HK1178440A - Association between ferroquine and an artemisinine derivative for treating malaria - Google Patents

Description

Combination of ferroquine and artemisinin derivatives for treating malaria

The application of the invention is a divisional application based on a patent application with application date of 18.04.2006, application number of 200680013013.6 (international application number of PCT/FR2006/000842) and title of 'composition of ferroquine and artemisinin derivative for treating malaria'.

Technical Field

The present invention relates to a novel combination of active ingredients against malaria, namely ferroquine and an artemisinin derivative, as well as to pharmaceutical compositions containing such a combination for the treatment and/or prevention of malaria.

Background

Malaria is one of the leading infectious causes of massive deaths in the world, involving 5 million people each year, with 3 million people dying each year. Such disasters are mainly related to africa, southeast asia and latin america, in the sub-saharan desert.

Anopheles mosquito carries four classes of plasmodium parasites (plasmodium falciparum, plasmodium malariae, plasmodium vivax and plasmodium ovale) that transmit malaria. Plasmodium falciparum, which is widely spread in africa, is the most virulent parasite and is responsible for the lethal forms of the disease.

The high recurrence of this disease has been observed for many years due to a number of factors, including resistance of many plasmodium falciparum species to commonly used drugs (e.g., chloroquine, flumequine hydroxypiperidine, carbamoquine, or pyrimethamine).

Artemisinin, isolated from the plant Artemisia annua (artemisinin) used in centuries of China in 1972, has a strong antimalarial activity. Derivatives with improved pharmacological properties, such as artemether, fleabane and artesunate, have also been commercialized.

Artemisinin and its derivatives are part of the most potent active components against plasmodium falciparum today. However, the use of artemisinin or its derivatives in monotherapy may be a causal factor in the selection of drug-resistant parasitic varieties.

The scientific community now advocates the use of combinations of active ingredients, in particular artemisinin or its derivatives with other antimalarial active ingredients. The world health Organization (OMS) recommended these multidrug therapies since 2002, called ACT (artemisinin-based combination therapy). They provide multiple benefits: the treatment efficiency of the drug-resistant variety is improved, the drug resistance of the two active components is prevented, and the disease transmission and the drug resistance spread are reduced.

For example, the trade nameCombinations of artemether and lum fantrine for sale, and combinationsArtesunate and carbamoquineIn line with the strategies supported by OMS, the search for new combinations of antimalarial active components should be continued.

Among the different antimalarial active components described in the literature, ferroquine is an active molecule against chloroquine-resistant plasmodium falciparum species. Ferroquine, also known as ferrocene-chloroquine or ferroquine, corresponds to 7-chloro-4 [ ({2- [ (N, N-dimethyl-amino) methyl ] ferrocenyl } methyl) amino ] quinoline. Relates to 4-aminoquinoline derivatives coupled to a ferrocene ring. Such molecules are described in particular in patents EP 0824536 and J.Med.chem., 1997, 40, 3715-3718, Antimicrob.Agents Chemother, 1998, 42, 540-544, journal of organic chemistry, 1999, 589, 59-65 and journal of organometallic chemistry, 2004, 689, 4678-4682.

While the combination of chloroquine and artesunate has not achieved satisfactory levels of efficacy (am.j. trop. med. hyg., 2003, 69(1), 19-25 and Transactions of the Royal Society of pharmaceutical Medicine and Hygi. ne, 2003, 97, 429 433), it has now surprisingly been found that the combination of ferroquine with an artemisinin derivative, in particular artesunate, artemether or arteether, is very effective for the treatment and/or prevention of malaria, in particular for the prevention and reduction, even the avoidance, of the development of parasite species resistant to both active components, when these drugs are administered as a single drug.

Disclosure of Invention

The object of the present invention is therefore a novel composition of ferroquine (molecule (I) below, represented in free base form, where Fe represents the ferrocene ring) and an artemisinin derivative.

In these compositions of the invention, the ferroquine may be in the form of the free base, but also in the form of a salt, hydrate or solvate (the latter being defined as an association or compound of ferroquine with one or more water or solvent molecules, respectively). Advantageously, the free base form of ferroquine is used.

The artemisinin derivative in these compositions of the invention is advantageously composed of artesunate (II) or artemether (III):

another subject of the invention is a pharmaceutical composition containing as active ingredients ferroquine (I) in combination with an artemisinin derivative, advantageously artesunate (II) or artemether (III).

Such a pharmaceutical composition comprises a therapeutically effective dose of ferroquine, or a pharmaceutically acceptable salt, hydrate or solvate of ferroquine, and at least one artemisinin derivative, and at least one pharmaceutically acceptable excipient. The excipients are selected from the general excipients known to those skilled in the art depending on the pharmaceutical dosage form and the desired mode of administration.

