsi yeni aktif ajan gelistirilmesi gerekmektedir. In recent years, farmers all over the world have been using traditional methods to control plant diseases. They reported that there was a decrease in the effectiveness of fungicides used as Same At the same time, improper use of fungicides also contributes to the increase of fungal resistance. caused and currently used fungicides, benzimidazole and dicarbomosides Resistance to conventional antifungals such as This resilience new control strategies that can replace current control strategies to overcome The discovery of antifungal agents is extremely important. For these reasons important plant pathogens, Fusarium moniliforme, Fusarium culmorum, Botrytis to microbial pathogens, mainly fungi such as cinerea and Candida albicans. It is necessary to develop a new active agent against it.
Sonuç olarak, yukarida bahsedilen tüm sorunlar, ilgili teknik alanda bir yenilik yapmayi zorunlu hale getirmistir. As a result, all the above-mentioned problems are a novelty in the relevant technical field. does not make it mandatory.
BULUSUN KISA AÇIKLAMASI Mevcut bulus yukarida bahsedilen dezavantajlari ortadan kaldirmak ve ilgili teknik alana yeni avantajlar getirmek üzere, bir farmasötik veya nutrasötik preparat/ ajan üretimi ile ilgilidir. BRIEF DESCRIPTION OF THE INVENTION The present invention is designed to eliminate the above mentioned disadvantages and a pharmaceutical or nutraceutical preparation/agent to bring new advantages to the field relates to production.
Bulusun bir amaci antimikrobiyal ve antifungal özelligine sahip bir farmasötik veya nutrasötik preparat/ ajan ortaya koymaktir. An object of the invention is a pharmaceutical or pharmaceutical product with antimicrobial and antifungal properties. to reveal a nutraceutical preparation/agent.
Bulusun bir amaci antimikrobiyal ve antifungal özelligine sahip bir mikroemülsiyon formda farmasötik veya nutrasötik preparat/ ajan üretimi ortaya koymaktir. An object of the invention is a microemulsion with antimicrobial and antifungal properties. to produce pharmaceutical or nutraceutical preparations/agents in the form of
Yukarida bahsedilen ve asagidaki detayli anlatimdan ortaya çikacak tüm amaçlari gerçeklestirmek üzere mevcut bulus, bir farmasötik veya nutrasötik preparat/ ajan üretimi ile ilgilidir. Buna göre bahsedilen farmasötik veya nutrasötik preparat/ ajan, Rosmarinus officinalis uçucu yagini içermektedir. Bu sayede antimikrobiyal ve antifungal özelligine sahip birfarmasötik veya nutrasötik ajan elde edilebilmektedir. All the above-mentioned purposes that will emerge from the detailed description below. present invention, a pharmaceutical or nutraceutical preparation/agent relates to production. Accordingly, the pharmaceutical or nutraceutical preparation/agent mentioned, Contains Rosmarinus officinalis essential oil. In this way, antimicrobial and A pharmaceutical or nutraceutical agent with antifungal properties can be obtained.
Bulusun mümkün yapilanmasinda, Rosmarinus officinalis uçucu yagina sahip farmasötik veya nutrasötik ajan, mikroemülsiyon formunda olmaktadir. Bu sayede uçucu yagin mikrobiyal inhibisyonunun arttirilmasi saglanmaktadir. In the possible embodiment of the invention, the essential oil of Rosmarinus officinalis The pharmaceutical or nutraceutical agent is in the form of a microemulsion. In this way It is provided to increase the microbial inhibition of essential oil.
Bulus, ayrica Rosmarinus officinalis uçucu yagini içeren bitkilerin. The invention also includes plants containing essential oil of Rosmarinus officinalis.
. Kurutulmasi, o Toz edilmesi o Distilasyonla uçucu yaginin elde edilmesi islem adimlarini içeren bir mikroemülsiyon formda farmasötik veya nutrasötik preparat/ ajan üretimi ile ilgilidir. Bahsedilen mikroemülsiyon formunda farmasötik veya nutrasötik preparat/ ajan üretimi asagidaki; 0 Agirlikça %O,1-5 araligindaki oranda uçucu yag maddesinin, agirlikça karistirilmasi, o Üzerine agirlikça %5-30 araligindaki oranlarda sürfaktan madde olarak; Span 60 ve Kromofor EL maddelerinden seçilen bir maddenin veya ikisinin kombinasyonunun ilave edilmesi, . kosürfaktan olarak agirlikça %35-50 araliginda bir oranda etanol ve su olarak distile su maddelerinden biri veya her ikisinin kombinasyonunun ilave edilmesi o hazirlanan mikroemülsiyon formülasyonunun karistirilmasi islem adimlari ile istenilen mikroemülsiyon formülasyonunun elde edilmesi ile karakterize edilmektedir. Böylece antimikrobiyal ve antifungal özelligine sahip mikroemülsiyon formunda farmasötik veya nutrasötik ajanin üretim metodu saglanmaktadir. . drying, o Dusting o Obtaining essential oil by distillation pharmaceutical or nutraceutical in the form of a microemulsion containing the processing steps relates to preparation/agent production. Pharmaceutical in the aforementioned microemulsion form or nutraceutical preparation/agent production as follows; 0 by weight of essential oil in the range of 0.1-5% by weight mixing, o Surfactant in the ratio of 5-30% by weight aspect; A substance selected from Span 60 and Chromophore EL or the addition of a combination of the two, . ethanol and water as cosurfactant in the range of 35-50% by weight distilled water, or a combination of both. adding o Mixing the prepared microemulsion formulation By obtaining the desired microemulsion formulation with the process steps is characterized. Thus, it has antimicrobial and antifungal properties. Production method of pharmaceutical or nutraceutical agent in microemulsion form is provided.
