WO2019004969A2 - Use of a vegetal formulation containing pomegranate seed oil as an assistive agent in reduction of blood lipids known as triglyceride - Google Patents

Abstract

The invention relates to the use of a vegetal formulation in the pharmaceutical industry prepared in capsule form by using pomegranate seed oil rich in essential fatty acids such as punicic acid (Omega 5) as an assistive agent in reduction of blood lipids known as triglyceride and whose formulation comprises pomegranate seed oil, dl a-tocopherol, dl a- tocopheryl acetate, glycerine, purified water and gelatin that procures the integrity of the content and the capsulated formation.

Description

DESCRIPTION

USE OF A VEGETAL FORMULATION CONTAINING POMEGRANATE SEED OIL AS AN ASSISTIVE AGENT IN REDUCTION OF BLOOD LIPIDS KNOWN AS

TRIGLYCERIDE

TECHNICAL FIELD

The present invention relates to the usage of a vegetal formulation consisting of pomegranate (Punica granatum) seed oil in pharmaceutical and chemical industries.

The invention particularly relates to the use of a vegetal formulation prepared in capsule form by using pomegranate seed oil rich in essential fatty acids such as punicic acid (Omega 5) as an assistive agent in reduction of blood lipids known as triglyceride.

PRIOR ART

Essential fatty acids are the type of acids that cannot be synthesized within human body and must be obtained extrinsically in order for an organism keep on living. These essential fatty acids ought to be obtained through certain nutrients or supplements. Essential fatty acids such as Omega 5 (Punicic acid) which is specific to pomegranate seed oil obtained extrinsically and are mainly used as immune system enhancers due to their antioxidant function, as supplements for the preservation of overall health and as agents in reducing blood lipids also known as triglyceride.

In the current state of the art; products used for enhancing the immune system and for lowering blood lipids known as triglyceride, usually contain synthetic or chemical materials in certain amounts. The use of synthetic or chemical materials results in many side effects in the organism and affects bodily systems adversely.

Furthermore, in most of the available embodiments, these products are in liquid form which constitutes certain difficulties for controlled dosing. Therefore, the required dosage cannot be taken in exact and distinct amounts. One of the related patents in the literature is the application bearing number J P2016027023. The said patent application refers to a vegetal and cosmetic formulation that in addition to the pomegranate seed oil, contains many other mixtures which is good for facial and dermal skin sagging when applied externally.

Patent application bearing number D E 102009019103 refers to a beverage in which the pomegranate seed oil is dissolved with an emulsifier.

Another patent application bearing number J P20031 13028 refers to a component used as a cosmetic and chemically basic hygiene product consisting of pomegranate and other vegetal seed essences.

Abovementioned disadvantages, technical problems and inadequacy of available solutions related to this particular issue necessitated an improvement in the related technical field regarding the use of pomegranate seed oil as an assistive agent in reduction of blood lipids known as triglyceride.

O BJ E CTS O F T H E INV E NTIO N

It is an object of this invention to make sure that the vegetal formulation prepared in capsule form with pomegranate seed oil that is rich in essential fatty acids such as P unicic acid (Omega 5) is used as an assistive agent in reduction of blood lipids known as triglyceride.

P unicic acid (Omega 5) which is the main constituent of this particular formulation, possesses preventive effects on lipid peroxidation both in vitro and in vivo. It shows antioxidative effects by preventing hydroperoxidation. F urthermore, punicic acid has anti- atherogenic effects. S tudies have demonstrated the effects of pomegranate seed oil treatment on serum lipid (fat) profiles. T hese studies have shown that at the end of the 4 week administration period, when the group of pomegranate seed oil was compared to basal values, there has been a significant amount of reduction in both TG (triglyceride) and TG :H DL (High Density Lipoprotein -C). Pomegranate seed oil also plays a regulative role in blood lipid values by reducing the levels of LDL (Low Density Lipoprotein).

Another object of the present invention is to ensure that the prepared formulation has 100% natural and vegetal content without comprising any chemical substances. Even during the extracting process of the pomegranate seed oil used in the content of the formulation, no thermal or chemical treatment is utilized. Thus, active substances, nutritional values, vitamins and mineral within the plant are transmitted straight to the product without any losses.

Another object of the present invention is to minimize the side effects by creating a substance without using any chemical materials that can affect human health adversely.