Unit dosage forms for appropriate administration include oral dosage forms, such as tablets, soft or hard capsules, powders, granules and oral liquids or suspensions, sublingual, oral, tracheal, intraocular, intranasal administration by inhalation, topical, transdermal, subcutaneous, intramuscular or intravenous administration, rectal and implant administration. For topical application, the compounds of the invention may be used in creams, gels, ointments or lotions.

Preferred routes of administration are oral, rectal and injectable.

For example, in the preparation of solid compositions in the form of tablets, the active ingredient is mixed with one or more pharmaceutical excipients, such as gelatin, starch, lactose, magnesium stearate, talc, silicon dioxide, acacia, mannitol, microcrystalline cellulose, hydroxypropyl-methylcellulose, croscarmellose and the like. These tablets may be encapsulated with sucrose, cellulose derivatives or other materials suitable for sugar coating. These tablets may be prepared using different techniques, for example using direct compression, dry granulation, wet granulation or hot melt techniques.

The preparation of the capsule dosage form can also be achieved by mixing the active ingredients with a diluent and pouring the resulting mixture into soft or hard gelatin capsules.

For parenteral administration, aqueous suspensions, isotonic saline solutions or sterile injection solutions are used, which contain pharmacologically compatible dispersing and/or wetting agents, such as ethylene glycol or butylene glycol.

When administered orally, the daily dosage of each of the two active ingredient combinations of the invention is as follows:

-ferroquine: 50-1600mg, preferably 200-1200mg, more preferably 400-800mg per person per day;

-an artemisinin derivative: 1-10 mg/kg/day, preferably 2-6 mg/kg/day, more preferably about 4 mg/kg/day.

There may be special cases where either high or low dosages are appropriate; such dosages are not outside the scope of the present invention. According to common practice, the appropriate dosage for each patient is determined by a physician, depending on the mode of administration, the weight and the response of said patient.

The active ingredient combinations of the invention can be administered for 3 consecutive days, with each of the two active ingredients being administered in one or more divided doses per day, preferably once daily. This treatment period, limited to 3 days, is particularly advantageous compared to the recommended single-drug treatment of artemisinin derivatives for 7 days, in that it allows these patients to better observe the treatment, thus avoiding premature cessation of treatment, which would induce resistance in the parasite over a long period of time.

Administration of each of the two active ingredients can be carried out simultaneously or separately or at the time of the development (sequential administration).

When administered simultaneously, the two active ingredients may be combined in a single pharmaceutical dosage form (fixed combination), for example in tablets or capsules suitable for oral administration.

The two active ingredient combinations of the invention can also be in different pharmaceutical dosage forms, whether or not they are administered simultaneously. For this purpose, the combination according to the invention may be in the form of a kit containing on the one hand ferroquine or the ferroquine salt, hydrate or solvate, and on the other hand at least one artemisinin derivative, such as artesunate or artemether, said ferroquine and said artemisinin derivative being in different compartments and intended for simultaneous, separate or simultaneous spread (sequential administration) administration.

By way of example, a unit dosage form of the tablet ferroquine may contain the following components:

also by way of example, a unit dosage form of artesunate tablet may contain the following components: 50 or 100mg artesunate and usual excipients, such as lactose, croscarmellose, anhydrous colloidal silicon dioxide, microcrystalline cellulose and magnesium stearate.

A further object of the invention is a method for the treatment and/or prevention of malaria, which method comprises administering to a patient a therapeutically effective dose of ferroquine or a pharmaceutically acceptable salt, hydrate or solvate thereof, and a therapeutically effective dose of at least one artemisinin derivative, said doses being administered to the patient simultaneously or sequentially, as described above.

The combination of active ingredients of the invention is the subject of biochemical tests in vivo in mice infected with plasmodium falciparum species (vinckei vinckei plasmodium species), and therefore can demonstrate the efficacy of treatment of this malaria.

The invention comprises the following steps:

a composition comprising as active ingredients chloroquine in free base, salt, hydrate or solvate form and an artemisinin derivative.

Item 2. the composition according to item 1, characterized in that the artemisinin derivative is composed of artesunate, arteether or artemether.

Item 3. the composition according to any one of items 1 or 2, characterized in that the artemisinin derivative is composed of artesunate or artemether.

Composition according to item 1, characterized in that the daily dose of ferroquine is 50-1600mg, preferably 200-1200mg, more preferably 400-800mg per person per day.

Item 5. the composition according to any one of items 1 to 3, characterized in that the daily dose of the artemisinin derivative is 1-10 mg/kg/day, preferably 2-6 mg/kg/day, more preferably about 4 mg/kg/day per person per day.

