CZ18413U1 - Composition to prevent and suppress coccidia - Google Patents

Description

The invention relates to a composition for the prevention and control of coccidiosis, in which formurindicarboxylic acid and its derivatives are newly used as a highly effective anticoccidial.

BACKGROUND OF THE INVENTION

Coccidia parasitize in a number of species, especially economically important animals, where they cause significant economic losses in the form of mortality, morbidity, reduced performance and increased treatment and prevention costs. The most common are coccidia of the genus Eimeria parasitic in both birds and mammals, and especially coccidia Cryptosporidium and Isospora. Cryptosporidim parvum, typically found in the small intestine in suckling ruminant pups, is also an important human pathogen. C. parvum and a genetically closely related C. hominis species are most commonly found in the human population in individuals with immune system disorders, particularly in AIDS patients who cause debilitating chronic watery diarrhea. In immunocompetent individuals, cryptosporidia causes dysentery, which usually subsides after a few days, but severe diarrhea and dehydration can cause serious complications in young children and the elderly.

Today, two different approaches can be used in the treatment and prevention of coccidiosis: 1) vaccination and 2) administration of coccidiostats and anticoccidials. Vaccination against a number of Eimeria species is practically applicable in poultry. For other animal species, it is necessary to use anticoccidial agents, the spectrum of which now covers all commonly occurring coccidia. However, the problem of using these substances, which were mostly synthesized as early as the 1950s to 1970s, is the emergence of resistance, the need to follow rotational programs and, last but not least, their residency, which is a serious problem in terms of food chain security.

The situation is specific for cryptosporide infections in humans and animals, where there is virtually no fully reliable preparation yet and the possibilities of immunotherapy are very limited (Harp et Goff, 1995;

Jenkins, 2004). In humans, paromomycin or azithromycin, nitazoxanide or lethrazuril may be used for cryptosporidiosis chemotherapy where partial anticryptosporidial activity has been confirmed (Blanshard et al., 1997; Giacometti et al., 1998). To date, only halofuginone lactate has been authorized for use in animals.

In view of the above, it is clear that there is a need to find new, highly effective anticoccidials that do not endanger the subjects treated with toxicity and leave no residues.

The essence of the technical solution

The above-mentioned drawbacks are eliminated by the coccidiosis prevention and suppression agent according to the invention, which consists in that 1 kg of suitable biologically acceptable matrix contains at least 0.1 pmol of formurindicarboxylic acid and its derivatives of the general formula I

In which R1 to R3 denote

R1 R2 R3 Formmaurindicarboxylic acid H COOH COOH Formaurindicarboxylates (salts of formurindicarboxylic acid) H COO ' coo · Aurintricarboxylic acid -OH COOH COOH COOH Aurintricarboxylates (salts of aurintricarboxylic acid) -OH οοσ COO ‘ COO ’ Aluminon -OH COO-NH / COO-NH / COO-NH /

The composition of the invention is further characterized in that the suitable biologically acceptable matrix is an aqueous alkaline solution or mere water.

The above-mentioned derivatives are known and, in particular, the most important of which is aurintricarboxylic acid, remarkable effects on living organisms have been described. Among the most important properties is that aurintricarboxylic acid is a very potent nuclease inhibitor, namely DNAse I, RNAase A, SI nuclease, exonuclease ΙΠ or restriction endonucleases Sal I, Bam HI, Pst I and Sma I (Hallick et al., 1977) . Other biological effects of aurintricarboxylic acid are derived, for example, from stimulation of tyrosine phosphorylation (Okada et Koizumi, 1995). Aurintricarboxylic acid also prevents blood clotting by inhibiting the binding of von Willebrand factor to platelets (Owens et Holme, 1996) and has a number of other biological properties. Known effects of aurintricarboxylic acid on pathogenic organisms include inhibition of YopH factor, which determines the virulence of Yersinia (Liang et al., 2003).

It is surprising that the use of formurindicarboxylic acid and its derivatives as an antiprotozoic drug has not been described in the literature. In the literature these substances are listed as aurintricarboxylic acid (CAS # 4431-00-9), aurintricarboxylic acid triammonium salt, the so-called aluminone (CAS # 569-58-4), formurindicarboxylic acid (CAS # 62051-06-3).

From the mechanism of biological action of these substances, it can reasonably be expected that their use could also be considered as medicaments against other protozoan infections, in particular in the form of suitable solutions, gels, ointments and solid dosage forms.

The following embodiment illustrates the device according to the invention without limiting it in any way. FIG. 1 is a structural formula with active compound derivatives; and FIG. Figure 2 shows the results from the use of aurintricarboxylic acid in a laboratory model, in newborn C57B1 / 6 mice experimentally infected with Cryptosporidium parvum.