Another object of the invention is to provide ease of use by capsulating the created formulation in terms of patient compliance and patient interaction. F urthermore, the capsulated form of the formulation constitutes further advantages in obtaining of appropriate dosing in aforementioned indications.

DIS C L OS U R E O F T H E INV E NTIO N

The preferred embodiment of the subject matter formulation obtained by using pomegranate seed oil to achieve invention's objective, is only elaborated for the further clarification of the subject and in a way that does not constitute any restrictive effects.

Pomegranate S eed Oil: The punicic acid (Omega 5) which is the main component of the pomegranate seed oil has preventive effects on lipid peroxidation as in vitro and in vivo. P unicic acid bears antioxidative effects by preventing hydroperoxidation. F urthermore, punicic acid has anti-atherogenic effects. Studies have demonstrated the effects of pomegranate seed oil treatment on serum lipid (fat) profiles. T hese studies have shown that at the end of the 4 week administration period, when the group of pomegranate seed oil was compared to basal values, there has been a significant amount of reduction in both TG (triglyceride) and TG :H DL (High Density Lipoprotein -C). P omegranate seed oil also plays a regulative role in blood lipid values by reducing the levels of LDL (Low Density Lipoprotein). Moreover, pomegranate seed oil blocks inflammatory cytokines, increases insulin sensitivity and boosts immune system. Also, it further strengthens the immune system by significantly increasing C D3 (Mature T-Lymphocyte) and total leucocyte values.

Pomegranate seed oil also contains phyto-estrogenic components (17 alpha-estradiol, estrone, estriol and testosterone). Therefore, it can be used as an assisting agent for relieving symptoms (sleep disorders, hot flashes, night sweats etc.) observed during the period of menopause due to the mild estrogenic effects of the said components.

Pomegranate seed oil capsules can also relieve sleep disorders in women observed during the period of menopause. P omegranate seed oil compounds increase bone density in ovariectomic female rats and also relieves depressive symptoms observed during the period of menopause.

Pomegranate seed oil increases insulin sensitivity in Type-Π diabetic patients.

Administration of pomegranate seed oil in rats reduces weight gain induced with diet obesity, insulin resistance and the risk rate of type 2 diabetes.

It is known for a fact that in experimental diabetes models created for Zucker rats with streptozosine and alloxane, compounds obtained from various fractions of the pomegranate influence diabetes parameters positively.

Caps ulated Formulation C ontent:

In this particular formulation, pomegranate seed oil is used as the active substance. As for excipients, the formulation comprises dl a-tocopherol, dl a-tocopheryl acetate, glycerine, purified water and gelatin. In this context dl a-tocopherol and dl a-tocopheryl acetate are used as preservative substances. For the said formulation, dl a-tocopherol and dl a- tocopheryl acetate are added on ppm level. For the gelatin added for the integrity of the content bovine gelatin is preferred. However, as a substitute for the bovine gelatin, animal and vegetal originated other gelatin products can also be utilized.

As it is demonstrated in Table 1 , content percentage by weight is as follows; 48,1 50% pomegranate seed oil; 0,004% dl a-tocopherol; 0,024% dl a-tocopheryl acetate; 9,690% glycerine; 21 ,065% purified water; 21 ,065% gelatin.

Table 1. Capsule's content and percentage by weight

G lycerine 9,690 80,5

Water 21 ,065 175

Gelatin 21 ,065 175

Caps ule's P roduction Method:

P roduction steps of the capsule's formulation can be described as below; pomegranate seed which is one of the raw materials is put to a sifting process. P ursuant to the sifting, crushing step is carried out if necessary. After pomegranate seeds are put through these processes and desired size is obtained, seeds are sent to a cold isostatic pressing unit This unit carries out the oil extraction process from the raw materials using the cold pressing method. While the said method is being put to practice, the temperature must not exceed 55¾J . E xtracted oil are preserved in mixing and settling tanks prior to the filtration process. P reservation of extracted oil in mixing and settling tanks facilitates the separation of occurring particles. Following its preservation in mixing and settling tanks, extracted oil is sent to filtering unit which is held indoors and room temperature. In the filtering unit if any, particles within the content of the oil are filtered. Following the filtering stage, dl a-tocopherol and dl a-tocopheryl acetate are added to the acquired oil as preservative substances. T hus, the mixture composing the content is obtained.

Meanwhile these production steps are carried out, in the gelatin preparation unit the gelatin that is used for the capsule integrity is prepared.