Bulusun mümkün yapilanmasinda, Rosmarinus officinalis bitkisinden elde edilen uçucu yagin mikroemülsiyon formülasyonu antimikrobiyal ve antifungal preparat/ ajan olarak kullanilmaktadir. In the possible embodiment of the invention, it is obtained from the plant Rosmarinus officinalis. essential oil microemulsion formulation antimicrobial and antifungal preparation/ used as an agent.
SEKILIN KISA AÇIKLAMASI Sekil 1' de uçucu yag ve mikroemülsiyon formülasyonunun karsilastirmali % miselyal büyüme oranlarina dair Poisoned PDA teknigi ile gerçeklestirilen test sonuçlarina iliskin grafik verilmistir. BRIEF DESCRIPTION OF THE FIGURE In Figure 1, the comparison of essential oil and microemulsion formulation % mycelial the test results with the Poisoned PDA technique on growth rates. A related graph is given.
Sekil 2' de saf uçucu yagin, Fusarium culmorum patojenine karsi poisoned PDA teknigi ile fungusun miselyal büyümesi ölçülerek gerçeklestirilen antifungal aktivite deney sonucu verilmistir. Figure 2 shows pure essential oil, poisoned PDA against Fusarium culmorum pathogen. Antifungal activity by measuring the mycelial growth of the fungus with the technique test results are given.
Sekil 3' de mikroemülsiyon formülasyonunun, Fusarium culmorum patojenine karsi poisoned PDA teknigi ile fungusun miselyal büyümesi ölçülerek gerçeklestirilen antifungal aktivite deney sonucu verilmistir. Figure 3 shows the microemulsion formulation against the pathogen Fusarium culmorum. by measuring the mycelial growth of the fungus with the poisoned PDA technique. The antifungal activity was given as a result of the experiment.
Sekil 4' de saf uçucu yagin, Botrytis cinerea patojenine karsi poisoned PDA teknigi ile fungusun miselyal büyümesi ölçülerek gerçeklestirilen antifungal aktivite deney sonucu verilmistir. Figure 4 shows pure essential oil, poisoned PDA technique against Botrytis cinerea pathogen. antifungal activity assay by measuring the mycelial growth of the fungus with result is given.
Sekil 5' de mikroemülsiyon formülasyonununi Botrytis cinerea patojenine karsi poisoned PDA teknigi ile fungusun miselyal büyümesi ölçülerek gerçeklestirilen antifungal aktivite deney sonucu verilmistir. Figure 5 shows the microemulsion formulation against Botrytis cinerea pathogen. by measuring the mycelial growth of the fungus with the poisoned PDA technique. The antifungal activity was given as a result of the experiment.
Sekil 6” da saf uçucu yagin, Fusarium muniliforme patojenine karsi poisoned PDA teknigi ile fungusun miselyal büyümesi ölçülerek gerçeklestirilen antifungal aktivite deney sonucu verilmistir. Figure 6” shows pure essential oil, poisoned PDA against Fusarium muniliforme pathogen. Antifungal activity by measuring the mycelial growth of the fungus with the technique test results are given.
Sekil 7' de mikroemülsiyon formülasyonunun, Fusarium muniliforme patojenine karsi poisoned PDA teknigi ile fungusun miselyal büyümesi ölçülerek gerçeklestirilen antifungal aktivite deney sonucu verilmistir. Figure 7 shows the microemulsion formulation against the pathogen Fusarium muniliforme. by measuring the mycelial growth of the fungus with the poisoned PDA technique. The antifungal activity was given as a result of the experiment.
BULUSUN DETAYLI AÇIKLAMASI Bu detayli açiklamada bulus konusu antimikrobiyal ve antifungal özelliklere sahip bir farmasötik veya nutrasötik preparat/ ajan ile ilgili olup özelligi; sadece konunun daha iyi anlasilmasina yönelik hiçbir sinirlayici etki olusturmayacak örneklerle açiklanmaktadir. DETAILED DESCRIPTION OF THE INVENTION In this detailed description, the subject of the invention is a pesticide with antimicrobial and antifungal properties. It is related to a pharmaceutical or nutraceutical preparation/agent and its feature is; more than just the subject with examples that will not have any limiting effect for better understanding. is explained.
