ES2489298B1 - USE OF EREMOFILAN SESQUITERPENOS AS GARRAPATICIDAS - Google Patents

Abstract

Use of eremofilanes sesquiterpenes as ticks. # Use of at least one eremofilane sesquiterpene of formula (I) as ticks, where R {sup, 1} is H or OH; [1] is a single or double bond; X is selected from the group consisting of absent, CH {sub, 3}, OH, OOH and together with Z forms an epoxy of structure (Z-IV); and Z is selected from the group consisting of (ZI), (Z-II), (Z-III) and together with X form (Z-IV), preferably sesquiterpene (I) is dehydrofukinone, 1,1-hydroxyremophilyl-6 (7), 9 (10) -dien-8-one, 7-11- epoxieremophil-9 (10) -en-8-one, or a mixture. Preferably, sesquiterpene acts against ticks of the genus Hyalomma spp. and / or Rhipicephalus spp. In particular, use to treat or prevent a disease caused by ticks in a human or animal; and use to prevent, control or eliminate the presence of ticks in the environment in contact with an animal or human. As well as a product comprising said sesquiterpene of formula (1).

Description

DESCRIPTION

Use of eremofilanes sesquiterpenes as ticks.

SECTOR OF THE TECHNIQUE 5

The present invention is within the livestock sector. In particular, it provides a phytosanitary product to combat ticks that especially affect cattle, sheep and goats. Additionally, the present invention is also within the pharmaceutical and veterinary sector, since said phytosanitary product, sesquiterpene 10 eremophilane, can also be used to treat or prevent a disease caused by ticks.

STATE OF THE TECHNIQUE

 fifteen

The tick is one of the most economically important ectoparasites worldwide, due to the decrease it causes in the production of cattle, goats, as well as throwing and riding horses. Direct consequences of tick parasitization are the least amount of food ingested by cattle, weight loss due to toxins and irritation, anemias caused by blood loss and transmission of 20 hemoparasites (Babesia bigemina, B. bovis, Anaplasma marginale) and the considerable depreciation of the skins because of the perforations produced by the bites. In addition, these perforations allow the access of bacteria, dermal mycoses and fly larvae (myiasis). By injuring the skin to suck blood, many species of ticks can also transmit the most diverse pathogens such as viruses, bacteria, riches and protozoa. This can lead to acute, chronic or even death.

From the point of view of damage and the potential disease transmitter, seven genera of Ixodidae 30 ticks (Amblyomma spp., Boophilus spp., Dermacentor spp., Haemaphysalis spp., Hyalomma spp., Lxodes spp.) Acquire veterinary importance worldwide . and Rhipicephalus spp.) and two of Argasidae (Ornithodorus spp. and Otobius spp.)

Rhipicephalus (Boophilus) microplus is considered the most important parasite of cattle, mainly infests cattle, reducing livestock production, causing loss of weight in animals and decreasing milk production. Its greatest importance lies in being a vector of limiting diseases in the tropics such as babesiosis (Babesia bovis, Babesia bigemina) and anaplasmosis (Anaplasma marginale).

Hyalomma tick species are of great economic importance in several 40 tropical and subtropical countries in Africa, Asia and Europe. Typical species are Hyalomma anatolicum anatolicum, Hy. marginatum rufipes, Hy. Truncatum and Hy. detritum detritum, which feed as adults in cattle, sheep and goats. Hyalomma dromedarii is specialized to feed on the dromedaries. On the other hand, the genus Hyalomma can act as a vector for the transmission of diseases in humans such as the 45 Hemorrhagic Fever of Crimea-Congo.

Hyalomma lusitanicum is a species of strictly Mediterranean distribution (Estrada et al. Ticks of Domestic Animals in the Mediterranean Region. A guide to Identification of Species. ICTTD. University of Zaragoza) so it is little known and studied. However, in Spain it is the most abundant exophyll tick species in the Center of the Iberian Peninsula (86%) (Barandika et al, Differences in Questing Tick Species Distribution Between Atlantic and Continental Climate Regions in Spain, Journal of Medical Entomology,

2011, 48 (1): 13-19). In addition to the direct pathogenic action derived from their feeding, this species is considered a vector of Theileria annulata, a protozoosis of endemic cattle in its area of distribution for which there is no commercialized curative treatment and that produces numerous economic losses in our livestock. In addition, recent studies have demonstrated the presence in this species of tick 5 of bacterial agents that affect man, such as Anaplasma phagocytophilum, responsible for anaplasmosis; Borrelia spp (Borrelia burgdorferi ss and B. garinii) agents of borreliosis, Francisella tularenisis producing tularemia, or Coxiella burnetii the agent of Q fever. (
Toledo et al, Tick-Borne Zoonotic Bacteria in Ticks Collected from Central Spain. Am J Trop Med Hyg July 2009 vol. 81 no. 1 67-74; Toledo et al. Detection of Coxiella 10 burnetii in ticks collected from Central Spain.
Vector Borne Zoonotic Dis. 2009 Oct; 9 (5): 465-8.).