S ubsequent to the creation of the mixture and the preparation of the gelatin, both the mixture and the gelatin are sent to the capsule production unit and capsules with desired dosage are prepared. This said capsule is namely soft gelatin capsule. Following the preparation of capsules, drying process is carried out and dried capsules are packed.

Caps ule's Package Insert: Therapeutic Indications

Capsule is used as an immune system enhancer for the protection of health due to its antioxidant functions and also used collaterally in reduction of blood lipids also known as triglyceride. Dos ing and Method of Administration

I Dosing administration interval and period: Unless advised otherwise by a physician, capsules can be taken by adults up to 1 to 2 capsule(s) (400-800 mg/day) per day.

I Method of Administration:

Capsules should be swallowed preferably on a full stomach with sufficient water and without being crushed or chewed.

S upplementary Information R elated To S pec ific P opulations:

I R enal/liver failure

S ince safety studies conducted in kidney and liver patients aren't adequate, capsules should not be used in these particular patient groups.

I P ediatric Population

S ince safety studies conducted in this particular population aren't adequate, capsules should not be used in patients under 12 years of age.

I G eriatric Population

There are no studies carried out in geriatric population. However, it is thought that a dose adjustment is unnecessary.

C ontraindications

Capsules should not be used by

persons who might develop allergic reactions to the active substance or any other component of the capsules ^"content

- women who are in pregnancy and lactation period

patients with liver and kidney failure

S pecial P recautions and Warnings For Use Capsule contains glycerine, however, this does not necessitate any warnings because of its amount.

It is advised that use of the capsule longer than 2 months is to be continued in consultation with a physician.

P regnancy and Lactation

I G eneral Advice

P regnancy category: C

I Women with childbearing potential/Birth control

Capsule should not be used since there are no studies related to this subject.

I P regnancy P eriod

There are no studies related to the use of pomegranate (P unica granatum) seed oil during the pregnancy period, therefore, use of the capsule is not advised.

I Lactation P eriod

There are no studies conducted for this particular patient group, therefore, use of the capsule is not advised.

Undesirable Effects

Even though the incidence is unclear in the use of the capsule, gastrointestinal side effects including nausea, vomiting and diarrhea could be observed. Apart from the abovementioned effects, no other undesirable effects have been reported.

Overdose

No overdose incidents have been reported to date with regard to pomegranate (P unica granatum) seed oil. In case of an overdose, the treatment must be discontinued and a physician should be consulted.

Storage C onditions The shelf life of the product is determined to be 2 years. The capsule whose content and production method is the subject matter above, has been licensed by the authority of the Ministry of Health of the R epublic of Turkey as "C onventional Herbal Medicinal P roduct". T he said license bears the date of 21.1 1 .201 6 and the sequence number of 201 6 829 and goes under the name of ZADE VITAL P U MININ 400 mg soft capsule. The capsule is a LIC E NS E D C onventional Herbal Medicinal P roduct by the Ministry of Health of the R epublic of Turkey in accordance with the abovementioned indications and the R egulation for C onventional Herbal Medicinal P roducts. T here are a number of licensed products containing a large number of synthetic chemicals in the market for the stated indications. However, in this particular field there are no conventional herbal medicinal products licensed for these indications by the health authorities.

While in the majority of the available embodiments the production is carried out under bad hygiene conditions, however, related formulation is manufactured in accordance with G MP (G ood Manufacturing P ractises) norms required by the Ministry of Health from a pharmaceutical factory.

As is known, it is rather difficult to maintain standardization in vegetal products.

R egarding the corresponding formulation;

I Microbiological controls and analyses of pesticides, aflatoxins, heavy metal and active substances of the seeds have been carried out.

I Microbiological controls, validation and method development for the analysis of active substances have been conducted in the cold pressed oil.

I P roducts, indications, dosing and form of dosing have been calculated in the design.

I Analyses of active substances/excipients, microbiological controls, quality control parameters and stability studies to be carried out were considered and the system was developed accordingly in the preformulation.

I C onformity to determined specifications during the predicted shelf life has been proven by stability studies. A 24-month study has been carried out in 25¾] e2, % 60 e5 humidity and another 6-month study has been conducted in 40¾J e2, % 75 e5 humidity throughout the stability studies. Validations and data related to analytic testing methods, packaging specifications, interactions, methods for analyses and controls have been determined.