Bahsedilen farmasötik veya nutrasötik preparat/ ajan bünyesinde Rosmarinus officinalis bitkisinden elde edilen uçucu yagi bulunmaktadir. Bulus konusu uçucu yag tercihen Rosmarinus officinalis yapraklarindan elde edilmektedir. Rosmarinus within the aforementioned pharmaceutical or nutraceutical preparation/agent It has essential oil obtained from officinalis plant. The subject of the invention is essential oil preferably obtained from the leaves of Rosmarinus officinalis.
Bulusta elde edilen farmasötik veya nutrasötik preparat/ ajan mikroemülsiyon formundadir. Bu sayede mikroemülsiyon formunda elde edilen ajanin vücutta hizli ve yüksek verimde emilimi saglanmaktadir. Mikroemülsiyon formülasyonu kontrollü ilaç salinimi saglamasi, ilaçlarin sistemik ve topikal emilimlerini arttirmasi gibi çesitli avantajlarindan dolayi gelistirilmektedir. Mikroemülsiyonlar, diger ilaç sekillerine göre oldukça avantajli sistemlerdir. Mikroemülsiyon formülasyonu ile ilacin emilimi tablet, kapsül gibi diger ilaç sekillerine göre çok daha yüksek ve hizli olmaktadir. Ayrica, yapisinda bulunan yüzey etkin madde sayesinde membran akiskanligini, dolayisiyla permeabiliteyi arttirmakta, böylece ilaç emilimi artmaktadir. Pharmaceutical or nutraceutical preparation/agent microemulsion obtained in the invention is in the form. In this way, the agent obtained in the form of microemulsion is rapidly absorbed in the body. and high efficiency absorption is provided. Microemulsion formulation controlled It provides drug release, increases systemic and topical absorption of drugs. developed for its advantages. Microemulsions compared to other drug forms They are very advantageous systems. Absorption tablet with microemulsion formulation, It is much higher and faster than other forms of medicine such as capsules. Moreover, Thanks to the surfactant in its structure, the membrane fluidity, therefore, increases permeability, thus increasing drug absorption.
Bulusta yer alan farmasötik veya nutrasötik preparat/ ajan, Rosmarinus officinalis bitkilerinden elde edilen yapraklarin kurutulmasi ve toz haline getirilmesi islemleri sonrasinda, uçucu yagin mikroemülsiyon formülasyonu haline getirilmektedir. The pharmaceutical or nutraceutical preparation/agent of the invention, Rosmarinus officinalis The processes of drying and pulverizing the leaves obtained from the plants Afterwards, the essential oil is turned into a microemulsion formulation.
Bahsedilen mikroemülsiyon form bir yagli faz içeren bilesen, bir sulu faz içeren bilesen, bir veya iki surfaktan ve bir kosurfaktan bilesen içermektedir. Said microemulsion form component containing an oily phase, an aqueous phase containing The component includes one or two surfactants and a cosurfactant component.
Mikroemülsiyon formda farmasötik veya nutrasötik preparat/ ajan üretimi asagidaki; i. Toz haline getirilen bitkiden uçucu yagin elde edilmesi, ii. (i) adiminda elde edilen uçucu yagin, baska bir yag ile karistirilmasi, iii. Kosürfaktan, sürtaktan ve suyun ilave edilmesi islem adimlarini içermesi ile karakterize edilmektedir. (ii) adiminda bahsedilen yag olarak belli agirlikça oranlarda bitkisel yaglar, sentetik yaglar, trigliseritler, yag asitlerinin esterlerinden en az bir veya belli oranlarda karisimlari olabilmektedir. Bulusun mümkün bir yapilanmasinda yag fazi olarak isopropil miristat tercih edilmistir. Böylece, toksik olmayan, biyolojik olarak kullanimi uygun olan bir yag kullanilmistir. (iii) adiminda bahsedilen sulu faz olarak belli agirlikça oranlarda su, sodyum klorür çözeltisi, tamponlar, propilenglikol, polietilen glikoller veya bunlarin kombinasyonlari kullanilabilmektedir. Bulusun mümkün bir yapilanmasinda sulu faz olarak Rosmarinus officinalis hidrosolü kullanilmistir. (iii) adiminda bahsedilen kosurfaktan madde olarak belli agirlikça oranlarda alkoller, glikollerin türevleri, poligliseroller ve propilen glikollerden en az birinin kullanilmasidir. Production of pharmaceutical or nutraceutical preparation/agent in microemulsion form is as follows; I. Obtaining essential oil from the powdered plant, ii. mixing the essential oil obtained in step (i) with another oil, iii. Addition of cosurfactant, surfactant and water It is characterized by including process steps. (ii) as the oil mentioned in step (ii) vegetable oils in certain weight ratios, synthetic fats, triglycerides, esters of fatty acids at least one or in certain proportions may have mixtures. In a possible embodiment of the invention, as the oil phase isopropyl myristate is preferred. Thus, non-toxic, biological use A suitable oil is used. As the aqueous phase mentioned in step (iii) certain weight ratios of water, sodium chloride solution, buffers, propylene glycol, polyethylene glycols or combinations thereof can be used. As the aqueous phase in one possible embodiment of the invention Rosmarinus officinalis hydrosol was used. Alcohols in certain weight ratios as cosurfactant substance mentioned in step (iii), derivatives of glycols, polyglycerols and at least one of propylene glycols.