Finally, the maximum alarm regarding this species has been triggered when Dr. Oteo's team publishes the finding of a Hyalomma lusitanicum infected with the virus 15 producing the Crimean Congo hemorrhagic fever in La Rioja (Crimean-Congo Hemorrhagic Fever Virus in Ticks , Southwestern Europe, 2010. Agustín Estrada-Peña, Ana M. Palomar, Paula Santibáñez, Nely Sánchez, Miguel A. Habela, Aránzazu Portillo, Lourdes Romero, and José A. Oteo. Emerg Infect Dis. 2012 January; 18 (1) : 179-180). To date, no product had been tested specifically for this species, whose importance in veterinary and health is demonstrated by the aforementioned.

The active substances most used in the market as synthetic ticks belong mainly to the following chemical classes: organophosphates, amidines, pyrethroids, phenylpyrazoles, endectocides (macrocyclic lactones) and benzoylureas. 25

Among the organophosphates is chlorfenvinfos and cumaphos. Currently its use is quite low in cattle. The resistance of ticks to organophosphates is widespread.

 30

Within the pyrethroid class there are compounds such as cypermethrin, a non-systemic insecticide of the family of synthetic pyrethroids that acts by contact and ingestion; Flumethrin used in cattle, sheep, goats, pigs and birds against ticks, mites, scabies, flies, lice; deltamethrin, contact tick also mosquito, sarnicide and piojicide. Pyrethroid resistance is widespread and is often marketed mixed with organophosphates, phenylpyrazoles, neonicotinoids, etc.

Amitraz, a triazapentadiene belonging to the amidine family that is used to control the red spider, leaf miners, scale insects, aphids, as well as ticks, mites, lice and other animal pests. It is a product of widespread use 40 in countries like Mexico. The resistance is on the rise.

The main endectocides (macrocyclic lactones) against ticks are: abamectin, doramectin, eprinomectin, ivermectin and moxidectin. The use of ivermectin, in countries like Mexico, has been greatly increased in recent years. Resistance to these active ingredients has been observed.

Fipronil is an active substance in phenylpyrazoles.

Benzoylureas (or dimyloids) are developmental inhibitors, preventing immature stages 50 from completing their development. The active substance fluazuron is distinguished.

However, in many cases these synthetic products present toxicity problems at the environmental, animal and human levels; resistance development; toxicity of some compounds; and contamination of the meat and milk of cattle, so in some cases their use is not authorized or waiting times are required for the use of the meat and milk of treated cattle. All this means that they are not suitable for use in 5 organic livestock. (
Brooke W. Bissinger et al.
Pesticide Biochemistry and Physiology.
Volume 96, Issue 2, February 2010, Pages 63–79).

In addition, the inappropriate and indiscriminate use of chemical synthesis substances has caused multiple problems in the control of ticks, since resistance to several of these products has been developing for several 10 years. (J-F Graft et al. Parasitology, 129 (S1), 2004, S427-S442 and J. E. George et al Parasitology, 129 (S1), 2004, S353 S366).

As previously mentioned, the main problem of the indiscriminate use of 15 conventional antiparasitic agents has been the emergence of resistant strains. That is the reason why currently Rhipicephalus (Boophilus) microplus, one of the species, in theory, easier to fight, because its entire cycle is completed in the host, is currently one of the main problems in the livestock of countries Latin America and Africa (Jonsson, NN, Hope M. 2007. Progress in the epidemiology and diagnosis of amitraz 20 resistance in the cattle tick Boophilus microplus-Vet. Parasitol. 146: 193-198; Miller, RJ, BD Ronald and JE George 2007. First Report of Permethrin-Resistant Boophilus microplus (Acari: Ixodidae) Collected within the United States. J. Med. Entomol 44 (2): 308-315.).

