OA17154A - Dronedarone for use in leishmaniasis, formulations and associations for use in leishmaniasis - Google Patents
Dronedarone for use in leishmaniasis, formulations and associations for use in leishmaniasis Download PDFInfo
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- OA17154A OA17154A OA1201400516 OA17154A OA 17154 A OA17154 A OA 17154A OA 1201400516 OA1201400516 OA 1201400516 OA 17154 A OA17154 A OA 17154A
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- 229960002084 dronedarone Drugs 0.000 title claims abstract description 96
- ZQTNQVWKHCQYLQ-UHFFFAOYSA-N dronedarone Chemical compound C1=CC(OCCCN(CCCC)CCCC)=CC=C1C(=O)C1=C(CCCC)OC2=CC=C(NS(C)(=O)=O)C=C12 ZQTNQVWKHCQYLQ-UHFFFAOYSA-N 0.000 title claims abstract description 95
- 239000000203 mixture Substances 0.000 title claims abstract description 54
- 238000009472 formulation Methods 0.000 title claims abstract description 52
- 208000004554 Leishmaniasis Diseases 0.000 title claims abstract description 38
- 238000011282 treatment Methods 0.000 claims abstract description 40
- 150000003839 salts Chemical class 0.000 claims abstract description 13
- XOGYVDXPYVPAAQ-SESJOKTNSA-M meglumine antimoniate Chemical group O[Sb](=O)=O.CNC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO XOGYVDXPYVPAAQ-SESJOKTNSA-M 0.000 claims description 41
- 229940005559 meglumine antimoniate Drugs 0.000 claims description 27
- 229960003775 miltefosine Drugs 0.000 claims description 24
- 241000222732 Leishmania major Species 0.000 claims description 12
- APKFDSVGJQXUKY-KKGHZKTASA-N Amphotericin-B Natural products O[C@H]1[C@@H](N)[C@H](O)[C@@H](C)O[C@H]1O[C@H]1C=CC=CC=CC=CC=CC=CC=C[C@H](C)[C@@H](O)[C@@H](C)[C@H](C)OC(=O)C[C@H](O)C[C@H](O)CC[C@@H](O)[C@H](O)C[C@H](O)C[C@](O)(C[C@H](O)[C@H]2C(O)=O)O[C@H]2C1 APKFDSVGJQXUKY-KKGHZKTASA-N 0.000 claims description 11
- 229960003942 amphotericin b Drugs 0.000 claims description 11
- 206010011668 Cutaneous leishmaniasis Diseases 0.000 claims description 10
- 239000003814 drug Substances 0.000 claims description 10
- 241000222724 Leishmania amazonensis Species 0.000 claims description 8
- 241000222727 Leishmania donovani Species 0.000 claims description 8
- 239000003795 chemical substances by application Substances 0.000 claims description 7
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 7
- 239000001866 hydroxypropyl methyl cellulose Substances 0.000 claims description 6
- 235000010979 hydroxypropyl methyl cellulose Nutrition 0.000 claims description 6
- 229920003088 hydroxypropyl methyl cellulose Polymers 0.000 claims description 6
- UFVKGYZPFZQRLF-UHFFFAOYSA-N hydroxypropyl methyl cellulose Chemical group OC1C(O)C(OC)OC(CO)C1OC1C(O)C(O)C(OC2C(C(O)C(OC3C(C(O)C(O)C(CO)O3)O)C(CO)O2)O)C(CO)O1 UFVKGYZPFZQRLF-UHFFFAOYSA-N 0.000 claims description 6
- CPKOXUVSOOKUDA-UHFFFAOYSA-N 1-bromo-5-fluoro-2-iodo-4-methylbenzene Chemical group CC1=CC(I)=C(Br)C=C1F CPKOXUVSOOKUDA-UHFFFAOYSA-N 0.000 claims description 4
- 229960002919 dronedarone hydrochloride Drugs 0.000 claims description 4
- 238000011200 topical administration Methods 0.000 claims description 4
- 244000045947 parasite Species 0.000 description 27
- PQLXHQMOHUQAKB-UHFFFAOYSA-N miltefosine Chemical compound CCCCCCCCCCCCCCCCOP([O-])(=O)OCC[N+](C)(C)C PQLXHQMOHUQAKB-UHFFFAOYSA-N 0.000 description 23
- 208000015181 infectious disease Diseases 0.000 description 15
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- 210000005069 ears Anatomy 0.000 description 9
- 238000000338 in vitro Methods 0.000 description 9
- 229940079593 drug Drugs 0.000 description 8
- 206010003658 Atrial Fibrillation Diseases 0.000 description 7
- 238000012360 testing method Methods 0.000 description 7
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- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- 230000000996 additive effect Effects 0.000 description 6
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- 230000000699 topical effect Effects 0.000 description 4
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- 230000001988 toxicity Effects 0.000 description 4
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 3
- 210000004207 dermis Anatomy 0.000 description 3