Bulusun mümkün bir yapilanmasinda kosurfaktan olarak etanol kullanilmistir. In one possible embodiment of the invention, ethanol is used as the cosurfactant.
Böylece formülasyonda hem çözücü, hem koruyucu özellik saglanmistir. (iii) adiminda bahsedilen surfaktan madde olarak belli agirlikça oranlarda nan-iyonik ve iyonik surfaktanlar, lesitin, poligliserol, yag asidi esterleri kullanilabilmektedir. Thus, both solvent and preservative properties are provided in the formulation. As the surfactant mentioned in step (iii) certain weight ratios of nano-ionic and ionic surfactants, lecithin, polyglycerol, fatty acid esters can be used.
Bulusun mümkün bir yapilanmasinda, sürfaktan madde olarak span 60 ve kromofor EL kullanilmistir. Span 60 ve Kromofor-EL toksik olmayan maddelerden olup formülasyonda tercih edilmistir. In one possible embodiment of the invention, span 60 and chromophore are used as surfactants. EL is used. Span 60 and Chromophore-EL are non-toxic materials. formulation is preferred.
Bulusun mümkün bir yapilanmasinda bahsedilen Rosmarinus officinalis uçucu yagi içeren mikroemülsiyon formunda farmasötik veya nutrasötik preparat/ ajan üretimi asagidaki; Agirlikça %0,1- 5 araligindaki oranda uçucu yag maddesinin, agirlikça karistirilmasi, . üzerine agirlikça %5-50 araligindaki oranlarda sürfaktan madde olarak; Span 60 ve Kromofor EL maddelerinden seçilen bir maddenin veya ikisinin kombinasyonunun ilave edilmesi, i kosürfaktan olarak agirlikça %25-50 araliginda bir oranda etanol ve su olarak distile su maddelerinden biri veya her ikisinin kombinasyonunun ilave edilmesi o hazirlanan mikroemülsiyon formülasyonunun karistirilmasi islem adimlari ile istenilen mikroemülsiyon formülasyonunun elde edilmesi prosesi ile saglanmaktadir. Rosmarinus officinalis essential oil mentioned in a possible embodiment of the invention Production of pharmaceutical or nutraceutical preparation/agent in microemulsion form containing the following; Essential oil at a rate of 0.1-5% by weight, by weight mixing, . as a surfactant in the range of 5-50% by weight; A substance selected from Span 60 and Chromophore EL or adding a combination of the two, i 25-50% by weight ethanol and water as cosurfactant distilled water, or a combination of both. adding o Mixing the prepared microemulsion formulation The process of obtaining the desired microemulsion formulation with processing steps is provided with.
Bulusun mümkün bir yapilanmasinda Rosmarinus officinalis mikroemülsiyon formülasyonunda; 100 gr mikroemülsiyon formülasyonunda Rosmarinus officinalis uçucu yagi agirlikça %0,1-%5 araliginda, isopropil miristat 5 gr- 45 gr araliginda, gr araliginda, distile su 0.5 gr- 20 gr araliginda bir degerde kullanilarak mikroemülsiyon hazirlanmistir. In one possible embodiment of the invention, Rosmarinus officinalis microemulsion in its formulation; Rosmarinus officinalis in 100 g microemulsion formulation essential oil is in the range of 0.1%-5% by weight, isopropyl myristate is in the range of 5 gr- 45 gr, g, by using distilled water at a value between 0.5 gr and 20 gr. microemulsion was prepared.
R. officinalis toprak üstü kisimlari 2018 yilinda Beykoz/Istanbuldan toplanmis ve gölgede uygun sartlar altinda kurutulmustur. Herbaryum örnekleri Istanbul Medipol Üniversitesi Herbaryumunda saklanmaktadir. Kurutulan örnekler toz haline getirilerek, Clevenger cihazinda R. officinalis uçucu yagi elde edilmistir. The above-ground parts of R. officinalis were collected from Beykoz/Istanbul in 2018 and dried under suitable conditions in the shade. Herbarium specimens Istanbul Medipol It is stored in the Herbarium of the University. The dried samples are turned into powder. R. officinalis essential oil was obtained in the Clevenger device.
Rosmarinus officinalis uçucu yagi içeren mikroemülsiyon formülasyonu; baska bir yag ve Rosmarinus officinalis uçucu yagi tartilip üzerine sürfaktan ve cosurfaktan ilave edilip en son su ilavesi ile karistirilip hazirlanmasi ile elde edilmistir. Microemulsion formulation containing Rosmarinus officinalis essential oil; another oil and Rosmarinus officinalis essential oil were weighed and added surfactant and cosurfactant. It is obtained by adding water and mixing it with the last addition of water.