 25

Aware that this situation can be repeated with other tick species and in other areas, the current trend of treatment involves the search for new substances that allow treatment rotation (Thullner, F., Willadsen P., and Kemp, D. 2007. Acaricide Rotation Srategy for Managing Resistance in the Tick Rhipicephalus (Boophilus) microplus (Acarina: Ixodidae): Laboratory Experiment with a Field Strain from Costa Rica. J. 30 Med. Entomol. 44 (5): 817-821) and, to the extent possible, using natural products compatible with organic farming and livestock (Olmeda et al, 2008. Toxicity of Oxalic Acid against Adult Hyalomma lusitanicum ticks (Ixodida: Ixodidae) in Laboratory Conditions: LD50, J. Med.Entomol. 45 (4): 715-719.) By establishing an integrated control plan. 35

In the state of the art the activity against Hyalomma spp ticks is known. and Rhipicephalus (Boophilus) of various compounds of natural origin. In particular, Maupin, Gary, O. et al describe the ability of eremophilic sesquiterpenes as tick control (PCT / US01 / 47736), among which are named Valencian, Nootkatol, Epinootkatol, 40 and Nootkatone. Additionally, Sardá Ribeiro LV et al. describe the use of an essential oil of Drimys brasiliensis Miers against larvae of Rhipicephalus (Boophilus) microplus cattle ticks and Rhipicephalus sanguineus dog ticks (Sardá Ribeiro LV et al., Parasitol. Res., 102, 2008, 531-535 ). On the other hand, Dolan M. C. et al. has described the ability of nootkatone and carvacrol to suppress Ixodes scapularis and Amblyomma 45 americanum in lyme disease (Dolan MC et al. Journal of Econoic Entomology, 102 (6), 2009, 2316-2324). Additionally, Macchioni et al. He has identified several sesquiterpenes and their acaricidal activity against Psoroptes cuniculi as major components of leaf oils of Laurus nobilis and L. novocanariensis (Macchioni et al., Journal of Essential Oil Research. Vol 18 (1), 2006). Also, Sevki Coskun et al. names the 50 tick capacity of carvacrol of essential oil over Rhipicephalus turanicus (Sevki Coskun et al Parasitol Res (2008) 103: 259-261).

With regard to the genus Hyalomma,
H. Cetin et al. has described the acaricidal capacity of carvacrol from the essential oil of Satureja thymbra L. on Hyalomma marginatum (
H. Cetin et al
Veterinary Parasitology.
Volume 170, Issues 3–4, June 24, 2010, 287-290).

 5

However, compounds called sesquiterpenes comprise a wide range of different skeletons, with very different properties and activities. Thus, for a person skilled in the art it is not evident that a sesquiterpene different from those described in any of the aforementioned documents shows acaricidal or tick-active activity.

 10

DESCRIPTION OF THE INVENTION

In a first aspect, the present invention relates to the use of at least one eremofilane sesquiterpene of formula (I) as a tick,

         (I) 15 O R 1 CH 3 CH 3 [1] X Z C 7

where

R1 is selected from the group consisting of H and OH;

[1] is selected from the group consisting of single bond and double bond;

X is selected from the group consisting of absent, CH3, OH, OOH and together with Z forms an epoxy of structure (Z-IV); and 20

Z is selected from the group consisting of (Z-I), (Z-II), (Z-III) and next to X form (Z-IV), provided that

a) when [1] is a double bond, X is absent and Z is (Z-I),

 (Z-I) 25 C7 R4 R3 R2

 where R2, R3 and R4 are independently selected from the group consisting of CH3, OH and OOH;

b) when [1] is a simple link and X is absent, Z is (Z-II), 30

 (Z-II) R4 R2 C7

where R2 and R4 are independently selected from the group consisting of CH3, OH and OOH;

 5

c) when [1] is a simple bond and X is selected from the group consisting of CH3, OH and OOH, Z is (Z-III),

 (Z-III) R 4 CH 2 C 7

where R4 is selected from the group consisting of CH3, OH and OOH; and 10

d) when [1] is a single bond, X and Z form an epoxide of formula (Z-IV),

 (Z-IV) C7 R4 R2 O

 fifteen

where R2 and R4 are independently selected from the group consisting of CH3, OH and OOH.

Eremofilane sesquiterpene of formula (I) as described in this patent application comprises any one of its isomers or mixtures thereof. Additionally, the eremofilane sesquiterpene of formula (I) also comprises any solvate thereof, in particular, any hydrate, alkanolate or mixtures.

In a preferred embodiment, the present invention relates to the use as tick of at least one eremophilane sesquiterpene of formula (I), wherein R1 is hydrogen, R2 and R4 are methyl, and R3 is OH.

In an even more preferred embodiment, the present invention relates to the use as tick of at least one eremofilane sesquiterpene of formula (I) selected from the group consisting of dehydrofukinone (compound 1), 11-hydroxyremophil-6 (7), 9 ( 10) -dien-8-30 one (compound 4) and 7-11-epoxieremophil-9 (10) -en-8-one (compound 8), as well as mixtures of isomers of at least any one of them.