I During the preparation of intra-materials and capsule basis certain tests such as content designation (analysis of oil components), viscosity and optical prospect etc. have been carried out.

I Throughout the production stage, quality control procedures such as content designations, microbiological analyses and optical controls, also in-process controls such as optical control, qualitative and quantitative content designation have been carried out. Moreover, H P LC analyses of the sample product and active substance were conducted and fatty acid compositions were calculated. F urthermore, calibration tables of the active substance were drawn.

Abovementioned processes are crucial in terms of standardization of vegetal products and these processes have been carried out at full length. The said product has obtained certification from a health authority such as the Ministry of Health through licensing of the pharmacodynamic characteristics of a C onventional Herbal Medicinal P roduct. Again, this particular product has obtained certification from a health authority such as the Ministry of Health by having its summary of product characteristics (spc) and its Package Insert (PI) licensed. T he said product obtained certification from a health authority such as the Ministry of Health through standardizing the method of administration and dosing of a C onventional Herbal Medicinal P roduct. In this context, dosing of a Ministry of Health licensed conventional herbal medicinal product was put in writing pertinent to the clinical trials, was officialized through the approval of health authorities and was put at the disposal of the global medicine.

PUBLISHED LITERATURE

Alam MS, Alam MA, Ahmad S, Najmi AK, Asif M, J ahangirT. Protective effects of Punica granatum in experimentally-induced gastric ulcers. Toxicol Mech Methods. 2010 Nov;20(9):572-8.

Al-Muammar MN, Khan F. Obesity: the preventive role of the pomegranate (Punicagranatum). Nutrition.2012 J un;28(6):595-604.

Asghari G, S heikholeslami S, Mirmiran P, Chary A, Hedayati M, Shafiee A, Azizi F. Effect of pomegranate seed oil on serum TNF- level in dyslipidemic patients. IntJ Food Sci Nutr.2012 May;63(3):368-71.

Auerbach L, Rakus J, Bauer C, Gerner C, Ullmann R, Wimmer H, Huber J. Pomegranate seed oil in women with menopausal symptoms: a prospective randomized, placebo-controlled, double-blinded trial. Menopause.2012 Apr;19(4):426- 32.

Aviram M, Dornfeld L. Pomegranate juice consumption inhibits serum angiotensin converting enzyme activity and reduces systolic blood pressure. Atherosclerosis. 2001 Sep;158(1 ):195-8.

Aviram M, Rosenblat M, Gaitini D, Nitecki S, Hoffman A, Dornfeld L, Volkova N, Presser D, Attias J , Liker H, Hayek T. Pomegranate juice consumption for 3 years by patients with carotid artery stenosis reduces common carotid intima-media thickness, blood pressure and LDL oxidation. Clin Nutr.2004J un;23(3):423-33.

Aviram M, Rosenblat M. Pomegranate Protection against Cardiovascular Diseases. Evid Based Complement Alternat Med.2012.

Azadzoi KM, Schulman RN, Aviram M, Siroky MB. Oxidative stress in arteriogenic erectile dysfunction: prophylactic role of antioxidants. J Urol.2005 J ul;174(1):386-93.

Bell C, Hawthorne S . E Magic acid, pomegranate and prostate cancer ~ a mini review. J Pharm Pharmacol.2008 Feb;60(2):139-44. Borochov-Neori H, J udeinstein S, Harari M, Bar-Yaakov I, Patil BS, Lurie S, Holland D. Climate effects on anthocyanin accumulation and composition in the pomegranate (Punicagranatum L.) fruit arils.J Agric FoodChem.2011 May 25;59(10):5325-34.

Borochov-Neori H, J udeinstein S, Tripler E; Harari M, Greenberg A, S homer I, Holland D. Seasonal and cultivar variations in antioxidant and sensory quality of pomegranate (Punica granatum L.) fruit J ournal of Food Composition and Analysis 2009; 22(3): 189- 195.

Bouroshaki MT, Sadeghnia HR, Banihasan M, Yavari S. Protective effect of pomegranate seed oil on hexachlorobutadiene-induced nephrotoxicity in rat kidneys. Ren Fail.2010J un;32(5):612-7.

Boussetta T, Raad H, Lett^ron P, Gougerot-Pocidalo MA, MarieJ C, Driss F, EI-BennaJ . Punicic acid a conjugated linolenic acid inhibits TNF alpha-induced neutrophil hyperactivation and protects from experimental colon inflammation in rats.PLoS One. 2009 J ul 31;4(7):e6458.