Item 6. the composition according to any one of items 1 to 5, characterized in that it is used for continuous administration for 2 to 4 days.

The composition according to any one of items 1 to 6, characterized in that each active ingredient is intended to be administered simultaneously or sequentially.

A pharmaceutical composition comprising a therapeutically effective amount of ferroquine or a pharmaceutically acceptable salt, hydrate or solvate of ferroquine, and at least one artemisinin derivative, and at least one pharmaceutically acceptable excipient.

Item 9. the pharmaceutical composition according to item 8, characterized in that the artemisinin derivative is composed of artesunate, arteether or artemether.

Item 10. the pharmaceutical composition according to any one of items 8 or 9, characterized in that the artemisinin derivative is composed of artesunate or artemether.

The pharmaceutical composition according to any one of items 8 to 10, characterized in that it is suitable for oral, rectal or injectable administration.

Item 12. the pharmaceutical composition according to any one of items 8 to 11, characterized in that it is for the treatment and/or prevention of malaria.

Item 13. use of the composition of any one of items 1-7 in the manufacture of a medicament for treating or preventing malaria.

Item 14 a kit for the treatment or prevention of malaria, which kit comprises on the one hand ferroquine or a salt, hydrate or solvate of such ferroquine and on the other hand at least one artemisinin derivative, said ferroquine and said artemisinin derivative being in different compartments and intended for simultaneous or sequential administration.

Drawings

Figure 1 is the percent survival starting on day 5 after infection of the animals.

Detailed Description

The following tests with artesunate are given as examples. Since these artemisinin derivatives have all the same metabolites (dihydroartemisinin) and short half-lives, the experimental results obtained for artesunate are generalizable to other artemisinin derivatives such as artemether or arteether.

Living mouse infected by Vinckei plasmodium, ferroquine, artesunate and two compound groups Assay for Complex Activity

1. Description of the use of the Living body test

Eight weeks old, Swiss, female like mice were vaccinated with the Vinckei vinckei Plasmodium parasite (Rodhain, 1952). These mice were acclimated to the new environment for two weeks in advance. These mice were fed ad libitum with drinking water.

The rats were kept weekly for 10 suspended in saline (0.9%) phosphate buffer⁷Each parasitic red blood cell infects a strain of plasmodium vinckei vinckei.

The first day of treatment (JO), one hour after infection (107 parasitic red blood cells suspended in saline (0.9%) phosphate buffer), according to which the animals were given oral ferroquine, artesunate or a mixture of the two active ingredients. This administration was then repeated for three days (J1 to J3) (Peter, 1987). When the first two artesunate products were administered in combination, the second administration of ferroquine was 45 minutes later. On the fourth day, blood smears were drawn at the tail of the rat. The sample was fixed on a plate. The number of parasitic blood cells was counted microscopically. The percentage of infected erythrocytes in this specimen, calculated as 1000 cell samples, represents parasitemia, six or seven mice per dose being used.

These mice, which did not show any parasite traces on the J4 smear, were again subjected to controls on days 10, 17, 24, 31, 38, 45, 52 and 59 in order to detect possible parasite recurrences.

Preparation of Compound dilution and administration suspensions

-Preparation of ferroquine suspension

(ferroquine from Sanof-Synth é labo, batch No. MY18.0088)

The ferroquine was mixed with methylcellulose (0/5(w/w)) and polysorbate 80(0/5 (w/w)). The formulation was stable for at least 7 days in the dark and cold (4 ℃) conditions and for 4 hours at room temperature. The concentration of the final suspension of ferroquine is 0.1-100 mg/mL.

-Artesunate suspension preparation

(Artesunate from Sanofi-Synthelabo, batch No. 1.04).

Artesunate was mixed with methylcellulose (0/5(w/w)) and polysorbate 80(0/5 (w/w)). The formulation was stable for 4 hours in the dark and at ambient temperature. The concentration of the final suspension of artesunate is 0.8-20 mg/mL.

2. Determination of CI for separate administration of Artesunate and Ferro-chloroquine ₅₀ And a therapeutically effective amount CI ₅₀ Method of measurement of

CI₅₀Defined as the concentration expressed in mg/kg/day of 50% inhibition of blood parasitemia on day 4 after infection (JO) (J4) and four days of treatment (J0, J1, J2, J3). 0% inhibition corresponds to the mean value of observed parasitemia in these untreated infected mice. 100% inhibition corresponds to very low or zero parasitemia below 0.1%. These CI's were determined by linear interpolation of response dose curves expressed as log concentration₅₀。

CI of ferroquine₅₀Is measured after administration at a concentration of 1-10 mg/kg/day. Concentrations of 0, 1, 1.47, 2.1, 3.2, 4.6, 6.8 and 10 mg/kg/day were used for 4 days.