In a particularly preferable embodiment, the eremofilane sesquiterpene of formula (I) is dehydrofukinone since it has an activity superior to the rest of the compounds, this being 35

even higher than other eremofilanes sesquiterpenes, such as nootkatone, as shown in table 1.

The compound known as dehydrofukinone, (4aR, 5S) -4a, 5-dimethyl-3- (propan-2-ylidene) -4,4a, 5,6,7,8-hexahydronaphthalen-2 (3H) -one, is an isolated Eremofilane sesquiterpene of S. 5 adenotrichius DC., (Astereceae), a plant that can be located in the central mountains of Chile, Las Coimas, Putaendo (V Region Chile). The structure of the active compound is known and has been previously published in Eremophil-7 (11) -dien-8-one (Dehidrofukinone) Journal Natural Products Vol 54, No. 1, pp 271-272 1991, Phytochemistry, 1973, Vol. 12 , pp. 2931-2933), Liebigs Ann. Chem. 1979, 470-472. However, its potent activity as a tick has not been described to date.

Eremofilanes sesquiterpenes of formula (I) referred to in the present invention can be isolated from plant extracts, for example S. adenotrichius DC, by extraction methods and, optionally, purification widely known to those skilled in the art. On the other hand, the eremofilanes sesquiterpenes of formula (I) of the present invention can also be obtained by chemical synthesis by conventional reactions known to an expert in the field of organic chemistry.

Additionally, the use of sesremiterpene eremophilane of formula (I), preferably dehydrofukinone, as a major component of a plant extract, for example from S. adenotrichius DC (Asteraceae), should be considered included within the scope of the present invention.

The present invention relates to the use as a tick of at least one of the 25 eremophilanes sesquiterpenes of formula (I) described in this patent application, "tick" being understood as a compound useful for preventing, controlling or eliminating ticks where said compound is applied . In particular, theremofilane sesquiterpenes of the present invention cause the death of the larvae of said ticks. 30

In another preferred embodiment, the present invention relates to the use of at least one of the eremophilanes sesquiterpenes of formula (I) as described in this patent application, preferably dehydrofukinone, compound 4, compound 8 or any combination thereof, as a tick against ticks belonging to at least one genus selected from the group consisting of Hyalomma spp. and Rhipicephalus spp .. Preferably, from Hyalomma lusitanicum or Rhipecephalus sanguineus ticks.

In this patent application, "any one of its combinations" means both the combination of more than one of the compounds of formula (I) to which reference is being made, and the combination of more than one of the isomers of a compound of specific formula (I), or a combination of more than one isomer of several of the compounds of formula (I) to which reference is being made.

As previously mentioned in the state of the art, ticks of the genus 45 Hyalomma spp. or Rhipicephalus spp. cause serious economic problems in livestock, problems aggravated by the development of resistance to some commercially available tick compounds. Additionally, both genders can be a disease transmission vector. Therefore, the present invention provides new tick agents, the sesquiterpenes of formula (I) described in this patent application and, in particular, dehydrofukinone, to prevent, control or eliminate ticks preferably from the Hyalomma spp. and Rhipicephalus spp .. In this way the economic losses mentioned above are reduced or avoided, to the

while the use of the Eremofilanes sesquiterpenes of formula (I), compounds of plant origin or hemi-synthetic derivatives thereof, allows their use in organic farming. Additionally, the development of new tick agents facilitates the rotation of different treatments, thus hindering the appearance of resistance.

 5

In another preferred embodiment, the present invention relates to the use of an eremofilane sesquiterpene of formula (I) as described in this patent application, where the mortality of tick larvae is at least 60% when said sesquiterpene is applied. formula (I) at a concentration of 5 μg / μl, it being especially preferred that the mortality of the larvae be at least 70% at said concentration. 10

In another preferred embodiment, the present invention relates to the use of at least one eremofilane sesquiterpene of formula (I) as described in this patent application for manufacturing a medicament for treating or preventing a disease caused by ticks in a human or animal . Preferably, the eremofilane sesquiterpene of formula (I) is selected from the group consisting of dehydrofukinone (compound 1), 11-hydroxyremophilyl-6 (7), 9 (10) -dien-8-one (compound 4), 7- 11-epoxieremophil-9 (10) -en-8-one (compound 8) and any one of its combinations, it being especially preferable that said sesquiterpene is dehydrofukinone.

 twenty

In an even more preferred embodiment, when the medicament is applied to a non-human animal, it is selected from the group consisting of livestock and domestic animal. Preferably, the cattle are selected from the group consisting of cattle, sheep and goats. Preferably, the pet is selected from the group consisting of dog and cat.