Braga LC, Leite AA, Xavier KG, Takahashi J A, Bemquerer MP, Chartone-Souza E, Nascimento AM. Synergic interaction between pomegranate extract and antibiotics against Staphylococcus aureus. Can J Microbiol.2005 J ul;51(7):541-7.

Caliskan O, Bayazit S. Phytochemical and antioxidant attributes of autochthonous Turkish pomegranates. Scientia Horticulturae 2012; 147: 81 ^"88.

Cayy K, Karadeniz A, SimCek N, Y tdyyn S, KarakuOE, Kara A, Akkoyun HT, Sengad E. Pomegranate seed extract attenuates chemotherapy-induced acute nephrotoxicity and hepatotoxicity in rats. J Med Food.2011 Oct;14(10):1254-62.

Celik I, Temur A, Isik I. Hepatoprotective role and antioxidant capacity of pomegranate (Punica granatum) flowers infusion against trichloroacetic acid-exposed in rats. Food C hem Toxicol.2009 J an;47(1):145-9.

Coursodon-Boyiddle CF, Snarrenberg CL, Adkins-Rieck CK, Bassaganya-Riera J, Hontecillas R, Lawrence P, Brenna J T, J ouni ZE, Dvorak B. Pomegranate seed oil reducesintestinal damage in a rat model of necrotizing enterocolitis. Am J Physiol Gastrointest Liver Physiol.2012 Sep 15;303(6):G744-51.

Dahham SS, AN MN, Tabassum H, Khan M. Studies on Antibacterial and Antifungal Activity of Pomegranate (Punica granatum L). Amer. Eurasian J. Agri. Environ. Sci., 2010; 9(3):273-281.

Das AK, Mandal SC, Banerjee SK, Sinha S, Das J, Saha BP, Pal M. Studies on antidiarrhoeal activity of Punica granatum seed extract in rats.J Ethnopharmacol. 1999 Dec 15;68(1-3):205- 8. de Nigris F, Balestrieri ML, Williams-Ignarro S, D;Armiento FP, Fiorito C, Ignarro LJ , Napoli C. The influence of pomegranate fruit extract in comparison to regular pomegranate juice and seed oil on nitric oxide and arterial function in obese Zuckerrats. Nitric Oxide.2007 Aug;17(1):50-4.

Dellagli M, Galli GV, Bulgari M, Basilico N, Romeo S, Bhattacharya D, Taramelli D, Bosisio E. Ellagitannins of the fruit rind of pomegranate (Punica granatum) antagonize in vitro the host inflammatory response mechanisms involved in the onset of malaria. Malar J .2010 J ul 19;9:208.

DemirezerLO. FFD Monograflary. Tedavide kullanlan Bitkiler.2011; pp.547-558.

El-Boghdady NA. Protective effect of ellagic acid and pumpkin seed oil against methotrexate-induced small intestine damage in rats. Indian J Biochem Biophys. 2011 Dec;48(6):380-7.

Elfalleh W, Ying M, Nasri N, Sheng-Hua H, Guasmi F, Ferchichi A. Fatty acids from Tunisian and Chinese pomegranate (Punica granatum L.) seeds.lntj Food Sci Nutr. 2011 May;62(3):200-6.

El-Sherbini GM, Ibrahim KM, El Sherbiny ET, Abdel-Hady NM, Morsy TA. Efficacy of Punica granatum extract on in-vitro and in-vivo control of Trichomonas vaginalis. J Egypt Soc Parasitol.2010 Apr;40(1):229-44. Endo EH, Cortez DA, Ueda-Nakamura T, Nakamura CV, Dias Filho BP. Potent antifungal activity of extracts and pure compound isolated from pomegranate peels and synergism with fluconazole against Candida albicans. Res Microbiol. 2010 Sep;161(7):534-40.

Faria A, Monteiro R, Azevedo I, Calhau C. Pomegranate juice effects on cytochrome P450S expression: in vivo studies. J Med Food.2007 Dec;10(4):643-9.

Forest CP, Padma-Nathan H, Liker HR. Efficacy and safety of pomegranate juice on improvement of erectile dysfunction in male patients with mild to moderate erectile dysfunction: a randomized, placebo-controlled, double-blind, crossover study. IntJ Impot Res.2007 Nov-Dec;19(6):564- 7.