Mikroemülsiyon formülasyonunu olusturabilmek için titrasyon metodu kullanilmistir. Titration method was used to form the microemulsion formulation.
Yag, sürfaktan ve kosürfaktan karisimlari 25°C'de 750 rpmide hidrosol kullanilarak titre edilmistir. Faz diyagrami olusturmak Için Triangle Phase Diagram Analysis Software kullanilmistir. Mikroemülsiyon yag fazi olarak isopropil miristat kullanilarak formüle edilmis ve sürfaktan olarak span 60 ve kromofor EL kullanilirken kosürfaktan olarak etanol tercih edilmis ve su olarak distile su tercih edilmistir. R. officinalis uçucu yagi formülasyon içerisindeki yag ile karistirilarak çözünmüs ve karistirilarak eklenmistir. Çalismada Rosmarinus officinalis yapraklarindan elde edilen uçucu yaglarin mikroemülsiyon formülasyonlarinin C. albicans, B. Cinerea ve farkli Fusarium türlerine karsi antifungal etkinligi tespit edilmistir. Mixtures of oil, surfactant and cosurfactant using hydrosol at 750 rpm at 25°C. titrated. Triangle Phase Diagram Analysis to create a phase diagram software has been used. Microemulsion using isopropyl myristate as oil phase. formulated and using span 60 and chromophore EL as surfactants, cosurfactant Ethanol was preferred as the water and distilled water was preferred as the water. R. officinalis volatile The oil is dissolved by mixing with the oil in the formulation and mixed. has been added. In the study, essential oils obtained from the leaves of Rosmarinus officinalis microemulsion formulations of C. albicans, B. Cinerea and different Fusarium Its antifungal activity has been determined against species.
R. officinalis uçucu yagi ve R. officinalis mikroemülsiyon formülasyonunun karsilastirmali olarak önemli bitki patojenleri olan Fusarium moniliforme NRRL 2374, Fusarium culmorum NRRL 3288 ve Botrytis cinerea AHU 9424 suslarina karsi miselyal büyümeyi inhibe edici aktivitesi incelenmistir. R. officinalis essential oil and R. officinalis microemulsion formulation comparatively important plant pathogens, Fusarium moniliforme NRRL 2374, against Fusarium culmorum NRRL 3288 and Botrytis cinerea AHU 9424 strains. Mycelial growth inhibitory activity was studied.
Candida albicans ATCC 64548 susu RPMI broth içerisinde çogaltilmis ve antifungal aktivite deneyleri mikrodilüsyon yöntemiyle yapilmistir. Bitki patojeni funguslarin antifungal aktivitesi ise miselyal büyümeleri ölçülerek gerçeklestirilmistir. Bunun için Poisoned PDA teknigi kullanilmistir. Candida albicans strain ATCC 64548 amplified in RPMI broth and antifungal Activity experiments were carried out by microdilution method. Plant pathogenic fungi its antifungal activity was carried out by measuring mycelial growth. For this Poisoned PDA technique was used.
Tablo 1'de R. officinalis uçucu yaginin maddedeki farkli bilesiklerin tespit edilmesi amaciyla bir analitik kimya yöntemi olan Gaz Kromatografisi - Kütle Spektrometresi (GC-MS) ile gerçeklestirilen analiz sonuçlari verilmistir. R. officinalis uçucu yaginin RRI Compound % 1014 Tricyclene 0.1 1032 oi-Pinene 6.5 1072 oi-Fenchene tr 1076 Camphene 1.5 1118 ß-Pinene 0.4 1159 ö-3-Carene 1.1 1174 Myrcene 0.9 1176 oi-Phellandrene 0.1 1183 p-Mentha-1,7(8)-diene (=Pseudolim0nene) tr 1 187 o-Cymene 0.1 1188 oi-Terpinene 0.2 Limonene 1,8-Cineole (2)-3-Hexenal 2-Pentyl furan (Z)-ß-Ocimene v-Terpinene 3-Octan0ne (E)-ß-Ocimene p-Cymene Terpinolene (Z}3-Hexen0l Filifolone trans-Linalool oxide (Furanoid) a,p-Dimethylstyrene 1-Octen-3-ol cis-Linalool oxide (Furanoid) oi-Ylangene oi-Copaene oi-Campholene aldehyde Camphor Pinocamphone Benzaldehyde Linalool trans-p-Menth-Z-en-1-0I Pinocarvone Bornyl acetate Terpinen-4-0I