 25

In another even more preferred embodiment, the present invention relates to the use of at least one of the eremophilanes sesquiterpenes of formula (I), preferably dehydrofukinone, compound 4, compound 8 or any combination thereof, to make a medicament for treating or treating prevent a disease selected from the group consisting of Anaplasma phagocytophilum, Coxiella burnetii, Borrelia burgdorferi, hemorrhagic fever virus 30 Crimea Congo and Theileria spp.

In another even more preferred embodiment, the medicament referred to in this patent application is a formulation suitable for topical, oral or injectable administration. Formulations suitable for topical administration may be, for example, solution, emulsion, gel, suspension or cream. For example, these formulations may be suitable for immersion baths, spray baths or pour-ons. On the other hand, formulations suitable for oral administration may be liquid, for example, solution or syrup; or they can be solid such as powders or tablets. Additionally, formulations suitable for oral administration are also considered as solid foods or liquids supplemented with the eremophilane sesquiterpene of formula (I) described in this patent application. In addition, formulations suitable for use as a medicament according to the present patent application may also be injectable solutions.

In a particularly preferred embodiment, the medicament referred to in this patent application is a formulation suitable for topical administration. Preferably, the eremofilane sesquiterpene of formula (I) is selected from the group consisting of dehydrofukinone (compound 1), 11-hydroxyremophilyl-6 (7), 9 (10) -dien-8-one (compound 4), 7-11 -epoxyieremophil-9 (10) -en-8-one (compound 8) and any one of its combinations, it being especially preferable that said sesquiterpene is dehydrofukinone.

The formulations referred to in this invention may comprise one or more conventional excipients known to the skilled person, such as, for example, cellulose. As well as they can be obtained by conventional techniques widely known in the veterinary, pharmaceutical or supplemented food industry.

 5

The present invention also refers to an eremophilane sesquiterpene of formula (I) as described in this patent application, as a medicament. Preferably, the eremofilane sesquiterpene of formula (I) is selected from the group consisting of dehydrofukinone (compound 1), 11-hydroxyremophilyl-6 (7), 9 (10) -dien-8-one (compound 4), 7-11 -epoxieremophil-9 (10) -en-8-one (compound 8) and any one of its combinations, 10 being especially preferable that said sesquiterpene is dehydrofukinone.

It should be understood that the present invention also refers to at least one eremophilane sesquiterpene of formula (I) as described in this patent application, both in its broadest definition and in the different preferred embodiments mentioned above, to treat or prevent a disease caused by ticks in a human or animal. Preferably, ticks of at least one genus selected from the group consisting of Hyalomma spp. and Rhipicephalus spp.

In a preferred embodiment, the present invention also relates to at least one eremophilane sesquiterpene of formula (I) as described in this patent application, both in its broadest definition and in the different preferred embodiments mentioned above, for treating or prevent a disease selected from the group consisting of Anaplasma phagocytophilum, Coxiella burnetii, Borrelia burgdorferi, Crimean Congo hemorrhagic fever virus and Theileria spp. 25

In another preferred embodiment, the present invention describes the eremofilane sesquiterpene of formula (I) for treating or preventing a disease caused by ticks as described in this patent application, wherein said treatment or prevention takes place by topical administration of the sesquiterpene of the invention. 30

Additionally, it should be understood that the present invention also refers to a method for treating or preventing a disease caused by ticks in a human or animal, wherein said method comprises administering an eremophilane sesquiterpene of formula (I) as described in this application for patent, both in its broadest definition and in the different preferred embodiments mentioned above. Preferably, to treat or prevent a disease caused by ticks of at least one genus selected from the group consisting of Hyalomma spp. and Rhipicephalus spp.

In a preferred embodiment, the present invention relates to the method of treating or preventing a disease as described in this patent application, wherein the disease is selected from the group consisting of Anaplasma phagocytophilum, Coxiella burnetii, Borrelia burgdorferi, virus Hemorrhagic fever Crimea Congo and Theileria spp.

In another preferred embodiment, the method of the present invention for treating or preventing a disease caused by ticks as described in this patent application, comprises administering the eremofilane sesquiterpene of formula (I) of the present invention topically.

In another preferred embodiment, the present invention describes the use of at least 50 sesremiterpene eremophilane of formula (I) as described in this patent application to prevent, control or eliminate the presence of ticks in the environment of an animal or human . Preferably, the eremofilane sesquiterpene of formula (I) is selected from

group consisting of dehydrofukinone (compound 1), 11-hydroxieremophile-6 (7), 9 (10) -dien-8-one (compound 4), 7-11-epoxieremophile-9 (10) -en-8-one (compound 8) and any one of its combinations, it being especially preferable that said sesquiterpene is dehydrofukinone.