- 230000002401 inhibitory effect Effects 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
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- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 206010014025 Ear swelling Diseases 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- 241000222722 Leishmania <genus> Species 0.000 description 2
- MBBZMMPHUWSWHV-BDVNFPICSA-N N-methylglucamine Chemical compound CNC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO MBBZMMPHUWSWHV-BDVNFPICSA-N 0.000 description 2
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- 125000000400 lauroyl group Chemical group O=C([*])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
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Abstract
The invention relates to dronedarone or one of its pharmaceutically acceptable salts for the treatment of leishmaniasis, formulations and associations comprising dronedarone or one of its pharmaceutically acceptable salts for the treatment of leishmaniasis.
Description
DRONEDARONE FOR USE IN LEISHMANIASIS, FORMULATIONS AND
ASSOCIATIONS FOR USE IN LEISHMANIASIS.
The présent invention relates to dronedarone or one of its pharmaceutically acceptable salts for the treatment of leishmaniasis, in particular cutaneous leishmaniasis with its various straîns around the worid, and/or leishmaniasis issued from Leishmania amazonensis, Leishmania donovani.or Leishmania major as well to formulation in particular topical formulation comprising dronedarone or one of its pharmaceutically acceptable salts, to their préparation and to their therapeutic application.
2-n-Butyl-3-[4-(3-di-n-butylaminopropoxy)benzoyl]-5-methylsu!phonamidobenzofuran, or dronedarone, and pharmaceutically acceptable salts thereof, in particular its hydrochloride salts, are described in Européen Patent EP 0 471 609 B1.
Moreover, dronedarone is indicated to reduce the risk of hospitalization for atrial fibrillation In patients ln sinus rhythm with a history of paroxysmal or persistent atrial fibrillation (AF) or Is Indicated for the maintenance of sinus rhythm after successful cardioversion in adult clinically stable patients with paroxysmal or persistent atrial fibrillation (AF).
Surprisingly, the applicant has now showed that dronedarone may be used to treat leishmaniasis.
Especially, the applicant proposed formulations for topical administration that are suitable to treat leishmaniasis.
In fact, to be effective, such formulation should allow the penetration/liberation of the active principle In the layer of the skin where the parasites are located.
It would then be possible to obtain high concentrations of the active drug locally ln the dermîs, and avoiding high plasma concentration of the drug and associated systemic side effects.
One additional feature of such formulation is to avoid/reduce toxicity reactions of the skin on contact with the formulation.
Furthermore, association of dronedarone with others anti-leishmaniasis agents is possible and has several advantages such as decreasing the dose of administrated drugs to avoid side effects and avoid apparition of résistances to the chosen treatment with time.
Thus, the présent invention relates to formulation in particular topical formulation comprising dronedarone or one of its pharmaceutically acceptable salts and at least a pharmaceutically acceptable excipient, to their préparation and to their therapeutic application such as treatment of leishmaniasis, in particular cutaneous leishmaniasts and/or leishmaniasis issued from Leishmanie amazonensis, Leishmanie donovani or Leishmanie major strains.
The présent invention relates to a pharmaceutical composition in particular for topical administration (topical pharmaceutical composition) comprising dronedarone or one of its pharmaceutically acceptable salts and at least a pharmaceutically acceptable excipient for topical administration, to their préparation and to their therapeutic application such as treatment of leishmaniasts, in particular cutaneous leishmaniasis and/or leishmaniasis issued from Leishmanie amazonensis, Leishmanie donovani or Leishmania major strains.