ß-Caryophyllene cis-p-Menth-Z-en-1-0I cis-Verbenol trans-Pinocarveol trans-Verbenol a-Humulene p-Mentha-1,8-dien-4-0I (=Lim0nen-4-0I) y-Muurolene a-Terpineol Borneol Verbenone Carvone ö-Cadinene a-Campholene alcohol p-Mentha-1(7),5-dien-2-0I (E)-ß-Damascen0ne trans-Carveol p-Cymen-8-0I lsopiperitenone (E)-Geranyl acetone cis-Myrtanol trans-Myrtanol oi-Calacorene 1949 Piperitenone 0.2 2008 Caryophyllene oxide 0.6 2029 Perilla alcohol 0.1 2030 Methyl eugenol 0.2 2045 Humulene epoxide-I tr 2071 Humulene epoxide-II 0.2 2073 ß-Caryophyllene alcohol tr 2088 1-epi-Cubenol 0.1 2186 Eugenol 0.1 2187 T-Cadinol 0.1 2209 T-Muurolol tr 2211 Clovenol 0.1 2239 Carvacrol 0.1 2232 oi-Bisabolol 0.1 2255 a-Cadinol 0.1 (=Cary0phylladien0l ll) -Methyl jasmonate 0.1 2384 Farnesyl acetone tr 2931 Hexadecanoic acid tr Oxygenated Monoterpenes 49.2 Sesquiterpene Hydrocarbones 12.4 Oxygenated Sesquiterpenes 2.3 Fatty acids tr Others 1.4 RRI Relative retention indices calculated against n-alkanes tr Trace (< 0.1 %) Tablo 1. R. Officinalis uçucu yaginin GC-MS analiz sonuçlari Asagida verilen Tablo 2'de ise R. offioinalis uçucu yaginin ve mikroemülsiyon formülasyonunun eseyli çogalan, diploit, maya tipi bir mantar türü olan ve insanlarda oral ve vajinal firsatçi enfeksiyonlarin etmeni olan C. albicans mantari üzerinde antimikrobiyal özellikleri belirtilmistir. Determination of different compounds in the substance of R. officinalis essential oil in Table 1 Gas Chromatography - Mass Spectrometry, which is an analytical chemistry method for The results of the analysis performed with (GC-MS) are given. R. officinalis essential oil RRI Compound % 1014 Tricyclene 0.1 1032 oi-Pinene 6.5 1072 oi-Fenchene en 1076 Camphene 1.5 1118 ß-Pinene 0.4 1159 d-3-Carene 1.1 1174 Myrcene 0.9 1176 oi-Phellandrene 0.1 1183 p-Mentha-1,7(8)-diene (=Pseudolim0nene) tr 1 187 o-Cymene 0.1 1188 oi-Terpinene 0.2 Limonene 1,8-Cineole (2)-3-Hexenal 2-Pentyl furan (Z)-ß-Ocimene v-Terpinene 3-Octan0ne (E)-ß-Ocimene p-Cymene terpinolene (Z}3-Hexen0l filifolone trans-Linalool oxide (Furanoid) a,p-Dimethylstyrene 1-Octen-3-ol cis-Linalool oxide (Furanoid) oi-Ylangene oi-Copaene oi-Campholene aldehyde Camphor pinocamphone Benzaldehyde linalool trans-p-Menth-Z-en-1-0I pinocarvone bornyl acetate Terpinen-4-0I ß-Caryophyllene cis-p-Menth-Z-en-1-0I cis-Verbenol trans-Pinocarveol trans-Verbenol a-Humulene p-Mentha-1,8-diene-4-0I (=Lim0nen-4-0I) y-Muurolene a-Terpineol Borneo verbena carvone d-Cadinene a-Campholene alcohol p-Mentha-1(7),5-diene-2-0I (E)-ß-Damascen0ne trans-Carveol p-Cymen-8-0I lsopiperitenone (E)-Geranyl acetone cis-Myrtanol trans-Myrtanol oi-Calacorene 1949 Piperitenone 0.2 2008 Caryophyllene oxide 0.6 2029 Perilla alcohol 0.1 2030 Methyl eugenol 0.2 2045 Humulene epoxide-I tr 2071 Humulene epoxide-II 0.2 2073 ß-Caryophyllene alcohol tr 2088 1-epi-Cubenol 0.1 2186 Eugenol 0.1 2187 T-Cadinol 0.1 2209 T-Muurolol en 2211 Clovenol 0.1 2239 Carvacrol 0.1 2232 oi-Bisabolol 0.1 2255 a-Cadinol 0.1 (=Cary0phylladien0l ll) -Methyl jasmonate 0.1 2384 Farnesyl acetone en 2931 Hexadecanoic acid en Oxygenated Monoterpenes 49.2 Sesquiterpene Hydrocarbones 12.4 Oxygenated Sesquiterpenes 2.3 Fatty acids en Others 1.4 RRI Relative retention indices calculated against n-alkanes tr Trace (< 0.1 %) Table 1. GC-MS analysis results of R. Officinalis essential oil Table 2 below shows that R. offioinalis essential oil and microemulsion which is a sexually reproducing, diploid, yeast type fungus of its formulation and on C. albicans fungus, which is the causative agent of oral and vaginal opportunistic infections. antimicrobial properties are noted.