 5

By "environment" means in this patent application, any surface in contact with the animal or human susceptible to harbor ticks, such as grass, weeds, stable, corral, garden, park, etc.

In a second aspect, the present invention provides a tick product comprising at least one eremophilane sesquiterpene of formula (I) as described in the first aspect of this patent application. Preferably, the tick-shaped product comprises at least one sesquiterpene eremophilane selected from the group consisting of dehydrofukinone, 11-hydroxyremophil-6 (7), 9 (10) -dien-8-one and 7-11-epoxieremophil-9 (10) - en-8-one, as well as mixtures of isomers of at least one of them. fifteen

In an even more preferred embodiment, the present invention provides a tick product comprising dehydrofukinone.

In another preferred embodiment, the tick product object of this invention may comprise other active compounds, preferably other compounds of plant origin with tick activity. For example, nootkatone or a derivative thereof.

In another preferred embodiment, the tick product comprising at least sesremiterpene eremophilane of formula (I) as described in this patent application 25 is a medicament. In particular, a medicament for treating or preventing a disease caused by ticks in a human or animal as described in the first aspect of this invention.

The medicament of the present invention can be administered by any route of usual administration in the pharmaceutical or veterinary industry. Preferably, this medicament can be administered topically, orally or by injection. Suitable formulations in both cases may be the same as described above in the first aspect of this invention.

 35

In an even more preferred embodiment, the tick product as described in this patent application is suitable for topical administration on a human or animal.

In another preferred embodiment, the tick product comprising at least sesremiterpene eremophilane of formula (I) as described in this patent application 40 is a product suitable for application in the environment of a human or animal. In particular, this product may be a liquid solution since these are easy to apply by spray or immersion bath.

EXAMPLES 45

EXAMPLE 1: Ethanolic extraction from S. adenotrichius DC (Asteraceae) and chromatographic purification of the extract obtained

The plants were dried, pulverized (1,185 kg) and extracted with ethanol at room temperature for 72 hours. They were subsequently filtered and the solvent was removed by concentration in vacuo. The ethanolic extract (102 g, 8.6% yield with respect to the weight of the dried plant) was chromatographed on a silica gel column (Si gel) of chromatography of

Flash column (FCC), using a gradient of hexane-ethyl acetate (100: 0-0: 100) and methanol (100%), obtaining six main fractions (identified from 0 to 6).

A portion (15.0 g) of fraction 1 (1.27% rend) was purified by silica gel column 5 flash column chromatography (hexane-AcOEt 90:10) and chromatographed through Sephadex LH-20 (hexane-CH2Cl2-MeOH 3: 1: 1) to obtain fractions 5-6 (2.80 g), 8-13 (76.7 mg), 14-18 (761.6 mg), 19-24 ( 944.1 mg) and 30-49 (698.1 mg).

All fractions were chromatographed by silica gel, from fractions 5-6, 1.5 g of compound 1 (126.6 x 10-3% yield) were obtained. The compounds were separated by semi-preparative normal phase HPLC on a silica column at a flow rate of 3ml / min and an isocratic mixture of hexane-AcOEt. From fractions 8-13, 6.0 mg of compound 4 (5.1 x 10-4% yield) was obtained, from fractions 19-24, 1.8 mg of compound 5 (1.5 x 10-4) was obtained. % rend). fifteen

 Compound 1 = Dehydrofukinone (Eremofil-7 (11), 9 (10) -dien-8-one) O145678101112131415

Majority Compound

HREIMS [M +] 218.1671 calculated for C15H22O. twenty

  Compound 4 = 11-Hydroxieremophil-6 (7), 9 (10) -dien-8-one OOH

HREIMS [M +] 234.1620 calculated for C15H22O2.

 25

 Compound 5 = 11-Hydroperoxieremophil-6 (7), 9 (10) -dien-8-one OOOH

HREIMS [M + -O] = 234.1620 calc. for C15H22O2

EXAMPLE 2: Obtaining compound 8 by epoxidation of dehydrofukinone 30

A mixture of compound 1 (500 mg, 1,798 mmol), CHCl3 (10 ml) and meta-chloroperoxybenzoic acid (465.2 mg, 2,696 mmol) was stirred for 5 hours at room temperature. Subsequently, the mixture was extracted at pH 10 with CH2Cl2 to obtain a crude extract (503.2 mg, 95% yield). Said extract was purified by SiO2 chromatographic column.

(hexane / AcOEt 80:20) obtaining compound 8 as a mixture of enantiomers (374.9 mg, 71% yield).