The présent invention also relates to the use of dronedarone or one of its pharmaceutically acceptable salts for the préparation of a medicine for the treatment of leishmaniasis particularly cutaneous leishmaniasis and/or leishmaniasis issued from Leishmania amazonensis, Leishmania donovani or Leishmania major strains.
The présent invention also relates to dronedarone or one of its pharmaceutically acceptable salts for use in the treatment of leishmaniasis particularly cutaneous leishmaniasis and/or leishmaniasis issued from Leishmania amazonensis, Leishmania donovani or Leishmania major strains.
Another objet of the invention is also an association or one of its pharmaceutically acceptable salts and an anti-leishmaniasis agent Said association is used for the treatment of leishmaniasis particularly cutaneous leishmaniasis and/or leishmaniasis issued from Leishmania amazonensis, Leishmania donovani or Leishmania major strains.
ln one embodiment, said anti-leischmaniasis agent is selected among the following agents:
Miltefosine or one of its pharmaceutically acceptable dérivatives,
- Amphotericin B or one of its pharmaceutically acceptable dérivatives,
- pentavalent antimonials dérivatives such as meglumine antimoniate.
Said pharmaceutically acceptable sait of dronedarone is the hydrochloride sait.
In one embodiment, said leishmaniasis is issued from Leishmanie strains résistant to pentavalent antimonials dérivatives , and in particular Leishmanie amazonensis strains, résistant to pentavalent antimonials dérivatives in particular résistant to meglumine antimoniate.
It may be mentioned that formulations of meglumine antimoniate are notably commercialized under the trademark Glucantime®.
Said topical formulation or pharmaceutical composition may be a hydro-alcoholic gel, a semi-solid hydrophilic waxy formula, an otl In water or a water in oil émulsion particulariy a hydro-alcoholic gel.
Said topical formulation or pharmaceutical composition may comprise an excipient such as hydroxypropyl methylcellulose (HPMC) in particular HPMC at 2% of the total weight of the formulation.
In one embodiment, said topical formulation or pharmaceutical composition may be a hydro-alcoholic gel comprising at least hydroxypropyl methylcellulose as excipient
Said topical formulation or pharmaceutical composition wherein dronedarone or one of its pharmaceutically acceptable sait may be used in a proportion of 10% by weight of the active principle in base form.
Higher or lower dosages may be appropriate; these dosages are comprised within the scope ofthe présent invention.
It may be mentioned that the term formulation or composition may be used Indifferently.
The examples which foilow describe the préparation of certain formulations in accordance with the invention. These examples are not limitative, and merely lllustrate the présent invention.
EXAMPLE 1
Tests against cutaneous leishmaniasis were performed using the experimental model of BALB/c mouse infection with Leishmanie amazonensis.
Formulations:
Formulations are prepared using techniques well known by one skilled in the art.
a) Dronedarone.
| Formulation 5 | Dronedarone | aqueous gel, [Dronedarone] =10%, in 20g vial |
| Formulation 6 | Dronedarone | semi solid hydrophilic waxy formula, [Dronedarone] = 10%, ln 20g vial |
| Formulation 7 | Dronedarone | oil in water émulsion, [Dronedarone] = 10%, in 20g vial |
Formulations 5,6 and 7 are detailed below:
| Formulation 5 (hydro-alcoholic gel = aqueous gel) | % |
| citrate buffer 0.5M pH 4.0 | 44 |
| Ethanol | 44 |
| HPMC | 2 |
| dronedarone | 10 (eq.base) |
| Formulation 6 (semi-solid hydrophilic waxy) | % |
| Lauroyl polyoxylglycerides (Gelucire 44/14) | 75 |
| Propylene glycol | 15 |
| dronedarone | 10 (eq.base) |
| Formulation 7 (oil in water émulsion*) | % |
| Anionic self-emulsifying base based on palmitostearate dérivatives (SEDEFOS 75) | 8.1 |
| Soya oil | 22.5 |
| Glycerin | 59.4 |
| dronedarone | 10 (eq. Base) |
b) Control Glucantime formulations :
* Water is replaced by an hydrophilic excipient. Thus, the émulsion is constituted by droplets of oil dispersed in a hydrophilic matrix.
• Glucantime- PBS formulation #8 : injectable formulation of Glucantime mixed with stérile PBS (20 mg / ml), stored at 4*C.