Madde 0. albicans (ug I mL) Uçucu Yag 62-5 Mikroemülsyion Formülasyonu 15-5 Ketokonazol 15'6 Tablo 2. R. officinalis uçucu yaginin ve mikroemülsiyon formülasyonunun antimikrobiyal aktivitesi (MIK degerleri) Sonuçlar karsilastirmali olarak incelendiginde mikroemülsiyon formülasyonunun C. albicans'a karsi antimikrobiyal etkinligi MIK degeri olarak verilmistir. MIK degeri; minimal inhibitör konsantrasyonu olup hücre sayisinin artisini engelleyen en düsük antimikrobiyal madde konsantrasyonu olarak degerlendirilmektedir. Tablo 2'de verilen R. officinalis uçucu yaginin ve mikroemülsiyon formülasyonunun antimikrobiyal aktivite sonuçlari incelendiginde MIK olarak mikroemülsiyon formülasyonunun saf uçucu yaga göre C. albiçans'a karsi oldukça anlamli sekilde artmis oldugu görülmektedir. Saf uçucu yagin C. albicans'a karsi MIK degeri 62.5 ug / mL iken mikroemülsiyon formilasyonu haline getirildiginde 15.6 iJg / mL olarak bulundugu görülmektedir. Dolayisi ile mikroemülsiyon formulasyonu haline getirilmis Rosmarinus officinalis maddesi ile uçucu yaga göre çok daha etkili bir antimikrobiyal antifungal ajan saglanmaktadir. Saf R. officinalis uçucu yaginin antikandidal aktivitesi çalisilmis olmasina ragmen bu örnekte görüldügü üzere mevcut bulus ile uçucu yag mikroemülsiyon formülasyonu haline getirilerek MIK degeri 4 kat arttirilmistir ve antikandidal bir preparat/ ajan olarak kullanim için daha etkili bir formülasyon haline getirilmistir. Item 0. albicans (µg I mL) Essential Oil 62-5 Microemulsion Formulation 15-5 Ketoconazole 15'6 Table 2. R. officinalis essential oil and microemulsion formulation antimicrobial activity (MIK values) When the results were examined comparatively, the microemulsion formulation was C. The antimicrobial activity against albicans was given as the MIC value. MIC value; has a minimal inhibitory concentration and is the lowest that inhibits the increase in cell number. evaluated as the antimicrobial substance concentration. in Table 2 of R. officinalis essential oil and microemulsion formulation given When the antimicrobial activity results are examined, microemulsion as MIC significantly significantly against C. albicans based on pure essential oil. appears to have increased. MIC value of pure essential oil against C. albicans 62.5 ug / mL when it is turned into microemulsion formulation as 15.6 iJg / mL appears to exist. Therefore, it has been turned into a microemulsion formulation. Rosmarinus officinalis is a much more effective antimicrobial than essential oil. Antifungal agent is provided. Anticandidal essential oil of pure R. officinalis As seen in this example, although the activity of the present invention has been studied, The essential oil is turned into a microemulsion formulation and its MIC value is 4 times. increased and a more effective alternative for use as an anticandidal preparation/agent. has been formulated.
Bulusun mümkün bir yapilanmasinda R. officinalis uçucu yaginin ve mikroemülsiyon formülasyonunun bitki patojenlerine karsi miselyal büyüme inhibisyonu da karsilastirmali olarak tespit edilmistir. Sekil 1ide verildigi üzere yaklasik %50 oraninda bir seçici inhibisyon saglandigi görülmektedir. F. culmorum, B. cinerea, ve F. muniliforme patojenlerinin saf uçucu yag kullanildiginda miselyal büyümeleri sirasiyla 50%, 33%, ve 42% iken mikroemülsiyon seklinde formüle edilmis uçucu yag kullanildiginda bu oranin sirasiyla 12%, 15%, ve 33% e geriledigi görülmüstür. In one possible embodiment of the invention, essential oil of R. officinalis and microemulsion formulation also inhibits mycelial growth against plant pathogens. determined by comparison. Approximately 50% as given in Figure 1 A selective inhibition appears to be achieved. F. culmorum, B. cinerea, and Mycelial growth of F. muniliforme pathogens when pure essential oil is used. essential oil formulated as microemulsion at 50%, 33%, and 42%, respectively used, this rate decreased to 12%, 15%, and 33%, respectively.
Sonuçlarda tespit edildigi üzere R. officinalis uçucu yaginin mikroemülsiyon formülasyonu uçucu yaga göre çok daha etkili antimikrobiyal ve antifungal özelliklere sahip bir farmasötik, nutrasötik preparat/ ajan veya pestisit olarak kullanilabilecektir. As determined in the results, the microemulsion of R. officinalis essential oil Its formulation has much more effective antimicrobial and antifungal properties than essential oil. It can be used as a pharmaceutical, nutraceutical preparation/agent or pesticide.