G0a_KcaeM, Tokg!Zfe H, nelikyurt MA. Nar ^'ekirde/linin bazy !Zkellikleri ve nar ^'ekirde/li ya/lyiyi ya/1 asiti bileQmi. Derim 2005; 22(2): 33-40.

G0a_KcaeM, Tokg!Zfe H. - Ikemizde YetiQirilen ^a nemli Nar(Punica granatum) nedtlerine Ait n ekirdeklerin Bazy^a zellikleri. GyJa 2008; 33 (6): 281-290.

Grossmann ME, Mizuno NK, Schuster T, Cleary MP. Punicic acid is an omega-5 fatty acid capable of inhibiting breast cancer proliferation. IntJ Oncol.2010;36(2):421-6.

Haidari M, AN M, Ward Casscells S 3rd, Madjid M. Pomegranate (Punica granatum) purified polyphenol extract inhibits influenza virus and has a synergistic effect with oseltamivir.Phytomedicine.2009 Dec;16(12):1127-36.

Hanley MJ , Masse G, HarmatzJS, Court MH, Greenblatt DJ . Pomegranate juice and pomegranate extract do not impair oral clearance of flurbiprofen in human volunteers: divergence from in vitro results. Clin Pharmacol Ther.2012 Nov;92(5):651-7.

Hartman RE, Shah A, Fagan AM, Schwetye KE, Parsadanian M, Schulman RN, Finn MB, Holtzman DM. Pomegranate juice decreases amyloid load and improves behavior in a mouse model of Alzheimer's disease. Neurobiol Dis.2006 Dec;24(3):506-15. Hidaka M, Okumura M, F ujita K, Ogikubo T, Yamasaki K, Iwakiri T, S etoguchi N, Arimori K. Effects of pomegranate juice on human cytochrome p450 3A (CY P3A) and carbamazepine pharmacokinetics in rats. Drug Metab Dispos. 2005 May;33(5):644-8.

Hong MY, S eeram NP, Heber D. P omegranate polyphenols down-regulate expression of androgen-synthesizing genes in human prostate cancer cells overexpressing the androgen receptor.J Nutr Biochem. 2008 Dec;19(12):848-55.

Hora J J , Maydew E R , Lansky E P, Dwivedi C . C hemopreventive effects of pomegranate seed oil on skin tumor development in C D1 mice. J Med Food. 2003 Fall;6(3):1 57-61 .

J urenka J S . Therapeutic applications of pomegranate (P unica granatum L): a review. Altern Med R ev. 2008; 13(2):128-44.

Khalil EAM. A hepatoprotective effect of an aqueous extract of pomegranate (P unica granatum L. ) rind against acetaminop hen treated rats. The Egyptian J ournal of Hospital Medicine Vol., 16 : 1 12 ^" 1 18

Khan SA. The role of pomegranate (P unica granatum L.) in colon cancer.Pak J P harm S ci. 2009 J ul;22(3):346-8.

Khuu T, Hickey A, Deng MC . P omegranate-containing products and tacrolimus: Apotential interaction.] Heart Lung Transplant. 2012 Nov 29. dokpii: S 1053- 2498(12)01388-5. 10.1016 j.hea lun.2012.10.015

Kohno H, S uzuki R, Yasui Y, Hosokawa M, Miyashita K, Tanaka T. Pomegranate seed oil rich in conjugated linolenic acid suppresses chemically induced colon carcinogenesis in rats. Cancer S ci. 2004; 95(6):481 -6.

Kumar S , Maheshwari KK, S ingha V. C entral nervous system activity of acuteadministration of ethanol extract of P unica granatum L. seeds in mice. Indian J E xp Biol. 2008 Dec;46(12):811 -6.

Lansky E P, Newman RA. P unica granatum (pomegranate) and its potential for prevention and treatment of inflammation and cancer.J Ethnopharmacol. 2007 J an 19;109(2):177-206. Lei F, Zhang XN, Wang W, Xing DM, Xie WD, Su H, Du LJ . Evidence of anti-obesity effects of the pomegranate leaf extract in high-fat diet induced obese mice. IntJ Obes (Lond).2007 J un;31(6):1023-9.