Exemplary embodiment

To verify the in vivo activity of aurintricarboxylic acid, the Tzipori (1998) model was used with the following modification: three-day-olds of C57B1 / 6 mice were randomized and groups of 8 to 9 pups were formed. Each group was bred with mat-2C 18413 UL in its own sterile micro-isolator under standard conditions for the duration of the experiment. The day after randomization, mice were orally infected with bovine C. parvum isolate. The infectious dose was 100,000 oocysts. Immediately after the experimental infection, the mice were orally administered a solution prepared to the desired concentrations by dissolving the aurintricarboxylic acid crystals in a 1: 3 molar ratio of sodium and potassium hydroxide or in a 100% dimethylsulfoxide solution.

Treatment with doses of 50 and 100 pmol / kg bw / day lasted 8 days and the treatment was tolerated without apparent signs of toxicity. The drug was administered orally regularly at 12-hour intervals. The control group received PBS buffer instead of aurintricarboxylic acid, the paromomycin-treated group (100 mg / kg / day) served as a positive control. The intensity of infection was evaluated by counting C. parvum oocysts in the intestinal tract homogenate on day 9 post infection.

The experiment was repeated four times between 2006 and 7, each with a very similar result. The attached Graphs 1 show the results in the order of the last verification when sodium hydroxide was used to dissolve the aurintricarboxylic acid. It is clear from the above that the efficacy of formurindicarboxylic acid and its derivatives is more than likely in other hosts, also against other coccidia (including Eimeria, isospora).

References:

Blanshard C, Shanson DC, Gazzard BG: Pilot studies of azithromycin, letrazuril and paromomycin in the treatment of cryptosporidiosis. Int J STD AIDS 1997; 8 (2): 124-9.

Giacometti A, Burzacchini F, Cirioni O, Barchiesi F, Dini M, Scalise G: Efficacy of treatment with paromomycin, azithromycin, and nitazoxanide in a patient with disseminated cryptosporidiosis. Eur J Clin Microbiol Infect Dis 1998; 18: 885-889.

Harp JA, Goff JP: Protection of calves with a vaccine against Cryptosporidium parvum. J Parasitol. 1995; 81 (1): 54-7.

Jenkins MC: Present and future control of cryptosporidiosis in humans and animals. Expert Rev Vaccines. 2004; 3 (6): 669-71.

Liang F, Huang Z, Lee J, Ivanov MI, Alonso A, Bliska JB, Lawrence DS, Mustelin T, Zhang ZY: Aurintricarboxylic acid blocks in vitro and in vivo activity of YopH, an essential virulent factor of Yersinia pestis, The agent of plague. J Biol Chem. 2003; 278 (43): 41734-41.

Okada N, Koizumi S: A neuroprotective compound, aurine tricarboxylic acid, stimulates the tyrosine phosphorylation Cascade in PC 12 cells. J Biol Chem. 1995; 270 (27): 16464-9.

Owens MR, Holme S: Aurin tricarboxylic acid inhibits adhesion of platelets to subendothelium. ThrombRes. 1996; 81 (2): 177-85.

Tzipori S: Cryptosporidiosis: laboratory investigations and chemotherapy. Adv Parasitol. 1998;

40: 187-221.

-3EN 18413 Ul

Claims (2)

A composition for the prevention and control of coccidiosis, characterized in that a suitable biologically acceptable matrix comprises at least 0.1 pmol of the formurindicarboxylic acid and its derivatives of the general formula I in which R1 to R3 denote R1 R2 R3 Formmaurindicarboxylic acid H COOH COOH Fomnaurindicarboxylates (formurindicarboxylic acid salts) H COO ’ coo- Aurintricarboxylic acid -OH COOH COOH COOH Aurintricarboxylates (salts of aurintricarboxylic acid) -OH οοσ coo · coo- Aluminon -OH COONH / COO-NH / COO-NH / The composition of claim 1, wherein the suitable biologically acceptable matrix is an aqueous alkaline solution or water. 2 drawings -4GB 18413 Ul Formmaindinedicarboxylic acids and derivatives thereof of the general formula I in which R 1 to R 3 denote R1 R2 R3 Formmaurindicarboxylic acid H COOH COOH Formaurindicarboxylates (salts of formurindicarboxylic acid) H COO ' COO ’ Aurintricarboxylic acid —OH COOH COOH COOH Aurintricarboxylates (salts of aurintricarboxylic acid) -OH coo- coo · COO ’ Aluminon COQNH / COONH / COO-NH / Fig.1 -5GB 18413 Ul Figure 1: Inhibitory effect of aurintricarboxylic acid on experimental Cryptosporidium parvum infection in sucking C57BI / 6 mice Fig.2 End of document