Compound 8 = 7-11-epoxieremophil-9 (10) -en-8-one (both 5 OO epoxides

       and its enantiomer

Bioassays:

 10

1. Maintenance of Hyalomma lusitanicum in the laboratory

Gravid females of H. lusitanicum were captured at the farm “La Garganta”, located in the south of the province of Ciudad Real at 37º24’78’’N 42º59’101’E at 669 meters above sea level. There are two procedures for obtaining copies. On the one hand, its capture directly from the soil or vegetation using the trawling technique or the flag (Sonenshine DE. 1993. Biology of ticks, Vols II. New York (USA): Oxford University Press), applied for a while 30 minutes at each sampling point recording temperature, humidity and soil moisture, height of vegetation, soil condition, weather and presence or not of animals in the area. On the other hand, obtaining 20 copies directly from the host (deer, wild boar and rabbit) by collecting them manually. For the identification of ticks the specific keys of Gil-Collado (1979) (Gil-Collado J, Guillén JL, Zapatero LM. 1979 were used. Keys for the identification of Spanish ixodoidea (adults). Iberian Journal of Pasarithology, 39 : 107-111); and Estrada et al. (2004) (Estrada Peña A, Bouattour A, Camicas JL, Walker AR. 2004. 25 Ticks of Domestic Animals in the Mediterranean Region: A Guide to Identification of Species. University of Zaragoza). Once the ticks were identified, the gravid females were separated from the rest and placed in glass test tubes with a piece of zig-zag filter paper inside and a cotton plug. The tubes were incubated at 26 ° C with an RH greater than 70% until oviposition took place. Once the oviposition was produced, they were left around 23-24 days to allow time for the eggs to hatch and the larvae to ripen. The larvae of H. lusitanicum require 15-16 days to complete the chitinization, that is, to mature (Ouhelli H, Pandey VS. 1984. Development of Hyalomma lusitanicum under Laboratory Conditions. Veterinary Parasitology, 15: 57-66). 35

2. Toxicity tests against Hyalomma lusitanicum

To evaluate the tick activity of compounds 1, 4, 5 and 8, three 40 eppendorf tubes were prepared for each eremofilane sesquiterpene to be tested with 25 mg of cellulose. 50 µl of the stock solution was deposited on the cellulose at the desired concentration (see table 1) and on the other hand the same was done with the solvent used. In this test the acetone solvent was used as a negative control and acibelte (0.5% cypermethrin) as a positive control. The cellulose was then allowed to dry with the products under vacuum and at room temperature.

Next, batches of 20 larvae of H. lusitanicum were prepared in glass test tubes. To do this, by using a brush the larvae were deposited in water and small groups of larvae were placed on filter paper and finally deposited one by one in the test tube. Finally, the cellulose was poured with the product to be tested in the tube with the larvae, it was covered with cotton and placed face down. 5

The tubes were left incubating for 24 hours at 26 ° C with an RH greater than 70% and after that time the observation was carried out, recording the number of live and dead larvae of each test tube.

 10

Subsequently the percentage of mortality was calculated with the following equation:

% Mortality = (% T -% C / 100 -% C) x 100

% T: Percentage of mortality in treatment. fifteen

% C: Percentage of mortality in the control.

3. Toxicity tests against R. sanguineus

To perform this bioassay, the conditions described above with respect to bioassay 2 were used. The activity of compounds 1, 4 and 5 at a concentration of 5 µg / µl was analyzed (see results in table 1).

Table 1: Results obtained against H. lusitanicum and R. sanguineus ticks

 25

 Concentration

 Product H. lusitanicum R. sanguineus

 5µg / µl

   O145678101112131415 (Comp. 1) 88 72

 5µg / µl

   OOH (Comp. 4) 74 29

 5µg / µl

   OOOH (Comp. 5) 14 41

 10µg / µl

      OO (Comp. 8) 72 -

 5µg / µl

    64 -

 2µg / µl

    17 -

 10µg / µl

    Nootkatona 83.3 -

As a comparative example, the tick-killing activity of nootkatone (Sigma-Aldrich) against H. lusitanium was also evaluated under the conditions described in bioassay 2 (see results in table 1).

 5

Additional References:

-Barandika Iza JF. 2010. Exophilic ticks as vectors of zoonotic agents: study on the abundance and activity of ticks in vegetation and investigation of the presence of pathogens in ticks and micromamifers. Doctoral thesis. 10 University of León, Faculty of Veterinary Medicine, Department of Animal Health.