McFarlin BK, Strohacker KA, Kueht ML. Pomegranate seed oil consumption during a period of high-fat feeding reduces weight gain and reduces type 2 diabetes risk in CD-1 mice. BrJ Nutr.2009J ul;102(1):54-9.

Meerts IA, Verspeek-Rip CM, Buskens CA, Keizer HG, Bassaganya-Riera J , J ouni ZE, van Huygevoort AH, van Otterdijk FM, van de Waart EJ . Toxicological evaluation of pomegranate seed oil. Food C hem Toxicol.2009 J un;47(6):1085-92.

Mena P, Girones-Vilaplana A, Moreno DA, Garcia-Viguera C. Pomegranate fruit for health promotion: Myths and realities. Functional Plant Science and Biotechnology 2011; 5(2):33-42

Mertens-Talcott SU, J ilma-Stohlawetz P, Rios J , Hingorani L, Derendorf H. Absorption, metabolism, and antioxidant effects of pomegranate (Punica granatum I.) polyphenols after ingestion of a standardized extract in healthy human volunteers. J Agric Food Chem.2006 Nov 15;54(23):8956-61.

Miguel MG, Neves MA, Antunes MD. Pomegranate (Punica granatum L.): A medicinal plant with myriad biological properties - A short review. Journal of Medicinal Plants Research 2010; 4(25): 2836^"2847

Mirmiran P, Fazeli MR, Asghari G, Shafiee A, Azizi F. Effect of pomegranate seed oil on hyperlipidaemic subjects: a double-blind placebo-controlled clinical trial. BrJ Nutr.2010 Aug;104(3):402-6.

Misaka S, Nakamura R, Uchida S, Takeuchi K, Takahashi N, Inui N, Kosuge K, Yamada S, Watanabe H. Effect of 2 weeks consumption of pomegranate juice on the pharmacokinetics of a single dose of midazolam: an open-label, randomized, single- center, 2-period crossover study in healthy Japanese volunteers. Clin Ther. 2011 Feb;33(2):246-52. Mori-Okamoto J , Otawara-Hamamoto Y, Yamato H, Yoshimura H. Pomegranate extract improves a depressive state and bone properties in menopausal syndrome model ovariectomized mice. J Ethnopharmacol.2004 May;92(1):93-101.

Nagata M, Hidaka M, Sekiya H, Kawano Y, Yamasaki K, Okumura M, Arimori K. Effects of pomegranate juice on human cytochrome P450 2C9 and tolbutamide pharmacokinetics in rats. Drug Metab Dispos.2007 Feb;35(2):302-5.

Neurath AR, Strick N, Y.Y. Li and Debnath AK: Punica granatum (pomegranate) juice provides an HIV-1 entry inhibitor and candidate topical microbicide. Annals of New York Academy of S cience 2005; 1056:311 -327. ^a zgen M, Durga^' C, Serce S, Kaya C. Chemical and antioxidant properties of pomegranate cultivars grown in the Mediterranean region of Turkey. Food Chemistry 2011; 111:703^"706.

Pantuck AJ , LeppertJT, Zomorodian N, Aronson W, Hong J , Barnard RJ , Seeram N, Liker H, Wang H, E lashoff R, Heber D, Aviram M, Ignarro L, Belldegrun A. Phase II study of pomegranate juice for men with rising prostate-specific antigen following surgery or radiation for prostate cancer.Clin Cancer Res.2006 J ul 1;12(13):4018-26.

PDR for Herbal Medicine, 2008. Sayfa 605-606.

Qnais EY, Elokda AS, Ghalyun YY, Abdulla FA. Antidiarrheal Activity of the Aqueous Extract of Punica granatum (Pomegranate) Peels. Pharmaceutical Biology (Formerly International J ournal of Pharmacognosy), 2007; 45(9): 715-720.

Rosenblat M, Hayek T, Aviram M. Anti-oxidative effects of pomegranate juice (PJ) consumption by diabetic patients on serum and on macrophages. Atherosclerosis.2006 Aug;187(2)363-71.

Seeram NP, Zhang Y, McKeever R, Henning SM, Lee RP, Suchard MA, Li Z, Chen S, Thames G, Zerlin A, Nguyen M, Wang D, Dreher M, Heber D. Pomegranate juice and extracts provide similar levels of plasma and urinary ellagitannin metabolites in human subjects.J Med Food.2008 J un;11(2):390-4. Shravan Kumar Y, Adukondalu D, Bhargavi Latha A, Vamshi Vishnu Y, Ramesh G, Shiva Kumar R, Madhusudan Rao Y, Sarangapani M. Effect of pomegranate pretreatment on the oral bioavailability of buspirone in male albino rabbits. Daru. 2011;19(4):266-9.