This préparation is équivalent to the commercial Glucantime®, identified by WHO as a first intention treatment for cutaneous leishmaniosis.
Procedures
a)
Protocol timelines:
Day
+
41
| Ph | JtO | Photo Photo | Ph | oto K | |||
| Clinlcal sIg ns | |||||||
| -t | 'mest | ||||||
| J* | Infection | 1 J Fiuorlmetry | |||||
| *--- | Tteatment |
LDA
Lésion growth data analysis & report
b) Infection
Mice at 8 weeks of âge were infected with 10 μΙ of 10e promastigotes in the right ear pinna under light ether anesthésia.
c) Lésion growth
Infected and non-infected contralatéral ear thicknesses were measured every 3-4 days with a caliper gauge, and the lésion sizes were expressed as the différence between the thickness of infected and non-infected ears.
d) Treatment with intra lesional glucantime and topical dronedarone
On day 7 of infection, animais were randomly separated at 5 animais / mouse cage (1 group of animais for each dronedarone or glucantime formulation). Another infected group was only for infection follow-up.
i) Topical treatment: Infected ears were treated with approx 20 mg of the appropriate topical dronedarone formulation once a day for 30 days. The formulations were collected from their original récipient with the tip of a disposable plastic spatula, and spread In the inner side of the infected pinna under a 10 secs-massage using the same spatula:
Ii) Intralesional treatment: Infected ears were s.c. injected în the inner side of the infected ear with 10 μΙ of PBS containing 200 pg of powder Glucantime, twice a week from days 7 to 37 of infection (8 doses):
e) Cutaneous sensitization (MEST):
Mouse Ear Swelling Test (MEST) can detect reliably moderate to strong sensitizers, as Indicated by OECD guidelines for testing of chemicals. On day 34 of infection the contralatéral non-infected ears were treated (challenged) with the correspondent Dronedarone formulation. Ear swelling was measured at various times after challenge (0h, 1h, 5h. 24h, 48 h and 72 h).
f) Clinical signs:
Animais were weighted on days 0 of infection and at weekly péri ods, subsequently. The animais were observed daily, for signs of morbidity and death. Abnormal clinical signs during these routine checks were recorded. Clinical observations including mortality, monitoring of convulsions, lethargy, sleep, coma, salivation.diarrhea, cage side examination, skin color, fur, eyes and mucous membrane,spontaneous and voluntary motor activity and necropsy — in case the animal dies were recorded. Any unusual aspect of the treated lésions such as vesicles, complété ulcération, crust, darkening were also recorded. according to the international guidelines (WHO /OECD).
g) Pictures
Pictures from infected ears (treated and untreated) were taken with a digital caméra at weekly intervals.
h) Parasite load by Limiting Dilution Assay (LDA)
On day 39 of infection, animais were sacrificed under anesthésia and the infected ears were aseptically cleaned with iodinated éthanol followed by éthanol alone. The ears were eut off along its base and aseptically weighted. They were then indïvîdually eut into pièces, and single-cell suspensions were prepared in PBS containing antibiotics (1 ml / ear) using a stainless steel mesh (Sigma). The sieved tissues were gently pipetted updown 8 x for cell dissociation. The cell suspensions were pre-diluted 500x in medium M199 supplemented with 10% heat inactivated foetal calf sérum, antibiotics 100 U/ml Penicilin /100 gg/ml streptomycin, 5 ug/ml hemine and 2% human urine), and then serially diluted 3-fold (50 μΐ + 100μ1) in triplicates in flat-bottom micropates for a total of 24 dilutions. Microplates were incubated at 26*C in a humidified BOD Incubator. Wells were checked every 3-4 days for the presence of promastigotes for up to 20 days. The number of parasites in each infected ear was calculated according to the tissue mass and the last dilution theoretically containing 1 tissue amastigote.
Results on effîcacy of dronedarone formulations:
Dronedarone: Lésion growth: Formulation # 5 (aqueous gel) was very effective throughout treatment, close to the reference treatment # 8 (intralesional Glucantime) (Fig 9). Formulations # 6 (hydrophilic waxy semi-solid) and # 7 (émulsion o/w) induced increased lésion slzes, This was due to skin irritancy (see MEST Fig 11), not to promoted parasite growth (see Fig 10). Besides, suspension of # 6 and # 7 treatments on day 32 led to rapid decrease of ear thicknesses: afterwards:
LESION GROWTH - Dronedarone formulations
day* after Infection
Figure 9: Lésion growth in Dronedarone-treated mice. Mice were infected on
Day 0. Topical daily treatment started on Day 7 and continued untii Day 37 with ail the indicated Dronedarone formulations, except for Groups # 6 and #7 which 5 treatment was suspended on Day 32 due to toxicity (skin inritancy). The lésion sizes were measured at the indicated times (n=5).