-González-Coloma A. 2009. Testing with insects: experimental design. In: Burillo J, González-Coloma A (Eds.) Insecticides and repellents of natural origin. Food Research and Technology Center, Zaragoza. fifteen

Claims (15)

1. Use of at least one eremofilane sesquiterpene of formula (I) as a tick,          (I) O R 1 CH 3 CH 3 [1] X Z C 7  5 where R1 is selected from the group consisting of H and OH; [1] is selected from the group consisting of single bond and double bond; X is selected from the group consisting of absent, CH3, OH, OOH and together with Z forms an epoxy of structure (Z-IV); Y Z is selected from the group consisting of (Z-I), (Z-II), (Z-III) and next to X form (Z-IV), provided that a) when [1] is a double bond, X is absent and Z is (Z-I), 15  (Z-I) C7 R4 R3 R2  where R2, R3 and R4 are independently selected from the group consisting of CH3, OH and OOH;  twenty b) when [1] is a simple link and X is absent, Z is (Z-II),  (Z-II) where R2 and R4 are independently selected from the group consisting of CH3, OH and OOH; 25 c) when [1] is a simple bond and X is selected from the group consisting of CH3, OH and OOH, Z is (Z-III),  (Z-III) where R4 is selected from the group consisting of CH3, OH and OOH; Y d) when [1] is a single bond, X and Z form an epoxide of formula (Z-IV),  (Z-IV) C7 R4 R2 O where R2 and R4 are independently selected from the group consisting of CH3, OH and OOH. 10 2. Use of at least one eremofilane sesquiterpene according to claim 1, wherein R1 is hydrogen, R2 and R4 are methyl, and R3 is OH. 3. Use of at least one eremofilane sesquiterpene according to claim 2, wherein the eremofilane sesquiterpene is selected from the group consisting of dehydrofukinone, 11-hydroxyremophilyl-6 (7), 9 (10) -dien-8-one and 7- 11-epoxieremophil-9 (10) -en-8-one, or a mixture of isomers of at least any one of them. 4. Use of at least one eremofilane sesquiterpene according to any one of 20 claims 1 to 3, against ticks belonging to at least one genus selected from the group consisting of Hyalomma spp. and Rhipicephalus spp. 5. Use of an eremofilane sesquiterpene according to any one of claims 1 to 4, wherein the mortality of tick larvae is at least 60% when applying the compound of formula (I) at a concentration of 5 μg / μl. 6. Use of at least one eremofilane sesquiterpene according to any one of claims 1 to 5, to manufacture a medicament for treating or preventing a disease caused by ticks in a human or animal. 30 7. Use of at least one eremofilane sesquiterpene according to claim 6, wherein the disease is selected from the group consisting of Anaplasma phagocytophilum, Coxiella burnetii, Borrelia burgdorferi, Crimean Congo hemorrhagic fever virus and Theileria spp.  35 8. Use of at least one eremofilane sesquiterpene according to any one of claims 6 to 7, wherein the medicament is administered topically. 9. Use of at least one eremofilane sesquiterpene according to any one of claims 1 to 5, to prevent, control or eliminate the presence of ticks in the environment in contact with an animal or human. 10. Tick product comprising at least one eremofilane sesquiterpene of formula (I)    (I) where 5 R1 is selected from the group consisting of H and OH; [1] is selected from the group consisting of single bond and double bond; X is selected from the group consisting of absent, CH3, OH, OOH and together with Z forms an epoxy of structure (Z-IV); Y Z is selected from the group consisting of (Z-I), (Z-II), (Z-III) and next to X form (Z-IV), 10 whenever a) when [1] is a double bond, X is absent and Z is (Z-I),  (Z-I)  fifteen  where R2, R3 and R4 are independently selected from the group consisting of CH3, OH and OOH; b) when [1] is a simple link and X is absent, Z is (Z-II),  (Z-II) 20 where R2 and R4 are independently selected from the group consisting of CH3, OH and OOH; c) when [1] is a simple bond and X is selected from the group consisting of CH3, OH and 25 OOH, Z is (Z-III),  (Z-III) R4 CH2 C7 where R4 is selected from the group consisting of CH3, OH and OOH; Y d) when [1] is a single bond, X and Z form an epoxide of formula (Z-IV),  (Z-IV) where R2 and R4 are independently selected from the group consisting of CH3, OH and OOH. 10 11. Product according to claim 10, which comprises at least one sesquiterpene eremophilane selected from the group consisting of dehydrofukinone, 11-hydroxyremophil-6 (7), 9 (10) -dien-8-one and 7-11-epoxieremophil-9 (10) -en-8-one, or a mixture of isomers of at least any one of them. fifteen 12. Product according to claim 11, comprising dehydrofukinone. 13. Product according to any one of claims 10 to 12, wherein said product is a medicament. twenty 14. Product according to claim 13, wherein the medicament is suitable for topical administration on a human or animal. 15. Product according to any one of claims 10 to 12, wherein said product is suitable for application in the environment of a human or animal.