Sumner MD, Elliott-Eller M, Weidner G, DaubenmierJJ , Chew MH, Marlin R, Raisin CJ , Ornish D. Effects of pomegranate juice consumption on myocardial perfusion in patients with coronary heart disease. Am J Cardiol.2005 Sep 15;96(6):810-4.

Syed DN, Afaq F, Mukhtar H. Pomegranate derived products for cancer chemoprevention.Semin Cancer Biol.2007 Oct;17(5)377-85.

HimCek N, Karadeniz A, Bayraktaro/llu AG. Ratlarda Periferal Kan Haacreleri - zerineL^" karnitin, ArySaiaeve Nar n ekirde/linin Etkileri. Kafkas - niv.Vet Fak Derg,15(1): 63-69, 2009.

Toklu HZ, Dumlu MU, Sehirli O, Ercan F, Gedik N, G!Zkmen V, SenerG. Pomegranate peel extract prevents liver fibrosis in biliary-obstructed rats. J Pharm Pharmacol.2007 Sep;59(9):1287-95.

Toklu HZ, Sehirli O, Ozyurt H, Mayada/lli AA, EkOo/llu-Demiralp E, Cetinel S, Sahin H, Ye/len BC, Ulusoylu Dumlu M, G!Zkmen V, Sener G. Punica granatum peel extract protects against ionizing radiation-induced enteritis and leukocyte apoptosis in rats. J Radiat Res.2009J ul;50(4):345-53.

Tark G, S0nmez M, Aydin M, Yaace A, GarS, Ya_xsel M, Aksu EH, Aksoy H. Clin Nutr. Effects of pomegranate juice consumption on sperm quality, spermatogenic cell density, antioxidant activity and testosterone level in male rats.2008 Apr;27(2):289-96. van Elswijk DA, Schobel UP, Lansky EP, Irth H, van der Greef J . Rapid dereplication of estrogenic compounds in pomegranate (Punica granatum) using on-line biochemical detection coupled to mass spectrometry. Phytochemistry.2004 J an;65(2):233-41.

Vioda-Martos M, Fernandez-Lopez J, Perez-Alvarez J A. Pomegranate and its Many Functional Components as Related to Human Health: A Review. Comprehensive Reviews in Food Science and Food Safety 2010; 9(6): 635^"654 Voruganti S, Rapolu K, Tota S, Yamsani SK, Yamsani MR. Effect of pomegranate juice on the pharmacokinetics of nitrendipine in rabbits. Eur J Drug Metab Pharmacokinet. 2012 J un;37(2):77-81

Vroegrijk 10, van Diepen J A, van den Berg S, Westbroek I, Keizer H, Gambelli L, Hontecillas R, Bassaganya-Riera J , Zondag GC, RomijnJA, Havekes LM, Voshol PJ . Pomegranate seed oil, a rich source of punicic acid, prevents diet-induced obesity and insulin resistance in mice. Food C hem Toxicol.2011 J un;49(6):1426-30.

Wang R, Ding Y, Liu R, Xiang L, Du L. Pomegranate: Constitituents, bioactivities and pharmacokinetics. Fruit, Vegetable and Cereal Science and Biotechnology 4(2): 77-87

Yamasaki M, Kitagawa T, Koyanagi N, Chujo H, Maeda H, Kohno-Murase J , ImamuraJ, Tachibana H, Yamada K. Dietary effect of pomegranate seed oil on immune function and lipid metabolism in mice. Nutrition.2006 J an;22(1):54-9.

Claims

C LAIMS

1. The invention is the use of a vegetal formulation comprising pomegranate seed oil as an assistive agent in reduction of blood lipids known as triglyceride in manufacturing of a medication.

2. The use according to claim 1 , characterized in that, the said vegetal formulation comprises pomegranate seed oil, dl a-tocopherol, dl a-tocopheryl acetate, glycerine, purified water and gelatin that procures the integrity of the content and the capsulated formation.

3. The use according to claim 2, characterized in that it is used in adults in the dosing of 400- 800 mg/day.

4 . T he use according to claim 2, characterized in that it is administered through the oral route.