Bitki patojeni funguslara karsi mikroemülsiyon formülasyonunun ve saf uçucu yagin antifungal aktivitesi poisoned PDA teknigi ile funguslarin miselyal büyümeleri ölçülerek arastirilmistir. Bunun için besiyeri ile madde karistirilarak petrilere dökülmüstür. Ardindan donmus ve ilaçla muamele edilmis besiyerine taze gelismis fungus ortamindan 4 mm çapinda alinarak petri kabinin ortasina yerlestirilmistir. Bir hafta boyu Inkübasyona birakilarak bir hafta sonunda funguslarin miselyal büyüme çaplari ölçülmüstür. Miselyal büyümeyi en çok inhibe eden maddenin bitki patojenlerine karsi antifungal ajan olarak kullanilabilecegi düsünülmüstür. Miselyal büyüme inhibisyonu söz konusu fungusun yayilmasinin önlenmesi anlami tasimaktadir. Pozitif kontrol olarak ketokonazol kullanilirken negatif kontrol olarak DMSO (Dimetil sülfoksit) kullanilmistir. Ketokonazol de miselyal büyüme hiç olmazken DiVISO da miselyal büyüme %100 olarak belirlenmistir. Microemulsion formulation and pure essential oil against plant pathogenic fungi Mycelial growth of fungi by poisoned PDA technique with antifungal activity measured and investigated. For this, the medium is mixed with the substance and poured into petri dishes. spilled. Then freshly grown on frozen and drug-treated medium The fungus was taken from the medium with a diameter of 4 mm and placed in the middle of the petri dish. A After a week of incubation, the mycelial growth of the fungi diameters were measured. The substance that most inhibits mycelial growth It is thought that it can be used as an antifungal agent against pathogens. mycelial growth inhibition means preventing the spread of the fungus in question carries. When using ketoconazole as a positive control, as a negative control DMSO (Dimethyl sulfoxide) was used. No mycelial growth in ketoconazole. While not, mycelial growth was determined as 100% in DiVISO.
Sekil 2 ve Sekil 3'te görüldügü üzere R. officinalis mikroemülsiyon formülasyonunun, R. officinalis saf uçucu yagina göre, Fusarium culmorum patojenine karsi daha etkili antifungal aktivite gösterdigi ve R. officinalis mikroemülsiyon formülasyonunun, saf R. officinalis uçucu yagina göre fungus miselyal büyümesini önemli ölçüde inhibe ettigi tespit edilmistir. As seen in Figure 2 and Figure 3, R. officinalis microemulsion formulation, It is more effective against Fusarium culmorum pathogen than R. officinalis pure essential oil. showed antifungal activity and R. officinalis microemulsion formulation, pure Significantly inhibited fungal mycelial growth compared to R. officinalis essential oil. it has been detected.
Sekil 4 ve Sekil 5'de görülebilecegi üzere R. officinalis mikroemülsiyon formülasyonunun, R. officinalis saf uçucu yagina göre, Botrytis cinerea patojenine karsi daha etkili antifungal aktivite gösterdigi tespit edilmistir. As can be seen in Figure 4 and Figure 5, R. officinalis microemulsion formulation, according to R. officinalis pure essential oil, against Botrytis cinerea pathogen. It was determined that it showed more effective antifungal activity against
Sekil 6 ve Sekil 7'de verildigi üzere R. officinalis mikroemülsiyon formülasyonunun, R. officinalis saf uçucu yagina göre, Fusarium muniliforme patojenine karsi ayni sekilde daha etkili antifungal aktivite gösterdigi ve fungusun miselyal büyümesini saf uçucu yaga göre daha fazla inhibe ettigi tespit edilmistir. As given in Figure 6 and Figure 7, the R. officinalis microemulsion formulation, According to the pure essential oil of R. officinalis, it is the same against the pathogen Fusarium muniliforme. Thus, it showed more effective antifungal activity and pure mycelial growth of the fungus. It has been found that it inhibits more than essential oil.
Bulusun koruma kapsami ekte verilen istemlerde belirtilmis olup kesinlikle bu detayli anlatimda örnekleme amaciyla anlatilanlarla sinirli tutulamaz. Zira teknikte uzman bir kisinin, bulusun ana temasindan ayrilmadan yukarida anlatilanlar isiginda benzer yapilanmalar ortaya koyabilecegi açiktir.The scope of protection of the invention is stated in the attached claims and this detailed for exemplification purposes, it cannot be limited to what is told. Because technical expert similar in the light of what has been described above without departing from the main theme of the invention. It is clear that structuring can occur.
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| EP2192887A1 (en) * | 2007-08-29 | 2010-06-09 | Agency for Science, Technology And Research | Sugar-based surfactant microemulsions containing essential oils for cosmetic and pharmaceutical use |
| US9687002B2 (en) * | 2015-05-21 | 2017-06-27 | IndusCo, Ltd. | Botanical antimicrobial microemulsions (BAMM) |
| FR3055544A1 (en) * | 2016-09-08 | 2018-03-09 | Actina | ASSOCIATION OF OLEA EUPOPAEA AND ROSMARINUS OFFICINALIS TO MODULATE THE ACTIVITY OF MITROCHONDRIA |
-
2020
- 2020-03-31 TR TR2020/05078A patent/TR202005078A2/en unknown
-
2021
- 2021-03-30 WO PCT/TR2021/050283 patent/WO2021201803A1/en active Application Filing
Also Published As
| Publication number | Publication date |
|---|---|
| WO2021201803A1 (en) | 2021-10-07 |
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