Dronedarone: Parasite loads In the ear. On day 39 of infection (30 days of treatment), 10 the parasite loads in the ears were signrficantly lower in the group topically treated with # (aqueous gel). Formulations # 6 (hydrophilic waxy semi-solid) and # 7 (émulsion o/w) did not alter the parasite loads (Fig 10):
___ «S (Prefo 03-120)
E3 WO (Prefo 03- 130)
M *7 (Prefo 03 - 132) «8 (GkJcentime - IL)
M «10 (Untreated)
5 6 7 8
Formulation
Figure 10: Parasite loads in Dronedarone-treated mice. On day 39 of infection, the parasite loads in the ears of mice treated with the indicated formulations as for Fig 9 were determined by Limiting Dilution Assay. Means ±
SD(n=5). **p< 0.005, ***p<0.001
Conclusions
The formulation 5 was the most promising in the control of lésion growth throughout the 10 infection, the low parasite load at the end of the experiment was compatible with the controlled lésion growth, similar to achieved with 8 doses of 200 pg intralesional Glucantime. The animais looked happy throughout the treatment, gained weight normally and did not mount cutaneous sensitivity upon challenge.
EXAMPLE 2
In vitro study of the efficacy of dronedarone on a strain of Lelshmanla amazonensls In comparison with the reference treatments meglumlne antlmonlate, miltefoslne, and amphotericln Bln similar laboratorv conditions..
A strain of Leishmania amazonensis (MHOM / BR / 73 / M2269) were used. Dronedarone hydrochloride and meglumine antimoniate were supplied by Sanofi. Amphotericin B (désoxycholate) and miltefbsine were bought at Sigma-Aldrich.
The study concemed two cellular models:
-1/Axenic amastigotes of Lamazonensis ( isolated parasites)
- 21 Intramacrophagic amastigotes of L. amazonensis (parasites hosted in macrophages, as found in dermis, during the cutaneous Leishmaniosis infection).
The first cellular model which cultivâtes in a medium based on M199, allows defining the intrinsic activity of a substance on the parasite itsetf while the second cellular model intégrâtes the capacity of the substance to cross the membrane of the macrophage and that of the phagolysosome. The used macrophages are cells RAW 264.7, which cultivate in medium based on DMEM with 10 % of foetal sérum of veal.
• Détermination of the IC50 of each of the moiecules
Solutions of dronedarone in the DMSO, meglumine antimoniale and miltefosine in the water, and amphotericin B in an isotonie glucose solution (G5) were prepared before every experiment The évaluations in vitro were the object of three independent experiments duplicate. The assays were realized by quantifying the parasitic DNA with the SYBRGREEN as described by Audisio and al. (Eur. J. Med. Chem 52: 44-50, 2012) as well as by qPCR for intramacrophagic amastigotes with amplification of the alpha-tubuline ofthe parasite. The results obtained with both methods are similar.
The inhibition concentration of 50 % of the growth of the parasites (IC50) was determined after 72 hours of contact with dronedarone compared with meglumine antimoniale, miltefosine and amphotericin B, used as reference products.
On axenic amastigotes (test conditions 1/), dronedarone has a IC50 of 0,34±0,06 μΜ, while meglumine antimoniate has a IC50 of 9081159 μΜ, miltefosine a IC50 of 0,910,2 μΜ and amphotericin B a IC50 of 0,03110,002 μΜ.
On intramacrophagic amastigotes (test conditions 2/) , dronedarone has a IC50 of 0,5010,22 μΜ, while meglumine antimoniate has a IC50 of 133,63110,41 μΜ, miitefosine a IC50 of 1,8710,032 μΜ and amphotericin B a IC50 of 0,04710,005 μΜ.
• Evaluation of the potential toxicity of dronedarone on human host cells (macrophages) compared to the reference drugs.