CI of artesunate₅₀Is measured after administration at a concentration of 1-15 mg/kg/day. Concentrations of 0, 1, 1.6, 2.5, 3.9, 6.1, 9.5 and 15 mg/kg/day were used for 4 days.

Obtained CI₅₀Listed in table I below:

	CI50(mg/kg/day)
		Ferro-chloroquine	3.32
Artesunate ester	2.79

The curative dose of the ferroquine is 10 mg/kg/day.

No therapeutic doses of artesunate were achieved in this study, so the dose was higher than 15 mg/kg/day.

Importantly, the interaction studies were performed using non-therapeutic (sub-optimal) doses of artesunate and ferroquine, thus approaching Cl achieved using two separately studied compounds₅₀. The curative effect dose of the ferroquine (total survival rate of treated mice) is close to Cl₅₀. For artesunate, the therapeutic dose is in combination with Cl₅₀Is higher, so that it is possible to use higher Cl than₅₀The dosage of (a). In the combined study, it was then considered that ferroquine was administered at 3 mg/kg/day together with artesunate at 6 mg/kg/day for 4 days.

3. Measuring ferroquine/Artesunate using Vinckei Plasmodium Living parasite species and using Antimalarial Activity

3.1. Determination of percent parasitemia

Mice were infected and administered separately for 4 days with 3 mg/kg/day ferroquine and 6 mg/kg/day artesunate, compared to a batch of mice that received no treatment. Table II shows the mean parasitemia (percentage of infected erythrocytes) observed on day four post-infection.

TABLE II

As shown in table II, the administration of 3 mg/kg/day of ferroquine and 6 mg/kg/day of artesunate in combination with 4 days of administration significantly reduced parasitemia in infected animals compared to separate administration of both products.

3.2. Determination of mortality in mice

To determine the percent mortality (number of dead mice compared to the number of the batch studied), the number of dead mice was counted daily starting on day 5 post-infection. The therapeutic dose was the first dose at which all mice in the batch survived.

Figure 1 is the percent survival starting on day 5 after infection of these animals.

As indicated in 1, these compounds were administered at sub-optimal doses and in combination (3 mg/kg/day for iron chloroquine and 4 days for artesunate) to improve animal survival compared to separate administration (4 days for iron chloroquine or 6 mg/kg/day for artesunate).

The results obtained from the infection of living mouse bodies with the Vinckei Plasmodium species clearly indicate an antagonistic effect between these two active ingredients and demonstrate that the combination of artesunate (or artemisinin derivatives in general) with ferroquine according to the invention is advantageous for the treatment of malaria.

Claims (14)

1. A composition comprising as active ingredients ferroquine in free base, salt, hydrate or solvate form and an artemisinin derivative.

2. The composition according to claim 1, characterized in that the artemisinin derivative is composed of artesunate, arteether or artemether.

3. Composition according to any one of claims 1 or 2, characterized in that the artemisinin derivative is composed of artesunate or artemether.

4. Composition according to claim 1, characterized in that the daily dose of ferroquine is 50-1600mg, preferably 200-1200mg, more preferably 400-800mg per person per day.

5. A composition according to any of claims 1-3, characterized in that the daily dose of artemisinin derivative is 1-10 mg/kg/day, preferably 2-6 mg/kg/day, more preferably about 4 mg/kg/day per person per day.

6. Composition according to any one of claims 1 to 5, characterized in that it is intended for 2 to 4 consecutive days of administration.

7. A composition according to any one of claims 1 to 6, characterised in that each active ingredient is intended for simultaneous or sequential administration.

8. A pharmaceutical composition comprising a therapeutically effective amount of ferroquine or a pharmaceutically acceptable salt, hydrate or solvate thereof, and at least one artemisinin derivative, and at least one pharmaceutically acceptable excipient.

9. Pharmaceutical composition according to claim 8, characterized in that the artemisinin derivative is composed of artesunate, arteether or artemether.

10. Pharmaceutical composition according to any of claims 8 or 9, characterized in that the artemisinin derivative is composed of artesunate or artemether.

11. Pharmaceutical composition according to any one of claims 8 to 10, characterized in that it is suitable for oral, rectal or injectable administration.

12. Pharmaceutical composition according to any one of claims 8 to 11, characterized in that it is used for the treatment and/or prevention of malaria.

13. Use of a composition according to any one of claims 1-7 in the manufacture of a medicament for the treatment or prevention of malaria.

14. A kit for the treatment or prevention of malaria, which kit comprises on the one hand ferroquine or a salt, hydrate or solvate of said ferroquine and on the other hand at least one artemisinin derivative, said ferroquine and said artemisinin derivative being in different compartments and intended for simultaneous or sequential administration.