Study of the cytotoxlcity on macrophages (cells RAW 264.7)
The non-toxic maximal concentration (CMA) was determined by the technique to the Blue Trypan according to has method described in Audisio and al. (Eur. J. Med. Chem 52:44-50,2012).
The CMA of the dronedarone is 12,5 μΜ
The CMA of the meglumine antimoniate is > 200 μΜ.
The CMA of the amphotericin B is 6,25 μΜ
The CMA of the miltefosine ls 50 μΜ
Then, the toxicity of dronedarone on the dermis macrophages ls lower than most of the référencé drugs, and will not Impede its therapeutic use for cutaneous leishmaniasis.
• Study of the association of dronedarone with meglumine antimoniate, miltefosine and amphotericin B
Association of dronedarone with each of the référencé products was studied according to the method described by Odds and al., J. Antimicrob. Chemother., 52:1, 2003, to Identify a synergie or additive or antagonistic action. The study comprised three independent experiments which allowed the calculation of the Fractional Inhibitory Concentration Index (FICI):
- If FICI < 0,5: Synergy
- If 0,5 < FICI < 4: additive Effect, no interaction
- If FICI < 4: antagonism
- Interaction between dronedarone and meglumine antimoniate:
On axenic amastigotes , FICI = 0,27. A synergie effect between dronedarone and meglumine antimoniate was obtained.
On intramacrophagic amastigotes, FICI = 0,79. An additive effect between dronedarone and meglumine antimoniate was obtained.
- Interaction between the dronedarone and the miltefosine:
On axenic amastigotes , FICI = 0,39. A synergy between dronedarone and miltefosine was obtained.
On intramacrophagic amastigotes , FICI = 0,46. A synergy between dronedarone and miltefosine was obtained.
- Interaction between dronedarone and amphotericin B:
On axenic amastigotes, FICI = 0,63. An additive effect between dronedarone and amphotericin B was obtained.
On întramacrophagic amastigotes, FICI = 0,54. An additive effect between dronedarone and arnphotericin B was obtained.
Conclusions
A / activity of dronedarone on the parasites, as compared with reference drugs.:
Dronedarone présents a strong activity anti-leishmaniasis in vitro on both models axenic amastigotes and întramacrophagic amastigotes of Leishmanîa amazonensis, with lower IC50 within the mlcromolar range, what places it between arnphotericin B and miltefosine, in term of activity. The intrinsic activity on the parasite itself is thus maintained when this one Is protected in the phagolysosome inside the macrophage.
B/ activity of dronedarone on the parasites, in association with reference drugs:
The interaction of dronedarone with meglumine antimoniate or with arnphotericin B is additive type. No antagoniste effect was demonstrated, what suggests that a concomitant use of dronedarone with meglumine antimoniate or arnphotericin B may be possible.
The interaction of dronedarone with miltefosine is of synergie type suggests that a concomitant use of dronedarone with miltefosine may be of Interest for example to decrease the administrated doses.
EXAMPLE 3
In vitro study on Leishmanîa amazonensis strains résistant to meglumine antimoniate (Glucantlme®)
On Leishmanie amazonensis strains résistant to_meglumine antimoniate (IC50 = 200mM) under promastigote form, chemosensitivity to dronedarone Is similar that one observed on strains that are not résistant to meglumine antimoniate (active principle of Glucantime®).
That means that the mechanism of action of dronedarone on the parasites differs from the one of glucantlme. Dronedarone has a pot en liai to treat Glucantime résistant strains as found eg. in Latin America.
EXAMPLE 4 ln vitro atudv on building of résistance to Dronedarone with time, using Lelshmania amazonensis strain,
The test consists in long term culture of a L amazonensis strain ( promastygote form), in contact with increasing doses of dronedarone, starting with initial concentrations lower than the IC 50.
At regular timepoints , the viability and pathogenicity of parasites is confirmed with moving the culture to axenic amastigotes and intramacrophagic amastigotes,and calculating the new inhibitive concentration for 50% of the population (IC50).
After six months of culture and exposition of the parasites to dronedarone, no significant modification of the inhibitory concentration (IC 50) was observed.
As a reference: a similar exposition of the strain to Glucantime has brought to an increase of the IC 50 x 200.
While the study will be continued for a while, it can already be stated that the molecuie does not generate a résistance in exposed parasites.
EXAMPLE 5 ln vitro study on Lelshmania donovanl and Lelshmania malor
As to evaluate the polyvalence of dronedarone as a treatment on the various strains of Leishmania spread around the worid, with known various sensitivities to the reference drugs, the previous tests were complétée! with strains considered as représentative of the various zones of endemy.
Strains of Leishmania donovanl (MHOM / ET / 67 / HU3) and Leishmania major (MHOM / SU / 73 / 5-ASKH) were used for this study.
Dronedarone hydrochloride and the meglumine antimoniate were supplied by Sanofl. Miltefosine was bought at Slgma-Aldrich.
The study in vitro concemed four cellular models:
-1/ Axenic amastigotes of L donovani and L major ( isolated parasites)
- 2/ Intramacrophagic amastigotes of L. donovani and L. major (parasites hosted in macrophages, as found in dermis, during the cutaneous Leishmaniosis infection).
The model of axenic amastigotes is cultivated in a medium based on M199 and allows defining the intrinsic activity of a substance on the parasite itself while the model of intramacrophagic amastigotes intégrâtes the capacity of the substance to cross the membrane of the macrophage and that of the phagolysosome.
The used macrophages are cells RAW 264.7, which cultivate in a medium based on DMEM with 10 % of foetal sérum of veal.
Détermination of the IC50 of dronedarone, Glucantime ® and of miltefosine:
The solutions of dronedarone in the DMSO, meglumine antimoniate and miltefosine in the water, were prepared before every experiment. The évaluations in vitro were the object of three independent experiments.
The assays were realized by quantifying the parasitic DNA with the SYBRGREEN as described by Audisio and al. (Eur. J. Med. Chem ., 52:44-50, 2012).
The inhibition concentration of 50 % of the growth of the parasites (IC50) was determined after 72 hours of contact with dronedarone compared with meglumine antimoniate and miltefosine, used as reference products.
- On the axenic amastigotes of L donovani, dronedarone has a IC50 of 47,38±6,27 μΜ, while meglumine antimoniate has a IC50 > 1000 mM, and miltefosine a IC50 of 13,011,25 μΜ.
- On L. majoras axenic amastigotes, dronedarone has a IC50 of 2,55±0,51 μΜ, while meglumine antimoniate has a IC50 > 1000 μΜ, and miltefosine a IC50 of 5,57±1,50 μΜ.
- On the intramacrophagic amastigotes of Ldonovani, dronedarone has a 1C50 of 1,67±0,10 μΜ, while meglumine antimoniate has a IC50 of 390,36140,46 μΜ, and miltefosine a IC50 of 0,88±0,10 μΜ.
- On L majoris intramacrophagic amastigotes , dronedarone has a IC50 of 1,3210,12 μΜ, while meglumine antimoniate has a IC50 of 347,98179,87 μΜ, and miltefosine a IC50of 0,7210,11 μΜ.
Besides, dronedarone does not présent toxicity for macrophages to the concentration of 12,5 μΜ. Its cytotoxic concentration 50 % (CC50) is thus superiorto 12,5.
Conclusions
1- in vitro actîvity of the dronedarone on Lelshmania donovani (as a model of Indian/African strains)
The dronedarone présents a strong actîvity on intramacrophagic amastigotes of L. donovani similarto that of miltefosine, ofthe order of 1-2 μΜ, while it Is weakly active on axenic amastigotes. The therapeutic index of dronedarone defined as the CC50 on intramacrophagic amastigotes of L donovani/ IC50is thus superiorto 7.
2- in vitro actîvity of the dronedarone on Leishmania major (as a model of middle east strains)
The dronedarone présents a strong actîvity in vitro on the models of axenic amastigotes and Intramacrophagic amastigotes of Leishmania major, with IC50 lower than 3 μΜ, what places It at a level of actîvity similar to that of miltefosine. The intrinsic actîvity on the parasite itself is thus maintained when this one is protected in the phagolysosome inside the macrophage. The therapeutic index of the dronedarone defined as the CC50 on intramacrophagic amastigotes of L. major / IC50 is thus superior to 9.
These results tend to demonstrate that dronedarone has a capacity to bring a therapeutic effect on various strains (3 models tested from around the worid), at concentrations similar to miltefosine, and much lower then glucantime.
Claims (31)
1. Dronedarone or one of its pharmaceutically acceptable salts for use in the treatment of leishmaniasis.
2. Dronedarone for use according to claim 1 in the treatment of cutaneous leishmaniasis.
3. Dronedarone for use according to anyone of claims 1 and 2 in the treatment of leishmaniasis issued from Leishmania amazonensis strains.
4. Dronedarone for use according to anyone of claims 1 to 3 in the treatment of leishmaniasis issued from Leishmania amazonensis strains résistant to pentavalent antimonials dérivatives.
5. Dronedarone for use according to claim 4 wherein said pentavalent antimonials dérivative is meglumine antimoniate
6. Dronedarone for use according to claim 1 in the treatment of leishmaniasis issued from Leishmania donovani strains.
7. Dronedarone for use according to claim 1 in the treatment of leishmaniasis issued from Leishmania major strains.
8. Dronedarone for use according to anyone of claims 1 to 7 wherein said pharmaceutically acceptable sait is dronedarone hydrochloride.
9. Formulation comprising dronedarone or one of Its pharmaceutically acceptable salts for topical administration.
10. Formulation according to claim 9 wherein said formulation comprises at least a pharmaceutically acceptable excipient
11. Formulation according to anyone of claims 9 or 10 wherein said pharmaceutically acceptable sait is dronedarone hydrochloride.
12. Formulation according to any one of claims 9 to 11 wherein said formulation is a hydro-alcoholic gel.
13. Formulation according to anyone of claims 9 to 12 wherein said pharmaceutically acceptable excipient is hydroxypropyl methylcellulose.
14. Formulation according to anyone of claims 9 to 13 for use as a medicine.
15. Formulation according to anyone of claims 9 to 14 for use in the treatment of leishmaniasis.
16. Formulation according to anyone of claims 9 to 15 for use in the treatment of cutaneous leishmaniasis.
17. Formulation according to anyone of the claims 9 to 16 for use in the treatment of leishmaniasis issued from Leishmanie amazonensis strains.
18. Formulation according to anyone of the claims 9 to 17 for use in the treatment of leishmaniasis issued from Leishmanie amazonensis strains résistant to pentavalent antimonials dérivatives.
19. Formulation according to claim 18 wherein said pentavalent antimonials dérivative is meglumine antimoniate.
20. Formulation according to anyone of the claims 9 to 16 for use in the treatment of leishmaniasis issued from Leishmanie donovani.
21. Formulation according to anyone of the claims 9 to 16 for use in the treatment of leishmaniasis issued from Leishmanie major.
22 Formulation according to anyone of daims 9 to 16 wherein dronedarone or one of its pharmaceutically acceptable sait may be used in a proportion of 10% by weight of the active principle in base form.
23. Association of dronedarone or one of its pharmaceutically acceptable salts and an anti-leischmaniasis agent.
24. Association according to claim 23 for use in the treatment of leishmaniasis.
25. Association according to anyone of claims 23 and 24 for use in the treatment of cutaneous leishmaniasis.
26. Association according to anyone of claims 23 to 25 for use in the treatment of leishmaniasis issued from Leishmanie amazonensis strains.
27. Association according to anyone of claims 23 to 26 for use in the treatment of leishmaniasis issued from Leishmanie amazonensis strains résistant to pentavalent antimonials dérivatives.
28. Association according to anyone of claims 23 to 27 wherein said antMeischmaniasis agent is selected among the foliowing agents:
- Miltefosine or one of its pharmaceutically acceptable dérivatives,
- Amphotericin B or one of its pharmaceutically acceptable dérivatives,
- pentavalent antimonials dérivatives such meglumine antimoniate.
29. Association according to anyone of claims 27 and 28 wherein said said pentavalent antimonials dérivative is meglumine antimoniate.
30. Association according to anyone of claims 23 to 25 for use in the treatment of leishmaniasis issued from Leishmania donovani strains.
31. Association according to anyone of claims 23 to 25 for use in the treatment of leishmaniasis issued from Leishmania major strains.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US61/650182 | 2012-05-22 | ||
| EP12306362.0 | 2012-10-31 | ||
| EP12306472.7 | 2012-11-28 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| OA17154A true OA17154A (en) | 2016-03-28 |
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